bluuebunny/crossref_metadata_2025 huggingface dataset:
Stage‐specific immunolabeling for oviductin in the secretory granules of the oviductal epithelium of the golden hamster during the estrous cycle
Authors: Roux Emmanuelle , Kan Frederick W. K.
Published: February 2005

Background: We have previously localized an antigen of oviductal origin in the zona pellucida of postovulatory hamster ova. This antigen is a high molecular weight glycoprotein secreted by the non‐ciliated secretory cells of the oviduct and is later transferred to the zona pellucida of the oocyte during oviductal transit. This glycoprotein is rich in N‐acetyl‐D‐galactosamine residues and has been designated Hamster Oviductin‐1. In the present study, a monoclonal antibody (MAb) raised against this oviductin was used to detect the presence of this antigen in oviductal tissue during the estrous cycle.

Methods: Twenty mature female golden hamsters were used and were divided into five groups of five animals each according to the five different stages of the estrous cycle. Quantitative immunocytochemistry was performed on MAb‐labeled thin sections of Lowicryl‐embedded ampullary region of hamster oviducts. Control experiments were also carried out to assess the specificity of the immunolabeling.

Results: Quantitative analysis of the immunogold labeling indicated that maximum labeling for oviductin in the secretory granules of oviductal epithelial secretory cells was found around the time of ovulation, i.e., at estrus. The intensity of immunolabeling decreased from metestrus to diestrus 1, was at a minimum at diestrus 2, and started to increase at proestrus.

Conclusion: Together, these quantitative results indicate that expression of oviductin in the secretory granules of the hamster oviductal secretory cells is stage specific. Maximum labeling for the antigen coincides with the time of ovulation suggesting an important role for the oviductal epithelium in contributing its secretory product to the zona pellucida of oocytes freshly released from the ovary. Since the oviduct is the site of sperm‐egg interaction and where fertilization and early embryo development take place, the maximal production of oviductin at the time of ovulation may facilitate some of these crucial steps during the intricate process of reproduction. © 1995 Wiley‐Liss, Inc.
Expression of renin in coagulating glands is regulated by testosterone
Authors: Kon Yasuhiro , Endoh Daiji , Murakami Kazuo , Yamashita Tadashi , Watanabe Tomomasa , Hashimoto Yoshiharu , Sugimura Makoto
Published: February 2005

Background: The presence of extrarenal or local renin‐angiotensin system (RAS) has been noted in several tissues, although its functions have not yet been clarified. Renin from the coagulating gland (CG) is the most recently discovered local RAS and is a significant subject for investigation because large amounts of both mRNA and proteins are detected in this organ. Recently, it has been reported that testosterone influences renin synthesis in several extrarenal tissues, although it has no effect on intrarenal renin. Therefore, it is possible that CG renin is also regulated by testosterone.

Methods: Forty‐four male C57BL/6 mice, aged 3 wk to 6 mo, were used in studies on the ontogeny and androgen regulation of the RAS in the CG. The tissues were fixed with Bouin's solution and paraffin sections were stained with immunohistochemical methods using antirenin antiserum. In each immunostained section, the relative number of renin‐containing cells in terminal portions of the CG were counted.

Results: Immunoreactivity for renin was first detected at 6 wk after birth. After that time, the number of renin‐containing cells gradually increased throughout the experiment. In adults, several patterns of renin immunoreactivity were demonstrated in almost all epithelial cells of CGs, specifically; (1) basolateral granular reaction, (2) diffuse immunoreactivity throughout the cytoplasm, and (3) restricted nuclear reaction. Excretory products of some terminal lumina were also found to be positive for renin. At 10 days after castration, renin‐containing cells in ductal termini were decreased and remained at low levels until at 4 wk after castration. After testosterone injection, numerical values of renin‐containing cells were high at 1 wk and then decreased at 2–3 wk.

Conclusion: It is suggested that CG renin of the mouse is expressed together with sexual maturation during development and that it depends on the testis, possibly the male sex hormone. © 1995 Wiley‐Liss, Inc.
Femoral peak bone mass and osteoclast number in an animal model of age‐related spontaneous osteopenia
Authors: Okamoto Yutaka , Takahashi Kenshirou , Toriyama Kazuhiro , Takeda Norikazu , Kitagawa Kaori , Hosokawa Masanori , Takeda Toshio
Published: February 2005

Background: SAMP6 was developed as a murine model of age‐related spontaneous osteopenia characterized by low peak bone mass. A morphometric study of the growing femur in SAMP6 and sex‐matched SAMP2 at 10 days to 4 months of age was done to examine the pathogenic process related to osteopenia.

Methods: Age‐related changes in cortical bone thickness, femur score, trabecular bone volume, thickness of epiphyseal growth plate, number of osteoclasts, and osteoclast surface were measured with a computerized image analyzer. Osteoclasts were examined cytomorphometrically after TRAP (tartrate resistant acid phosphatase) staining of the femoral sections.

Results: Cortical bone thickness and femur score increased significantly with age, while trabecular bone volume decreased significantly. Comparing mean values of cortical bone thickness, femur score and trabecular bone volume, we noted significantly lower mean values in SAMP6 than in SAMP2 mice. These significant inter‐stain differences first became evident in 20–40‐day‐old mice, but there was no significant difference in thickness of the epiphyseal growth plate between the two strains. The mean values of the number of osteoclasts per unit none surface length and of the osteoclast surface in SAMP6 were significantly greater than in age‐ and sex‐matched SAMP2. Histograms of distribution of size of osteoclasts of 40‐day‐old male mice revealed that larger ones were more frequently seen in SAMP6. Furthermore, the ratio of osteoclasts/TRAP positive cells free in the bone marrow cavity was significantly higher in SAMP6 than in SAMP2.

