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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Annals of surgical oncology 6 (1999), S. 315-321 
    ISSN: 1534-4681
    Keywords: Melanoma ; Locoregional cutaneous recurrence ; In-transit metastases ; Lymphatic vascular invasion ; Microscopic satellites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: In-transit metastases and satellite lesions are manifestations of locoregional cutaneous recurrence that are characteristic of malignant melanoma. They are the result of tumor cell emboli entrapped in the dermal lymphatics between the primary tumor and the regional lymph node basin. Histopathological features of lymphatic invasion were investigated to determine the possibility of predicting locoregional cutaneous metastases in melanoma patients. Methods: In a prospective study, 258 patients with clinical stage I melanoma underwent wide local excision and sentinel node biopsy. Nodal metastases were found in 53 (21%) patients. Of 29 patients (11.2%) who had developed recurrences to date, 17 (6.6%) had locoregional cutaneous metastases. All surgical specimens were examined with particular attention to histopathological signs of lymphatic vascular invasion or microscopic satellites. Results: Unequivocal signs of lymphatic invasion were observed in 14 of 258 patients (5.4%), and 13 (93%) of these patients subsequently developed in-transit metastases, after a median interval of 10 months. The primary melanoma was located on the extremities in seven patients. The median Breslow thickness was 2.5 mm, and 5 showed ulceration. In 244 of 258 patients (94.6%), there were no signs of lymphatic invasion. To date, only four patients (1.6%) have had a locoregional cutaneous recurrence, occurring after a median interval of 29 months. All four of these patients had ulcerative melanomas on an extremity, with a median thickness of 4.0 mm. The presence of lymphatic invasion was significantly related to early locoregional cutaneous relapse (P _ .0001). Conclusions: Locoregional cutaneous recurrence appears to be highly predictable in the presence of histopathological signs of lymphatic invasion. Lymphatic invasion is an important prognostic parameter and should be included as a stratification criterion when selecting patients for adjuvant (locoregional) therapy.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1534-4681
    Keywords: Melanoma ; Sentinel node procedure ; Follow-up ; False-negative ; Success
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: The sentinel node (SN) concept assumes that early lymphatic metastases, if present, always are found first in the SN. The aim of this study was to determine the reliability ofthis procedure by establishing the success rate and number of failed procedures during a follow-up period of at least 2 years. Methods: From August 1993 to November 1996, 204 consecutive patients with stage I and II cutaneous melanoma underwent SN biopsy by a triple technique. Preoperatively, all patients underwent (dynamic) lymphoscintigraphy. A gamma probe and blue dye helped localize the SN(s) during surgery, and these nodes subsequently were excised. These lymph nodes were step-sectioned and examined by routine and immunohistochemical staining. If the SN contained tumor cells, a lymphadenectomy was performed at a later date. Results: The median follow-up time was 42 months. The success rate was 99%. Three patients developed a recurrence in the negative SN basin during follow-up, without simultaneous appearance of (locoregional) metastases. Conclusions: With a 99% success rate and a 1.5% rate of failed SN procedures (7% falsenegative rate) after a median follow-up of 3.5 years, we concluded that the combined triple technique approach of detecting the SN was a reliable method to accurately identify and retrieve the SN.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1534-4681
    Keywords: Melanoma ; Sentinel node ; Pathology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Sentinel lymph node (SLN) biopsy can accurately predict the presence of metastatic melanoma (MM) and has been used to identify patients with occult metastases. We present an analysis of the sensitivity and specificity of standard pathological techniques including intraoperative frozen section, permanent section, and immunohistochemistry in diagnosing MM within the SLN. Methods: Sixty-nine consecutive patients with primary malignant melanoma thickness of .1.0 mm or thinner lesions invading the reticular dermis (Clark level IV) who underwent SLN biopsy were reviewed. Lymph nodes were examined intraoperatively by frozen section (FS), permanent section (H&E), and by immunohistochemistry (IH) for S-100 protein and HMB45. Results: MM was found in 14 of 69 cases (20%). Permanent section H&E was performed in all cases, FS in 64 cases, and IH in 65 cases. FS analysis diagnosed MM in 4 of 14 cases (29%), was suspicious in 2 of 14 (14%), and falsely negative (FN) in 8 of 14 (57%) ultimately found to be positive with further workup. Within the FN group, MM was identified on review of the original FS slides in 3 of 8 cases (38%). Furthermore, within the FN group, the remaining 5 cases were identified as positive for MM by either permanent and/or deeper H&E sections and IH. IH alone with permanent H&E sections would have diagnosed MM in only 8 of 10 cases (80%) that were FS negative or suspicious. In no cases was MM identified by IH alone with the permanent and deeper H&E sections being negative. It is noteworthy that no false-positive cases were identified. Conclusions: Intraoperative FS has low sensitivity in identifying MM within the SLN. IH alone does not increase the diagnostic yield. A combination of permanent H&E sections with deeper levels and S-100 and HMB45 IH dramatically increases the overall diagnostic sensitivity of SLN biopsy. Definitive diagnosis should await permanent H&E sections and IH staining.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1534-4681
