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  • Electronic Resource  (187)
  • 1995-1999  (187)
  • 1890-1899
  • 1999  (187)
  • apoptosis  (112)
  • nitrogen  (75)
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  • Electronic Resource  (187)
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  • 1995-1999  (187)
  • 1890-1899
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  • 1
    ISSN: 1436-2813
    Keywords: thyroid tumor ; apoptosis ; TUNEL ; MIB-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To clarify the growth mechanisms of thyroid tumors, we examined apoptotic cells in 61 thyroid tumors, consiting of 14 adenomas, 35 papillary carcinomas, 4 follicular carcinomas, and 8 undifferentiated carcinomas, using terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate digoxigenin-nick end labeling (TUNEL). The proliferative activity was also evaluated immunohistochemically using the monoclonal antibody to Ki-67 antigen (MIB-1) in the same tumors. The apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells, and a proliferation index (PI), being the percentage of Ki-67 positive cells, was calculated for each tumor. The overall level of AI was very low in all histotypes of the thyroid tumors analyzed, the mean AI being 0.5±0.4 in adenoma, 0.4±0.3 in differentiated carcinoma, and 1.8±1.5 in undifferentiated carcinoma. The PI in the thyroid tumor subtypes was significantly lower in adenoma and differentiated carcinoma, at 0.5 ±0.7 and 1.1±0.7, respectively, than that in undifferentiated carcinoma at 14.5±3.7 (P〈0.05). There was no correlation between clinicopathological factors and AI or PI in differentiated thyroid carcinoma. Our findings suggest that apoptosis occurs infrequently in thyroid tumors, and that proliferative activity markedly differs according to the thyroid tumor subtypes. Moreover, the ratio between proliferating cells and apoptotic cells may reflect thyroid tumor progression.
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  • 2
    ISSN: 1436-2813
    Keywords: Key Words: thyroid tumor ; apoptosis ; TUNEL ; MIB-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: P 〈 0.05). There was no correlation between clinicopathological factors and AI or PI in differentiated thyroid carcinoma. Our findings suggest that apoptosis occurs infrequently in thyroid tumors, and that proliferative activity markedly differs according to the thyroid tumor subtypes. Moreover, the ratio between proliferating cells and apoptotic cells may reflect thyroid tumor progression.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 92 (1999), S. 205-216 
    ISSN: 1570-7458
    Keywords: Samea multiplicalis ; Spodoptera pectinicornis ; Pistia stratiotes ; waterlettuce ; nitrogen utilization efficiency ; compensatory feeding ; nitrogen ; biological control of weeds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Compensatory responses of caterpillars fed low quality food include increased consumption and utilization of essential nutrients. Information about an insect's responses to nutritional challenges from their host plants could benefit weed biological control efforts in the selection and establishment of new agents. The target weed, Pistia stratiotes L. (Araceae) is a floating aquatic plant that has relatively low nitrogen levels which are further diluted with high water content. Efforts to establish the insect Spodoptera pectinicornis (Hampson) (Lepidoptera: Noctuidae) for biological control of P. stratiotes could benefit by examining the nutritional responses of a similar widely established lepidopteran species, Samea multiplicalis (Guenèe) (Lepidoptera: Pyralidae). Larvae of this species were fed leaves of P. stratiotes plants that had been fertilized (NPK) at high and low rates. The leaves of the fertilized plants had a 4.3-fold increase in nitrogen (dry weight) and a 1.6-fold increase in water content. The results suggest that no compensatory increases occurred in larvae fed leaves from the low fertilized plants as no changes were found in fresh mass consumption or nitrogen utilization efficiency. Consequently, development time from second-third instars to pupation was delayed about 3 days compared with larvae fed the high nitrogen leaves. Furthermore, consumption of nitrogen was only 30% and its accumulation into larval tissues was only 60% compared with the larvae fed the high fertilized leaves. The resulting larvae had both a final biomass and a growth rate that were reduced by 40%. Regardless of plant fertilizer level, the larvae fed at a rate 5–10 times greater than that of similar lepidopteran species consuming either low or high quality diets, suggesting that the S. multiplicalis larvae may be functioning at their biological limit for ingesting food.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 92 (1999), S. 165-177 
    ISSN: 1570-7458
    Keywords: Chrysomelidae ; herbivory ; Asteraceae ; life history ; nitrogen ; plant quality ; season
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytophagous insects which feed on the leaves of herbaceous host plants have to adapt their life histories to the fact that protein nitrogen is usually highest in growing tissues in spring. We monitored field populations of larvae and adults of three chrysomelid species (Galeruca tanaceti (L.) (main host Achillea millefolium (L.) Yarrow), Cassida rubiginosa (Mueller) (main host Cirsium arvense (L.) Scop.) and Oreina luctuosa (Suffrian) (host Centaurea scabiosa (L.)) together with the amount of protein nitrogen of their food resources and host plant biomass. As expected, the development of host quality, measured as concentration of protein nitrogen, and host plant biomass showed inverse trends during the season. The euryphagous G. tanaceti attacks Achillea early and profits from high nitrogen concentrations in the leaves. Occasional overexploitations of local populations of Achillea are compensated by the capacity to move to other host species. In C. rubiginosa, a species with a host range restricted to the Cardueae, the main larval feeding activity is postponed to a period when the nitrogen content of the host leaves had dropped to 50% of its initial value, but when host plant biomass had increased by 30%. In the monophagous O. luctuosa the larval development is synchronized with a still later phase of host phenology, at which the nitrogen content is below 50% but plant biomass has reached its maximum. There seem to be selection factors, which oppose the use of high quality food in spring and which force the latter two species to postpone their larval development to a later time in the year. This could be caused by numerous factors like, for example, mean daytime temperature. Later in the season the larvae have to cope with the low quality of their host plants. They have, however, the advantage of large quantities of food available. A laboratory study with adults and mature larvae of O. luctuosa shows that this species can overcome low levels of protein nitrogen either by selecting younger leaves with higher nitrogen concentrations or by increasing the daily food consumption rate (RCR) on leaves with a low level of nitrogen and by a prolongation of the feeding period. In this way the larvae compensate the effect of lower daily growth rates (RGR) and a lower food conversion index (ECI) on poor food quality: Regardless of the level of protein nitrogen there was no statistically significant difference in total gain of weight during the third-instar feeding period and in the weight at the end of the third larval stage. The three investigated chrysomelids show that there exists a broad spectrum of adaptations to overcome the dilemma of variable food quality.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of neural transmission 106 (1999), S. 1-21 
    ISSN: 1435-1463
    Keywords: Keywords: Paraquat ; Parkinson's disease ; transcription factor ; AP-1 ; apoptosis ; cycloheximide ; genistein ; SOD ; catalase ; oxidative stress.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary. Drugs and certain environmental toxins may be responsible for the pathogenesis of Parkinson's disease. We have used paraquat as a model toxin for this study since paraquat has been shown to make its way to the nerve terminals and cause cell death of dopamine neurons by oxidative injury. We have shown by the electrophoretic mobility shift assay that paraquat, together with low concentrations of chelated iron (Fe++/DETAPAC), induced the activation of transcription factor AP-1 binding activity to DNA. Under similar conditions we also found by both a DNA laddering assay procedure and by terminal deoxynucleotidyl transferase assay (TUNEL assay) that paraquat also induces apoptotic cell death. Interestingly, both apoptotic cell death and AP-1/DNA binding activity induced by paraquat were blocked by cyclohexamide and genistein, indicating that both the AP-1/DNA binding activation and apoptosis induced by paraquat are closely related. Moreover, cells were also protected from paraquat toxicity in the presence of antioxidant defense enzymes SOD and catalase. The results support the hypothesis that oxidative stress may be contributing to the apoptotic cell death of dopaminergic neurons, leading to the manifestation of Parkinson's disease. Since paraquat was an important herbicide in the mid 20th Century, our results have the important implication that exposure to environmental toxins such as paraquat may induce Parkinson's disease.
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  • 6
    ISSN: 1572-8773
    Keywords: cadmium ; apoptosis ; RT-PCR ; p53 gene expression ; testes ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Reverse transcription (RT) PCR technique was used to investigate the mechanism of apoptosis induced by Cd and the change of its related genes in testes and prostate of rats. Adult male rats were given a single (s.c.) injection of CdC l2 0, 2.5, 5.0, 10 μmol/kg. 48 h and 72 h after administration of Cd, animals were sacrificed. The results indicated that Cd can induce apoptosis in testes via p53-independent pathway. No apoptosis occurred in prostate in any of the Cd-exposed groups. There was a clearly negative relationship in testes between p53 gene expression and Cd exposure and this dose-response relationship was observed both at 48 h and 72 h. There was a very small increase of this gene expression in the dorsolateral lobe of the prostate in Cd exposed groups. The other apoptosis related gene, bcl-x, was not detectable in either control or Cd-exposed group in testes and dorsal prostate. Although the MT-I gene was expressed in testes or dorsal prostate both in control and exposed groups, no overexpression of MT-I gene was found after administration of Cd . The expression of MT-I in the ventral prostate was not detected in the control group, but a weak expression was found after Cd exposure. Since p53 is a tumo r suppressor gene which can inhibit tumorigenesis, the consequence of a Cd-induced decrease of p53 in testes may have a relation to the known risk of Cd tumorigenesis in this tissue.
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  • 7
    ISSN: 1572-879X
    Keywords: adsorption ; adsorption isotherms ; dinitrogen ; FTIR spectroscopy ; geminal species ; NaY ; nitrogen ; zeolites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Adsorption of N2 on NaY zeolites at 85 K and equilibrium pressures higher than 1 kPa results in the formation of geminal dinitrogen complexes characterized by an IR band at 2333.5 cm−1 (2255.4 cm−1 after adsorption of 15N2). With decreasing equilibrium pressure the complexes tend to loose one N2 ligand, thus forming linear species characterized by an IR band at 2336.8 cm−1 (2258.7 cm−1 after adsorption of 15N2). All species disappear completely after evacuation. Co-adsorption of N2 and CO revealed that the dinitrogen complexes are formed on Na+ cations. The changes in the concentrations of the linear and geminal N2 species with the changes in the equilibrium pressure are excellently described by equations of adsorption isotherms proposed earlier for mono- and di-carbonyls.
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  • 8
    ISSN: 1572-8757
    Keywords: kinetics ; isotope-exchange ; nitrogen ; adsorption ; methane ; zeolite ; equilibria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Physics , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The Isotope Exchange Technique (IET) was used to simultaneously measure pure and binary gas adsorption equilibria and kinetics (self-diffusivities) of CH4 and N2 on pelletized 4A zeolite. The experiment was carried out isothermally without disturbing the adsorbed phase. CH4 was selectively adsorbed over N2 by the zeolite because of its higher polarizability. The multi-site Langmuir model described the pure gas and binary adsorption equilibria fairly well at three different temperatures. The selectivity of adsorption of CH4 over N2 increased with increasing pressure at constant gas phase composition and temperature. This curious behavior was caused by the differences in the sizes of the adsorbates. The diffusion of CH4 and N2 into the zeolite was an activated process and the Fickian diffusion model described the uptake of both pure gases and their mixtures. The self-diffusivity of N2 was an order of magnitude larger than that for CH4. The pure gas self-diffusivities for both components were constants over a large range of surface coverages (0 〈 θ 〈 0.5). The self-diffusivities of CH4 and N2 from their binary mixtures were not affected by the presence of each other, compared to their pure gas self-diffusivities at identical surface coverages.
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  • 9
    ISSN: 1569-8041
    Keywords: AML ; apoptosis ; etoposide ; γ-GCS ; glutathione ; oxidative stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Etoposide mediates its cytotoxicity by inducing apoptosis. Thus, mechanisms which regulate apoptosis should also affect drug resistance. Oxidants and antioxidants have been shown to participate in the regulation of apoptosis. We were interested in studying whether responsiveness of acute myeloblastic leukemia (AML) cells to etoposide is mediated by oxidative stress and glutathione levels. Patients and methods: Two subclones of the OCI/AML-2 cell line which are etoposide-sensitive (ES), and etoposide-resistant (ER), were established by the authors at the University of Oulu, and used as models. Assays for apoptosis included externalization of phosphatidylserine (as evidenced by annexin V binding), and caspase activation as indicated by cleavage of poly(ADP-ribose)polymerase (Western blotting). Peroxide formation was analyzed by flow cytometry. Glutathione and gamma-glutamylcysteine synthetase (γ-GCS) levels were determined spectrophotometrically and by Western blotting, respectively. Results: Etoposide-induced apoptosis was evident 12 hours after treatment in the ES subclone, but was apparent in the ER subclone only after 24 hours. The basal glutathione and γ-GCS levels were higher in the ER than the ES subclone. Etoposide increased peroxide formation in both subclones after 12-hour exposure. Significant depletion of glutathione was observed in the ES subclone during etoposide exposure, while glutathione levels were maintained in the ER subclone. In neither of the subclones was induction of γ-GCS observed during 24-hour exposure to etoposide. Furthermore, the catalytic subunit of γ-GCS was cleaved during apoptosis, concurrent with depletion of intracellular glutathione. When glutathione was depleted by treatment with buthionine sulfoximine, a direct inhibitor of γ-GCS, the sensitivity to etoposide was increased, particularly in the ER subclone. Conclusions: The results underline the significance of glutathione biosynthesis in the responsiveness of AML cells to etoposide. The molecular mechanisms mediating glutathione depletion during etoposide exposure might include the cleavage of the catalytic subunit of γ-GCS.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Annals of oncology 10 (1999), S. 495-503 
    ISSN: 1569-8041
    Keywords: antisense ; apoptosis ; bcl-2 ; lymphoma ; leukaemia ; phase I
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Annals of oncology 10 (1999), S. 1011-1021 
    ISSN: 1569-8041
    Keywords: apoptosis ; chemosensitivity ; cytotoxicity ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Although hematologic malignancies and some solid tumors such as germ cell tumors and pediatric malignancies can be cured by cytotoxic treatment, the most prevalent solid tumors are relatively resistant to these interventions. Apoptosis is involved in the cell kill of anticancer drugs and p53 is believed to be of principal importance in this process. However p53 also plays a role in cell cycle arrest and DNA repair, cellular processes that can decrease the sensitivity to chemotherapy. Therefore, p53 may play a dual role after exposure to cytotoxic treatment, activating either mechanisms that lead to apoptosis or launching processes directing to DNA repair and survival of the cell. Design: In this article, we review in details the p53functions involved in the mediation of chemosensitivity. The preclinical and clinical data published in the recent years about the relation between p53 and chemosensitivity are discussed and the potential pitfalls associated to most of these studies, and that may account for the contradictory results produced so far are also mentioned.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Annals of oncology 10 (1999), S. 31-38 
    ISSN: 1569-8041
    Keywords: apoptosis ; lanreotide treatment ; neuroendocrine gastrointestinal tumors ; octreotide ; somatostatin analogs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuroendocrine gastrointestinal tumors express somatostatin receptors (ssts) in 80%–90% of cases and somatostatin analogs have become increasingly important in the management of these patients. Most of the presently available somatostatin analogs (octreotide, RC-160, and lanreotide) bind to the sst2 and sst5, and in higher doses to sst3 of the ssts 1–5 described. Clinical improvement during somatostatin analog therapy is mainly mediated via a direct inhibitory effect on hormone production from the tumors, seen in 30%–70% of the patients. Also indirect non-tumor mediated effects on peripheral target organs contribute to the subjective improvement, achieved in 30%–70% of patients. Recently, significant improvement of quality of life has been demonstrated with long-acting depot formulations. There is little or no effect on tumor growth during octreotide therapy; tumor shrinkage has been reported in 10%–20% of patients, but stabilization of tumor growth can be achieved in about half of the patients with a duration of 8–16 months. Recently, induction of apoptosis has been described with high doses of lanreotide (12 mg/d). Eventually, however, all patients escape from somatostatin analog therapy with regard both to hormonal production and tumor growth, and the mechanism behind the tachyphylaxis is not yet known. Studies of optimal dosage and modes of administration, development of new slow release formulations, the potential value of high-dose somatostatin analog therapy and novel somatostatin receptor subtype specific analogs are important directions for the use of somatostatin analogs in the future. In addition, assessment of somatostatin receptor status for each patient and studies of tumor biology, e.g., inhibition of exocytosis, antiproliferative effects and induction of apoptosis during treatment will help to optimize treatment and provide new insights into mechanisms of action of somatostatin analogs.
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  • 13
    ISSN: 1436-2813
    Keywords: Key Words: nitric oxide ; DNA damage ; apoptosis ; tumor necrosis factor-α ; mitochondrial respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: S -nitroso-N-acetylpenicillamine protected cul-tured L929 cells from apoptosis induced by tumor necrosis factor-α (TNF-α) plus actinomycin D, as determined by the detection of DNA fragmentation and morphological changes. NO also prevented an enhancement of the production of reactive oxygen intermediates by TNF-α plus actinomycin D, as assessed by the oxidation of dihydrorhodamine 123 and hydroethidine. Because the inhibition of mitochondrial respiration by rotenone or antimycin A suppressed the increased oxidation of both dihydrorhodamine 123 and hydroethidine, it was suggested that TNF-α accelerated the leakage of reactive oxygen intermediates from the mitochondrial electron transport system. Polarography showed that NO reversibly inhibited mitochondrial respiration at either complexes I–III, II–III, or IV, thus suggesting the inhibition of cytochrome oxidase. Taken together, these findings indicate that the decreased mitochondrial formation of reactive oxygen intermediates in the presence of NO might have a protective effect against TNF-α plus actinomycin D-induced apoptosis.