Conclusion: Activated bone resorption may play a role in the osteopenia seen in SAMP6. © 1995 Wiley‐Liss, Inc.
Synthesis and expression of laminin during human foetal lung development
Authors: Lallemand Aude V. , Ruocco Sandrine M. , Gaillard Dominique A.
Published: February 2005

Backgrounds: The lung develops by epithelial tubes budding and branching into a flexible mesenchyme. This growth is associated with the remodelling of the epithelial basement membrane, of which laminin is a major component. Methods: Both the synthesis and expression of laminin were studied in the human lung between 10 and 31 weeks of gestation, using in sity hybridization and immunohistochemistry. Results: The synthesis of the β chain was active in the epithelial and surrounding mesenchymal cells. The mRNAs coding for the γ chain were less abundant and mainly found in the epithelium. The synthesis of these two chains continued throughout gestation, and no significant difference in the density of hybridization grains could be detected between the tips of the expanding buds and the proximal portions. Immunohistochemical localization of laminin showed important modifications of the basement membrane during gestation. In the first part of the pseudoglandular stage the epithelial basement membrane stained continuously for laminin. Later, the basement membrane was labelled in a graded fashion: at the apex of the growing buds the staining became weak with focal disruptions. Both epithelial and mesenchymal synthesis of laminin remained active, while the polypeptide was undetectable using immunohistochemistry. Conclusions: These findings suggest that the remodelling of the basement membrane during human lung morphogenesis is probably not related to a decreasing synthesis of laminin, but to either a proteolytic degradation or the assembly of an inadequate complex undetectable with the polyclonal antibody antilaminin. © 1995 Wiley‐Liss, Inc.
Trans‐Golgi network (TGN) of different cell types: Three‐dimensional structural characteristics and variability
Authors: Clermont Y. , Rambourg A. , Hermo L.
Published: February 2005

Background: The trans‐Golgi network (TGN) is generally considered as a distinct and permanent structural compartment of the Golgi apparatus of various cell types. To verify this postulate we examined and compared the three‐dimensional characteristics of the TGNs of 14 different mammalian cell types as presented in our various publications since 1979 when we initially described the trans‐tubular network of Sertoli cells.

Methods: In all these studies we used low and high voltage electron microscopes on thin or thick sections of tissues fixed with glutaraldehyde and postfixed with reduced osmium. The section were stained with uranyl acetate and lead citrate. Stereopairs, prepared from photographs of tilted specimens, permitted a direct observation of the three‐dimensional structure of the various elements of the Golgi apparatus.

Results: The TGNs are multilayered and extensive in cells which do not form large typical secretory granules (Sertoli cells, nonciliated cells of ductuli efferentes, spinal ganglion cells) but have an extensive lysosomal system. The TGN is absent in cells forming very large secretory granules (secretory cells of seminal vesicles and lactating mammary glands). The TGNs are small in cells producing small to medium‐size secretory granules and/or appear as residual fragments on the trans aspect of the Golgi stacks (e.g., mucous cells of Brunner's gland, pancreatic acinar cells, etc.). In cells with multiple and extensive TGNs, a continuity of these tubular networks with the two or three transmost saccules of the stack is observed but there are seemingly no connections between the TGNs. Whenever the TGNs are present, they do not form a continuous structure along the Golgi ribbon. However, they do present, in all cases, configurations suggestive of desquamation and renewal.

Conclusions: The structure of the TGN varies considerably from one cell type to another, being extensive in cells not showing typical secretory granules but having an extensive lysosomal system, while in secretory cells showing small or large secretory granules the TGN is either small or even entirely absent. © 1995 Wiley‐Liss, Inc.
Distribution of epithelioid cells in the wall of the chicken aorta and their functional role as chemoreceptors
Authors: Miyoshi Masakazu , Hashimoto Yoshiharu , Kon Yasuhiro , Sugimura Makoto
Published: February 2005

Background: The ultrastructural characteristics of epithelioid cells in the wall of the chicken aorta have been studied by several investigators. Their characteristics were homologous to those of carotid body type I cells and are considered to be one of the peripheral chemoreceptors. However, there are few descriptions about their location, distribution, and how they react to chemical signals, nor have there been many reports about the localization of bioactive substances in the epithelioid cells. Therefore we designed this investigation to address these problems.

Methods: Wholemount immunohistochemistry using antiserotonin antiserum, scanning (SEM), and transmission (TEM) electron microscopy were used to observe the epithelioid cells and the lumen of the chicken aorta. The localizations of bioactive substances in the epithelioid cells were immunohistochemically investigated using 11 antisera.

Results: Epithelioid cells were dispersed in the wall of the aorta, forming a band ∼1 mm in width, located 10 mm proximal to the confluence of the right and left ligamenta arteriosa. Serotonin, chromogranin, and neuron specific enolase immunoreactivities were detected in the epithelioid cells. SEM observations clearly demonstrated intraendothelial fenestrations, 1–3 μm in diameter, on the endothelial surface of the region of the band of epithelioid cells. TEM observations revealed that these fenestrations corresponded to endothelial gaps, directly beneath which epithelioid cells were sometimes located.

