Scanning electron microscopy of the endometrium of mares infused with gentamicin.

Scanning electron microscopy (SEM) was used to study the endometrium of nine 1-year-old thoroughbred mares after twice intrauterine infusions of gentamicin, on 2 consecutive days. Five mares were infused on 2 consecutive days with 40 ml gentamicin (50 mg/ml) mixed with 80 ml of normal saline. Four mares served as controls and were infused with 120 ml of saline on 2 consecutive days. Endometrial biopsies were obtained from all mares 3 days after the second intrauterine infusion. Each biopsy was processed for SEM by standard methods. The endometrial epithelium of the gentamicin-infused mares had more cellular perforations than the saline-infused mares. The gentamicin-infused mares had less and shorter microvilli. The ciliated cells were fewer and some ciliated cells had disrupted and some had drooping cilia. The endometrial epithelium of the gentamicin-infused mares had a considerable number of endometrial cells that lost their luminal surfaces and some that lost their microvilli, compared to the saline-infused mares. We suggest that the information gathered in this pilot study should be used as basis for further investigation, on a larger scale basis, of the effects of repeated intrauterine infusion of gentamicin on the endometrial mucosa of mares.

Surgically induced cryptorchidism-related degenerative changes in spermatogonia are associated with loss of cyclic adenosine monophosphate-dependent phosphodiesterases type 4 in abdominal testes of rats.

The present study was undertaken to investigate the role of phosphodiesterase type 4 (PDE4) enzymes in cryptorchidism-induced apoptosis of the germ cells. Regulation of expression of PDE4 enzymes was studied in the abdominal and scrotal testes of surgically induced cryptorchid rats for 10, 20, and 30 days. In some cases orchidopexy was performed after 30 days of cryptorchidism, and rats were allowed to recover for an additional 50 days. Upon histological examination, marked degenerative changes in the epithelial lining of the seminiferous tubules within abdominal testes were observed compared with contralateral control or age-matched sham-operated rats. These changes included degeneration of some spermatogonia, apoptosis of the secondary spermatocytes, incomplete spermatogenesis, and lack of spermatozoa in the lumen. In contrast, contralateral scrotal testes exhibited normal histology. Significant improvement in the regeneration of spermatogonia was observed in rats after 50 days of recovery following orchidopexy. Immunocytochemical examination suggested the presence of PDE4A in germ cells while PDE4B was predominantly expressed on somatic cells. Western blotting using PDE4 subtype-selective antibodies showed the presence of two PDE4A variants (a 109-kDa PDE4A8 and a previously uncharacterized 88-kDa PDE4A variant) and two PDE4B (78-kDa PDE4B2 and 66-kDa PDE4B variant) bands. In unilaterally cryptorchid animals, the abdominal testis showed a time-dependent decrease in both PDE4A8 and 88-kDa PDE4A variants. In contrast, the expression of 66-kDa PDE4B was markedly increased in a time-dependent fashion in abdominal testes of cryptorchid rats. Animals surgically corrected for cryptorchidism and allowed to recover for 50 days exhibited normal expression of both PDE4A and PDE4B variants compared with aged-matched, sham-operated controls. In conclusion, this study suggests that down-regulation of PDE4A variants in cryptorchid testes may play an important role in the degeneration of spermatogonia and increased apoptotic activity in the germ cells.

Degenerative changes in spermatogonia are associated with loss of glucose transporter (Glut 3) in abdominal testis of surgically induced unilateral cryptorchidism in rats.

Expression of facilitative glucose transporters (Glut 1, 2 and 3) was examined by Western blot analyses 10, 20 and 30 days following surgically induced unilateral abdominal cryptorchidism. The cryptorchid testes exhibited marked degenerative changes in the seminiferous tubules and spermatogonia, impaired and incomplete spermatogenesis and lack of spermatozoa in the lumen. Immunoblotting of testis proteins with Glut transporter antibodies revealed only the presence of Glut 2 and 3 proteins. Glut 2 expression in abdominal testis was increased (45%, 67%, and 40% at 10, 20 and 30 days, respectively) but no significant change was observed in contralateral scrotal testis. Glut 3 expression was reduced by 85-95% compared with contralateral scrotal testis. A significant decrease in Glut 3 levels in abdominal testis was accompanied by an increase in scrotal testis Glut 3 content (80%, 144% and 212% at 10, 20 and 30 days, respectively) compared to age matched control rats. These results suggested that the degenerative changes in abdominal testis may be associated with decreased Glut 3 mediated glucose transport in seminiferous tubules and spermatogonia.

