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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Digestive diseases and sciences 44 (1999), S. 1349-1355 
    ISSN: 1573-2568
    Keywords: ANTITHROMBIN III ; THROMBIN ; HEPARIN ; BLOOD COAGULATION ; ACETALDEHYDE ; ALCOHOL ; ALCOHOLISM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Acetaldehyde (AcH) at preincubationconcentrations of 447, 89.4, and 17.9 mM potentiates theeffects of heparin on the clotting time of plasma. Whilecontrol plasma clotted in the range of 12.6 ± 0.1 to 13.8 ± 0.1 sec, and heparin-treatedplasma clotted in a range from 131.5 ± 2.5 to168.2 ± 1.2 sec, heparin that was preincubated atroom temperature for 30 min with 89.4 or 447 mM AcH didnot clot plasma in 300 sec. Heparin exposed to 17.9 mMAcH clotted plasma in 193 ± 1.1 sec. Ethanol ata 404 mM concentration also prolonged the clotting timeof heparin-treated plasma 〉300 sec, while 202 mM ethanol prolonged the clotting time ofheparin-treated plasma from 149.0 ± 2.0 sec to219.5 ± 1.7 sec. It is suggested that AcH altersthe tertiary structure of heparin by adduct formation,possibly by formation of cyclic acetals with iduronicand glucuronic acids, thereby more readily affectingbinding of the glycosaminoglycan to antithrombin IIIand/or thrombin, prolonging clotting time. Ethanol, which does not react covalently with heparin,might affect its conformation as a consequence of anorganic solvent effect. Protamine sulfate prolonged theclotting time of plasma from 13.6 ± 0.1 sec to 17.9 ± 0.2 sec. Protaminesulfate-treated heparin clotted plasma in 21.0 ±0.4 sec relative to heparin-treated plasma (160.4± 1.7 sec). In subsequent experiments,AcH-treated protamine sulfate extended the clotting time of protamine sulfate from17.9 ± 0 sec to 33.7 ± 0.6 sec. Prioraddition of protamine sulfate to AcH- heparin mixturesor heparin to protamine sulfate-AcH mixtures beforeaddition to plasma resulted in clotting times of 22.0± 0.4 sec and 24.1 ± 0.5 sec,respectively, relative to control clotting times of162.3 ± 2.6 sec for plasma-heparin mixtures.These results confirm both the reduction in coagulation time ofheparin-treated plasma by protamine sulfate and theprolongation of clotting time of plasma by protaminesulfate. Furthermore, and importantly, they indicatethat acetaldehyde-treated protamine sulfate is a more effectiveanticoagulant than protamine sulfate. It is suggestedthat reversible adduct formation between acetaldehyde,heparin, and protamine sulfate may occur as a meansexplaining the essentially identical coagulation time ofthese mixtures when added to plasma regardless of theorder of premixing. Ethanol (404 mM) did not influenceprotamine sulfate effects. Lastly, the potentiation of the anticoagulant function of heparin byacetaldehyde suggests that a structural modification ofthe glycosaminoglycan may occur in alcoholics.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-2568
    Keywords: ALCOHOL ; COAGULATION ; FACTOR VII ; FACTOR IX ; ACETALDEHYDE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The first metabolite of ethanol, acetaldehyde,has the ability to form adducts with proteins and altertheir function. It has been shown that acetaldehydereacts with various proteins of the blood coagulation pathway and, subsequently, produces aprolongation of the clotting time. This study evaluatedthe function of clotting proteins from the extrinsiccoagulation pathway (factor VII) and the intrinsiccoagulation pathway (factor IX) when preincubated withacetaldehyde as compared to a control and compared topreincubation with ethanol. Prior to use in a clottingassay, incubation times with acetaldehyde, ethanol, and the control were the same for both factorsVII and IX. An automatic fibrometer measured theclotting times. Factor VII preincubated withacetaldehyde prolonged the clotting time. However,factor IX preincubated with acetaldehyde actuallydecreased the clotting time. Of interest, both factorsVII and IX preincubated with acetaldehyde producedstatistically significant results when compared to thecontrol and ethanol. This experiment indicates thatacetaldehyde, in forming an adduct with proteins of theblood coagulation pathway, may induce a conformationalchange of factors VII and IX so as to either increase or decrease the clotting time. Therefore, it ispossible that some of the deranged coagulation inalcohol abusers may be a final net result of theinteraction of acetaldehyde and proteins of thecoagulation pathway.