Conclusions: The epithelioid cells are in direct access to the aortic lumen through endothelial fenestrations. Thus they may be able to perceive chemical signals from arterial blood directly. © 1995 Wiley‐Liss, Inc.
Anatomy of the sinus node, av node, and his bundle of the heart of the sperm whale (Physeter macrocephalus), with a note on the absence of an os cordis
Authors: James Thomas N. , Kawamura Keishiro , Meijler Frits L. , Yamamoto Shoji , Terasaki Fumio , Hayashi Tetsuya
Published: February 2005

Background: Atrioventricular (AV) conduction time in large whales is only slightly greater than in smaller mammals even though their hearts are enormously larger. Little is known of the detailed histology or cytology of the conduction system of large whales. Such knowledge could be useful in defining the nature of cardiac rhythm and conduction of the whale as well as smaller mammals including humans.

Methods: We studied hearts from seven sperm whales. After fixation in formaldehyde and later dissection, specimens were prepared for histological examination.

Results: Cell size, histological organization, and innervation of the sperm whale's sinus node, AV node, and His bundle are similar to most mammalian hearts, except the sinus node is substantially larger. There is no central fibrous body between the atrial and ventricular septa, and the whale has no os cordis. Only the upper quarter of the interventricular septum is fully formed; below that there is only a thin layer of fatty connective tissue between the two ventricles.

Conclusions: Given our morphological findings, we believe that the whale's comparatively short AV conduction time may be best explained by the sinus node and AV node functioning as coupled relaxation oscillators. Absence of an os cordis or central fibrous body or strong attachment between the two ventricles may pose both electrophysiological and hemodynamic hazards when the whale is no longer in its normally buoyant aquatic environment. © 1995 Wiley‐Liss, Inc.
Histamine release from Weibel‐Palade bodies of toad aortas induced by endothelin‐1 and sarafotoxin‐S6b
Authors: Doi Yoshiaki , Ozaka Takatoshi , Katsuki Masatsugu , Fukushige Hiroshi , Toyama Eiichiro , Kanazawa Yosuke , Arashidani Keiichi , Fujimoto Sunao
Published: February 2005

Background: Endothelin‐I (ET‐1) and sarafotoxin‐S6b (STX) induce a remarkable degranulation of Weibel‐Palade (WP) bodies prior to the vasocontraction of toad aortas. As WP bodies play the role of a reservoir site of the histamine in the endothelial cells, there is the possibility that ET‐1 and STX evoke the release of histamine from WP bodies of this vessel.

Methods: Histamine concentrations were assayed by high‐performance liquid chromatography (HPLC) from the perfusate after being perfused with a solution containing ET‐1 and STX. Each vessel was fixed and embedded for conventional electron microscopy and immunoelectron microscopy using antihistamine sera.

Results: The appreciable concentrations of histamine were assayed by HPLC from the perfusate after the toad aortas were perfused with a solution containing ET‐1 and STX. The immunoelectron microscopy revealed that histamine immunoreactive gold particles in the WP bodies remarkably decreased in number in the treated samples when compared to the control ones. Our immunoelectron micrographs indicated that the release of histamine from the endothelial cells occurred in association with the degranulation and the exocytosis of the WP bodies after treatment with ET‐1 and STX.

Conclusions: The present study clearly shows that ET‐1 and STX induce the histamine release from WP bodies of the toad aortas by means of HPLC and immunoelectron microscopy. Histamine discharged from the WP bodies may be involved in the vasocontraction evoked by ET‐1 and STX. © 1995 Wiley‐Liss, Inc.
Corrosion casting study on the vasculature of nasal mucosa in the human fetus
Authors: Składzień J. , Litwin J. A. , Nowogrodzka‐Zagórska M. , Miodoński A. J.
Published: February 2005

Background: The vasculature of the nasal mucosa in the human fetus so far has not been investigated morphologically by modern techniques.

Methods: Nasal blood vessels were studies in 18–21‐week‐old human fetuses by corrosion casting and scanning electron microscopy.

Results: The general vascular architecture was similar to that described for adult humans, with (1) a dense network of subepithelial capillaries, (2) a mucosal layer of larger vessles, predominantly veins, and (3) large arteries located near the perichondrium. Specific vessel types characteristic of the nasal mucosa and important for its functions in adults, such as cavernous veins or cushion veins, were, however, absent in the fetuses, although probable precursor forms of cushion veins could be observed. The arteriovenous anastomoses had the form of short bridges, and no tortuous or glomerular anastomoses present in the mature nasal mucosa were found.

Conclusions: The underdevelopment of some vascular areas is postulated to reflect the functional immaturily of the respiratory system in the fetus. © 1995 Wiley‐Liss, Inc.
Morphology of the dorsal phalangeal connective tissue body
Authors: Bade Holger , Schubert Markus , Koebke Jürgen
Published: February 2005

Background: The connective tissue body of the dorsal finger is partitioned by multiple horizontal lamellae derived from expansions of intercellular spaces. These connective fibres are involved in providing gliding spaces for the extensor apparatus, and they formed spreading rooms of infection processes.

Methods: In order to describe the topographical histological and histochemical behavior of cellular and intercellular elements of the border lamellae of the dorsal aponeurosis of the fingers, a modified method of Epoxid resin embedding technique without pre‐infiltration is used.