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  • 14
    ISSN: 1436-2813
    Keywords: angiogenesis ; p53 ; apoptosis ; lung adenocarcinoma ; hematogenous metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The purpose of this study was to clarify which factors are important as predictors not only of patient survival but also of hematogenic metastasis in 15 patients with stage I lung adenocarcinoma who underwent curative operation. The relationship between tumor angiogenesis, apoptosis, and p53 oncogene was also studied. A total of 15 patients were divided into two groups: surviving group (n=7) and nonsurviving (metastasis) group (n=8). We studied the medical charts, operative records, pathologic reports, and tumor specimens taken at surgical resection. We measured the apoptotic index using the ApopTag kit and the intratumoral microvessel count using an anti-CD34 monoclonal antibody. In addition, immunohistochemical staining for the expression of p53 was conducted simultaneously. The clinicopathological characteristics, including age, sex, tumor size (pT), and histological differentiation, were not significantly different between the surviving and the nonsurviving group. The microvessel count was significantly higher in nonsurviving group than in the surviving group. The apoptotic index and the expression of p53 was not significantly different between the two groups. An inverse correlation between the apoptotic index and microvessel count, and a positive correlation between the expression of p53 and microvessel count, were observed. Angiogenesis may be an important prognostic factor in patients with stage I lung adenocarcinoma.
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  • 15
    ISSN: 1436-2813
    Keywords: Key Words: angiogenesis ; p53 ; apoptosis ; lung adenocarcinoma ; hematogenous metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: n = 7) and nonsurviving (metastasis) group (n = 8). We studied the medical charts, operative records, pathologic reports, and tumor specimens taken at surgical resection. We measured the apoptotic index using the ApopTag kit and the intratumoral microvessel count using an anti-CD34 monoclonal antibody. In addition, immunohistochemical staining for the expression of p53 was conducted simultaneously. The clinicopathological characteristics, including age, sex, tumor size (pT), and histological differentiation, were not significantly different between the surviving and the nonsurviving group. The microvessel count was significantly higher in nonsurviving group than in the surviving group. The apoptotic index and the expression of p53 was not significantly different between the two groups. An inverse correlation between the apoptotic index and microvessel count, and a positive correlation between the expression of p53 and microvessel count, were observed. Angiogenesis may be an important prognostic factor in patients with stage I lung adenocarcinoma.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Nutrient cycling in agroecosystems 53 (1999), S. 209-218 
    ISSN: 1573-0867
    Keywords: cattle slurry ; fertilizer splitting ; nitrogen ; recovery ; residual nitrogen ; Zea mays L
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The recovery of soil mineral nitrogen (N) by crops, and its subsequent utilisation for dry matter (DM) production may be increased when the application of N is postponed until after crop emergence. The significance of this strategy for silage maize was studied in nine field experiments on Dutch sandy soils from 1983 to 1988. In five experiments the effect of slurry applied before planting at a rate of circa 66 m3 ha-1, was compared to the effect of a similar rate of which half was applied before planting and half at the 4–6 leaf stage. In the 4-6 leaf stage slurry was either injected or banded. In four other experiments the effect of mineral fertilizer-N splitting was studied. In these experiments, 30 m3 ha-1 cattle slurry, applied before planting, was supplemented with mineral fertilizer-N at rates ranging from 40 to 160 kg ha-1, either fully applied before crop emergence or split. When split, 40 kg ha-1 of the mineral fertilizer-N rate was banded at the 4–6 leaf stage. According to balance sheet calculations, substantial losses of slurry N and mineral fertilizer-N occurred during the growing season. Losses were compensated for, however, by apparent mineralization, ranging from 0.34 to 0.77 kg N ha-1 day-1. Split applications of cattle slurry had a significant positive effect on the DM yield in two out of five experiments compared to the conventional non-split application, but only when the post-emergence slurry application was banded which is no longer in accordance with present legislation. Split applications of mineral fertilizer-N had a significant positive effect in one experiment where rainfall was excessive but not in the others. The results provide insufficient evidence to recommend farmers to split applications. Soil mineral N sampling at the 4–6 leaf stage should hence be considered a control on the appropriateness of early N applications after exceptional weather conditions rather than a routine observation on which the post-emergence N dressing is to be based in a deliberate splitting strategy. Our data suggest that the financial return of a 40 kg ha-1 supplementation with mineral fertilizer-N, was questionable when more than 175 kg N ha-1 were found in the upper 0.6 m soil layer at the 4–6 leaf stage.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Nutrient cycling in agroecosystems 53 (1999), S. 259-267 
    ISSN: 1573-0867
    Keywords: farming systems ; nitrogen ; nitrogen budgets ; sustainability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Three approaches to nitrogen budgeting were developed and their ability to quantitatively describe nitrogen cycling in a fertilizer based and a grass–clover based beef system tested. Budgets ranged in complexity from the Economic Input:Output (EIO) budget, which accounted simply for purchases and sales of nitrogen over the farmgate, through the Biological Input:Output (BIO) budget, which included estimates of biological nitrogen fixation and attempted to partition losses into leaching and gaseous forms, to the Transfer:Recycle:Input:Output (TRIO) budget, which also accounted for key soil processes. Nitrogen unaccounted for in the fertilized system decreased with increasing budget complexity (285, 212 and 188 kg ha-1 yr-1 unaccounted for by the EIO, BIO and TRIO budgets, respectively). In the legume based grass–clover system, the EIO budget did not accurately describe total nitrogen inputs as it did not include 146 kg ha-1 yr-1 from symbiotic nitrogen fixation. In the grass–clover system, nitrogen unaccounted for was again greater using the BIO than the TRIO budget (103 and 79 kg ha-1 yr-1, respectively). In conclusion, the most complex budgeting approach (TRIO) was able to account for the fate of a greater proportion of nitrogen inputs than the simpler approaches. However, the perceived success of the different approaches was strongly dependent on the precise objective.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Nutrient cycling in agroecosystems 55 (1999), S. 1-6 
    ISSN: 1573-0867
    Keywords: chlorophyll meter ; irrigation ; LAI ; nitrogen ; Spring Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A Spring Wheat crop was grown using three irrigation levels and nitrogen rate applications to evaluate chlorophyll meter measurements as a possible nitrogen nutrition index for modelling. These measurements yielded the most reliable indications at Zadoks GS45. The lower limit indicating severe nitrogen deficiency in the leaves was approximately 35 SPAD units while the upper limit of 45 SPAD units, indicated an excess consumption.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Nutrient cycling in agroecosystems 55 (1999), S. 89-94 
    ISSN: 1573-0867
    Keywords: algae ; flooded soils ; N cycling ; nitrogen ; 15N ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Algal N labelled with 15N added to a flooded soil in laboratory columns without plants was studied to determine the changes over time in the fate of N assimilated by algae and to study how its fate is affected by (a) exclusion of light simulating complete closure of the rice canopy, and (b) addition of fertilizer-NH4 *. In the light but with no added fertilizer-N there was little net mineralization of the added algal N during the first 4 weeks, but after 8 weeks 42% had been mineralized, of which 95% was denitrified. Exclusion of light caused net mineralization to proceed more rapidly in the first 4 weeks due to the death of algal cells and lowered reassimilation. After 8 weeks 51% had been mineralized, of which 54% was denitrified, 16% volatilized and 30% was present as KCl exchangeable NH4 +-N. Application of fertilizer-NH4 + apparently caused mineralization of 25% of the algal N within one week but the results were probably affected by pool substitution in which labelled N mineralized to NH4 +-N was diluted with fertilizer – NH+ 4 and then immobilized leaving more labelled NH4–N in the mineral pool. After 8 weeks, 42% of algal N had been mineralized, of which 69% was estimated to have been denitrified, 19% lost through NH3 volatilization and 12% remained as extracted NH4 ++NO- 3. Uptake of N by a rice crop would reduce the gaseous losses. Algal N was mineralized quickly enough to be available during the growing season of a rice crop and, depending on field conditions, algae may have a role in assimilating N and protecting it from loss as well as being a major driving force for NH3 volatilization through diurnal increases in pH.
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  • 20
    ISSN: 1572-9702
    Keywords: tomato plants ; susceptibility ; mite ; tridecan-2-one ; nitrogen ; potasium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The objective of this work was to study the effect of NK fertilization levels and canopy height on the resistance of Lycopersicon hirsutum and Lycopersicon esculentum to Aculops lycopersici (Acari: Eriophydae). The effects of NK fertilization levels and canopy height in the leaf size and density of trichomes and their effects on tridecan-2-one (2-TD) and undecan-2-one (2-UD) limiting the attack of A. lycopersici on tomato plants were assessed. Different NK fertilization levels had no effect on the resistance of L. hirsutum to A. lycopersici. No significant differences were found in attack rates of this mite on leaves of the top and median parts of L. hirsutum canopy. The type and density of trichomes were the main determining factor of A. lycopersici attack on tomato plants. High trichome densities and type VI glandular trichomes which produce tridecan-2-one are important resistance factors on tomato plants. L. hirsutum showed a high resistance level to A. lycopersici due to high densities of type VI glandular trichomes and consequently higher levels of tridecan-2-one in its leaves.
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  • 21
    Electronic Resource
    Electronic Resource
    Springer
    Diabetologia 42 (1999), S. 711-718 
    ISSN: 1432-0428
    Keywords: Keywords Malnutrition ; ageing ; beta-cell mass ; apoptosis ; glucose tolerance.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Aims/hypothesis. In a recently developed rat model, maternal food restriction from day 15 of pregnancy until weaning induced low birth weight and a 70 % reduction of beta-cell mass in the offspring at day 21 after birth. Subsequent renutrition from weaning was insufficient to fully restore beta-cell mass in young adult rats. The aim of this study is to investigate the long-term consequences of early malnutrition on beta-cell mass and function. Methods. Oral glucose tolerance tests were done in 3- and 12-month-old animals and beta-cell mass and apoptosis were determined by morphometrical measurements on pancreatic sections. The specific impact of postnatal malnutrition was studied by comparing control animals (C group) with animals malnourished during their fetal life only (R/C group), and animals malnourished during fetal life and until weaning (R group). Results. In 3-month-old R/C animals beta-cell mass reached 8.0 ± 1.5 mg with no further increase until 12 months (8.1 ± 1.5 mg), compared with 9.3 ± 1.9 mg in control rats. Twelve-month-old R/C animals showed normal plasma insulin responses and borderline glucose tolerance. In R animals, apoptosis reached 1.9 ± 0.4 % of the beta cells at 3 months, compared with 0.7 ± 0.5 % in control rats, and beta-cell mass did not increase between 3 and 12 months (4.7 ± 0.8 mg at 12 months). In aged control and R animals, apoptosis affected 8 % of the beta cells. At 12 months only, R animals showed profound insulinopenia and marked glucose intolerance. Conclusion/interpretation. In conclusion, perinatal malnutrition profoundly impairs the programming of beta-cell development. In animals with decreased beta-cell mass the additional demand placed by ageing on the beta cells entails glucose intolerance since beta-cell mass does not expand and apoptosis is increased. [Diabetologia (1999) 42: 711–718]
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  • 22
    ISSN: 1432-0428
    Keywords: Keywords Type I diabetes ; interferon-γ ; transgenic mice ; apoptosis ; insulin secretion ; tumour necrosis factor.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Aims/hypothesis. To examine whether interferon-γ destroys islet beta cells directly or indirectly through lymphocyte activation, or whether direct action of interferon-γ on beta cells by itself induces diabetes without insulitis. Methods. To avoid possible nonspecific breakdown of beta cells by transgenic overexpression of interferon-γ by the insulin promoter, we generated transgenic mice expressing interferon-γ under the control of rat glucagon promoter (RGP-IFN-γ-Tg mice). Results. The absence of insulitis in RGP-IFN-γ-Tg mice enabled us to investigate the direct effects of paracrine interferon-γ. In RGP-IFN-γ-Tg mice, serum concentrations of interferon-γ and tumour necrosis factor-α (TNF-α) were 50 and 6 times higher than those in their littermates, respectively, and glucose-responsive insulin secretion decreased to one-half the level of that in the littermates. Transgenic interferon-γ induced remodelling of beta cells where apoptosis of many beta cells was compensated by their vigorous regeneration and diabetes did not occur in most of the RGP-IFN-γ-Tg mice. Conclusion/interpretation. Interferon-γ alone is insufficient for the complete destruction of beta cells in vivo, and factors other than interferon-γ including activated lymphocytes or other cytokines, are necessary in addition to interferon-γ for the development of Type I (insulin-dependent) diabetes mellitus. [Diabetologia (1999) 42: 566–573]
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  • 23
    ISSN: 1432-0428
    Keywords: Keywords Nicotinamide ; cytokine ; islet ; insulin ; apoptosis ; diabetes.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Nicotinamide intervention trials are presently undertaken to prevent Type I (insulin-dependent) diabetes in high risk subjects. They are based on studies in rodents reporting nicotinamide protection against beta-cell injury in vitro and in vivo. This study examines whether nicotinamide can protect human beta cells in vitro. At concentrations (2 and 5 mmol/l) to protect rat beta cells against necrosis by streptozotocin or hydrogen peroxide, nicotinamide prevents hydrogen peroxide-induced necrosis of human beta cells. As with rat beta cells, nicotinamide fails to protect human beta cells against apoptosis induced by a combination of the cytokines interleukin-1β , interferon-γ and tumour necrosis factor-α. In rat beta cells, nicotinamide (2 to 20 mmol/l) was also found to induce apoptosis, in particular during the days following its protection against necrosis; this cytotoxic effect was not observed with human beta cells. These data demonstrate that nicotinamide can protect human beta cells against radical-induced necrosis, but not against cytokine-induced apoptosis. This effect is not associated with a delayed apoptosis as in rat beta cells. [Diabetologia (1999) 42: 55–59]
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  • 24
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    Annals of oncology 10 (1999), S. 139-150 
    ISSN: 1569-8041
    Keywords: apoptosis ; chemotherapy ; clinical trials ; gene therapy ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nearly twenty years after the initial discovery of p53, we are now in an ideal position to exploit our vast knowledge of p53 biology in the creation of novel cancer therapies. Disruption of p53 function through mutation, or other means, occurs very frequently in human cancer. Loss of p53 function has been linked with unfavourable prognosis in a large number of tumour types, as indicated by more aggressive tumours, early metastasis and decreased survival rates. Many different avenues of research have converged upon p53 to highlight this protein as being one of the foremost cellular responders to stress, in particular to DNA damage. Huge advances have been made in understanding the complex role p53 plays in the regulation of apoptosis and cell cycle arrest. This review is not meant to be a comprehensive description of p53 biology, but rather serves to highlight current progress in the development of p53- oriented cancer therapies. These may be categorised into three basic strategies: gene replacement therapy using wild-type p53, restoration of p53 function by other means and, finally, targeting of the p53 dysfunction itself. Rapid progress is expected to be made regarding the identification of conventional pharmaceutical agents which either work in a p53-independent manner or act preferentially in p53 defective cells. Gene replacement therapy with wild-type p53 also holds considerable potential for obtaining clinically relevant results quickly. The other forms of cancer therapies based around p53 are much further behind in the developmental process, but may prove to more efficacious in the long run, especially in terms of specificity. As with many other fields, the innovation of successful p53-oriented cancer therapies is only limited by our understanding of p53 biology and the creative use of such knowledge.
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  • 25
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    Annals of oncology 10 (1999), S. 122-126 
    ISSN: 1569-8041
    Keywords: apoptosis ; biliary carcinogenesis ; cholangiocarcinoma ; genotoxicity ; risk factors ; therapeutic strategies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Carcinomas of the biliary tract are rare cancers developing from the epithelial or blast-like cells lining the bile ducts. A variety of known predisposing factors and recent experimental models of biliary carcinogenesis (e.g., infection with the liver fluke Opisthorchis viverrini, models of chemically induced carcinogenesis and experimental models of pancreaticobiliary maljunction) have elucidated different stages of this complex system of biliary tumorigenesis. Chronic inflammatory processes, generation of active oxygen radicals, altered cellular detoxification mechanisms, activation of oncogenes, functional loss of tumor-suppressor genes and dysregulation of cell proliferation and cell apoptotic mechanisms have been identified as important contributors in the development of cholangiocarcinomas. In this review, the known mechanisms involved in the carcinogenesis of biliary epithelium are addressed. We will divide the topic into four stages: 1) Predisposition and risk factors of biliary cancer, 2) Genotoxic events and alterations leading to specific DNA damage and mutation patterns. 3) Dysregulation of DNA repair mechanisms and apoptosis, permitting survival of mutated cells and 4) Morphological evolution from premalignant biliary lesions to cholangiocarcinoma. Finally, established and hypothetical future therapeutic strategies directed towards specific pathogenetic events during biliary carcinogenesis will be addressed.
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  • 26
    ISSN: 1569-8041
    Keywords: acute myelogenous leukemia ; apoptosis ; ara-CTP ; cytosine arabinoside ; HL-60 ; retinoic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Accumulation of the cytosine arabinoside (ara-C) metabolite ara-C-triphosphate (ara-CTP) in leukemic blast cells is considered to be the main determinant of ara-C cytotoxicity in vitro and in vivo. Retinoids such as all-trans-retinoic acid (ATRA) have been shown to increase the sensitivity of acute myelogenous leukemic (AML) blast cells to ara-C. To investigate the mechanism of this sensitisation, the hypothesis was tested that ATRA augments cellular ara-CTP levels in human-derived myelogenous leukemia HL-60 cells. Materials and methods: The effect of ATRA and 13-cis-retinoic acid on ara-CTP accumulation and ara-C-induced apoptosis was studied. Ara-CTP levels were measured by high-performance liquid chromatography (HPLC), cytotoxicity by the tetrazolium (MTT) assay, and apoptosis by occurrence of DNA fragmentation (gel electrophoresis), cell shrinkage and DNA loss (flow cytometry). Results: Pretreatment of HL-60 cells with ATRA (0.01–1 µM) caused a significant decrease in intracellular ara-CTP levels; e.g., incubation for 72 hours with ATRA 1 µM prior to one hour ara-C 10 µM reduced ara-CTP levels to 41% ± 4% of control. Similar results were obtained after preincubation with 13-cis-retinoic acid. In spite of decreased ara-CTP levels, the cytotoxicity of the combination was supraadditive and ATRA augmented ara-C-induced apoptosis. Conclusions: At therapeutically relevant concentrations ATRA increased ara-C cytotoxicity and ara-C induced apoptosis but this augmentation is not the corollary of elevated ara-CTP levels. The feasibility of ara-C treatment optimisation via strategies other than those involving elevation of ara-CTP levels should be investigated further.