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Digestive diseases and sciences 43 (1998), S. 1746-1751 
    ISSN: 1573-2568
    Keywords: ANTITHROMBIN III ; THROMBIN ; ACETALDEHYDE ; ALCOHOL ; ALCOHOLISM ; BLOOD COAGULATION
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The anticoagulant activity of antithrombin III(ATIII), as observed in a plasma-free system consistingof thrombin and fibrinogen, is readily reduced byacetaldehyde (AcH) at concentrations of 447, 89.4, and 17.9 mM. Whereas controlthrombin-fibrinogen mixtures clotted in 17.7 ±0.75 sec, ATIII prolonged clotting time to 55.0 ±1.75 sec on preincubation with thrombin for 30 min atroom temperature. On subsequent preincubation of ATIII with theAcH for 30 min at room temperature and passage of themixture through Sephadex G-25 minicolumns to removeexcess AcH, the eluates were tested for anticoagulant activity. Clotting times of 20.9 ± 1.0,32.3 ± 1.0, and 45.3 ± 1.6 sec wereobtained with 447, 89.4, and 17.9 mM AcH-ATIII mixtures,respectively. These data suggest that functional groupson ATIII, such as guanidiniums, aminos, and others aresusceptible to adduct formation with AcH, therebyaltering the shape and charge of the anticoagulant. Asa consequence of this type of reaction, an alteredmolecule of reduced biological activity may be produced.These experimental results may explain, in part, thereduction in ATIII levels reported by others in patientswith alcoholic liver disease.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 175-193 
    ISSN: 0886-1544
    Keywords: primary cilia ; connective tissues ; secretory organelles ; extracellular matrix ; cybernetic probe ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: More than 300 primary cilia have been identified electronmicroscopically in a variety of embryonic and mature connective tissue cells. To further define the enigmatic function of these cilia, we examined the interrelationships between the basal apparatus and cytoplasmic organelles and the ciliary shaft and the extracellular matrix. The basal diplosome was consistently associated with the secretory organelles including the maturing face of the Golgi complex, Golgi vacuoles and vesicles, the microtubular network, the plasma membrane, and coated pits and vesicles. Small vesicles and amorphous granules were also observed within the ciliary lumen and adjacent to the ciliary membrane. Microtubule-membrane bridges linked axonemal tubules to the ciliary membrane. The position, projection, and orientation of the axoneme were influenced by the structural organisation and mechanical properties of the matrix and frequently caused angulation of the ciliary shaft relative to the basal body. Located midway between the secretory apparatus and the extracellular matrix, primary cilia would appear ideally situated to mediate the necessry interaction between the cell and its surrounding environment prerequisite to the formation and maintenance of a functionally effective matrix. We propose that primary cilia in connective tissue cells could act as multifunctional, cellular cybernetic probes, receiving, transducing, and conducting a variety of extrinsic stimuli to the intracellular organelles responsible for effecting the appropriate homeostatic feedback response to changes in the extracellular microenvironment.