Results: The bordering elements of the connective tissue lamellae described in this investigation are especially well defined in the region of the dorsal aponeurosis. The morphological examinations show that both the expansions of the intercellular space as well as defined fibrocytic cellular elements function in a manner equivalent to a synovial membrane of tendon sheaths.

Conclusions: Saving or reconstructing these structures appears to be of importance in preventing for adhesive disorders of finger motion following surgical intervention in this region. © 1995 Wiley‐Liss, Inc.
Morphometric characterisation of the fine structure of human type II pneumocytes
Authors: Fehrenbach Heinz , Schmiedl Andreas , Wahlers Thorsten , Hirt Stefan W. , Brasch Frank , Riemann Dörte , Richter Joachim
Published: February 2005

Background: Pulmonary type II pneumocytes have been examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and morphometry in numerous mammals. Until now, the fine structure of the human type II pneumocyte has not been studied by means of morphometry.

Methods: Eleven human donor lungs, which could not be made available for a suitable recipient, were preserved with Euro Collins solution (ECS) according to clinical organ preservation techniques. The lungs were fixed via the airways. Systematic random samples were analyzed by SEM, TEM, and classical stereological methods.

Results: Type II pneumocytes showed normal fine structural characteristics. Morphometry revealed that although inter‐individual variation due to some oedematous swelling was present, the cells were in a normal size range as indicated by an estimated mean volume of 763 ± 64 μm3. The volume densities were: nucleus 21.9 ± 2.2%, mitochondria 5.8 ± 0.9%, lamellar bodies 9.8 ± 3.6%, and remaining cytoplasmic components 62.4 ± 2.9% of the cell volume. Since the inter‐individual variations in the volume densities referred to the cell may, to variable degrees, reflect the variation in the reference space, the volume densities referred to the constant test point system and the respective volume‐to‐surface ratios were used for interindividual comparisons. These parameters indicate that lamellar bodies were independent of cellular swelling, while mitochondria nucleus remaining cytoplasmic components increased in size with increasing cell size.

Conclusions: Two to 7.5 hours of cold ischemia following ECS preservation do not deteriorate the fine structure of type II pneumocytes of human donor lungs. For reliable assessment of fine structural variation, morphometric parameters are required that are independent of variations in cell size. © 1995 Wiley‐Liss, Inc.
Organization and cytology of lymphoid tissue in the cynomolgus monkey conjunctiva
Authors: Ruskell G. L.
Published: February 2005

Background: Knowledge of the structure of conjunctival lymph follicles is limited in comparison with those of other mucosa. Consequently a thorough morphological study of the follicles was undertaken to see if they are similar to others or whether they have any unique features and to provide an adequate background for future experimental studies.

Methods: Normal lymphoid tissue of the conjunctiva of one side of nine young adult cynomologus monkeys was examined using a whole mount method and light and electron microscopy.

Results: Only the peripheral tarsal and orbital regions of the upper and lower palpebral conjunctiva contained lymph follicles. Lymphocytes, reticulocytes, macrophages including tingible body macrophages, putative follicular dendritic cells, and plasmacytes were indentified in the follicles and M‐cells were present in the follicle associated epithelium. High endothelial venules (HEV) displaying evidence consistent with lymphocyte extravasation were present in both primary and secondary follicles. A majority were secondary follicles with a germinal center composed largely of lymphoblasts. Mitoses were found in secondary follicles, mostly confined to the germinal centers, and they were numerous in many. Increased cell density and virtual elimination of intercellular space adjacent to some high endothelial venules suggested a possible association of these vessels with germinal center development. Chains of follicles were linked by lymph vessels, an approaching lymphatic dividing to embrace a follicle and recombining beyond it and passing to the next in line; many conjunctival follicles there‐fore have both afferent and efferent lymphatics. The vessels were in direct contact with the follicle perimeters; lymph capillaries were not present within the substance of most follicles. Numerous valves indicated drainage radially from the tarsal conjunctiva then laterally or medially from the orbital conjunctiva through the follicle chains towards the nearest canthus.

Conclusions: Conjunctival lymph follicles are fundamentally similar in structure to those found elsewhere in the body. However, the beads on a chain‐like arrangement of lymph vessels and follicles may be unique. The possible association of HEV with follicle foramtion could be a general characteristic of mucosal follicles. ©1995 Wiley‐Liss, Inc.
Cytoskeletal architecture of rat calvarial osteoclasts: Microfilaments, intermediate filaments, and nuclear matrix as demonstrated by detergent perfusion
Authors: Watanabe Hiroki , Yanagisawa Takaaki , Sasaki Junzo
Published: February 2005

Background: There have been few detailed transmission electron microscopic studies of the cytoskeleton of osteoclasts to date because of the high electron density of their cytoplasmic matrix, and there is a controversy concerning the presence of stress fibers in these cells.

Methods: The cytoskeleton and nuclear matrix of osteoclasts in the rat cranium were studied by transmission electron microscopy after perfusing the organs with a mixture of Triton X‐100 and a low concentration of glutaraldehyde to reduce the electron density of the intracellular matrix and by immunoelectron microscopy using anti‐actin and anti‐vimentin antibodies.