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  • 27
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    Molecular and cellular biochemistry 196 (1999), S. 13-21 
    ISSN: 1573-4919
    Keywords: apoptosis ; DNA fragmentation ; GSHPx-1 knockout mice ; GSHPx-1 transgenic mice ; ischemia/repurfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Apoptosis, a genetically controlled programmed cell death, has been found to play a role in ischemic reperfusion injury in several animal species including rats and rabbits. To examine whether this is also true for other animals, an isolated perfused mouse heart was subjected to 30 min of ischemia followed by 2 h of reperfusion. Experiments were terminated before ischemia (baseline), after ischemia, and at 30, 60, 90 and 120 min of reperfusion. At the end of each experiment, hearts were processed for the evaluation of apoptosis and DNA laddering. The in situ end labeling (ISEL) technique was used to detect apoptotic cardiomyocyte nuclei while DNA laddering was evaluated by subjecting the DNA obtained from the cardiomyocytes to 1.8% agarose gel electrophoresis followed by photographing under UV illumination. The results of our study revealed that apoptotic cells appear only after 60 min of reperfusion as demonstrated by the intense fluorescence of the immunostained genomic DNA when observed under fluorescence microscopy. None of the ischemic hearts showed any evidence of apoptosis. These results were corroborated with the findings of DNA fragmentation showing increased ladders of DNA bands in the same reperfused hearts representing integer multiples of the internucleosomal DNA length (about 180 bp). Since our previous studies showed a role of glutathione peroxidase (GSHPx) in apoptotic cell death, we performed identical experiments using isolated hearts from GSHPx-l knockout mice and transgenic mice overexpressing GSHPx-l. GSHPx-l knockout mice showed evidence of apoptotic cell death even after 30 min of reperfusion. Significant number of apoptotic cells were found in the cardiomyocytes as compared to non-transgenic control animals. To the contrary, very few apoptotic cells were found in the hearts of the transgenic mice overexpressing GSHPx-l. Hearts of GSHPx-l knockout mice were more susceptible to ischemia/reperfusion injury while transgenic mice overexpressing GSHPx- 1 were less susceptible to ischemia reperfusion injury compared to non-transgenic control animals. The results of this study clearly demonstrate a role of GSHPx in ischemia/reperfusion-induced apoptosis in mouse heart.
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  • 28
    ISSN: 1573-4919
    Keywords: MKP-1 ; Fas ligand ; Fas ; apoptosis ; prostate cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Recent studies have suggested that MAP kinase phosphatase 1 (MKP-1) is overexpressed in prostate cancer. To evaluate the role of MKP-1 in regulating cell death and tumor growth in prostate cancer, MKP-1 was conditionally overexpressed in the human prostate cancer cell line DU145. Overexpression of MKP-1 in DU145 cells blocked activation of stress-activated protein kinase (SAPK/JNK). MKP-1 overexpression in DU-145 cells was also found to inhibit Fas ligand (FasL)-induced apoptosis, as well as block the activation of caspases by Fas engagement. In addition, MKP-1 blocked the activation of apoptosis by transfected MEKK-1 and ASK-1, presumably through its inhibition of the SAPK/JNK family of enzymes. MKP-1 blocked the ability of FasL to induce loss of mitochondrial transmembrane potential (Δγm), suggesting that MKP-1 acts upstream of mitochondrial pro-apoptotic events induced by FasL and that the SAPK/JNK pathway may form the signaling link between Fas receptor and mitochondrial dysfunction. Thus, MKP-1 overexpression in prostate cancer may play a role in promoting prostate carcinogenesis by inhibiting FasL-induced cell death.
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  • 29
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    Molecular and cellular biochemistry 199 (1999), S. 125-137 
    ISSN: 1573-4919
    Keywords: apoptosis ; ADP-ribosylation ; caspases ; PARP ; PARG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribosylation) is a post-translational modification playing a relevant role in DNA damage recovery, DNA replication and viral integration. Several reports also suggest a modulation of this process during cell death by apoptosis. The aim of this review is to discuss the possible involvement of poly(ADP-ribosylation) during apoptosis, by dealing with general considerations on apoptosis, and further examining the correlation between NAD consumption and cell death, the regulation of poly(ADP-ribose) metabolism in apoptotic cells, the effect of poly(ADP-ribose) polymerase inhibition on cell death occurrence and the use of enzyme cleavage as a marker of apoptosis. Finally, the future prospects of the research in this area will be addressed.
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  • 30
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    Journal of materials synthesis and processing 7 (1999), S. 311-319 
    ISSN: 1573-4870
    Keywords: Iron ; carbon ; nitrogen ; microstructure ; characterization ; HIP-drip
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract A new processing technique makes nitrogen alloying possible by adding nitrogen under elevated nitrogen pressure to prealloyed Fe-C ingots during continuous casting, producing a whole new class of precipitation-free, iron–carbon–nitrogen alloys. When both carbon and nitrogen bulk concentration levels exceeded 0.5 wt%, a duplex fcc-/(bcc-bct-) Fe microstructure resulted that is iron carbide- and nitride-free. With increasing carbon and nitrogen concentrations, there was an increase in the retained fcc-Fe phase. In cooling rate studies, increasing carbon and nitrogen concentrations shifted the knee of the fcc-Fe-to-bcc-Fe phase time–temperature–transformation (T–T–T) curve to longer times. Hardness, compression strength, and wear resistance increased with increasing carbon and nitrogen concentrations and were superior to iron–carbon alloys without the nitrogen addition.
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  • 31
    ISSN: 1573-4919
    Keywords: DNA binding protein ; NAD metabolism ; cellular response to DNA damage ; γ-rays ; alkylating agents ; genomic instability ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract A dual approach to the study of poly (ADP-ribose)polymerase (PARP) in terms of its structure and function has been developed in our laboratory. Random mutagenesis of the DNA binding domain and catalytic domain of the human PARP, has allowed us to identify residues that are crucial for its enzymatic activity. In parallel PARP knock-out mice were generated by inactivation of both alleles by gene targeting. We showed that: (i) they are exquisitely sensitive to γ-irradiation, (ii) they died rapidly from acute radiation toxicity to the small intestine, (iii) they displayed a high genomic instability to γ-irradiation and MNU injection and, (iv) bone marrow cells rapidly underwent apoptosis following MNU treatment, demonstrating that PARP is a survival factor playing an essential and positive role during DNA damage recovery and survival.
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  • 32
    ISSN: 1573-4919
    Keywords: PARP ; poly(ADP-ribosyl)ation ; apoptosis ; DNA replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have focused on the roles of PARP and poly(ADP-ribosyl)ation early in apoptosis, as well as during the early stages of differentiation-linked DNA replication. In both nuclear processes, a transient burst of PAR synthesis and PARP expression occurs early, prior to internucleosomal DNA cleavage before commitment to apoptosis as well as at the round of DNA replication prior to the onset of terminal differentiation. In intact human osteosarcoma cells undergoing spontaneous apoptosis, both PARP and PAR decreased after this early peak, concomitant with the inactivation and cleavage of PARP by caspase-3 and the onset of substantial DNA and nuclear fragmentation. Whereas 3T3-L1, osteosarcoma cells, and immortalized PARP +/+ fibroblasts exhibited this early burst of PAR synthesis during Fas-mediated apoptosis, neither PARP-depleted 3T3-L1 PARP-antisense cells nor PARP -/- fibroblasts showed this response. Consequently, whereas control cells progressed into apoptosis, as indicated by induction of caspase-3-like PARP-cleavage activity, PARP-antisense cells and PARP -/- fibroblasts did not, indicating a requirement for PARP and poly(ADP-ribosyl)ation of nuclear proteins at an early reversible stage of apoptosis. In parallel experiments, a transient increase in PARP expression and activity were also noted in 3T3-L1 preadipocytes 24 h after induction of differentiation, a stage at which ~95% of the cells were in S-phase, but not in PARP-depleted antisense cells, which were consequently unable to complete the round of DNA replication required for differentiation. PARP, a component of the multiprotein DNA replication complex (MRC) that catalyzes viral DNA replication in vitro, poly(ADP-ribosyl)ates 15 of ~40 MRC proteins, including DNA pol α, DNA topo I, and PCNA. Depletion of endogenous PARP by antisense RNA expression in 3T3-L1 cells results in MRCs devoid of any DNA pol α and DNA pol δ activities. Surprisingly, there was no new expression of PCNA and DNA pol α, as well as the transcription factor E2F-1 in PARP-antisense cells during entry into S-phase, suggesting that PARP may play a role in the expression of these proteins, perhaps by interacting with a site in the promoters for these genes.
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  • 33
    ISSN: 1573-4919
    Keywords: breast cancer cells ; anti-apoptotic genes ; apoptosis ; progesterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Progesterone inhibits the proliferation of normal breast epithelial cells in vivo, as well as breast cancer cells in vitro. But the biologic mechanism of this inhibition remains to be determined. We explored the possibility that an antiproliferative activity of progesterone in breast cancer cell lines is due to its ability to induce apoptosis. Since p53, bcl-2 and survivin genetically control the apoptotic process, we investigated whether or not these genes could be involved in the progesterone-induced apoptosis. We found a maximal 90% inhibition of cell proliferation with T47-D breast cancer cells after exposure to 10 μM progesterone for 72 h. Control progesterone receptor negative MDA-231 cancer cells were unresponsive to 10 μM progesterone. The earliest sign of apoptosis is translocation of phosphatidylserine from the inner to the outer leaflet of the plasma membrane and can be monitored by the calcium-dependent binding of annexin V in conjunction with flow cytometry. After 24 h of exposure to 10 μM progesterone, cytofluorometric analysis of T47-D breast cancer cells indicated 43% were annexin V-positive and had undergone apoptosis and no cells showed signs of cellular necrosis (propidium iodide negative). After 72 h of exposure to 10 μM progesterone, 48% of the cells had undergone apoptosis and 40% were annexin V positive/propidium iodide positive indicating signs of necrosis. Control untreated cancer cells did not undergo apoptosis. Evidence proving apoptosis was also demonstrated by fragmentation of nuclear DNA into multiples of oligonucleosomal fragments. After 24 h of exposure of T47-D cells to either 1 or 10 μM progesterone, we observed a marked down-regulation of protooncogene bcl-2 protein and mRNA levels. mRNA levels of survivin and the metastatic variant CD44 v7-v10 were also downregulated. Progesterone increased p53 mRNA levels. These results demonstrate that progesterone at relative high physiological concentrations, but comparable to those seen in plasma during the third trimester of human pregnancy, exhibited a strong antiproliferative effect on breast cancer cells and induced apoptosis.
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  • 34
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    Molecular and cellular biochemistry 200 (1999), S. 51-57 
    ISSN: 1573-4919
    Keywords: smokeless tobacco ; apoptosis ; nitric oxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Smokeless tobacco usage is, a growing public health concern in the United States. Lesions of the oral cavity have been clearly linked to smokeless tobacco use. The objective of this study was to determine the biochemical effects of smokeless tobacco extract (STE) exposure upon hamster cheek pouch cell (HCPC-1) cultures. HCPC-1 cells were exposed to a 5 -fold dose-range of STE (0.5, 1.0 and 2.5%) over a time-course of 24-96 h. Following each exposure we measured various biochemical parameters of cell proliferation and cell death. Cell viability, cell cycle progression and S-phase DNA synthesis were measured as markers of cell proliferation. We measured lactate dehydrogenase leakage as a marker of cell membrane damage and cell death due to necrosis. No significant alterations were observed in cell cycle progression and cell proliferation as a result of exposure to STE. LDH measured colorimetrically indicated no significant effect with the lower doses (0.5, 1.0 and 2.5% STE). Apoptosis measured as the A0 peak and by the TUNEL procedure revealed that STE caused significant rates of apoptosis. Maximal apoptosis was noted between 48-96 h. In order to probe the mechanism further we measured the levels of nitrites as an indicator of nitric oxide (NO) in the media. NO levels were significantly elevated at the doses that caused an induction of apoptosis. The results from this study indicate that STE causes a dose-dependent induction of apoptosis and that this is mediated by nitric oxide.
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  • 35
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    Molecular and cellular biochemistry 193 (1999), S. 37-42 
    ISSN: 1573-4919
    Keywords: Rho ; GTPase ; toxins ; Clostridium ; signal transduction ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The Rho family small GTPases are members of the Ras superfamily of small GTPases. Rho proteins were first determined to act as key regulators of many types of actin cytoskeletal-dependent cellular functions. Recent work by several investigators indicates that Rho GTPases are also critical modulators of several important intracellular and nuclear signal transduction pathways. Certain clostridial toxins and exoenzymes covalently modify, and thereby inactivate, specific types of Rho family GTPases. As such, these microbial enzymes have proven invaluable in helping to identify structural and functional attributes of Rho GTPases.
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  • 36
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    Molecular and cellular biochemistry 193 (1999), S. 103-108 
    ISSN: 1573-4919
    Keywords: Poly(ADP-ribose) polymerase ; Drosophila melanogaster ; alternative splicing ; apoptosis ; DNA repair ; development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribose) polymerase (PARP) is conserved in eukaryotes. To analyze the function of PARP, we isolated and characterized the gene for PARP in Drosophila melanogaster. The PARP gene consisted of six translatable exons and spanned more than 50 kb. The DNA binding domain is encoded by exons 1-4. Although the consensus cleavage site of CED-3 like protease during apoptosis is conserved from human to Xenopus laevis PARPs, it is neither conserved in the corresponding region of Drosophila nor Sarcophaga peregrina. There are two cDNAs species in Drosophila. One cDNA could encode the full length PARP protein (PARP I), while the other is a truncated cDNA which could encode a partial-length PARP protein (PARP II), which lacks the automodification domain and is possibly produced by alternative splicing. The expression of these two forms of PARP in E. coli demonstrated that while PARP II has the catalytic NAD-binding domain and DNA-binding domain it is enzymatically inactive. On the other hand PARP I is active. A deletion mutant of PARP gene could grow to the end of embryogenesis but did not grow to the adult fly. These results suggest that the PARP gene plays an important function during the development of Drosophila.
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  • 37
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    Molecular and cellular biochemistry 193 (1999), S. 119-125 
    ISSN: 1573-4919
    Keywords: benzamides ; nicotinamides ; apoptosis ; inflammation ; NF-kB ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Our laboratory has concentrated on the possible regulation the benzamides and nicotinamides may have on the processes of DNA repair and apoptosis. Recent reports [14-16] have suggested that both apoptosis and inflammation are regulated by the transcription factor NF-kB. We have initiated studies regarding the hypothesis that the benzamides and nicotinamides could inhibit the production of tumor necrosis factor alpha (TNFalpha) and the inflammatory response as well as induce apoptosis via inhibition of NF-kB. Our data have shown that nicotinamide and two N-substituted benzamides, metoclopramide (MCA) and 3-chloroprocainamide (3-CPA), gave dose dependent inhibition of lipopolysacharide induced TNFalpha in the mouse within the dose range of 10-500 mg/kg. Moreover, lung edema was prevented in the rat by 3 ï 50 mg/kg doses of 3-CPA or MCA, and 100-200 μM doses of MCA could also inhibit NF-kB in Hela cells. Taken together these data strongly support the notion that benzamides and nicotinamides have potent anti-inflammatory and antitumor properties, because their primary mechanism of action is regulated by inhibition at the gene transcription level of NF-kB, which in turn inhibits TNFalpha and induces apoptosis.
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  • 38
    ISSN: 1573-4919
    Keywords: antisense oligonucleotide ; apoptosis ; cAMP-dependent protein kinase ; cancer cells ; growth inhibition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The enhanced expression of the RIα subunit of cyclic AMP-dependent protein kinase type 1 (PKA-I) has been correlated with cancer cell growth. We have investigated the effects of sequence-specific inhibition of RIα gene expression on the growth of MCF-7 human breast cancer cells. We report that RIα antisense treatment results in a reduction in RIα expression at both mRNA and protein levels and inhibition of cell growth. The growth inhibition was accompanied by changes in cell morphology, cleavage of poly(ADP-ribose) polymerase (PARP) and appearance of apoptotic nuclei. In addition, bcl-2 protein level was reduced and p53 expression increased in growth arrested cells. Interestingly, RIα antisense inhibited cell viability and induced apoptosis in the absence of p53, suggesting that these actions of RIα antisense are exerted independent of p53. In contrast, two- and four-base mismatched control oligonucleotides had no effect on either cell growth or morphology. These results demonstrate that the RIα antisense, which efficiently depletes the growth stimulatory molecule RIα, induces cell differentiation and apoptosis, providing a new approach to combat breast cancer cell growth.
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  • 39
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    Molecular and cellular biochemistry 197 (1999), S. 97-108 
    ISSN: 1573-4919
    Keywords: neutrophil ; PKC ; TNF-α ; apoptosis ; DNA fragmentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In the present study we investigated the TNF-α induced signal transduction mechanism in human neutrophil. Exogenously added TNF-α affects both PKC activity and its translocation from cytosol to the membrane. Endogenous protein phosphorylation pattern is inhibited in TNF-α induced neutrophil in Ca-dependent and Ca-independent manner, including a major 47 and 66 kDa cytosolic proteins, which may be implicated in superoxide anion generation. However TNF-α dose dependently enhances the expression of ζ-PKC isotype but not the β-PKC. Morphology and cell cytotoxicity are studied in TNF-α treated neutrophil to understand the TNF-α induced cell death or apoptosis and these experiment is further confirmed by DNA fragmentation analysis. These results clearly demonstrate that TNF-α induces cellular death of human neutrophil at least in part by enhanced expression of Ca-independent ζ-PKC. These observations provide an insight towards understanding the function of ζ-PKC in apoptotic pathway.