    Keywords: Bryostatin ; Ionomycin ; Immunotherapy ; T-lymphocytes ; Vaccine ; Melanoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: For the relatively nonimmunogenic B16-F10 murine melanoma, it has been found that genetically engineered expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) but not interleukin (IL)-2, IL-4, or interferon-gamma (IFN-γ) resulted in a vaccine that could induce resistance to rechallenge. Because T cells from lymph nodes draining the sites of some progressive tumors can mediate tumor regression after in vitro activation, it seemed possible that even apparently nonimmunogenic melanoma cells might induce similar preeffector cells in the vaccine-draining lymph nodes (DLNs). Methods: C57BL/6 mice were vaccinated with B16-F10 cells that were either unmodified or genetically modified to produce IL-2, IL-4, GM-CSF, or IFN-γ. DLNs were harvested 10 days after vaccination for adoptive immunotherapy (AIT). The DLN cells were activated with bryostatin 1 and ionomycin (B/I), expanded for 10 days in culture, and transferred to mice with 3-day pulmonary metastases. Pulmonary nodules were counted 14 days after AIT. Results: Adoptive transfer of expanded DLN lymphocytes sensitized by inoculation of WT B16-F10, or IL-4, GM-CSF, or IFN-γ expressing cells significantly reduced pulmonary metastases. Despite the spontaneous regression of IL-2-transduced B16-F10 tumors, DLN from mice inoculated with IL-2 producing B16 cells had little or no antitumor activity. Conclusions: B16-F10 vaccination strategies that apparently do not induce systemic immunity can effectively sensitize DLN preeffector cells.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Psychopharmacology 96 (1988), S. 237-242 
    ISSN: 1432-2072
    Keywords: Methysergide ; Cyproheptadine ; Metergoline ; Ritanserin ; Food intake ; Satiety ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Two series of experiments examined whether 5-hydroxytryptamine (5-HT) antagonists induce feeding in rats. In the first series of experiments separate groups of rats were injected with various doses of methysergide, cyproheptadine, metergoline or ritanserin prior to a 2 h period of access to a wet mash diet which induced vigorous feeding under control conditions. None of the antagonists increased food intake in this paradigm. Rather, at certain doses, methysergide, cyproheptadine and ritanserin induced slight decreases in food intake. Since 5-HT may be involved in controlling satiety, it may be that a more appropriate test of the efficacy of these compounds involves administering them to maximally satiated rats. Consequently, the effects of these drugs were investigated in groups of rats which had fed to satiety immediately prior to drug treatment. In this paradigm methysergide, metergoline and ritanserin, but not cyproheptadine, induced definite increases in food intake. It is suggested that this effect occurs via a dissipation of satiety signals, and that these results further support the hypothesis that 5-HT is involved in controlling satiety. The possibility that these antagonists act on peripheral 5-HT systems is discussed.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2072
    Keywords: Key words Nicotinic receptor ; Dorsal hippocampus ; Anxiety ; Phobia ; Elevated plus-maze ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract   Rationale: The elevated plus-maze provides a test situation in which distinctive states of anxiety are elicited on trials 1 and 2 and the dorsal hippocampus has previously been shown to mediate the anxiogenic effects of (–)-nicotine in the social interaction test. Objective: To determine the effects of a wide dose range of (–)-nicotine on trial 1 and 2 in the plus-maze after systemic administration and whether the dorsal hippocampus is a site mediating the anxiogenic effect of nicotine. Methods: (–)-Nicotine (0.001, 0.005, 0.01, 0.05, 0.1, 0.5 and 1 mg/kg) was injected IP 30 min before testing for 5 min in the plus-maze. Rats receiving dorsal hippocampal infusions received bilateral infusions of 0.5 μl of artificial CSF or (–)-nicotine (0.1, 1, 4 or 8 μg). The needle was left in place for 50 s after injection and testing took place 3 min later. Rats tested on trial 1 were naive to the plus-maze, those tested on trial 2 had received a previous 5-min undrugged exposure to the maze 48 h earlier. Results: Low doses of (–)-nicotine (0.001, 0.005, 0.01, 0.05 and 0.1 mg/kg, IP) were without effect on either trial, but higher doses (0.5 and 1 mg/kg, IP) had anxiogenic effects on both trials, as shown by decreases in percentage time spent and percentage entries onto the open arms. Infusion of (–)-nicotine (0.1, 1, 4 and 8 μg) bilaterally into the dorsal hippocampus was without effect on trial 1, but 1 μg had an anxiolytic effect on trial 2, shown by an increased percentage time spent on the open arms. Conclusions: The results on both trials in the plus-maze after systemic administration of nicotine add to previous reports from the social interaction test that high doses of nicotine have anxiogenic effects. However, the effects of nicotine in the dorsal hippocampus are different in all three anxiety tests (anxiogenic in social interaction, ineffective on trial 1, anxiolytic on trial 2) showing that nicotinic cholinergic control in this brain region may vary depending on the state and/or type of anxiety generated by the test. The brain region(s) underlying the anxiogenic effects of IP nicotine on both trials in the plus-maze remain to be identified.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2072