Results: In osteoclasts, abundant microfilaments, 5 – 6 nm in diameter, extended across the cell organelles. These filaments were also observed just under the cell membrane facing the bone, but thick bundles of microfilaments were not noted. In the cytoplasm, there were fibers 10–12 nm in diameter, which were considered to be intermediate filaments distributed in the same pattern as microfilaments. Filamentous structures consisting both of the microfilaments and intermediate filaments connected the nuclei. Intermediate filaments linked nuclei at nuclear pores (near the euchromatin), and microfilaments connected nuclei at areas of the nuclear membrane containing nucleolus‐associated heterochromatin. In the nuclear matrix after detergent treatment, thin filaments and intermediate ‐type filaments were found connecting the heterochromatins.

Conclusions: Euchromatin is considered to be an active site of DNA and RNA synthesis. Our structural evidence has lead to the proposal that intermediate filaments in the cytoplasm may act as tracks for transmission of information between nuclei and guides for the transit of molecules through the pores. © 1995 Wiley‐Liss, Inc.
Morphomechanics of the humero‐ulnar joint: I. Joint space width and contact areas as a function of load and flexion angle
Authors: Eckstein Felix , Löhe Florian , Hillebrand Susanne , Bergmann Mathias , Schulte Erik , Milz Stefan , Putz Reinhard
Published: February 2005

Background: Previous studies have shown that the trochlear notch is deeper than necessary for an exact fit with the humerus. However, humero‐ulnar joint space width and contact areas have so far not been quantified for variations in the load and angle of flexion.

Methods: Six fresh cadaveric specimens were investigated at 30°, 60°, 90°, and 120° of flexion and at loads of 25 and 500 N, simulating resisted elbow extension. The joint space width and contact were determined, using polyether casting material.

Results: At 25 N all joints made contact in the ventral and dorsal aspects of the articular surfaces, whereas in the depth of the trochlear notch the joint space was on average between 0.3 and 2.8 mm wide, with some variation between individuals. At 500 N the joint space width was considerably reduced and the contact areas expanded towards the depth of the notch. The size of the dorsal contact areas was significantly smaller at 30° and that of the ventral ones at 120°, their ventro‐dorsal ratio decreasing considerably from 30° to 120° (P< 0.01).

Conclusion: These results indicate that the size of the contact areas depends to a slight extent on the joint position, but that at all loads and flexion angles a bicentric contact and an important central joint space width emerge because of the concave incongruity of the joint, These data may be used for numerical calculations, analysing the effects of incongruity on the joint stress and on the functional adaptation of the subarticular tissues. © 1995 Wiley‐Liss, Inc.
Culture of bovine oviduct epithelial cells (BOEC)
Authors: Walter I.
Published: February 2005

Background: Bovine oviduct epithelial cells are widely used in co‐culture experiments to improve early embryonic development and in vitro fertilization in embryo transfer programmes for domestic animals.

Methods: The present study compares different methods for harvesting and culture of bovine oviduct epithelial cells in order to optimize handling. Bovine oviduct epithelial cells were mechanically or enzymatically isolated and cultured on glass, on permeable membranes, or in suspension. Growth of the cells and their state of differentiation was examined by means of classical staining methods, immunohistochemistry and electron microscopy.

Results: Initial cell suspensions contained sheets of ciliated and nonciliated (secretory) cells; 24 h after seeding, free floating epithelial cells formed vesicles with cilia on their external surface. First adhesion of cells was seen 72 h after seeding. Later on, cells grew continuously and confluent monolayers were formed after 7 days. Results were identical after mechanical or enzymatical cell harvesting and were identical on both substrata tested, i.e., on glass and on permeable membranes. Light and electron microscopy proved the monolayers to resemble a polarized, simple, cuboidal to columnar epithelial membrane with intact junctional complexes and numerous apical microvilli. Their epithelial nature was established by immunostaining for cytokeratins. Cilia were missing and secretory granules were scarce. A layer of acidic glycoprotein material was demonstrated on the apical surface. Monolayers of bovine oviduct epithelial cells stored lipid droplets and large quantities of glycogen. About 50% of the seeded cells did not adhere but survived in the culture medium as free floating cells. These suspended cells maintained morphological criteria of differentiation (cilia and secretory granules) until day 12 of culture. Proliferation rates of cultivated cells were determined by counting mitoses and by immunostaining with MIB1 antibody. Results showed coincidence of rapid proliferation and morphological dedifferentiation of monolayers. Suspended cells, by contrast, did not proliferate but retained cellular differentiation under identical culture conditions.

Conclusions: The results strongly suggest that monolayers of bovine oviduct epithelial cells will not fully substitute for original oviduct epithelium when used in co‐culture experiments after in vitro fertilization. © 1995 Wiley‐Liss, Inc.
Comparative morphology and cytology of the olfactory organs in moray eels with remarks on their foraging behavior
Authors: Fishelson Lev
Published: February 2005

Background. This study compares the morphology and cytology of olfactory organs in moray eels (Muraenidae), particularly Siderea grisea and some species of the genera Echidna, Gymnothorax, and Lycodontis, fishes that are top predators in shallow‐water marine habitats. Some of the species search visually for food while others search by olfaction.

Methods. The morays were collected in the Red Sea; the nasal olfactory organs were dissected and fixed in Bouin's solution for light‐microscopy, and 3.5% glutaraldehyde for electron‐microscopy studies.