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  • 40
    ISSN: 1573-4919
    Keywords: rotenone ; apoptosis ; oncogenes ; liver cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Rotenone decreases the incidence of hepatocellular carcinoma and lowers rates of hepatocellular proliferation. In an effort to delineate mechanisms involved, the in vivo effect of rotenone on liver mitochondrial metabolism, apoptotic machinery as well as elements of the hepatic signal transduction pathways were investigated. Mitochondria from livers of male B6C3F1 mice fed a standard diet containing 600 ppm rotenone for 7 days were uncoupled or inhibited when succinate or glutamate plus malate were used as the substrate, respectively. These livers also showed a significant increase in apoptosis compared with control livers. Furthermore, rotenone increased the expression of c-myc mRNA to 5-fold of control values within 3 days, an effect which was still observed (3-fold) after 7 days. Levels of p53 mRNA were also increased 3-fold after 1 day, but declined to control levels by 7 days. Rotenone also caused a transient, yet marked increase in liver particulate glyceraldehyde phosphate dehydrogenase (GAPDH) protein expression, while it did not alter the expression of the cytosolic form of the enzyme. Conversely, mRNA of the proto-oncogene H-ras showed a decline of 35% after 3 days of rotenone treatment, and remained diminished for the duration of the experiment. These data suggest that rotenone may act as an anticancer agent by diminishing mitochondrial bioenergetics which prevents basal hepatocyte proliferation and lowers the threshold for liver cells with DNA damage to undergo apoptosis.
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  • 41
    ISSN: 1573-4943
    Keywords: Two-dimensional electrophoresis ; MALDI-MS ; apoptosis ; RNA polymerase B transcription factor 3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Apoptosis or programmed cell death is essential in the process of controlling lymphocyte growth and selection. We identified RNA polymerase B transcription factor 3 (BTF3), which is associated with anti-IgM antibody-mediated apoptosis, using a subclone of the human Burkitt lymphoma cell line BL60. To identify the transcription factor BTF3, which is expressed only in minor amounts, we used preparative high-resolution two-dimensional gel electrophoresis (2DE) employing carrier ampholytes for isoelectric focusing. Comparison of the 2DE protein patterns from apoptotic and nonapoptotic cells showed BTF3 as a predominantly altered protein spot. The characterization of the differentially expressed transcription factor and 13 marker proteins described in this study were performed by internal Edman microsequencing and/or by peptide mass fingerprinting using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The proteome analysis was significantly improved by performing the newly developed preparative high-resolution two-dimensional gels employing high protein concentrations.
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  • 42
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    Potato research 42 (1999), S. 559-568 
    ISSN: 1871-4528
    Keywords: plastic mulch ; transplanting ; nitrogen ; planting depth ; seed tuber weight ; physiological age ; radiation conversion efficiency ; harvest index
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In many seed potato producing areas, micro- and minitubers are too small for direct planting as seed tubers in the field. Such use of these propagules can, however, be feasible if the crop's growth and development can be advanced. Increasing light interception, harvest index and yield of useable progeny tubers has been proved possible with plastic mulch and pre-planting of small tubers in a greenhouse. High amounts of nitrogen (up to 180 kg ha−1) or deep planting (up to 9 cm) were less effective. Using older or pre-sprouted micro- or minitubers may be beneficial, because this might increase the number of sprouts per mother tuber (and thus stems per plant) or advance the growth of sprouts or stems. However, this would require even more careful management, due to the weakness of these sprouts and stems. Micro- and minitubers should be as large as feasible when used for direct planting in the field.
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  • 43
    ISSN: 1573-5125
    Keywords: denitrification ; nature restoration ; nitrogen ; phosphorus ; riparian areas ; sedimentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Denmark, as in many other European countries, the diffuse losses of nitrogen (N) and phosphorus (P) from the rural landscape are the major causes of surface water eutrophication and groundwater pollution. The export of total N and total P from the Gjern river basin amounted to 18.2 kg ha−1 and 0.63 kg P ha−1 during June 1994 to May 1995. Diffuse losses of N and P from agricultural areas were the main nutrient source in the river basin contributing 76% and 51%, respectively, of the total export. Investigations of nutrient cycling in the Gjern river basin have revealed the importance of permanent nutrient sinks (denitrification and overbank sedimentation) and temporary nutrient storage in watercourses. Temporary retention of N and P in the watercourses thus amounted to 7.2–16.1 g N m−2 yr−1 and 3.7–8.3 g P m−2 yr−1 during low-flow periods. Deposition of P on temporarily flooded riparian areas amounted from 0.16 to 6.50 g P m−2 during single irrigation and overbank flood events, whereas denitrification of nitrate amounted on average to 7.96 kg N yr−1 per running metre watercourse in a minerotrophic fen and 1.53 kg N yr−1 per linear metre watercourse in a wet meadow. On average, annual retention of N and P in 18 Danish shallow lakes amounted to 32.5 g N m−2 yr−1 and 0.30 g P m−2 yr−1, respectively, during the period 1989–1995. The results indicate that permanent nutrient sinks and temporary nutrient storage in river systems represent an important component of river basin nutrient budgets. Model estimates of the natural retention potential of the Gjern river basin revealed an increase from 38.8 to 81.4 tonnes yr−1 and that P-retention increased from −0.80 to 0.90 tonnes yr−1 following restoration of the water courses, riparian areas and a shallow lake. Catchment management measures such as nature restoration at the river basin scale can thus help to combat diffuse nutrient pollution.
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  • 44
    ISSN: 1573-5052
    Keywords: mineralization ; nitrogen ; phosphorus ; salinity ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant zonation patterns across New England salt marshes have been investigated for years, but how nutrient availability differs between zones has received little attention. We investigated how N availability, P availability, and plant N status varied across Juncus gerardii, Spartina patens, and mixed forb zones of a Northern New England high salt marsh. We also investigated relationships between several edaphic factors and community production and diversity across the high marsh. P availability, soil salinity, and soil moisture were higher in the mixed forb zone than in the two graminoid zones. NH+ 4-N availability was highest in the J. gerardii zone, but NO− 3-N availability and mid season net N mineralization rates did not vary among zones. Plant tissue N concentrations were highest in the mixed forb zone and lowest in the S. patens zone, reflecting plant physiologies more so than soil N availability. Community production was highest in the J. gerardii zone and was positively correlated with N availability and negatively correlated with soil moisture. Plant species diversity was highest in the mixed forb zone and was positively correlated with P availability and soil salinity. Thus, nutrient availability, plant N status, and plant species diversity varied across zones of this high marsh. Further investigation is needed to ascertain if soil nutrient availability influences or is a result of the production and diversity differences that exist between vegetation zones of New England high salt marshes.
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  • 45
    ISSN: 1573-5125
    Keywords: denitrification ; eutrophication ; estuary ; nitrogen ; sediment-water exchange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this review of sediment denitrification in estuaries and coastal ecosystems, we examine current denitrification measurement methodologies and the dominant biogeochemical controls on denitrification rates in coastal sediments. Integrated estimates of denitrification in coastal ecosystems are confounded by methodological difficulties, a lack of systematic understanding of the effects of changing environmental conditions, and inadequate attention to spatial and temporal variability to provide both seasonal and annual rates. Recent improvements in measurement techniques involving 15 N techniques and direct N2 concentration changes appear to provide realistic rates of sediment denitrification. Controlling factors in coastal systems include concentrations of water column NO 3 − , overall rates of sediment carbon metabolism, overlying water oxygen concentrations, the depth of oxygen penetration, and the presence/absence of aquatic vegetation and macrofauna. In systems experiencing environmental change, either degradation or improvement, the importance of denitrification can change. With the eutrophication of the Chesapeake Bay, the overall rates of denitrification relative to N loading terms have decreased, with factors such as loss of benthic habitat via anoxia and loss of submerged aquatic vegetation driving such effects.
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  • 46
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    Investigational new drugs 17 (1999), S. 227-240 
    ISSN: 1573-0646
    Keywords: apoptosis ; protein kinase C ; sphingoid bases ; safingol ; diglyceride ; bryostatin 1 ; staurosporine ; 7-hydroxy staurosporine (UCN-01) ; 4′-N-benzoyl staurosporine (CGP-41251) ; calphostin C (UCN-1028c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Neoplastic cell survival is governed by a balance between pro-apoptotic and anti-apoptotic signals. Noteworthy among several anti-apoptotic signaling elements is the protein kinase C (PKC) isoenzyme family, which mediates a central cytoprotective effect in the regulation of cell survival. Activation of PKC, and subsequent recruitment of numerous downstream elements such as the mitogen-activated protein kinase (MAPK) cascade, opposes initiation of the apoptotic cell death program by diverse cytotoxic stimuli. The understanding that the lethal actions of numerous antineoplastic agents are, in many instances, antagonized by cytoprotective signaling systems has been an important stimulus for the development of novel antineoplastic strategies. In this regard, inhibition of PKC, which has been shown to initiate apoptosis in a variety of malignant cell types, has recently been the focus of intense interest. Furthermore, there is accumulating evidence that selective targeting of PKC may prove useful in improving the therapeutic efficacy of established antineoplastic agents. Such chemosensitizing strategies can involve either (a) direct inhibition of PKC (e.g., following acute treatment with relatively specific inhibitors such as the synthetic sphingoid base analog safingol, or the novel staurosporine derivatives UCN-01 and CGP-41251) or (b) down-regulation (e.g., following chronic treatment with the non-tumor-promoting PKC activator bryostatin 1). In preclinical model systems, suppression of the cytoprotective function(s) of PKC potentiates the activity of cytotoxic agents (e.g., cytarabine) as well as ionizing radiation, and efforts to translate these findings into the clinical arena in humans are currently underway. Although the PKC-driven cytoprotective signaling systems affected by these treatments have not been definitively characterized, interference with PKC activity has been associated with loss of the mitogen-activated protein kinase (MAPK) response. Accordingly, recent pre-clinical studies have demonstrated that pharmacological disruption of the primary MEK-ERK module can mimic the chemopotentiating and radiopotentiating actions of PKC inhibition and/or down-regulation.
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  • 47
    ISSN: 1573-0646
    Keywords: UCN-01 ; IL-2 receptor ; Fas ; Fas-ligand ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract 7-hydroxystaurosporine (UCN-01) is a new anticancer agentwhich exerts an inhibitory effect on cell cycle check points andis currently under phase I clinical trials in US and Japan.Preliminary clinical data indicated that UCN-01 remained inplasma at high concentrations for long periods of time. Thisunavoidable high plasma drug exposure is likely to lead tohematological toxicities in patients. In the present study,cultured human peripheral blood lymphocytes (PBLs) were used toevaluate the possible hematological toxicities of UCN-01treatment. UCN-01 induces apoptosis, and the induction ofapoptosis-related surface markers were also examined toinvestigate the involvement of these molecules in UCN-01-inducedapoptosis in PBLs. in vitroviability of PBLs wasdecreased by high dose of UCN-01 (25 μM, 3-day exposure). Thiseffect of UCN-01 was significantly suppressed by the presence ofhuman serum, suggesting that some specific inhibitory factor(s)in human serum may antagonize the lympholytic effect of UCN-01.The percentage of annexin V-positive PI-negative cells increasedwith exposure to UCN-01 in a time- and dose-dependent manner; byup to 30.3% after exposure to 25 μM UCN-01 for 3 days.At the same time, the expression of both interleukin-2 receptor(IL-2R, CD25) and Fas (CD95), analyzed by flow cytometry, wasinduced. Con A-stimulated PBLs were more sensitive toUCN-01-induced apoptosis than non-stimulated lymphocytes andUCN-01 increased the sFas-L released into culture medium from conA-stimulated PBLs. Therefore, lymphocyte depletion mediated byactivation-induced apoptosis is likely to occur in patientstreated with UCN-01 at high doses.
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  • 48
    ISSN: 1573-0778
    Keywords: apoptosis ; Bcl-2 ; fixed-bed ; hollow fibre ; hybridoma ; perfusion ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.
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  • 49
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    Aquatic ecology 33 (1999), S. 55-64 
    ISSN: 1573-5125
    Keywords: estuaries ; nitrogen ; oxygen depletion ; phosphorus ; regression model ; vertical mixing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a detailed analysis of oxygen saturation in a shallow Danish estuary it was possible to separate the effect of meteorological forcings (i.e. wind and solar radiation) and nutrient loads on oxygen depletion in bottom water. Regression analysis showed that oxygen saturation tied to nitrogen load rather than to phosphorus load. During summer periods of stratification the oxygen saturation could be attributed to the time elapsed after the onset of stratification and the accumulated nitrogen loading 10 month prior to measurement. Using a 10-year meteorological database and an empirical model it was calculated that a 25% reduction in nitrogen loading would reduce the number of days with severe oxygen depletion (i.e. 〈15% of saturation) by more than 50%.
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  • 50
    ISSN: 1573-0778
    Keywords: antibody productivity ; apoptosis ; BAG-1 ; Bcl-2 ; cell survival ; hybridoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human bcl-2 and bag-1 DNA were introduced into mouse hybridoma 2E3- O cells and expressed. The expression of bcl-2 in BCMGneo-bcl2 transfectants was confirmed by ELISA and that of bag-1 in pZeo-bag1 was confirmed by western blotting. In batch cultures, the over-expression of bcl-2 prolonged the culture period by 2 days and co-expression of bcl-2 and bag-1 prolonged the culture period by 3 days. The delayed increase in the dead cell number in culture of the bcl-2 and bag-1 cotransfectant indicated the additional antiapoptosis effect of bcl-2 and bag-1 cotransfection in comparison with the bcl-2 only transfection. The bcl-2 transfectants (2E3O-Bcl2) produced antibody twofold per batch culture in comparison with 2E3-O cells transfected with BCMGSneo (2E3O-Mock). Enhancement of this MoAb production was due to the improved survival of the cells and was not due to stimulation of antibody production rate per cell by Bcl-2 expression. And the bcl-2 and bag-1 co-transfectant (2E3O-Bcl2-BAG1) produced antibody approximately fourfold of 2E3O-Mock per batch culture. Enhancement of this MoAb production was due to the improved survival of the cells and was partly due to stimulation of MoAb production rate per cell in the non-growing phase by the cotransfection. The method to engineer hybridoma cells genetically with bcl-2 and bag-1 for increasing viability and productivity would be widely applied for improving antibody productivity of hybridoma cultures.
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  • 51
    ISSN: 1573-0778
    Keywords: annexin V ; Apo-2 ligand ; apoptosis ; Cytostar-T® scintillating microplates ; flow cytometry ; lymphotoxin (LT)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The translocation of phospholipids across the plasma membrane has been widely documented as one of the earliest measurable biochemical events of apoptosis. Using fluorescently labelled annexin V, which preferentially binds phosphatidylserine (PS) in the presence of Ca2+, the externalization of PS can be measured and apoptosis quantified using flow cytometry. Conventional detection methods utilizing annexin V, while faster than in situ DNA end-labelling or DNA laddering, require extensive sample preparation which may compromise samples and makes rapid, high volume screening prohibitive. This paper describes a novel assay for the measurement of apoptosis based upon binding of radiolabelled annexin V to apoptotic cells attached to the growth surface of a 96-well scintillating microplate (Cytostar-T®). We compared measurements of apoptosis made by flow cytometry to those obtained with the scintillating microplate in three model systems, treatment of: mouse connective tissue (L-M) cells with lymphotoxin (LT), human lung carcinoma (H460) cells with Apo-2 ligand and human umbilical vein endothelial (HUVE) cells with staurosporine. In this assay, we compare both direct and indirect labelling methods by utilizing either iodinated annexin V or biotinylated annexin V/[35S] streptavidin to radiolabel apoptotic cells. The signal detected is a direct consequence of the binding of annexin V to externalized PS on apoptotic cells and the proximity of the label to the base of the plate. Using this method, separation of bound and unbound radiolabel signal occurs directly within the well resulting in a sensitive assay that requires minimal manipulation and can accomodate a large number of samples.
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  • 52
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    Cardiovascular drugs and therapy 13 (1999), S. 289-294 
    ISSN: 1573-7241
    Keywords: heart failure ; apoptosis ; protein kinases ; caspases ; DNA damage ; cardiomyocytes ; β-blocks
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Apoptosis as defined by contemporary science describes a form of cell death that involves discrete genetic and molecular programs, de novo protein expression and unique cellular phenotype. Evidence for the existence of apoptosis in the human heart has been reported in various cardiac diseases, including ischemic and non-ischemic heart failure, myocardial infarction and arrhythmias. Among the most potent stimuli that elicit cardiomyocyte apoptosis are: oxygen radicals (including NO), cytokines, (FAS/TNFα family of cytokines) and growth factors/energy deprivation. Several complex signal transduction pathways have been implicated in execution of cardiomyocyte apoptosis, including: Fas/TNFα receptors signaling, stress or mitogen activated protein kinases (SAPK/MAPK), sphingolipids metabolites (ceramide), G-protein coupled receptor (GPCR) signaling (Gαi, Gαq) and NFkB activation. Apoptosis of cardiac myocytes may contribute to progressive pump-failure, arrhythmias and cardiac remodeling. The recognition of numerous molecular targets associated with cardiomyocyte apoptosis that are amenable for pharmacologic manipulation, may provide novel therapeutic strategies for diverse cardiac ailments, as recently suggested by pharmacologic studies in experimental animals.
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  • 53
    ISSN: 1573-7373
    Keywords: apoptosis ; cell death ; C6 glioma ; nerve growth factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nerve growth factor (NGF) plays an important physiological role in differentiation and survival of various types of neurons. Glial cells and glial tumor cells synthesize multiple neurotrophic factors including NGF and secrete them into the surrounding environment; however, the mechanisms of NGF and the significance of NGF receptors have not been studied in detail. The C6 glioma cell line can synthesize NGF, respond to exogenous application of NGF and stimulate the expression of NGF receptor in an autocrine manner. In order to determine the significance of such an NGF autocrine system, the effects of exposure to exogenous NGF and deprivation of endogenous NGF were examined in a C6 glioma cell line in vitro. Exogenous NGF significantly inhibited maintenance of the cell number and thymidine incorporation. Morphological changes, including the formation of growth cones, outgrowth of processes and cellular hypertrophy, were observed, concurrently, indicating that exogenous NGF stimulated differentiation and thereby inhibited proliferation of the cells. Deprivation of endogenous NGF with anti-NGF antibody elicited a rapid decrease in cell number and thymidine incorporation, and led almost all of the cells to death within 8 days. The protein synthesis inhibitor, cycloheximide, strongly inhibited the death of NGF-deprived cells, suggesting the involvement of an active process requiring synthesis of suicide proteins. These findings imply that the NGF autocrine system plays a significant role in regulating the differentiation and survival of C6 glioma cells, similarly to neuronal cells.