    Keywords: Key words Amphetamine ; Choice performance ; Stimulus detection ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Conditions under which amphetamine may facilitate stimulus detection task choice performance in rats were investigated. Rats (n=15) were trained in a two-choice, light-detection task to three successively more stringent criterion levels of task training (minimal, intermediate, and extended) and then tested after administration of saline, 0.25, 0.50, and 0.75 mg/kg d-amphetamine (AMP). For each training level, baseline levels of choice accuracy were maintained at approximately 82% by manipulating the animals’ cue duration. No aspect of performance was enhanced by any dose of AMP after minimal criteria training, and there was a dose-dependent decrease in the number of trials completed. After the intermediate level of training, the 0.25 mg/kg dose of AMP reliably increased choice accuracy, there was no reliable change in choice reaction time, and there was a dose-dependent decrease in the number of trials completed. After the extended training, the 0.25 mg/kg dose of AMP reliably increased choice response accuracy, the 0.25 and 0.50 mg/kg doses of AMP reliably decreased choice reaction time, and there was no reliable change in the number of trials completed at any dose of AMP. These results support the contention that psychostimulants can facilitate the choice performance of rats in stimulus detection tasks if an appropriately low dose is used and the animal’s behavior is strongly controlled by the stimulus-reinforcement contingencies of the task.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2013
    Keywords: Key words Mast cells ; Endothelial cells ; Cell adhesion molecules ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Mast cell (MC)-mediated induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) and of E-selectin was studied in cultures of rat heart endothelial cells (EC) and human umbilical vein EC (HUVEC) respectively. MC induced VCAM-1 and E-selectin, but hardly any ICAM-1 in EC. Induction was not dependent on MC degranulation, but seemed to be provoked by constitutively released substances, other than histamine, from MC. Co-incubation of MC and EC, allowing for direct contact between the two cell types, was more potent in induction than MC co-incubated separately from EC using a permeable membrane. MC were less potent in induction than exogenous added cytokines or LPS. Induction of cell adhesion molecules in rat heart EC was MC-specific, since EC incubations with either rat cardiomyocytes or heart fibroblasts had no effect. The data show that rat MC, independent of degranulation, secrete mediators relevant for the induction of a specific set of EC adhesion molecules in vitro. This suggests a (supportive) role for MC in cell-adhesion molecule induction in the endothelium in settings of early or mild inflammation. The results are discussed in the context of inflammatory processes in the heart in vivo.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-0738
    Keywords: Key words 5-Lipoxygenase inhibitors ; N-Hydroxyureas ; Nephrotoxicity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The N-hydroxyurea derivatives 70C ((E)-N-{3-[3-(4-fluorophenoxy)phenyl]-1-(R,S)-methylprop-2-enyl}-N-hydroxyurea) and its (R) 225C and (S) 404C enantiomers, which were being developed as 5-lipoxygenase inhibitors for the treatment of certain allergic and inflammatory conditions, were found to cause severe glomerulonephropathy in the rat. The lesion appeared to be of greater severity in female rats compared with male rats. In addition, 70C and 225C treated animals appeared more severely affected than 404C treated animals. Detailed examination of the lesion in animals dosed with 225C showed that there was a clear relationship between the onset of the lesion and the dose given, i.e. the higher the dose the sooner the lesion developed. The earliest changes detected in the kidney by transmission electron microscopy were noted in the glomeruli, in which the visceral cells appeared enlarged and showed varying degrees of foot process loss. In the more advanced lesion, the degree of foot process loss became more obvious and changes in the kidney tubules were seen by light microscopy. The morphological changes were mirrored by a dose-related increase in water consumption, an increased kidney to body weight ratio and gastrointestinal oedema, suggesting impaired renal function. Shortly after the onset of foot process loss, decreases in the total plasma protein and albumin and increases in the plasma cholesterol, triglycerides, urea and creatinine were recorded. These changes, particularly the foot-process loss, together with increased proteinuria, hypoalbuminaemia, hypercholesterolaemia and lipaemia, are all characteristic of “minimal change nephrotic syndrome”. Because of the serious nature of the kidney lesion caused by these N-hydroxyureas in the rat, it was considered that it precluded their development as therapeutic agents for use in man.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; virulence gene ; host range ; plant tumour induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Octopine and nopaline strains of Agrobacterium tumefaciens were found to differ in virulence on Nicotiana glauca. This difference is due to the absence of a functional virF locus, which is necessary for efficient tumorigenesis on N. glauca, from the nopaline Ti plasmids. Genetic studies and DNA sequence analysis of the virF locus revealed that virF embraces one open reading frame coding for a hydrophilic protein with a molecular mass of 22437 Da. Transcription of virF is directed from left to right, towards the T region, and is strongly induced by the phenolic compound acetosyringone. We established that virA and virG, two genes known to be essential for induction of the vir regulon, are necessary for acetosyringoneinduced virF expression, implying that virF is a member of this vir regulon. Agrobacterium virF mutants can be complemented for tumor induction by co-infection with avirulent Agrobacterium ‘helper’ strains. We found that such ‘helper’ strains must express not only the virF gene but also the vir operons virA, virB, virD and virG.
    Type of Medium: Electronic Resource
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