Results. In each studied species the olfactory rosettes are elongated structures situated in closed olfactory chambers between anterior tubular inlet nares and slit‐form posterior outlet openings. The double row of lamellae constituting these rosettes are round in Siderea and Echidna and elongated in the other species. They are attached at their base to a median raphe and range in number from 20 in the youngest observed Siderea to 168 in Gymnothorax of 1,500 mm total length. As in other teleosts, the lamellae are covered by a ciliated epithelium composed of three types of sensory cells: two of these, ciliated sensory neurons and ciliated supporting cells, differ in detail, length, and thickness of their cilia and intracellular rootlet system; the third type of sensory cells bears microvillae as well as cilia. Proximal, axonal extensions of the ciliated cells cross the basal lamina in bundles and combine to form fila olfactoria from which the two olfactory nerves extend towards the olfactory bulbs. Lateral extensions at the basal parts of these ciliated cells, the so‐called spines, cross the membranes of neighboring cells as dendrites, possibly changing part or all of the ciliated epithelium into an olfactory field. The density and number of sensory cells on the lamellae, as well as observed differences in their foraging behavior in nature and captivity, enable the morays to be divided into two groups: one group, in which the lamellae are richly covered with stereocilia, includes species of the genera Siderea and Echidna, that search for food by olfaction, and the second group, which has a great deal less cells with stereocilia and includes the studies species of Gymnothorax and Lycodontis, locates its food visually. © 1995 Wiley‐Liss, Inc.
Surface coat of sheep pulmonary intravascular macrophages: Reconstitution, and implication of a glycosyl‐phosphatidylinositol anchor
Authors: Singh Baljit , Ireland William P. , Minhas Kanwaljit , Atwal Onkar S.
Published: February 2005

Background: Pulmonary intravascular macrophages (PIMs) of sheep have a globular surface coat that facilitates endocytosis of tracer particles and Escherichia coli lipopolysaccharide, and is disrupted by the heparin and Brefeldin A treatments. The present study investigated the in vivo dynamics of the coat globules following heparin‐mediated removal, and the mechanism of globule organization on the plasma membrane of PIMs in vitro.

Methods: Sheep were administered heparin at a dose of 50 IU/kg body weight IV, and euthanised at 30 min, 3, 6, 12, 48, and 120 hr (n = 2 for each treatment) after the treatment. Control sheep (n = 2) were injected with normal saline solution. The tissues were processed for an ultrastructural examination and acid phosphatase (ACPase) cytochemistry. Heparintreated lungs were subjected to morphometric analysis of the coat globules. Lung tissues from normal sheep (n = 2) were incubated with phosphatidylinositol‐specific‐phospholipase C (PIPLC; 2 IU/ml PBS) in vitro for 30 and 75 min.

Results: Heparin study: The ultrastructural and morphometric data showed that the coat globules were removed at 30 min and reconstituted within 48 hr of the treatment. The PIMs showed priminent Golgi complexes associated with secretory vesicles, microtubules, and centriole between 3–12 hr of heparin treatment. Acid phosphatase cytochemistry also demonstrated secretory activity in the Golgi complexes of PIMs during the coat reconstitution. PIPLC study: The coat globules of PIMs were removed in a time‐dependent mode by the PIPLC treatment without damage to other cell organelles.

Conclusions: This study demonstrates a time‐dependent reconstitution of the coat of PIMs in conjunction with secretory activity following heparinmediated removal, probably through sequenstration of the globules from blood. This ability is of functional significance as the coat mediates particle endocytosis by the PIMs. The results also suggest the presence of a glycosyl‐phosphatidylinositol (GPI) anchor in tethering of globules on the plasma membrane of PIMs to offer a structural basis for their integrity in pulmonary vascular flow. © 1995 Wiley‐Liss, Inc.
Rat hippocampal mossy fibers contain cholecystokinin‐like immunoreactivity
Authors: Chandy Joby , Pierce Joseph P. , Milner Teresa A.
Published: February 2005

Background: The possibility that mossy fiber endings in the rat hippocampal formation may contain cholecystokinin (CCK) was reexamined.

Methods: For this, CCK‐immunoreactivity was examined by light and electron microscopy using the avidin‐biotin complex method.

Results: At the light level, the topographical distribution of perikarya and processes with CCK‐like immunoreactivity (CCK‐LI) was similar to that previously described by others. Ultrastructural analysis of the dentate gyrus and CA3 region of the hippocampus revealed that some mossy fiber terminals contained CCK‐LI most often affiliated with large, dense‐core vesicles (DCV). Quantitative analysis revealed that 4–8% of the mossy terminal profiles examined (n = 350) contained CCK‐labeled DCVs, which corresponded to 0.03–0.2 labeled DCVs per 100 μm2 of neuropil.