    Additional Material: 17 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Clinical Anatomy 4 (1991), S. 447-455 
    ISSN: 0897-3806
    Keywords: connective tissue ; gross anatomy ; histology ; pelvic fascia ; ligaments ; Life and Medical Sciences ; Miscellaneous Medical
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A preliminary report from this group described a unique layer of subperitoneal elastic fibers in the female pelvis. The current study was undertaken to better characterize this elastic layer in the female and determine if it exists in the male pelvis. Thirty-seven hemipelves were used in this study and were employed for one of two purposes: dissection and/or sampling for histologic study. The following histologic stains were employed: Verhoeff's hematoxylin for elastic fibers, Masson's trichrome for collagen, and hematoxylin and eosin for general morphology. In some areas (broad ligament) the subperitoneal elastic layer was very thin with dispersed fibers while in other areas (uterosacral fold) the layer was thick with tightly packed fibers. A subperitoneal layer of elastic fibers was not detected in samples from the greater omentum; however, a dense but poorly organized layer of elastic fibers was noted between the posterior layer of the rectus sheath and the peritoneum. While a similar layer of elastic fibers was found in the male pelvis, the layer was less dense and less well organized than in the female. These findings indicate that in addition to the well-accepted pelvic ligaments, in the female there may be a layer of elastic fibers that assists in supporting pelvic viscera in a more dynamic manner than the ligaments. Proper alignment or manipulation of this layer during reconstructive surgery may enhance postoperative results.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 37 (1994), S. 457-461 
    ISSN: 1040-452X
    Keywords: Canine sperm ; Pyospermia ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The red wolf (Canis rufus) is an endangered species with 194 individuals remaining in the wild and in various captive facilities. Breeding efforts at the Graham, WA site (Point Defiance Zoo and Aquarium) have involved artificial insemination with fresh or frozen semen in an effort to increase population and maximize the genetic potential of the stock. Electron microscopic observations were made in semen specimens obtained by electroejaculation from mature males prior to their use in an effort to determine semen parameters that might be useful in guiding breeding procedures. Sperm samples were either fixed immediately or treated with capacitating media and fixed after 4 to 7 hr of incubation. Many of the specimens examined were pyospermic (white cell in semen) and showed evidence of spermophagy, primarily by neutrophils. Of the six animals surveyed, only one showed little evidence of spermophagy, and three had extensive pyospermia and spermophagy but this finding was not correlated with fertility. Samples fixed immediately as well as those incubated for several hours showed evidence of spermophagy, indicating that the phagocytosis was not the result of culture. Gene pool restriction and/or captive stress may be contributing factors of reduced semen quality. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 34 (1993), S. 175-182 
    ISSN: 1040-452X
    Keywords: Fixation ; Artifact ; Ruthenium red ; Percoll ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Various fixation protocols were used in an attempt to improve preservation of rat epididymal sperm for high-resolution low-voltage scanning electron microscopy (HR-LVSEM). Wash solutions and fixatives of different composition and osmolarity were tested. Paraformaldehyde and glutaraldehyde concentrations were varied between 0.5% and 3%. Ruthenium red was tested as an additive in both primary fixation and postfixation, or in postfixation alone. HR-LVSEM revealed various degrees of ruffing, folding, blebbing, and peeling off of the plasma membrane, as well as holes of different sizes. The plasma membrane overlying the acrosome and the connecting piece proved to be particularly sensitive to varying fixation conditions. Consistent topographical differences were revealed among the different domains over the sperm head. Most of the differences were considered to be artifacts. Their consistency, however, suggests that structural and biochemical differences exist either within the membrane or in the structures subjacent to the membrane. Primary fixation turned out to be less critical than postfixation. Preservation of a smooth plasma membrane without holes could only be achieved when primary fixation in low aldehyde concentrations, with or without ruthenium red, was followed by postfixation with OsO4 and 1,000 ppm ruthenium red. Examination of thin sections of the same material confirmed that even a considerable number of small holes are difficult to detect in transmission electron microscopy. These results show that with the recent increase in resolution of LVSEM there is need for further effort to improve sample processing. © 1993 Wiley-Liss, Inc.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 411-431 
    ISSN: 1059-910X
    Keywords: Corrosion casts ; LM ; SEM ; TEM ; Microvasculature ; Ultrastructure ; Absorption ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The aim of the present study was to provide a comprehensive morphological analysis of the porcine epididymis in view of the specific functions being performed in different regions of this organ. Blood supply and microvasculature of efferent ductules and epididymal duct were investigated by means of corrosion casts which were analysed macroscopically and by scanning electron microscopy. This revealed blood supply to the testis and epididymis to be closely related. The capillary pattern was typical for the efferent ductules, the caput, corpus, and distal cauda epididymidis, respectively. Corrosion casts were also used to visualize the course of the efferent ductules themselves. Tissue samples from different regions of the efferent ductules and epididymal duct were examined by light microscopy and both scanning and transmission electron microscopy, with special attention being payed to transitional areas. Morphological criteria allowed the distinction of three segments within the efferent ductules and of the initial segment, proximal caput, distal caput, corpus, proximal cauda, and distal cauda regions of the epididymal duct. Components of the endocytic apparatus of efferent ductule principal cells were identified by ferritin uptake. Ultrastructural evidence of absorption in the epididymal duct was particularly prominent in proximal and distal caput. Extensive cisternae of rough endoplasmic reticulum and a well-developed Golgi apparatus were indicative of active protein synthesis and secretion especially in the distal caput and corpus regions. However, assignment of various organelles in principal cells of the epididymal duct to either absorptive or secretory pathways still remains tentative. © 1994 Wiley-Liss, Inc.