Proliferating cell nuclear antigen expression in sheep infected with bovine leukemia virus.

Proliferating cell nuclear antigen (PCNA) or cyclin (C), a major nuclear protein, has been shown to be associated with human leukemia and malignancies. PCNA protein was quantitated in this study, in lymphocytes from bovine leukemia virus (BLV) infected and non-infected sheep, using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and silver staining. The PCNA mean levels in lymphocytes of BLV-infected sheep (27 months post-infection) was significantly (P < 0.001) higher than in the lymphocytes of the non-infected sheep. The mean of PCNA levels in lymphocytes of sheep, 21 days after BLV infection, showed a two-fold increase compared with the non-infected sheep. Phytohemagglutinin (PHA) (3 days) treatment of lymphocytes from the non-infected and from the BLV-infected sheep resulted in a significant (P < 0.01) increase in the mean of PCNA levels only in the non-infected sheep. The mean lymphocyte counts of the BLV-infected sheep were not significantly different from the mean counts of the non-infected sheep at the time of lymphocyte protein analysis. Thus, these findings showed, similar to human leukemia and malignancies, that high levels of PCNA were found in lymphocytes from BLV-infected sheep compared with those from the non-infected sheep, and this was independent of high cell count. Our results also suggest that PCNA protein may play a role in the process of lymphoid transformation as a result of BLV infection of sheep.

The morphology of abdominal and inguinal cryptorchid testes in stallions: a light and electron microscopic study.

Eleven unilateral cryptorchid stallions, two to three years old, were castrated at Louisiana State University Veterinary Teaching Hospital. Five of these cryptorchid cases were abdominal and the rest were inguinal. This study was initiated to document the differences between the abdominal and inguinal equine cryptorchid testes. Specimens were obtained from the abepididymal side of each cryptorchid testes and processed for light and electron microscopic study. The cryptorchid testes were smaller than the scrotal testes, with the abdominal testes being one-fourth the size of the scrotal testes. Two of the abdominal testes had cysts filled with a straw-colored fluid. The seminiferous tubules of the abdominal testes were larger than those in the inguinal testes. The epithelial linings of the seminiferous tubules of the abdominal testes were vacuolated and did not contain more than two layers of undifferentiated cells. The interstitial collagen fibers of the abdominal testes were coarse and more abundant than those of the inguinal testes. The seminiferous tubules of the inguinal testes were smaller and contained many layers of epithelial cells at different stages of embryological differentiation, with scattered primordial germ cells. The necrotic, degenerative changes of the epithelial cells of the abdominal testes were distinct, while the inguinal testes had healthy cells at embryological arrest.

Use of antipeptide antibodies to probe the catalytic activity of p37v-mos.

Several site-directed antipeptide antibodies were generated to probe the kinase function of p37v-mos. Anti-mos(37-55) antibodies allowed autophosphorylation of p37v-mos, as well as transphosphorylation of exogenously added purified bovine vimentin. Rabbit antipeptide antibodies against v-mos residues 158-70, 194-206, 260-271, and 363-374 and a mouse monoclonal antibody against residues 344-359 completely inhibited p37v-mos protein kinase activity in vitro. p37v-mos autophosphorylation and vimentin transphosphorylation were affected similarly. These results suggest important roles for insert and the carboxy-terminal domains in the catalytic activity of p37v-mos.

Evidence for involvement of the protein kinase C pathway in the activation of p37v-mos protein kinase.