Conclusions: The presence of CCK‐LI within mossy fibers in the rat suggests that there is less species variability in peptide expression in this pathway than formerly believed. © 1995 Wiley‐Liss, Inc.
Potential neuroprotective effect of t‐butylhydroquinone against neurotoxicity—Induced by 1‐methyl‐4‐(2′‐methylphenyl)‐1,2,3,6‐tetrahydropyridine (2′‐methyl‐MPTP) in mice
Authors: Abdel‐Wahab M. H.
Published: February 2005

Dopaminergic damage inducing Parkinson's disease (PD) is ubiquitous neurodegenerative disorder, characterized by the progressive loss of dopaminergic neurons in the nigrostriatal pathways. The etiology and pathogenic factors implicated in dopaminergic damage are still unexplored to develop causal therapeutic strategies aimed to halt its progressive loss. The neurotoxicity induced by 1‐methyl‐4‐(2′‐methylphenyl)‐1,2,3,6‐tetrahydropyridine (2′CH3‐MPTP), which is more potent neurotoxic than MPTP in mice, is one of the most valuable models for analyzing pathological feature of dopaminergic damage. Herein, we investigated the neuroprotective effect of the potent antioxidant tertiary butylhydroquinone (TBHQ) against 2′CH3‐MPTP‐induced neurotoxicity in mice as well as the possible mechanism underlying such neurotoxicity. Male albino mice were injected with two doses of 2′CH3‐MPTP (20 mg/kg, i.p) for two consecutive days. Animals were killed after 2 weeks from the last dose of 2′CH3‐MPTP. Control animals received 10 mL/kg body weight i.p of distilled water. In both groups, brain stems containing the nigrostriatal pathways were dissected and reduced glutathione (GSH), malonyldialdehyde (MDA) contents, and superoxide dismutase (SOD) activity were estimated. Also, brain stem histopathological and histochemical changes were examined. The results of this study revealed that i.p injection of 2′CH3‐MPTP caused decrease in the brain stem content of GSH. On the other hand, the content of MDA and SOD activity was increased as compared with control groups. Also, 2′CH3‐MPTP showed severe histopathological changes including swelling of cytoplasm, interstitial edema, and complete loss of the neurons with reactive microglial proliferation and gliosis. Furthermore, histochemical examination of brain stem qualitatively showed depletion of dopaminergic neurons of nigrostriatum. Oral administration of TBHQ (100 mg/kg) prior to 2′CH3‐MPTP for 7 days caused normalization of GSH content and SOD activity and ameliorated the MDA content but still above the control value. Pretreatment with TBHQ slightly mitigated the histopathological and histochemical changes observed in 2′CH3‐MPTP‐treated mice. Based on these observations, it can be concluded that the antioxidant TBHQ has the ability to reverse the oxidative stress caused by 2′CH3‐MPTP in mice while failed to challenge the histopathological and histochemical changes induced by that toxicant. © 2005 Wiley Periodicals, Inc. J Biochem Mol Toxicol 19:32–41, 2005; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20053
Usefulness of studies on the molecular mechanism of action of herbals/botanicals: The case of St. John's wort
Authors: Choudhuri Supratim , Valerio Luis G.
Published: February 2005

The use of herbals/botanicals has been gaining wide popularity in recent years in the United States as well as in other parts of the world. The mechanism of action of most of these herbals/botanicals has not been subjected to thorough scientific investigations. St. John's wort (Hypericum perforatum) represents a useful case study in this sense. Traditionally, it is used as a natural treatment for depression; however, in recent years its molecular mechanism of action has been elucidated by a number of laboratories across the world. Such studies have helped understand potential interactions of St. John's wort with drugs and other xenobiotics. St. John's wort activates a nuclear receptor called pregnane X receptor (PXR). PXR is a ligand‐activated transcription factor that induces a number of xenobiotic‐metabolizing enzymes and transporters including cytochrome P4503A4 (CYP3A4) in humans. Because CYP3A4 alone metabolizes about 60% of all clinically relevant drugs, induction of CYP3A4 may result in the rapid elimination of these drugs and a consequent reduction in drug efficacy. Ironically, such enzyme‐inducing effects may not produce any immediate adverse symptomatology in the person taking St. John's wort. Therefore, the case of St. John's wort should serve as a good example of the usefulness and importance of studies on the mechanism of action of the herbals/botanicals, particularly those with widespread use. Scientists, physicians, and other health professionals can make use of the knowledge from such studies as an additional risk management tool. © 2005 Wiley Periodicals, Inc. J Biochem Mol Toxicol 19:1–11, 2005; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20057
The effect of bile salts on oxygen consumption, oxidative phosphorylation and ATP‐ase activity of mucosal homogenates from rat Jejunum and Ileum
Authors: Faust Robert G. , Wu Shih‐Min Liu
Published: February 2005

The conjugated bile salts, sodium taurocholate and glycocholate, inhibited oxygen consumption and uncoupled oxidative phosphorylation of mucosal homogenates from rat jejunum and ileum. These bile salts also were capable of increasing the ATP‐ase activity, in the presence of Na+ + K+ with Mg++, of both mucosal homogenates. Consequently, it was concluded from the results of this investigation that the previously observed decrease in ATP levels of rat jejunum and ileum, in the presence of bile salts, can be accounted for by both a complete uncoupling of oxidative phosphorylation and by an increase in ATP‐ase activity. Furthermore, the mechanism of bile salt inhibition of tissue ATP levels was discussed in relation to a regulatory role played by bile salts in the active transport of water soluble substances across the small intestine.
Possible role of Ca ions in the resting metabolism of frog sartorius muscle during potassium depolarization
Authors: Novotný I. , Vyskočil F.
Published: February 2005

Oxygen consumption and Ca exchangeability at different levels of potassium depolarization were studied in frog sartorius muscle. It was found that the changes in oxygen consumption parallel the changes in Ca exchangeability. Procaine (10−3 M) and CaCl2 (2.10−2 M) suppressed both extra oxygen consumption and Ca exchangeability at low values of depolarization. At higher values of depolarization procaine and CaCl2 differed in their action. Procaine favored inhibition of these processes, CaCl2 caused their activation. The effects of these compounds was not a result of a change in the membrane potential, since their effect on potassium depolarization was found to be small.