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  • 54
    ISSN: 1573-7373
    Keywords: glutamate ; glioblastoma ; apoptosis ; Bcl-2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glutamate has been shown to function as a toxic agent in neuronal and glial cells, as well as an excitatory neurotransmitter throughout the central nervous system. In the present study, we examined the effect of increasing glutamate concentration on the induction of apoptosis in the two human glioblastoma cell lines GB-4 and GB-12. Glutamate exposure caused cell death of GB-4 and GB-12 in a dose-dependent manner. The cells were found to die via apoptosis in response to glutamate based on the following criteria: propidium iodide (PI) staining, H–E staining, electron microscopic analysis, and the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. The glutamate-induced apoptosis appears to involve the modulation of Bcl-2 family gene products such as Bcl-2, Bcl-xL, and Bax-α. Both Bcl-2 and Bcl-xL were down-regulated by glutamate at 24 h and further at 48 h. The apoptosis-promoting product p21 Bax-α was also down-regulated in GB-12 but slightly up-regulated in GB-4, accompanied by generation of variant form of p18 Bax-α in both cell lines. These findings suggest that glutamate toxicity results in cellular death via an apoptotic mechanism which appears to involve the Bcl-2/Bax-α molecular complex.
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  • 55
    ISSN: 1573-7365
    Keywords: Cerebral ischemia ; focal ischemia ; mutant mice ; bcl-2 ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Neuronal death after brain ischemia is mainly due to necrosis but there is also evidence for involvement of apoptosis. To test the importance of apoptosis, we investigated the effect of targeted disruption of the apoptosis-suppressive gene bcl-2 on the severity of ischemic brain injury. Transient focal ischemia for 1 hour was induced by occlusion of the middle cerebral artery in homozygous (n=7) and heterozygous (n = 6) bcl-2 knockout mice as well as in their wildtype littermates (n=5). Bcl-2 ablation did not influence cerebral blood flow but it significantly increased infarct size and neurological deficit score at 1 day after reperfusion in a gene-dose dependent manner. The exacerbation of tissue damage in the absence of Bcl-2 underscores the importance of apoptotic pathways for the manifestation of ischemic injury after transient vascular occlusion.
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  • 56
    ISSN: 1573-7373
    Keywords: glioma ; immunohistochemistry ; MIB-1 ; apoptosis ; TUNEL
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract With the introduction of new (immuno-)histochemical techniques it is now possible to assess rates of proliferation and apoptosis in brain gliomas using archival paraffin embedded material. As proliferation and apoptosis are related to tumour growth rate quantification of these processes has prognostic value and is related to tumour grading. In this study we assessed the proliferation rate by measuring the Ki-67 labelling index using the MIB-1 antibody (MIB-LI) and the apoptotic rate using the in situ labelling of DNA strand breaks with TUNEL (TUNEL-LI) in 315 supratentorial gliomas. MIB-LI and TUNEL-LI in astrocytomas (A) where significantly lower compared to anaplastic astrocytomas (AA), glioblastomas (GBM) and oligodendroglial tumours [oligodendrogliomas (O) and anaplastic oligodendrogliomas (AO)]. MIB-LI and TUNEL-LI were significantly lower in AA compared to GBM. In astrocytic tumours MIB-LI and TUNEL-LI appeared to be correlated. As the distinction between A and AA is of clinical value but can be difficult histomorphologically we analysed the prognostic value of MIB-LI and TUNEL-LI in gliomas with particular emphasis on A and AA. MIB-LI below 10% was of prognostic value in A and AA, O and AO but not in GBM on univariate survival analysis. TUNEL-LI was of no prognostic value. With multivariate survival analysis MIB-LI lost prognostic significance in O and AO. Astrocytomas with a gemistocytic component (AG) are similar to A with respect to survival and MIB-LI and TUNEL-LI. MIB-LI is of independent prognostic value in A and AA. Assessment of MIB-LI in A and AA can be used as an aid in distinguishing A and AA.
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  • 57
    ISSN: 1573-7373
    Keywords: apoptosis ; astrocytoma ; glioma ; growth inhibition ; protein kinase C ; signal transduction ; UCN-01
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Recent studies in our laboratory have shown that UCN-01 (7-hydroxystaurosporine), which is a derivative of the non-selective protein kinase inhibitor staurosporine that exhibits relative selectivity for protein kinase C (PKC), is a potent inhibitor of glioma growth in in vitro and in vivo models. This agent exhibits both cytotoxic and cytostatic effects, depending on the time period of drug exposure. In the present study, we examined whether UCN-01-induced cytotoxicity correlated with the induction of apoptosis, and characterized further the time course of this process as a prelude to application of UCN-01 in clinical trials. We first demonstrated that the cytotoxic effects of UCN-01 were associated with the induction of morphological features of apoptosis. Secondly. we identified electrophoretic features of apoptosis semiquantitatively at a series of time points using field inversion gel electrophoresis. These studies showed a peak in the induction of high-molecular-weight DNA fragmentation after 3–6 days of drug treatment. Thirdly, we measured the percentage of cells undergoing apoptosis at various time points using a terminal transferase-catalyzed in situ end-labeling technique, which confirmed a time- and concentration-dependent increase in apoptotic cell numbers. This correlated with a progressive decrease in the percentage of cells that were viable as assessed by trypan blue exclusion. Cell killing peaked within 2–4 days after beginning UCN-01 treatment, but continued at a lower level in the ensuing days. Taken together, these studies demonstrated that extended periods of exposure to UCN-01 are needed for optimal manifestation of cytotoxic effects against glioma cells, a factor that must be taken into consideration in the design of future clinical trials with this agent for malignant gliomas.
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  • 58
    ISSN: 1573-7373
    Keywords: germinoma ; spontaneous regression ; multiple intracranial ; germ cell tumor ; immunological mechanism ; apoptosis ; MIB-1 index
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A case of multiple intracranial germ cell tumor in which a pineal tumor regressed spontaneously after resection of the cerebellar mass is reported. Immunohistochemical staining of the cerebellar mass showed that most of the infiltrating lymphocytes were positive for CD3 and CD8. The anti-Ki-67 monoclonal antibody MIB-1 staining of the resected tumor revealed a high MIB-1 positivity ratio (36.1%) among the large tumor cells, and TUNEL staining demonstrated that positivity in up to 6% of the tumor cells. Possible mechanisms responsible for this spontaneous regression including immunological responses and apoptosis induced by T lymphocytes are discussed.
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  • 59
    ISSN: 1573-7373
    Keywords: apoptosis ; cell cycle ; cell death ; medulloblastoma ; T cells ; tumour necrosis factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We studied the effect of the treatment of a medulloblastoma cell line by human T cells derived soluble factors. Medulloblastoma is one of the more common aggressive solid neoplasms in children for which there is no adequate therapy. Cell lines established from such tumours may be helpful to test the effect of various molecules on cell proliferation. Previous studies have suggested that T cell-derived factors may be toxic for the medulloblastoma cell line Dev. Cytokines were thought to mediate this effect. In this paper, we described changes in morphology, survival and cell cycle induced in Dev cells cocultured with human T cell lines chronically infected with a retrovirus (HTLV-I) and known to secrete high level of cytokines TNFα, IL1α and IL6. Such cocultures resulted in the death of a part of Dev cells and in decreased proliferation of surviving cells, associated with morphological changes and increase in vimentin expression. Treatment with conditioned medium from infected Dev cells, containing virus induced cytokines, triggered the same effect. Reduction of these effects by TNFα deprivation of conditioned medium suggested that this cytokine may be implicated. Direct treatment of Dev cells with recombinant cytokines indicated that TNFα, but not IL1 or IL6, is associated with Dev cell alterations. TNFα was shown to induce the death of Dev cells by an apoptotic pathway. Furthermore, TNFα had a bimodal effect on the cell cycle of surviving Dev cells. These differential effects of such cytokines on medulloblastoma cells could be therefore of interest for immunotherapy of these tumours.
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  • 60
    ISSN: 1573-7373
    Keywords: adenoviral gene transfer ; p53 ; apoptosis ; experimental malignant gliomas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Adenoviral-mediated gene transfer for the treatment of experimental intrinsic malignant brain neoplasms holds promise. The role, however, of intracellular, adenoviral-mediated p53 expression to inhibit growth of experimental human intracranial malignant gliomas remains largely unexplored. Using the AdCMV.p53 vector we measured the in vitro expression of p53 and the resultant effect upon U251 human malignant glioma cellular proliferation. We further measured the survival of nude mice after intracranial injection of the infected vs. control U251 cells. The growth of the infected U251 cells was inhibited when compared to both the uninfected cells and cells infected with the control vector (AdCMV.Null). Agarose gel electrophoresis confirmed the AdCMV.p53-dependent cellular apoptosis. Nude mice having intracranial injections of the U251 cells infected with the control (AdCMV.Null) vector showed diminished survival. In contrast, mice having intracranial injections of the cells infected with the AdCMV.p53 vector showed 100% survivorship measured 100 days after treatment. Gene therapy via the AdCMV.p53 viral vector holds promise for the clinical treatment of human malignant gliomas.
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  • 61
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    Journal of neuro-oncology 41 (1999), S. 159-166 
    ISSN: 1573-7373
    Keywords: neuroblastoma ; chemotherapy ; apoptosis ; dopamine receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract While neuroscientists are often involved in the assessment and care of patients with central nervous system tumors, they are only rarely involved in the case of peripheral nervous system neoplasia. Neuroblastoma is a childhood tumor of the primitive sympathetic nervous system. It is at once one of the most common and one of the most deadly tumors of childhood. The prognosis for children with this tumor has not changed in the past two decades. Clearly, a fresh approach to neuroblastoma is needed. The neuroscientist has much to add to our understanding and treatment of neuroblastoma and its sequelae. Conversely, neuroblastoma has much to teach us regarding the normal development of the neural crest and the aberrant loss of neurons in this lineage. A neuroscientist's approach to neuroblastoma, its biology and clinical features, is presented herein.
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  • 62
    ISSN: 1573-7373
    Keywords: apoptosis ; chemotherapy ; human cell lines ; lovastatin ; medulloblastoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Medulloblastoma is a malignant paediatric central nervous system tumor with a poor prognosis, stimulating the evaluation of improved treatment strategies. Lovastatin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, is currently used to treat patients with hypercholesterolemia. This compound also inhibits the production of non-steroidal mevalonate derivatives that are implicated in the control of cellular proliferation, and can induce cell-cycle arrest in vitro. We recently showed that lovastatin inhibited growth and promoted apoptosis of neuroblastoma, the peripheral nervous system ‘cousin’ of medulloblastoma. Therefore the potential of lovastatin as a possible anticancer drug against medulloblastoma was evaluated in vitro. Four medulloblastoma cell lines, Daoy, UW228, D341 Med and D283 Med, were treated with 1–40 µM of lovastatin in vitro. Analysis of cell morphologic changes, cell viability, DNA fragmentation and flow cytometry in all four cell lines showed growth inhibition and induction of apoptosis with lovastatin treatment. As little as 10 µM of lovastatin was sufficient to cause a marked reduction in cell numbers, and more than 20 µM of lovastatin induced 〉90% cells to undergo apoptosis, after intervals ranging between 36 and 96 h, depending on the cell line. Lovastatin induced apoptosis in these cell lines was concomitant with cell cycle arrest in G1. The attached cell lines UW228 and Daoy were more sensitive to lovastatin than D283 Med and D341 Med. Daoy cells which survived several cycles of lovastatin treatment could still be induced to undergo apoptosis after longer treatment times. The efficient induction of apoptosis by lovastatin favours this drug as a potential new avenue of therapeutic intervention for medulloablastoma.
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  • 63
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    Journal of neuro-oncology 43 (1999), S. 237-239 
    ISSN: 1573-7373
    Keywords: primary CNS lymphoma ; glucocorticoids ; cerebral edema ; apoptosis ; bcl-2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glucocorticoid therapy may result in the rapid resolution of cerebral mass lesions in patients with primary CNS lymphoma. Since glucocorticoids will obscure the histological diagnosis of primary CNS lymphoma upon biopsy, steroids should be withheld if primary CNS lymphoma is a likely diagnosis by neuroradiological criteria. The lympholytic effect of glucocorticoids is mediated by cytoplasmic steroid receptors which are translocated to the nucleus and signal apoptosis. Glucocorticoid-induced apoptosis of lymphoid cells does not require wild-type p53 activity, seems not to depend on caspase activation, but is attenuated by the bcl-2 protooncogene product. Long-term glucocorticoid therapy of primary CNS lymphoma is not recommended because relapse is probably inevitable and because of the prominent side effects of long-term glucocorticoid treatment. Further, long-term glucocorticoid treatment is contraindicated in immunocompromised patients with primary CNS lymphoma.
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  • 64
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    Journal of neurocytology 28 (1999), S. 149-159 
    ISSN: 1573-7381
    Keywords: prion disease ; transmissible spongiform encephalopathy ; cell culture ; toxicity ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aetiological agent of prion disease is proposed to be an aberrant isoform of the cell surface glycoprotein known as the prion protein (PrPc). This pathological isoform (PrPSc) is abnormally deposited in the extracellular space of diseased CNS. Neurodegeneration in these disease has been shown to be associated with accumulation of PrPSc in affected tissue. To investigate the possible uptake mechanisms that may be required for PrPSc-induced neurodegeneration we studied the cellular trafficking of the neurotoxic fragment, PrP106-126. We were able to detect, by fluorescence microscopy, PrP106-126 inclusions in murine neurones, astrocytes and microglia in vitro. These inclusions were abundant after 24 hour exposure and still present 48h post-exposure. Shorter exposure times yielded only occasional cells with inclusions. Large extracellular aggregates of PrP106-126 could also be detected, which appeared in a time dependent manner. The appearance of inclusions or aggregates was not dependent on PrPc expression as determined by exposure of peptides from PrP-null mice. Using transmission electron microscopy and gold particle detection, positively labelled osmiophilic inclusions of peptide could be detected in the cytoplasm of exposed cells. These results demonstrate that cultured cells are capable of sequestering PrP106-126 and may indicate uptake pathways for PrPSc in various cell types. Toxicity of PrP106-126 may thus be mediated via a sequestration pathway that is not effective for this peptide in PrP-null cells.
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  • 65
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    European journal of plant pathology 105 (1999), S. 61-76 
    ISSN: 1573-8469
    Keywords: core sampling ; foliar nutrient concentrations ; minirhizotrons ; nitrogen ; phosphorus ; potassium ; Rhizolab
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Potato-cyst nematodes (Globodera pallida) cause severe yield losses in potato. Plants infected with potato-cyst nematodes generally have reduced concentrations of nitrogen, phosphorus and potassium in the foliage. This study investigated whether reduced growth of nematode-infected potato is caused by nutrient limitation. Experiments in the field and in containers showed that phosphorus concentration correlated best with total crop biomass at early stages of growth. The role of phosphorus in nematode damage was further investigated in the field and in the Wageningen Rhizolab. The experimental field was infested with potato-cyst nematodes and two levels of nematode density were established by fumigation with a nematicide. Prior applications of calcium carbonate resulted in pHKCl levels of 4.8 and 6.1. Two levels of phosphorus fertiliser were applied: either 0 or 225 kg P ha−1. In the Wageningen Rhizolab, soil of both pH levels from the field was used after treatment with 1 MRad gamma irradiation to kill the nematodes. Subsequently, half of the soil was inoculated with cysts to give a nematode density of 30 viable juveniles per gram of soil. In the field, nine weeks after planting, the total crop biomass ranged from 107 g m−2 for the treatment with nematodes at pHKCl 6.1 without phosphorus fertiliser to 289 g m−2 for the fumigated treatment at pHKCl 4.8 with phosphorus fertiliser. The differences in total biomass for the various treatments were explained by differences in foliar phosphorus concentration. Nematodes induced or aggravated P deficiency and reduced total biomass. This was not the only damage mechanism as at high, non-limiting levels of foliar phosphorus concentration, nematodes still reduced total biomass. In the Wageningen Rhizolab, directly after planting, the number of roots visible against minirhizotrons was reduced by nematodes. However, the increase of root number in the nematode treatment continued longer than in the control, until root number was higher than that of the control. The compensary root growth of the nematode treatment was restricted to the top 30 cm and nematodes reduced rooting depth. High soil pH reduced growth, mainly by reducing the availability of phosphate. Both nematodes and high soil pH reduced nutrient uptake per unit root length. Our results lead us to suggest an interaction between nematodes and soil pH, with nematode damage being higher at pHKCl 6.1 than at pHKCl 4.8.
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  • 66
    ISSN: 1573-868X
    Keywords: Osaka Bay ; sediment ; carbon ; nitrogen ; organic matter ; stable isotope ratio ; terrestrial organic matter ; TOC ; POC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences
    Notes: Abstract Carbon and nitrogen stable isotope ratios (δ13C and δ15N) of surface sediments were measured within Osaka Bay, in the Seto Inland Sea in Japan, in order to better understand the sedimentation processes operating on both terrestrial and marine organic matter in the Bay. The δ13C and δ15N of surface sediments in the estuary of the Yodo River were less than −23‰ and 5‰ respectively, but increased in the area up to about 10 km from the river mouth. At greater distances they became constant (giving δ13C of about −20‰ and δ15N about 6‰). It can be concluded that large amounts of terrestrial organic matter exist near the mouth of the Yodo River. Stable isotope ratios in the estuary of the Yodo River within 10 km of the river mouth were useful indicators allowing study of the movement of terrestrial organic matter. Deposition rates for total organic carbon (TOC) and total nitrogen (TN) over the whole of the Bay were estimated to be 63,100 ton C/year and 7,590 ton N/year, respectively. The deposition rate of terrestrial organic carbon was estimated to be 13,200 (range 2,000–21,500) ton C/year for the whole of Osaka Bay, and terrestrial organic carbon was estimated to be about 21% (range 3–34) of the TOC deposition rate. The ratio of the deposition rate of terrestrial organic carbon to the rate inflow of riverine TOC and particulate organic carbon (POC) were estimated to be 19% (range 3–31) and 76% (range 12–100), respectively.