Protein kinases are known to undergo phosphorylation to regulate their activity. To determine whether the protein kinase activity of p37v-mos was similarly regulated, we investigated the influence of two well known protein kinases, namely protein kinase C and protein kinase A, on the activity of p37v-mos in vivo. NIH3T3 cells chronically transformed with Moloney murine sarcoma virus 124 were treated with high concentrations (200-400 nM) of phorbol 12-myristate 13-acetate (PMA) for 24-48 h, concentrations known to result in the total loss of protein kinase C by causing its translocation from the cytosol to cell membranes where it is downregulated. PMA treatment caused a drastic decrease in the protein kinase activity of p37v-mos without affecting its steady state level. Similar results were obtained with p85gag-mos expressed in ts110 Mo-MuSV transformed NRK cells. Control treatment with an inactive analogue of PMA, 4-alpha phorbol 12,13-didecanoate, had no effect on the p37v-mos protein kinase activity. Treatment of cells with a direct chemical inhibitor of protein kinase C, H-7 (1-(5-isoquinoline sulfonyl)-2-methylpiperazine dihydrochloride), approximately halved p37v-mos kinase activity, although the drug did not inhibit p37v-mos kinase activity directly in vitro. In contrast to the PMA effect, in vivo activation of protein kinase A by 8-(4-chlorophenylthio)-adenosine 3',5' cyclic monophosphate did not affect p37v-mos protein kinase activity levels. These findings indicate that the protein kinase C pathway but not the protein kinase A pathway modulates v-mos protein kinase activity.

SEM and TEM study of caprine superficial lymph nodes.

The splenic capsule was characteristic, having dense connective tissue. Smooth muscle cells and unmyelinated nerve fibers were observed. Smooth muscle cells were found to be independent of blood vessels in both the capsule and trabeculae. Littoral cells separated the capsule from the subcapsular sinus. Highly branched reticular cells were associated with the sinuses. The cellular components (large and small lymphocytes, plasma and mast cells, and macrophages) of the cortex and medulla were observed and described. No Golgi apparatus was observed in small lymphocytes and two surface types (rough and smooth) were observed on lymphocytes. Russell bodies were not observed in plasma cells. The paracortical postcapillary venule had cuboidal endothelium with microvilli. Two shapes of lymphocytes were seen associated with the endothelium of postcapillary venules.

The normal structure of regional feline gastric mucosae: scanning electron microscopic study.

Regions of cat's stomach can be identified by looking at the surface epithelial cells by scanning electron microscopy (SEM). The luminal surface of cells of the cardiac region were elongated, of the fundus rounded, of the corpus polygonal shaped, and of the pyloric region diamond shaped. The quantity and distribution of microvilli covering the epithelial cells varies, being abundant and evenly distributed in the cardiac region and gradually decreasing in number toward the gastro-duodenal junction, where they were confined to cell perimeters. The colliculi varied in shape and distribution from few in the fundus and corpus to numerous in the pyloric region. Large numbers of gastric pits were present in the corpus. They diminish toward both the cardia and gastro-duodenal junction. The cardiac and pyloric glands were coiled. The gastric glands (glandula gastrica propria) were straight tubules in the fundus and coiled in the corpus. All luminal surfaces of glandular epithelial cells were covered with microvilli, but the regional distribution of microvilli on the cell was variable. Parietal, mucous neck, and chief (zymogen) cells were identified by their cytoplasmic structure. Parietal cells had long apical microvilli, mucous neck cells contained large numbers of globular mucous granules, and chief cells were vacuolated. A few G cells (Endocrinocytus gastrointestinalis) were seen in the cardiac region, large numbers in the pyloric region, and not found in fundus or corpus.

Smooth muscle distribution in the capsule and trabeculae of the caprine superficial cervical lymph node.

This study centers around the dichotomy found in the literature concerning the presence of smooth muscle cells in the trabeculae and capsule of lymph nodes. Various superficial lymph nodes (mammary, mandibular, popliteal, subiliac, and superficial cervical) of the goat were collected and examined by light and electron microscopy. Smooth muscle cells were demonstrated in the capsule and trabeculae of lymph nodes independent of the blood and lymph vessels.

Correlative light and scanning electron-microscopic study of feline gastric mucosa: the cardiac region (pars cardiaca).