Relations between oxygen consumption and Ca exchangeability similar to those observed at potassium depolarization seem to exist under conditions where caffeine was applied. It is proposed that the extra oxygen consumption caused by potassium depolarization or on application of caffeine and unaccompanied by mechanical changes is related to the release of Ca from its bound form. Oxygen consumption in isotonic sucrose solution was also studied, but some different data from the above were obtained.
The in vitro response of thymic lymphocytes of the pig to phytohemagglutinin
Authors: Weber W. T.
Published: February 2005

The response of thymic lymphocytes of the pig to phytohemagglutinin was studied with H3 thymidine in cultures, from 0–72 hours. At the beginning of the culture period 6–18% of lymphocytes were in DNA synthesis. during the first 24 hours a sharp decrease in the number of DNA synthesizing cells was observed in both pha and control cultures, although pha cultures consistently showed small but significantly greater numbers of DNA synthesizing cells. this was followed by a definite peak in DNA synthesis and mitotic response of a minority of the cells in pha cultures between 48–54 hours, whereas in control cultures activity ceased. in addition, a small proportion of the progeny of initially DNA synthesizing medium sized lymphocytes was apparently stimulated by pha and found in mitosis by 48 hours.

It was concluded that the thymus contains a fraction of lymphocytes, not in the mitotic cycle, which are capable of being transformed by pha to mitotic activity. the data also suggests some stimulation of cells already in the mitotic cycle.
Isolation and characterization of an isoantigenic variant from a heterozygous mouse lymphoma in culture
Authors: Papermaster Ben W. , Herzenberg Leonard A.
Published: February 2005

A cell line from a mouse lymphoma heterozygous at the chromosome region for the H‐2d and H‐2k alleles was originally obtained from a transplantable lymphoma in the (C3H × DBA/2)F1 hybrid (H‐2d/H‐2k) and cultured in vitro. The original cultured line, termed parent line, was susceptible to the cytotoxic action of antibodies directed against antigenic components of both the d and k alleles. The parent line also absorbed hemagglutinins from both anti‐d anti‐k antisera. A resistant, variant subline was selected from the original population by immunoselection in vitro with anti‐H‐2d antibody and complement in a cytotoxic system. After one year in continuous culture in the absence of selecting antisera, the variant subline was still resistant to the cytotoxic action of anti‐H‐2d antibody. Serologic analysis of the variant indicated that it had lost the D antigenic component of the d allele, had a reduced amount of the H component, controlled by both the d and k alleles, and had retained the K component of the k allele. Possible genetic mechanisms that might account for the emergence of the variant line are discussed. While the results do not necessarily support an analysis based on mitotic recombination, ascribing other mechanisms is also difficult because of aneuploidy in the cell line. Finally, the experiments point out the advantages of using in vitro immunoselective methods in the genetics of mammalian somatic cells.
5‐Hydroxytryptamine in frog's skin
Authors: Welsh John H. , Zipf Joyce B.
Published: February 2005

The amounts of 5‐hydroxytryptamine (5‐HT) in the skin of nine species of anurans were estimated by the fluorescence method. Relatively large quantities of 5‐HT were found in the dorsal skin of five species of semi‐terrestrial frogs, while in the skins of four highly aquatic species none, or only a trace, was found. Using the histochemical fluorescence method of Falck, 5‐HT was located in the granules of the venom glands of Rana pipiens, a semi‐terrestrial species, while no fluorescing glands were found in R. catesbeiana, a highly aquatic species. The results of Erspamer and others are discussed. It is suggested that the most probable role of 5‐HT in frog skin is that of defense against land predators.
The effect of fluoride and calcium ions upon the insulin plus lactate‐stimulated K uptake and respiratory rate of frog muscles
Authors: O'Niell K. E. , Manery J. F. , Dryden E. E.
Published: February 2005

Using paired frog muscles it was shown that fluoride caused no observable change in the rate of oxygen consumption in Ca++‐free Ringer's solution, but stimulated the rate in Ca++‐containing Ringer's; the obvious explanation for this stimulation is the spontaneous activity of the muscle caused by the rapid lowering of Ca++ ion concentration when fluoride was added. Fluoride had no effect on potassium movement in these experiments. In a Ca++‐free solution fluoride (0.03 M) inhibited the insulin + lactate‐stimulated oxygen consumption of muscles which had been adapted to the absence of calcium by overnight soaking. In Ca++‐containing Ringer's there was no net effect of fluoride, demonstrating the fact that the direct inhibition of oxygen consumption was cancelled by the indirect stimulation caused by the fluoride‐lowering of the Ca++ ion concentration. Fluoride depressed the IL‐induced K uptake but for this effect some calcium, albeit a very low concentration, was necessary. Fluoride did not alter the muscle glycogen concentrations.

The only observable effect of calcium (in the absence of fluoride) was a rise in respiration caused by exposure to a sudden reduction in Ca++ ion concentration. Both the IL‐stimulated O2 rate and K uptake were independent of Ca++ ions since they occurred in Ca++‐free adapted muscles, in muscles exposed to a rapid reduction of Ca++ ions as well as in muscles in a Ca++‐containing Ringer's solution.