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  • 67
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    Water resources management 13 (1999), S. 303-314 
    ISSN: 1573-1650
    Keywords: economics ; irrigation ; nitrogen ; nutrients ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Architecture, Civil Engineering, Surveying , Geography
    Notes: Abstract The optimal wastewater treatment level is affected by costs, hazards and benefits. Lowering the wastewater treatment level decreases fertilization costs because of the increased levels of available nutrients left in the water, and irrigation costs decrease if water prices reflect the lower treatment costs. Agricultural yields and/or prices may decrease according to differences between levels of nutrients needed by crops and those available in wastewater. The present article focuses on determination of monthly optimal treatment levels and of the mix of crops calculated to maximize agricultural incomes, according to farmers' point of view. It does not reflect the national point-view focusing on maximization of net national benefits considering also environmental hazards. The methodology appears in Haruvy (1994) and application will be presented in another article (Haruvy et al., 1999).
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  • 68
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    Bioscience reports 19 (1999), S. 345-354 
    ISSN: 1573-4935
    Keywords: Glycolipid ; apoptosis ; intracellular traffic ; multidrug resistance ; ovarian carcinoma ; astrocytoma ; post transplant lymphoproliferative disease ; bone marrow purging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Verotoxin (VT) is involved in the etiology of both hemorrhagic colitis and the hemolytic uremic syndrome which are microvasculopathies of the colon and pediatric renal glomerulus respectively. Thus, VT can be considered a vasotoxin. Cell sensitivity in vitro varies according to the receptor glycolipid (globotriaosyl ceramide-Gb3) expression and also to intracellular trafficking of the receptor/toxin complex, such that in highly sensitive cells, the toxin is targeted to the endoplasmic reticulum and nuclear envelope. Such cells include tumor cells which have become drug resistant. Thus Gb3 is upregulated in certain tumors and when such tumor cells become drug resistant, their sensitivity to verotoxin increases. This may be due to a direct role of the MDR1 drug efflux pump in glycolipid biosynthesis. In addition to the tumor tissue, the toxin receptor may also be expressed in the tumor neovasculature suggesting that activated endothelial cells may be verotoxin sensitive. Thus VT may have both a direct and indirect antineoplastic potential. VT has proved highly effective in a xenograft cancer model and the possible therapeutic use of VT is discussed.
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  • 69
    ISSN: 1573-4935
    Keywords: ceramide glycanase ; cancer cells ; glycosphingolipid ; sphingosine ; ceramide ; apoptosis ; PPMP ; PDMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Ceramide glycanase (CGase) activities have been detected in different human tumor cells (colon, carcinoma Colo-205; neuroblastoma, IMR-32; breast cancer lines, SKBr3 and MCF7). However, the level of enzymatic activity is lower in these cells compared to that present in other mammalian tissues reported before (Basu, M., Kelly, P., Girzadas, M. A., Li, Z., and Basu, S. Methods Enzymol. (in press)). The majority of CGase activity was found in the 100,000g soluble supernatant fraction isolated from all these cell lines and tissues. Using the soluble enzyme, the requirement for optimum CGase activity was found to be consistent with previous observations found for rat and rabbit tissues (Basu, M., Dastgheib, S., Girzadas, M. A., O'Donnell, P. H., Westervelt, C. W., Li, Z., Inokuchi, J. I., and Basu, S. (1998) Acta Pol. Biochim. 42:327). The CGase activities from both Colo-205 and IMR-32 cells are optimum at a protein to detergent ratio of one. All the mammalian CGases, including human cancer cells, show an optimum pH between 5.5 and 5.8 in sodium acetate buffer. The CGase activities from cancer cells are found to be cation-independent; however, mercury, zinc, and copper ions seem to inhibit the enzyme activity substantially in both tumor cells lines. The mercury ion inhibition of CGase activities from all different sources indicates a possible structural homology in the CGase proteins. Radiolabeled substrates, labeled at the sphingosine double bond or at the 3-position of sphingosine without modifying double bond of sphingosine were used in this investigation. Both were active substrates with all enzyme preparations isolated from different cancer cells (apparent Km, 500 μM for nLcOse5[3H-DT]Cer and 350 μM for GgOse4[sph-3-3H]Cer with Colo-205 enzyme). Structural analogues of ceramide and sphingosine (L-PPMP, L-PDMP, alkylamines, and Tamoxifen) inhibited cancer cell CGase activities in vitro.
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  • 70
    ISSN: 1573-7403
    Keywords: apoptosis ; corticotroph adenomas ; heat shock proteins ; p53
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The pathogenesis of corticotroph adenomas is unknown. In a recent study accumulation of p53 protein was detected by immunohistochemistry in a substantial proportion of pituitary corticotroph adenomas, and it has been suggested that it may be causally related to their development. However, other immunohistochemical studies have not confirmed the high incidence of p53 accumulation in this tumor type. Therefore, in the present study, p53 protein accumulation was re-examined in a series of 31 cases of corticotroph adenomas, using different sets of well validated anti-p53 antibodies. Furthermore, in view of the known association of p53 protein with apoptosis, and the known property of p53 to form complexes with heat shock proteins (HSPs), the relationship of p53 accumulation in corticotroph adenomas with apoptosis and HSP-70 was also investigated. Tumor samples staining positively for ACTH from patients with Cushing's disease or Nelson's syndrome were studied. Accumulation of p53 protein was tested by the standard ABC method using two different sets of clone Pab1801 and DO-7 monoclonal antibodies, applied after incubation of sections in a microwave oven. Using the DO-7 antibody, nuclear accumulation of p53 protein was detected in a total of 15 cases, with cytoplasmic staining observed in only 3 tumors. In contrast, using the Pab1801 antibody nuclear staining was observed in only 5 adenomas, with 11 adenomas demonstrating focal cytoplasmic immunoreactivity. Parallel sections of all corticotroph tumors demonstrating cytoplasmic accumulation of p53 protein were tested for the immunohistochemical presence of heat shock protein HSP-70. A striking similar distribution pattern of these two proteins was observed. Apoptosis, identified by the in situ end labeling technique, was detected in a total of 15 out of 28 corticotroph adenomas tested. Calculation of the apoptotic labeling index (ALI) by image analysis showed a significantly lower ALI in those corticotroph adenomas demonstrating nuclear p53 accumulation compared to those with no nuclear p53 immunostaining (p〈0.05). There was no significant difference in the ALI between cytoplasmic p53 positive and negative tumors. It is concluded that depending on the antibody used there is a significant variation of p53 protein detection in corticotroph adenomas. Overall, a significant proportion of corticotroph adenomas studied expressed the p53 protein, which depending on the antibody used, was located either in the nucleus and/or the cytoplasm of tumorous corticotroph cells. Cytoplasmic accumulation of p53, as shown by our colocalization studies with HSP-70, may be due to p53/HSP-70 complex formation. Although such a complex-mediated cytoplasmic exclusion of p53 has no significant effect on apoptosis, nuclear accumulation of p53 protein is associated with a significantly lower apoptotic index indicating a failure of p53 protein to exert its apoptotic action in at least a subset of this tumor type.
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  • 71
    ISSN: 1573-675X
    Keywords: Adenosine ; apoptosis ; necrosis ; physiopathological implications.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Adenosine modulates the proliferation, survival and apoptosis of many different cell types, ranging from epithelial, endothelial and smooth muscle cells, to cells of the immune and neural lineages. In this review, we critically discuss the available in vitro and in vivo data which support a role for adenosine in both development-associated apoptosis, and in diseases characterized by either pathologically increased cell death (e.g., ischemia, trauma and aging-associated neurodegeneration) or abnormally reduced spontaneous apoptosis (e.g., cancer). Particular emphasis is given to the possible role of extracellular adenosine receptors, since these may represent novel and attractive molecular targets for the pharmacological modulation of apoptosis. In some instances, adenosine-induced cell death has been demonstrated to require entry of the nucleoside inside cells; however, in many other cases, activation of specific adenosine extracellular receptors has been demonstrated. Of the four G protein-coupled adenosine receptors so far identified, the A2A and the A3 receptors have been specifically implicated in modulation of cell death. For the A3 receptor, results obtained by exposing both cardiomyocytes and brain astrocytes to graded concentrations of selective agonists suggest induction of both cell protection and cell death. Such opposite effects, which likely depend on the degree of receptor activation, may have important therapeutic implications in the pharmacological modulation of cardiac and brain ischemia. For the A2A receptor, recent intriguing data suggest a specific role in immune cell death and immunosuppression, which may be relevant to both adenosine-deaminase-immunodeficiency syndrome (a pathology characterized by accumulation of adenosine to toxic levels) and in tumors where induction of apoptosis via activation of specific extracellular receptors may be desirable. Finally, preliminary data suggest that, in a similar way to the adenosine-deaminase-immunodeficiency syndrome, the abnormal accumulation of adenosine in degenerative muscular diseases may contribute to muscle cell death. Although the role of adenosine receptors in this effect still remains to be determined, these data suggest that adenosine-induced apoptosis may also represent a novel pathogenic pathway in muscular dystrophies.
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  • 72
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    Apoptosis 4 (1999), S. 317-319 
    ISSN: 1573-675X
    Keywords: Anti-tumor therapy ; Apoptin® ; apoptosis ; Bcl-2 ; p53.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptin, a protein encoded by an avian virus, induces apoptosis in various cultured human tumorigenic and/ or transformed cell lines, e.g. derived from breast and lung tumor, leukemia, lymphoma, osteosarcoma melanoma, cholangiocarcinoma, and hepatoma. In such cells, Apoptin induces p53-independent apoptosis, and the proto-oncogene Bcl-2 can accelerate this effect. The latter is surprising for, in general, Bcl-2 is known to inhibit e.g., p53-induced apoptosis. On the other hand, in normal non-transformed human cells, Apoptin is unable to induce apoptosis, even when Bcl-2 is over-expressed. In animal models Apoptin-induced apoptosis appears to be a safe and efficient anti-tumor agent. These data, in continuation with the observations that Apoptin is specifically stimulated by Bcl-2 in tumor cells, does not need p53, and is not inhibited by Bcr-Abl in these cells, imply that Apoptin is a potential anti-tumor therapy.
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  • 73
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    Apoptosis 4 (1999), S. 229-234 
    ISSN: 1573-675X
    Keywords: Adenovirus ; apoptosis ; Bcl-2 ; p53 ; Rb ; ventricular myocytes.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptosis or programmed cell death is an important physiologic event crucial for the selective removal of damaged or unwanted cells from body tissues. In the cardiovascular system, apoptosis has been observed in the vasculature and myocardium. Untimely or inappropriate myocardial cell loss through an apoptotic process may contribute to ventricular remodeling and the ultimate demise of ventricular function following injury. Therapeutic interventions designed to modulate or prevent myocardial apoptotic cell loss may therefore prove beneficial in maintaining cardiac function. Incite into the molecular mechanisms that govern apoptosis in mammalian cells has led to the identification of several key factors that promote or prevent the apoptotic process. In this report, we discuss putative regulators of cardiac cell apoptosis with specific reference to the tumor suppressor proteins, p53 and Rb. The interplay between these factors, as well as the anti-apoptotic molecules related to the Bcl-2 the family are discussed in the context of the heart under normal and disease conditions.
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  • 74
    ISSN: 1573-675X
    Keywords: Anti-microtubule agents ; apoptosis ; doxorubicin ; neuroblastoma ; tau ; taxoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Paclitaxel and docetaxel are potent anti-microtubule and antimitotic agents that induce apoptosis in bone marrow-derived cells and epithelial cells. This study examined apoptosis induced by anti-microtubule agents in the neuroblastoma SK-N-SH cell line with a special focus on tau protein which is one of the main Microtubule-Associated- Proteins (MAPs) in neuronal cells. In time, treatment with 1 μM paclitaxel successively induced formation of bundles, then pseudo-asters concomitantly with mitotic block and phosphorylation of bcl-2 (48 h), then phosphorylation of tau and externalization of phosphatidylserine at the early phase of apoptosis (72 h) and finally DNA fragmentation (96 h). Similar results were obtained with 0.5 μM vinorelbine. Paclitaxel induced a lower increase in tau phosphorylation in differentiated SK-N-SH/RA+ cells which are less sensitive to apoptosis. Moreover, doxorubicin whose mechanism of action is independent of microtubules also induced immunostaining of tau at 72 h treatment. In conclusion, our results on neuroblastoma cells show that overexpression of hyperphosphorylated tau is involved in the apoptotic process induced by anti-microtubule agents and may be extended to others cytostatic drugs. Thus, tau protein may play a role in the cellular events observed in neuroblastoma cells undergoing apoptosis.
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  • 75
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    Apoptosis 4 (1999), S. 71-80 
    ISSN: 1573-675X
    Keywords: abl ; apoptosis ; interleukin-3 ; oncogenes ; ras.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Growth factors promote cell survival and proliferation. Homeostasis is maintained by programmed cell death which occurs when the growth stimulus is withdrawn, in response to negative growth regulators such as interferons, TNF-α and CD95 ligand, or following differentiation. Although acutely-transforming oncogenes often overcome the need for growth factors, growth regulatory cytokines can influence proliferative responses of transformed cells. In this study we investigated the effects of IL-3 on the proliferative responses of parental bone marrow-derived 32D cells and cells transformed with ras and abl oncogenes. We show that treatment of ras-transformed 32D cells with IL-3 reduced proliferative responses and decreased colony-forming ability. These effects were exacerbated in the absence of serum and associated with inhibition of tyrosine kinase activity, down-regulation of RAS and MYC expression, and induction of apoptosis as indicated by DNA fragmentation. In contrast, treatment of parental 32D cells with IL-3, which is obligatory for cell survival and proliferation, increased tyrosine kinase activity, upregulated MYC and RAS expression and maintained DNA integrity. With abl-transformed cells, proliferation and colony-forming ability were also inhibited by IL-3. Tyrosine kinase activity and MYC expression were reduced, but early apoptosis was not evident. Calcium uptake however, was stimulated by IL-3 in both parental and oncogene-transformed cells. These results suggest that threshold levels of tyrosine kinase activity are necessary for cell survival and proliferation and that with ras-transformed cells, IL-3 treatment may result in this threshold being breached. We conclude that in some situations, growth-promoting cytokines can inhibit proliferation of transformed cells and induce cell death by apoptosis.
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  • 76
    ISSN: 1573-675X
    Keywords: ameloblasts ; amelogenesis ; apoptosis ; insulin-like growth factor.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are “hard wired” for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.
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  • 77
    ISSN: 1573-675X
    Keywords: AKR lymphoma malignancy variants ; apoptosis ; apoptosis-related gene expression ; tumor progression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Resistance to apoptosis may be related to tumor progression, due to the implications it might have on both tumor mass and genetic instability. We compared the tendency to spontaneous apoptosis and the proliferative capacity of metastatic growths of several AKR lymphoma variants (TAU-45, TAU-47, TAU-44, TAU-33, TAU-42 and TAU-46, in the order of increasing metastatic potential). We further compared the expression of several apoptosis-related genes. Cell proliferative capacity did not appear to determine malignant behavior since, on the whole, a decrease in S + G2M fraction was observed with increasing malignancy. Sensitivity to apoptotic cell death decreased with increasing malignancy when comparing the TAU-45, TAU-47, TAU-44 and TAU-33 variants, suggesting a role of reduced apoptosis in this T-cell lymphoma. An increase in Bcl-2 content with increasing aggressiveness among these variants, implicates this protein in this tumor progression-related resistance to apoptosis. However, the two variants of highest malignancy, TAU-42 and TAU-46, did not follow the same trend, since they displayed a relatively high content in apoptotic cells and a low Bcl-2 content. Fas receptor expression did not correlate with tendency to apoptosis, indicating that malignant behavior in the AKR lymphoma does not depend on CD95/Fas/APO1 downregulation. Overexpression of p53 was observed only in one of the variants of lowest malignancy.
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  • 78
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    Apoptosis 4 (1999), S. 419-427 
    ISSN: 1573-675X
    Keywords: Anti-oxidant defence ; apoptosis ; ether lipids ; glutathione ; ionising radiation ; stress.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have investigated the mechanisms of induction of apoptosis by the antineoplastic ether lipid ET-18-OCH3 (ALP) in sensitive S49wt mouse lymphoma cells and ALP-resistant S49ar variants, both with wild-type p53, and in related L1210 cells with mutated p53. Ether lipid-resistant S49ar cells were cross-resistant to extracellular stress factors (cold shock, heat shock, H2O2, dimethylsulfoxide) and to radiation-induced apoptosis but not to physiological apoptotic signals (dexamethasone, growth factor deprivation, thapsigargin, C2-ceramide) and expressed similar levels of the apoptosis-regulating proteins Bcl-2, Bcl-X, Bax, Bad and Bak as did the parent S49wt cells. The uptake of [3H]-ALP was strongly reduced in the stress-resistant cells but this was not associated with significant differences in membrane cholesterol:phospholipid content nor in membrane microviscosity. In S49ar cells the activity of the antioxidant enzyme glutathione peroxidase (GSH-Px) was increased 4-fold and depletion of glutathione with the drug L-buthionine-S-R-sulfoximine (L-BSO) lowered the resistance of S49ar cells to ALP, stress factors and ionising radiation. The results indicate that ether lipids induce apoptosis by imposing a special form of physico-chemical stress, mediated by reactive oxygen species but independent of p53 status. The capacity of glutathione-dependent anti-oxidant defence appeared an important and shared determinant of the sensitivity to ether lipids, several types of extracellular stress and ionising radiation.