The cardiac region (pars cardiaca) of the cat's stomach was examined with light and scanning electron microscopy. The glands are simple, coiled tubular, and contain mucus-secreting cells. Their surfaces are covered with microvilli which are concentrated on the boundaries of the mucus-secreting cells. A few cells interposed between the glandular cells are probably G cells. They are identified by apical projections of long microvilli into the lumen of the gland. The surface epithelial cells lining the cardiac region are covered by minute microvilli. The muscularis mucosae is not distinctly divided into two layers. However, a group of smooth muscle cells which are directed in a circular manner around the gastroesophageal junction is considered to be the distal esophageal sphincter.

Duodenal microanatomy of the domestic cat (Felis catus).

Duodenal samples were taken from similar locations in six cats, processed, stained, and examined via light microscope. There were no prominent circular folds (plicae circulares) or stratum compactum (lamina subglandularis). The 1072 microns x 201 microns villi were covered by 46 microns high columnar epitheliocytes proximally which decreased in height (41 microns) distally and displayed a 1.1-1.7 microns striated border. Globular leukocytes, mononuclear cells, and twenty-eight goblet cells (exocrinocytus calciformis) per villus were seen. The intestinal gland (crypt of Lieberkuhn) epithelium was 20 microns tall and had a less distinct striated border. The 515 microns simple straight tubular intestinal gland layer displayed distal branching. Many mitotic figures, 12 goblet cells per gland, and occasional columnar to triangular cells with red cytoplasmic granules were seen. The thickness of the lamina propria mucosa (glandular portion) decreased from proximal to distal (563-465 microns). The lamina muscularis mucosa had two layers and decreased in thickness distally (71-28 microns). The proximal muscularis mucosa was penetrated by the ducts of submucosal (Brunner's, duodenal) glands. The tela submucosa decreased in thickness distally (593-192 microns) and contained submucosal glands with 11.5-75 microns lumina for the first 1.5-2.5 cm. However, submucosal glands could be found to a distance of 8 cm. The glandular epithelium ranged from 7.5-22.5 microns in height. Only one type of secretory cell was observed, with both mucous and serous properties. The tunica muscularis ranged from 190-1425 microns (median thickness of 557 microns) and had two layers.

Scanning electron microscopic study of the surface of feline gastric epithelium: a simple method of removing the coating material.

Scanning electron microscopic examination of the gastric surface epithelial cells is often hindered by the presence of a coating material. Several methods for removal of coating material on feline gastric mucosa were utilized. The cleansed tissues were evaluated using the scanning electron microscope to assess damage caused by the use of various cleansing methods to surface epithelial cells. The stretched stomach washed several times, including rubbing the mucosal surface with gloved fingers, yielded the best results with no apparent damage to the surface epithelial cells. Flushing unstretched stomachs with saline only did not adequately remove coating material. Flushing unstretched stomachs with saline while stroking the surface with a cotton tipped applicator stick removed debris but damaged the surface epithelium.

Cerebellar disease in an adult cow.

This is the report of clinical signs and lesions of a cerebellar disorder in an adult four year old Limousin cow grazing perennial ryegrass (Lolium perenne). The most striking histopathological lesion was a marked paucity of Purkinje cells throughout the cerebellum.

Follicular cysts in sheep.

A flock of 188 sheep was surveyed for cutaneous lesions that were noticed 3 days after shearing. On the basis of histologic features of the cyst wall, ie, association with sebaceous glands and lack of follicular structures or rete pegs, the cysts were classified as follicular cysts.

A light and electron microscopic study of the periparturient bovine corpus luteum.

Corpora lutea were obtained surgically from fifteen mature Angus crossbred cows representing three experimental groups of five cows each. Cows in Group A were 180 days of gestation, cows in Group B had recently experienced parturition (

Medical records and the law.

Practitioners involved in the diagnosis and treatment of health-related conditions are vulnerable to litigation. Courts of law frequently award large settlements to plaintiffs who can convince a jury that the quality of service was less than could be reasonably expected. Court judgments are based on evidence supported by hard facts. Practitioners should maintain an accurate accounting of each case, not only as good business practice but also as evidence in case of litigation. The medical record is an excellent source of such evidence.