    Additional Material: 53 Ill.
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  • 9
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Since Euglena gracilis Klebs var. bacillaris Pringsheim contains a species of DNA unique to the chloroplast, an important question concerns the extent to which light unblocks the reading of the organelle's template to provide the informational RNA's necessary to construct the plastid proteins. Experiments with 32Pi labeling of chloroplast and nonchloroplast RNA's during light-induced chloroplast development show that both the RNA of the chloroplast and of the rest of the cell become labeled during this process, with the chloroplast RNA's displaying the higher specific activity. The fact that chloroplast RNA is not uniquely labeled indicates that process other than a simple reading of the chloroplast DNA are involved. If we are to preserve the concept of a reasonable degree of chloroplast informational autonomy, we may assume, from this and other data, that the light induction of chloroplast development involves not only the unblocking of chloroplast DNA to make information available, but also a concomitant unblocking of other sites of informational RNA synthesis (e.g., nuclear and mitochondrial DNA's). Such sites external to the developing chloroplast may be concerned with making available the building blocks and energy necessary for the synthesis of chloroplast constituents coded for by the chloroplast DNA. This model leads to the prediction that photosynthesis could be gratuitous for chloroplast development if these nonchloroplast sites were providing most of the building blocks and energy. Experiments are reported which show that chloroplast formation and the acquisition of photosynthetic competence can be achieved under conditions where photosynthesis is completely inhibited for the entire span of development by using the highly selective inhibitor 3, (3,4-dichlorophenyl) 1, 1-dimethyl urea (DCMU), in agreement with the proposed model. The fact that more than just the chloroplast responds to the inducing signals for chloroplast differentiation raises the problem of experimental measurement of interaction among cellular organelles. Since chloroplast development is usually carried out in resting cells to avoid complications due to cell division, we discuss the limitations imposed by turnover in such nondividing systems and present evidence that most of the RNA labeling observed, although actinomycin-D-sensitive, is due to turnover and/or the utilization of preexisting pools. Evidence obtained with mutants of Euglena that form only partial chloroplasts or that lack plastid DNA and plastid-related structures is reported. Such evidence indicates that the functional proplastid restrains overall RNA labeling in the uninduced cells and suggests that the proplastid might be the source of regulatory metabolic signals in the normal plastid-containing cells.
    Additional Material: 16 Ill.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 193 (1979), S. 23-41 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: To further characterize Sertoli cell-germ cell junctional specializations seminiferous tubules from sexually mature Sprague-Dawley rats were dissociated by enzymatic and mechanical methods. Ultrastructural analysis of cell suspensions prepared by incubation in collagenase alone or by mechanical methods revealed that spermatids remained attached to Sertoli cells or Sertoli cell fragments. Such cellular associations were found only between Sertoli cell fragments and spermatids in which the developing acrosome had made contact with the plasma membrane (step 8 and subsequent steps of spermiogenesis). Furthermore, the fragments were confined to that region of the plasma membrane over the acrosome. The Sertoli cell half of this adhesive site displayed the typical elements of Sertoli cell junctions, filamentous bundles and associated cisterna of endoplasmic reticulum, in apposition to the spermatids. The spermatids demonstrated no surface specializations at the attachment sites. In contrast, in cell suspensions prepared with trypsin, spermatids were free of attachments to Sertoli cells or their fragments. These results demonstrate that: (1) the junctions act to bind cells together, (2) adhesive type contact is established between Sertoli cells and spermatids at step 8 and subsequent steps of spermiogenesis, (3) contact is restricted to the spermatid plasma membrane over the acrosome, and (4) spermatids can be freed from the junctional specializations by treatment with trypsin.
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