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  • 79
    ISSN: 1573-675X
    Keywords: Annexin V ; apoptosis ; flow cytometry ; intracellular Ca2+ ; intracellular pH ; mitochondrial membrane potential.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The sequential occurrence of plasma and mitochondrial membrane alterations, intra-cellular pH shifts and changes in intracellular Ca2+ concentration after induction of cell death was monitored by flow cytometry in Jurkat and HSB2-cells. Cell death was induced by treatment with anti-Fas antibodies or by irradiation. Phosphatidylserine (PS) exposure and plasma membrane integrity were measured with FITC-Annexin V adhesion and by Propidium Iodide exclusion. Transition of the mitochondrial membrane potential was monitored by the occurrence of decay of DiOC6 fluorescence. Intracellular pH shifts were monitored by changes in the ratio of fluorescence at 575 nm and at 635 nm of SNARF-1-AM. Fluctuations in intracellular Ca2+ concentration were established by changes in Fura red quenching. The Jurkat cells were sensitive to anti-Fas treatment, while HSB-2 cells were not. HSB-2 cells appeared more sensitive to radiation damage than Jurkat cells. In all experiments the transition of mitochondrial membrane potential occurred first, almost immediately followed by PS exposure. Fluctuations in intracellular Ca2+ concentration occurred later and were less outspoken. A decrease in intracellular pH occurred not earlier than 24 hours after anti-Fas treatment. Chelation of intracellular Ca2+ concentration with BAPTA-AM had no effect on the time sequence of cell death related events.
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  • 80
    ISSN: 1573-515X
    Keywords: Baltic Sea ; cyanobacteria ; estuaries ; grazing ; iron ; lakes ; molybdenum ; nitrogen ; nitrogen fixation ; nitrogen limitation ; Zooplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Explaining the nearly ubiquitous absence of nitrogen fixation by planktonic organisms in strongly nitrogen-limited estuaries presents a major challenge to aquatic ecologists. In freshwater lakes of moderate productivity, nitrogen limitation is seldom maintained for long since heterocystic, nitrogen-fixing cyanobacteria bloom, fix nitrogen, and alleviate the nitrogen limitation. In marked contrast to lakes, this behavior occurs in only a few estuaries worldwide. Primary production is limited by nitrogen in most temperate estuaries, yet no measurable planktonic nitrogen fixation occurs. In this paper, we present the hypothesis that the absence of planktonic nitrogen fixers from most estuaries is due to an interaction of bottom-up and top-down controls. The availability of Mo, a trace metal required for nitrogen fixation, is lower in estuaries than in freshwater lakes. This is not an absolute physiological constraint against the occurrence of nitrogen-fixing organisms, but the lower Mo availability may slow the growth rate of these organisms. The slower growth rate makes nitrogen-fixing cyanobacteria in estuaries more sensitive to mortality from grazing by Zooplankton and benthic organisms. We use a simple, mechanistically based simulation model to explore this hypothesis. The model correctly predicts the timing of the formation of heterocystic, cyanobacterial blooms in freshwater lakes and the magnitude of the rate of nitrogen fixation. The model also correctly predicts that high Zooplankton biomasses in freshwaters can partially suppress blooms of nitrogen-fixing cyanobacteria, even in strongly nitrogen-limited lakes. Further, the model indicates that a relatively small and environmentally realistic decrease in Mo availability, such as that which may occur in seawater compared to freshwaters due to sulfate inhibition of Mo assimilation, can suppress blooms of heterocystic cyanobacteria and prevent planktonic nitrogen fixation. For example, the model predicts that at a Zooplankton biomass of 0.2 mg l−1, cyanobacteria will bloom and fix nitrogen in lakes but not in estuaries of full-strength seawater salinity because of the lower Mo availability. Thus, the model provides strong support for our hypothesis that bottom-up and top-down controls may interact to cause the absence of planktonic nitrogen fixation in most estuaries. The model also provides a basis for further exploration of this hypothesis in individual estuarine systems and correctly predicts that planktonic nitrogen fixation can occur in low salinity estuaries, such as the Baltic Sea, where Mo availability is greater than in higher salinity estuaries.
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  • 81
    ISSN: 1573-515X
    Keywords: N15 ; nitrogen ; nutrient cycling ; plants ; stable isotopes ; soil ; temperate forest ; tropical forest
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Several lines of evidence suggest that nitrogen in most tropical forests is relatively more available than N in most temperate forests, and even that it may function as an excess nutrient in many tropical forests. If this is correct, tropical forests should have more open N cycles than temperate forests, with both inputs and outputs of N large relative to N cycling within systems. Consequent differences in both the magnitude and the pathways of N loss imply that tropical forests should in general be more 15N enriched than are most temperate forests. In order to test this hypothesis, we compared the nitrogen stable isotopic composition of tree leaves and soils from a variety of tropical and temperate forests. Foliar δ15N values from tropical forests averaged 6.5‰ higher than from temperate forests. Within the tropics, ecosystems with relatively low N availability (montane forests, forests on sandy soils) were significantly more depleted in 15N than other tropical forests. The average δ15N values for tropical forest soils, either for surface or for depth samples, were almost 8‰ higher than temperate forest soils. These results provide another line of evidence that N is relatively abundant in many tropical forest ecosystems.
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  • 82
    ISSN: 1573-515X
    Keywords: estuaries ; lakes ; marine ; nitrogen ; phosphorus ; rivers ; streams ; temperate ; tropics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Published data and analyses from temperate and tropical aquatic systems are used to summarize knowledge about the potential impact of land-use alteration on the nitrogen biogeochemistry of tropical aquatic ecosystems, identify important patterns and recommend key needs for research. The tropical N-cycle is traced from pre-disturbance conditions through the phases of disturbance, highlighting major differences between tropical and temperate systems that might influence development strategies in the tropics. Analyses suggest that tropical freshwaters are more frequently N-limited than temperate zones, while tropical marine systems may show more frequent P limitation. These analyses indicate that disturbances to pristine tropical lands will lead to greatly increased primary production in freshwaters and large changes in tropical freshwater communities. Increased freshwater nutrient flux will also lead to an expansion of the high production, N- and light-limited zones around river deltas, a switch from P- to N-limitation in calcareous marine systems, with large changes in the community composition of fragile mangrove and reef systems. Key information gaps are highlighted, including data on mechanisms of nutrient transport and atmospheric deposition in the tropics, nutrient and material retention capacities of tropical impoundments, and N/P coupling and stoichiometric impacts of nutrient supplies on tropical aquatic communities. The current base of biogeochemical data suggests that alterations in the N-cycle will have greater impacts on tropical aquatic ecosystems than those already observed in the temperate zone.
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  • 83
    ISSN: 1573-515X
    Keywords: Baltic Sea ; cyanobacteria ; estuaries ; grazing ; iron ; lakes ; molybdenum ; nitrogen ; nitrogen fixation ; nitrogen limitation ; zooplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Explaining the nearly ubiquitous absence of nitrogen fixation by planktonic organisms in strongly nitrogen-limited estuaries presents a major challenge to aquatic ecologists. In freshwater lakes of moderate productivity, nitrogen limitation is seldom maintained for long since heterocystic, nitrogen-fixing cyanobacteria bloom, fix nitrogen, and alleviate the nitrogen limitation. In marked contrast to lakes, this behavior occurs in only a few estuaries worldwide. Primary production is limited by nitrogen in most temperate estuaries, yet no measurable planktonic nitrogen fixation occurs. In this paper, we present the hypothesis that the absence of planktonic nitrogen fixers from most estuaries is due to an interaction of bottom-up and top-down controls. The availability of Mo, a trace metal required for nitrogen fixation, is lower in estuaries than in freshwater lakes. This is not an absolute physiological constraint against the occurrence of nitrogen-fixing organisms, but the lower Mo availability may slow the growth rate of these organisms. The slower growth rate makes nitrogen-fixing cyanobacteria in estuaries more sensitive to mortality from grazing by zooplankton and benthic organisms. We use a simple, mechanistically based simulation model to explore this hypothesis. The model correctly predicts the timing of the formation of heterocystic, cyanobacterial blooms in freshwater lakes and the magnitude of the rate of nitrogen fixation. The model also correctly predicts that high zooplankton biomasses in freshwaters can partially suppress blooms of nitrogen-fixing cyanobacteria, even in strongly nitrogen-limited lakes. Further, the model indicates that a relatively small and environmentally realistic decrease in Mo availability, such as that which may occur in seawater compared to freshwaters due to sulfate inhibition of Mo assimilation, can suppress blooms of heterocystic cyanobacteria and prevent planktonic nitrogen fixation. For example, the model predicts that at a zooplankton biomass of 0.2 mg l−1, cyanobacteria will bloom and fix nitrogen in lakes but not in estuaries of full-strength seawater salinity because of the lower Mo availability. Thus, the model provides strong support for our hypothesis that bottom-up and top-down controls may interact to cause the absence of planktonic nitrogen fixation in most estuaries. The model also provides a basis for further exploration of this hypothesis in individual estuarine systems and correctly predicts that planktonic nitrogen fixation can occur in low salinity estuaries, such as the Baltic Sea, where Mo availability is greater than in higher salinity estuaries.
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  • 84
    ISSN: 1573-904X
    Keywords: MAPK ; caspases ; chemopreventive agents ; phase II drug metabolizing enzymes ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract In therapeutic response to drugs, the plasma concentration range leads to the establishment of a safe and effective dosage regimen. Our hypothesis is that by studying drug concentration-dependent effect on signal transduction mechanisms, a better understanding of the beneficial pharmacodynamic and adverse toxicodynamic responses elicited by the drug may be achieved. Using two classes of chemopreventive compounds (phenolic antioxidants and isothiocyanates), we illustrate the potential utility of two signal transduction pathways elicited by these agents to predict the pharmacodynamic effect (induction of Phase II drug metabolizing enzymes) and the potential toxicodynamic response (stimulation of caspase activity and cytotoxic cell death). At lower concentration, phenolic antioxidants and isothiocyanates activate mitogen-activated protein kinase (MAPK; extracellular signal-regulated protein kinase 2, ERK2; and c-Jun N-terminal kinase 1, JNK1) in a concentration-and time-dependent manner. The activation of MAPK by these compounds may lead to the induction of cell survival/protection genes such as c-jun, c-fos, or Phase II drug metabolizing enzymes. However, at higher concentrations, these agents activate another signaling molecule, ICE/Ced3 cysteine protease enzymes (caspases) leading to apoptotic cell death. The activation of these pathways may dictate the fate of the cells/tissues upon exposure to drugs or chemicals. At lower concentrations, these compounds activate MAPK leading to the induction of Phase II genes, which may protect the cells/tissues against toxic insults and therefore may enhance cell survival. On the other hand, at higher concentrations, these agents may activate the caspases, which may lead to apoptotic cell death, and have toxicity. Understanding the activation of these and other signal transduction events elicited by various drugs and chemicals may yield insights into the regulation of gene expression of drug metabolizing enzymes and cytotoxicity. Thus, the study of signaling events in cell survival (hemeostasis) and cell death (cytotoxicity) may have practical application during pharmaceutical drug development.
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  • 85
    ISSN: 1573-5036
    Keywords: ectomycorrhizae ; Eucalyptus regnans ; forest burns ; nitrogen ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This study was conducted to compare the effects on the growth of Eucalyptus regnans seedlings of unheated soil and soil heated to different extents (as indicated by soil colour–bright red or black) in burnt logging coupes, and to separate the effects of heating of the soil on direct nutrient availability and on morphotypes and effectiveness of ectomycorrhizae. Burnt soils were collected from three logging coupes burnt 2, 14 and 25 months previously and unbumt soil from adjacent regrowth forests. Compared to unburnt soil, the early seedling growth was stimulated in black burnt soil from all coupes (burnt 2, 14 and 25 months previously). Seedling growth was generally poor in red burnt soil, especially in soil collected 2 months after burning. However, the concentration of extractable P was extremely high in red burnt soil, especially in soil collected 2 months after burning. In black burnt soil, extractable P was increased in soil 2 months after burning, but not in the soils collected 14 or 25 months after burning. However, both total P content and concentration in seedlings were increased in all collections of black burnt soil. Frequency of ectomycorrhizae was high in seedlings grown in all black burnt soils, but the mycorrhizal mantles were poorly developed in seedlings in black burnt soil collected 2 months after burning. Seedlings were also ectomycorrhizal in red burnt soil, except in soil collected 2 months after burning. Fine root inocula from seedlings grown in black burnt soils collected 14 and 25 months after burning significantly stimulated both seedling growth and P uptake compared with the uninoculated control, whereas the fine root inocula from the seedlings grown in all the other soils did not. These results suggest that, in black burnt soil, both direct nutritional changes and changes in the ectomycorrhizae may contribute to seedling growth promotion after regeneration burns. The generally poor seedling growth in red burnt soils is likely to have been due to N deficiency as the seedlings in these soils were yellow-green and the tissue concentrations of N were significantly lower than in other treatments.
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  • 86
    ISSN: 1573-5036
    Keywords: crop residues ; isotope dilution ; 15N ; nitrogen ; organic matter ; pool substitution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Experiments were carried out to compare the direct approach for estimating crop N uptake from 15N labelled organic inputs, to two indirect approaches, 15N isotope dilution and A value. In the first experiment soils received 25, 50, 75, or 100 mg N kg soil−1 in the form of Casuarina equisitifolia residues in addition to ammonium sulphate fertiliser, to give a total of 100 mg N kg soil−1 added. This was a cross labelling design, thus two matching sets of treatments, were set up, identical in all but the position of the 15N label. Maize (Zea mays L.) plants were grown in the soils amended with residues for 11 weeks and N derived from residues (Ndfr) estimated using the A-value or the direct approach. The A-value approach appeared to significantly overestimate %Ndfr compared to the direct method. In the second experiment contrasting residues were added to soil, fababean (Vicia faba L. var. minor), alfalfa (Medicago sativa L.), soyabean fixing, (Glycine max (L.) Merrill), soyabean non-fixing, barley (Hordeum vulgare L.) and maize. This was also cross-labelling design, labelled and unlabelled residues were used. Maize plants were grown in these soils for 11 weeks and %Ndfr in the maize plants estimated using 15 N isotope dilution and the direct approach. The 15 N isotope dilution approach also overestimated %Ndfr compared to the direct method in this experiment. Pool substitution appeared to be responsible for the discrepancy between the direct and indirect techniques. It was concluded that 15N isotope dilution and A-value approaches as used in these experiments (i.e where residues and 15N label are added simultaneously) were not appropriate techniques for estimating N derived from organic residues in soils.
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  • 87
    ISSN: 1573-5036
    Keywords: nitrogen ; Prunus ; remobilisation ; storage ; uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two-year old nectarine trees (Prunus persica, Batsch, var. nectarina, cv. Starkredgold on GF305 rootstock) planted in pots each received five applications of 1.0 g 15N labelled urea either from mid May to mid July (early uptake) or from mid August to the beginning of October (late uptake). All trees were supplied with a corresponding amount of unlabelled urea when they did not receive the labelled N. In autumn, all abscised leaves were collected and during winter randomly selected trees were harvested and divided into main organs. The remaining trees were transplanted into similar pots filled with sand; they received no N fertiliser and were harvested in May to evaluate the remobilisation of N. Total N and 15N abundance were determined in each organ. Nectarine trees took up similar amounts of N in the 'early' and in the 'late' period; however, more labelled nitrogen was recovered in the perennial organs during the winter when trees received the labelled N in the 'late' than in the 'early' period. Some 73–80% of the N present in the dormant trees was stored in the roots, which contained almost twice the amount of labelled N taken up 'late' than that absorbed 'early'. Nitrogen for spring growth was remobilised predominantly from the roots and accounted for some 43–49% of the labelled N recovered in the tree during winter. Results suggest that the nitrogen taken up 'late' in the season is preferentially stored in roots and used by peach trees to sustain new growth the following spring.
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  • 88
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    Plant and soil 212 (1999), S. 207-217 
    ISSN: 1573-5036
    Keywords: legume ; nitrogen ; N2-fixation ; pea ; sulphur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A S-deficient soil was used in pot experiments to investigate the effects of S addition on growth and N2-fixation in pea (Pisum sativum L.). Addition of 100 mg S pot−1 increased seed yield by more than 2-fold. Numbers of pods formed were the most sensitive yield component affected by S deficiency. Sulphur addition also increased the concentration of N in leaves and stems, and the total content of N in the shoots. The amounts of N fixed by pea were determined at four growth stages from stem elongation to maturity, using the 15N dilution technique. Sulphur addition doubled the amount of N fixed at all growth stages. In contrast, leaf chlorophyll content and shoot dry weight were increased significantly by S addition only after the flowering and pod fill stage, respectively. Pea roots were found to have high concentrations of S, reaching approximately 10 mg g−1 dry weight and being 2.6–4.4 times the S concentration in the shoots under S-sufficient conditions. These results suggest that roots/nodules of pea have a high demand for S, and that N2-fixation is very sensitive to S deficiency. The effects of S deficiency on pea growth were likely to be caused by the shortage of N, due to decreased N2-fixation.
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  • 89
    ISSN: 1573-5036
    Keywords: fire ; nitrogen ; phosphorus ; soil nutrient heterogeneity ; tree effects ; tropical dry forest
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Individual trees are known to influence soil chemical properties, creating spatial patterns that vary with distance from the stem. The influence of trees on soil chemical properties is commonly viewed as the agronomic basis for low-input agroforestry and shifting cultivation practices, and as an important source of spatial heterogeneity in forest soils. Few studies, however, have examined the persistence of the effects of trees on soil after the pathways responsible for the effects are removed. Here, we present evidence from a Mexican dry forest indicating that stem-related patterns of soil nutrients do persist following slash-and-burn removal of trees and two years of cropping. Pre-disturbance concentrations of resin extractable phosphorus (P), bicarbonate extractable P, NaOH extractable P, total P, total nitrogen (N) and carbon (C), KCl extractable nitrate (NO3 -), and net N mineralization and nitrification rates were higher in stem than dripline soils under two canopy dominant species of large-stemmed trees with contrasting morphologies and phenologies (Caesalpinia eriostachys Benth. and Forchhammeria pallida Liebm.). These stem effects persisted through slash burning and a first growing season for labile inorganic and organic P, NaOH inorganic P, and plant-available P, and through a second growing season for labile organic P, NaOH organic P, and plant-available P. While stem effects for extractable NO3 -, net nitrification rates, total N and C disappeared after felling and slash burning, these stem effects returned after the first growing season. These results support the view that tree-influenced patterns of soil nutrients do persist after tree death, and that trees contribute to the long-term spatial heterogeneity of forest soils.
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  • 90
    Electronic Resource
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    Plant and soil 212 (1999), S. 173-181 
    ISSN: 1573-5036
    Keywords: nitrogen ; phosphatase activity ; phosphorus ; protease activity ; soil microbial biomass ; substrate-induced respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Estimating in situ N and P status of the soil microbiota is complicated because microbiological features reflect potentials rather than field conditions. Complementary microbiological assays were, therefore, combined to evaluate the N and P requirement of the microbiota in seven agricultural, grassland and forest topsoils of the Bornhöved Lake district as follows: (i) the sensitivity of the substrate-induced respiration (SIR) to supplemental addition of N and P was monitored during microbial growth and (ii) soil protease and phosphatase activities were analysed and related to soil mass and microbial biomass content. Nitrogen addition increased the maximal SIR rate in all except one soil indicating that the growth of organisms is restricted by this element when easily degradable C source is present. Supplemental N (and in some cases also P) retarded the respiratory response within the first 24 h which suggests microbial sensitivity and/or greater anabolic efficiency. With additional N the maximal SIR rate was most strongly enhanced in topsoils of the beech forest and the dystric alder forest. Thus, the microbial growth in these soils that were below litter horizons seems to be mostly restricted by N. Supplemental P positively affected respiratory response of soils under monoculture, wet grassland and dystric alder forest. In the dystric alder forest soil, high rates of alkaline and unbuffered phosphatase activity were observed when activity was related to either soil mass or microbial biomass content. The data of proteolytic and phospholytic enzymes are discussed with reference to nutrient deficiency and microbial strategy for N and P adsorption.
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  • 91
    ISSN: 1573-5036
    Keywords: effluent ; leaching ; 15N isotope ; nitrogen ; pasture ; uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The objective of this study was to compare the N leaching loss and pasture N uptake from autumn-applied dairy shed effluent and ammonium fertilizer (NH4Cl) labeled with 15N, using intact soil lysimeters (80 cm diameter, 120 cm depth). The soil used was a sandy loam, and the pasture was a mixture of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). The DSE and NH4Cl were applied twice annually in autumn (May) and late spring (November), each at 200 kg N ha-1. The N applied in May 1996 was labeled with 15N. The lysimeters were either spray or flood irrigated during the summer. The autumn-applied DSE resulted in lower N leaching losses compared with NH4Cl. However, the N applied in the autumn had a higher potential for leaching than N applied in late spring. Between 4.5–8.1% of the 15N-labeled mineral N in the DSE and 15.1–18.8% of the 15N-labeled NH4Cl applied in the autumn were leached within a year of application. Of the annual N leaching losses in the DSE treatments (16.0–26.9 kg N ha-1), a fifth (20.3–22.9%) was from the mineral N fraction of the DSE applied in the autumn, with the remaining larger proportion from the organic fraction of the DSE, soil N and N applied in spring. In the NH4Cl treatments, more than half (53.8–64.8%) of the annual N leaching loss (55.9–57.6 kg N ha-1) was derived from the autumn-applied NH4Cl. DSE was as effective as NH4Cl in stimulating pasture production. Since only 4.4–4.5% of the annual herbage N uptake in the DSE treatment and 12.3–13.3% in the NH4Cl treatment were derived from the autumn-applied mineral N, large proportions of the annual herbage N uptake must have been derived from the N applied in spring, the organic N fraction in the DSE, soil N and N fixed by clover. The recoveries of 15N in the herbage were similar between the DSE and the NH4Cl treatments, but those in the leachate were over 50% less from the DSE than from the NH4Cl treatment. The lower leaching loss of 15N in the DSE treatment was attributed to the stimulated microbial activities and increased immobilization following the application of DSE.
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  • 92
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    Plant growth regulation 27 (1999), S. 119-123 
    ISSN: 1573-5087
    Keywords: apoptosis ; carrot protoplast ; DNA ladder ; ethrel ; ethylene ; nucleus condensation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In recent years, apoptosis has been reported to exist in plants during normal development and in response to stress. However, little is known about the relation of hormones to this form of programmed cell death. Here, we report examination of characteristics of apoptosis in carrot protoplasts induced by ethylene evolved from ethrel (2-chloroethylphosphonic acid). Nucleus condensation and DNA ladders were observed, and neutral comet assay, which detects DNA cleavage, also provided evidence that ethrel treatment resulted in nuclear DNA fragmentation. Strikingly, a close correlation between the incidence of DNA comets and the percentage of apoptotic protoplasts was shown in ethrel-treated carrot protoplasts. In conclusion, this study demonstrated that ethylene is an active inducer of apoptosis in carrot protoplasts, and that ethylene-induced plant cell death showed characteristics similar to those of apoptosis in animals.
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  • 93
    Electronic Resource
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    Hydrobiologia 408-409 (1999), S. 375-387 
    ISSN: 1573-5117
    Keywords: eutrophication ; multi-lake studies ; phosphorus ; nitrogen ; chlorophyll-a ; transparency ; zooplankton ; macrophytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Monitoring data obtained from 231 freshwater lakes and ponds in the Netherlands, covering the period 1980–1996, were used to analyse the relationships between (a) transparency and chlorophyll-a, and the effect of system characteristics on this relationship, (b) chlorophyll-aand nutrient concentrations, and the effect of biological variables and (c) nutrient concentrations and nutrient loading. (a) Chlorophyll-aimposes a maximum on water transparency, but deviations from this maximum can be large. Reducing chlorophyll-a, therefore, does not guarantee a sufficient improvement of transparency. Soil type and the average depth of a lake were shown to influence the relationship between chlorophyll-aand transparency. (b) The maximum ratios of both chlorophyll-a: total-P and chlorophyll-a: total-N were higher in systems dominated by filamentous cyanobacteria than in systems dominated by other algae, indicating the efficiency of the former group with respect to nutrients. In systems with an areal coverage with submersed macrophytes above 5%, concentrations of chlorophyll-aand nutrients were lower than in systems with lower coverages. The ratios between chlorophyll-aand nutrients were lower at coverages larger than 10%. This indicates both bottom-up and top-down control of algae by macrophytes. Grazing pressure by zooplankton was also found to lower the chlorophyll-a: nutrient ratios. (c) System specific linear relationships were found between the average concentrations of total-P and total-N in the incoming water and the summer mean concentration in the lake. This allows the assessment of admissible loads for individual lakes, with narrower confidence limits compared to traditional relationships based on combined data from many lakes. From the analysis, it is concluded that the chain of relationships from nutrient loading to transparency is complex, and depends on biological variables as well as system characteristics.
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  • 94
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    Biogeochemistry 44 (1999), S. 93-118 
    ISSN: 1573-515X
    Keywords: Everglades National Park ; mangrove soils ; organic matter ; nitrogen ; phosphorus ; sedimentation ; simulation model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The distribution and accumulation of organic matter, nitrogen (N) and phosphorus (P) in mangrove soils at four sites along the Shark River estuary of south Florida were investigated with empirical measures and a process-based model. The mangrove nutrient model (NUMAN) was developed from the SEMIDEC marsh organic matter model and parameterized with data from mangrove wetlands. The soil characteristics in the four mangrove sites varied greatly in both concentrations and profiles of soil carbon, N and P. Organic matter decreased from 82% in the upstream locations to 30% in the marine sites. Comparisons of simulated and observed results demonstrated that landscape gradients of soil characteristics along the estuary can be adequately modeled by accounting for plant production, litter decomposition and export, and allochthonous input of mineral sediments. Model sensitivity analyses suggest that root production has a more significant effect on soil composition than litter fall. Model simulations showed that the greatest change in organic matter, N, and P occurred from the soil surface to 5 cm depth. The rapid decomposition of labile organic matter was responsible for this decrease in organic matter. Simulated N mineralization rates decreased quickly with depth, which corresponded with the decrease of labile organic matter. The increase in organic matter content and decrease in soil bulk density from mangrove sites at downstream locations compared to those at upstream locations was controlled mainly by variation in allochthonous inputs of mineral matter at the mouth of the estuary, along with gradients in mangrove root production. Research on allochthonouns sediment input and in situ root production of mangroves is limited compared to their significance to understanding nutrient biogeochemistry of these wetlands. More accurate simulations of temporal patterns of nutrient characteristics with depth will depend on including the effects of disturbance such as hurricanes on sediment redistribution and biomass production.
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  • 95
    ISSN: 1573-515X
    Keywords: Chaohu Lake ; chemical fertilizer ; cycling ; denitrification ; multipond system ; nitrogen ; nutrient budget ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract During a two-year field study, an annual nutrient budget and cycles were developed for a small agricultural watershed. The study emphasized the integrated unit of the watershed in understanding the biogeochemistry. It was found that the total nutrient input was 39.1× 104 kg nitrogen and 3.91×104 kg phosphorus in the year 1995, of which the greatest input of nutrients to the watershed was chemical fertilizer application, reaching 34.7×104 kg (676 kg/ha) nitrogen and 3.88×104 kg (76 kg/ha) phosphorus. The total nutrient output from the watershed was 13.55×104 kg nitrogen and 0.40×104 kg phosphorus, while the largest output of nitrogen was denitrification, accounting for 44.1% of N output; the largest output of phosphorus was sale of crops, accounting for 99.4% of P output. The results show that the nutrient input is larger than output, demonstrating that there is nutrient surplus within the watershed, a surplus which may become a potential source of nonpoint pollution to area waters. The research showed that both denitrification and volatilization of nitrogen are key ways of nitrogen loss from the watershed. This suggests that careful management of fertilizer application will be important for the sustainable development of agriculture. The research demonstrated that a multipond system within the watershed had high retention rate for both water and nutrients, benefiting the water, nutrient and sediment recycling in the terrestrial ecosystem and helping to reduce agricultural nonpoint pollution at its source. Therefore, this unique watershed system should be recommended due to its great potential relevance for sustainable agricultural development.
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  • 96
    ISSN: 1573-515X
    Keywords: Chihuahuan desert ; desert ; desertification ; grassland ; nitrogen ; nutrient budgets ; phosphorus ; runoff
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Rainfall simulation experiments were performed in areas of semiarid grassland (Bouteloua eriopoda) and arid shrubland (Larrea tridentata) in the Chihuahuan desert of New Mexico. The objective was to compare the runoff of nitrogen (N) and phosphorus (P) from these habitats to assess whether losses of soil nutrients are associated with the invasion of grasslands by shrubs. Runoff losses from grass- and shrub-dominated plots were similar, and much less than from bare plots located in the shrubland. Weighted average concentrations of total dissolved N compounds in runoff were greatest in the grassland (1.72 mg/1) and lowest in bare plots in the shrubland (0.55 mg/1). More than half of the N transported in runoff was carried in dissolved organic compounds. In grassland and shrub plots, the total N loss was highly correlated to the total volume of discharge. We estimate that the total annual loss of N in runoff is 0.25 kg/ha/yr in grasslands and 0.43 kg/ha/yr in shrublands — consistent with the depletion of soil N during desertification of these habitats. Losses of P from both habitats were very small.
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  • 97
    ISSN: 1573-5117
    Keywords: nitrogen ; phosphorus ; pore water ; macrophyte ; floodplain ; Paraná River
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inorganic nitrogen and soluble reactive phosphate (o-P) concentrations were measured in the water of a marsh and in its interstitial water at two sites, and in the river water of a floodplain marsh of the Lower Paraná River. These values were compared with the N and P concentration in sediments and macrophyte biomass in order to assess nutrient availability, fate and storage capacity. High variability was found in the interstitital water using a 1 cm resolution device. Nitrate was never detected in the pore water. Depth averaged NH4 + concentrations in the upper 30 cm layer often ranged from N = 1.5 to 1.8 mg l-1, but showed a pronounced minimum (0.5–0.7 mg l-1), close to (March 95), or relatively soon after (May 94) the end of the macrophyte growing season. Soluble phosphate showed a large variation between P = 0.1–1.1 mg l-1 without any discernible seasonal pattern. NH4 + depletion in the pore water concentration and low N/P ratios (3.7 by weight) within the macrophyte biomass at the end of the growing period suggest that available N limits plant growth. NH4 + and o-P concentrations were 35 and 7 times higher, respectively, in the pore water than in the overlying marsh, suggesting a permanent flux of nutrients from the sediments. o-P accumulate in the marsh leading to higher concentrations than in the incoming river. NH4 + did not accumulate in the marsh, and no significant differences were observed between the river and the marsh water, while the NO3 - contributed by the river water was depleted within the marsh, caused probably by coupled nitrification-denitrification at the sediment–water interface. Although an order of magnitude smaller, the pore water pool can supply enough nutrients to build up the macrophyte biomass pool, but only if a fast turnover is attained. The Paraná floodplain marsh retains a large amount of nutrients being stored mainly in the sediment compartment.
    Type of Medium: Electronic Resource
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  • 98
    ISSN: 1573-5117
    Keywords: shallow lake ; nutrient loading ; retention ; nitrogen ; phosphorus ; release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The shallow (mean depth 4.9 m), polymictic and eutrophic lake Müggelsee was highly loaded with phosphorus (6 gP m-2a-1) and nitrogen (170 gN m-2a-1) by the river Spree up to the end of the 1980s. Annual load declined by 40–50% during the last years (1991–97). Phosphorus retention fluctuated strongly during the seasonal cycle between −200 and +100 kgP d-1and from year to year between −44% and + 26% of the P import. At the end of the eighties, the P retention capacity of the sediment was exceeded and Müggelsee became a source of phosphorus. The lake regained its ability to retain P in the sediments after external load reduction in the 1990s. However, the internal load of P reached the level of the external one. The release of P during summer was strongly related to the import of nitrate. On long-term average (1979–1997), less than 1% of the P input was retained in Müggelsee. About 24% of the nitrogen load were removed in the lake on annual mean. This rate decreased during the last years.
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    Springer
    Hydrobiologia 395-396 (1999), S. 389-401 
    ISSN: 1573-5117
    Keywords: legislation ; eutrophication ; phosphorus ; nitrogen ; wildlife conservation ; SSSI
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract There is no single piece of legislation which comprehensively addresses the problem of eutrophication of freshwater lakes. An examination is made of the extent to which national and EU laws applicable in England and Wales may be used to reduce phosphorus inputs to eutrophicated lakes. This case study is then used to outline the shortcomings of existing legislation and the future challenges for lake eutrophication control. Applicable legislation may be divided into that relating to nature conservation, to water quality and to agricultural extensification. Nature conservation laws are applicable, in that lakes designated as Sites of Special Scientific Interest (SSSIs), are theoretically safeguarded from degradation. The main limitation of the SSSI system lies in the boundary definition process under which, as a rule, no other category of legally protected land may be designated. Thus, the use of surrounding buffer land to protect water quality at the catchment level is generally precluded. The introduction of consultation areas around SSSIs of international importance under the EU Directive on Habitats is unlikely to alleviate the problem, since the area involved remains minimal compared to the extent of the majority of catchments. Intensive agricultural practices are responsible for significant nutrient enrichment of rivers and lakes, so that legislation aimed at extensification is also relevant. Examples of agricultural extensification include the Environrnentally Sensitive Areas programme and various options available for Set-Aside land. However, the prime purpose of such initiatives is the alleviation of surplus and budgetary problems and, as such, they fall short of a fully integrated approach to the ecological management of farmland. Nutrient enrichment is, essentially, a water quality issue, but policy and legislation in this area are not yet sufficiently developed to address the problem comprehensively. The current regulatory process for water quality carries the potential to work comparatively well for point sources under the system of consents to discharge. This potential is limited, however, by the paucity of information available on ecologically acceptable concentrations of phosphorus in discharges. In addition, the consents system is not constructed to deal effectively with diffuse agricultural losses of phosphorus, since, unlike point sources, these tend to arise from the cumulative effect of many activities. The main legislative challenge for lake eutrophication control lies in the area of diffuse agricultural losses of phosphorus. In this respect, experience in the U.S.A. reveals that the use of comprehensive and catchment-wide ‘Best Management Practices’ is capable of producing significant water quality improvements, providing that some degree of mandatory compliance is incorporated.
    Type of Medium: Electronic Resource
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  • 100
    ISSN: 1573-5117
    Keywords: Integrated lake management ; biomanipulation ; lake recovery ; phosphorus ; nitrogen ; eutrophication ; restoration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The majority of Danish lakes are highly eutrophic due to high nutrient input from domestic sources and agricultural activities. Reduced nutrient retention, and more rapid removal, in catchments as a result of agricultural drainage of wetlands and lakes and channelisation or culverting of streams also play a role. Attempts have recently been made to reduce nutrient loading on lakes by intervening at the source level and by improving the retention capacity of catchment areas. The former measures include phosphorus stripping and nitrogen removal at sewage works, increased use of phosphate-free detergents, and regulations concerning animal fertiliser storage capacity, fertiliser application practices, fertilisation plans and green cover in winter. In order to improve nutrient retention capacity of catchments, wetlands and lakes have been re-established and channelised streams have been remeandered. In addition, cultivation-free buffer strips have been established alongside natural streams and there has been a switch to manual weed control. These measures have resulted in a 73% reduction of the mean total phosphorus concentration of point-source polluted streams since 1978; in contrast, there has been no significant change in the total nitrogen concentration. Despite the major reduction in stream phosphorus concentrations, lake water quality has often not improved. This may reflect a too high external or internal phosphorus loading or biological resistance. Various physico-chemical restoration measures have been used, including dredging and oxidation of the hypolimnion with nitrate and oxygen. Biological restoration measures have been employed in 17 Danish lakes. The methods include reducing the abundance of cyprinids, stocking with 0+ pike ( Esox lucius) to control 0+ cyprinids, and promoting macrophyte recolonization by protecting germinal submerged macrophyte beds against grazing waterfowl and transplanting out macrophyte shoots. In several lakes, marked and long-lasting improvements have been obtained. The findings to date indicate that fish manipulation has a long-term effect in shallow lakes, providing nutrient loading is reduced to a level so low as to ensure an equilibrium lake water phosphorus concentration of less than 0.05–0.1 mg phosphorus l−1. If nitrogen loading is very low, however, positive results may be obtained at higher phosphorus concentrations. Macrophyte refuges and transplantation seem to be the most successful as restoration measures in the same nutrient-phosphorus regime as fish manipulation.
    Type of Medium: Electronic Resource
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