Expression of cyclooxygenase genes in the jejunum of horses during low-flow ischemia and reperfusion.

OBJECTIVE: To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. ANIMALS: 5 healthy adult horses. PROCEDURES: Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20% of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. RESULTS: COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were ß-actin and hypoxanthine phosphoribosyltransferase, whereas glyceraldehyde 3-phosphate dehydrogenase and ß-2 microglobulin were the least stably expressed. CONCLUSIONS AND CLINICAL RELEVANCE: Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.

The normal genital tract of the female California sea lion (Zalophus californianus): cyclic changes in histomorphology and hormone receptor distribution.

Changes in reproductive tract histomorphology, and estrogen (ERalpha) and progesterone receptor (PR) expression throughout the breeding cycle were evaluated in free-ranging stranded female California sea lions (Zalophus californianus). Hormone receptor expression in the ovaries, uterus, cervix, and vagina was evaluated using an immunohistochemical technique with monoclonal antibodies. During a large portion of the cycle, ovaries contained both a corpora lutea (CL) and follicles in varying stages of development. In the periods of pupping and estrus during June and July, and in the spring morphologic features of the endometrium suggested estrogen influence. There were areas of squamous differentiation in the pseudostratified columnar epithelium of the cervix and vagina in some animals during estrus and in the spring. Estrogen receptor immunohistochemical scores were highest during pupping and estrus and in the spring and lowest during embryonic diapause. Cyclic changes in uterine PR expression throughout the cycle were minimal. Both ERalpha and PR were expressed in epithelial and stromal cells throughout the cervix and vagina, however, receptor expression was typically higher in the stroma. Stromal cell hormone receptors may play an important role in epithelial responses to circulating sex hormones. The results of this investigation add to the general knowledge of California sea lion reproduction and establish baseline information on reproductive tract hormone receptors that will aid in determining the factors involved in urogenital cancer development in sea lions.

Normal morphology and hormone receptor expression in the male California sea lion (Zalophus californianus) genital tract.

Histomorphology and estrogen alpha (ER alpha), and progesterone receptor (PR) expression were evaluated in free-ranging stranded male California sea lions (Zalophus californianus). Hormone receptor expression was evaluated using an immunohistochemical technique with monoclonal antibodies. Estrogen and PRs were identified in the efferent ductules, prostate gland, corpus cavernosa, corpus spongiosium, penile urethra, and in the epithelium and stroma of both the penis and prepuce. In some tissues, ER alpha expression was more intense in the stroma, emphasizing the importance of the stroma in hormone-mediated growth and differentiation of reproductive organs. To our knowledge, this is the first study to localize ER alpha and PR to the epithelium of the glans penis. The results of this investigation add to the general knowledge of male California sea lion reproduction and suggest that estrogens could have a role in the function of the male reproductive tract.

Lymphoma with cutaneous involvement in three domestic rabbits (Oryctolagus cuniculus).

Three domestic rabbits (Oryctolagus cuniculus) with cutaneous lymphoma are described. Two rabbits were young (7 weeks and 1 years) and were euthanized within 1 week of showing clinical signs. Lymphoma was found in the skin and internal organs. The third rabbit was 9.5 years of age, and lived for more than 1 year after diagnosis. No response was seen to either 2 months of alpha-interferon administration or a 2.5-week course of isotretinoin treatment. After 1 year the rabbit died suddenly; the owner refused necropsy. Immunologic stains of the tumour in all three rabbits showed T cells to be the lymphoma cell type.

Leukocyte differentiation antigens in canine cutaneous and oral plasmacytomas.

Seventeen cutaneous and oral tumours with light microscopic features of plasmacytomas from 16 dogs were studied. Clinically, most neoplasms were benign, although three recurred after excision and three were locally invasive. Tumours most often arose on the pinnae, digits, gingiva, and inguinal regions near areas of chronic inflammation and exhibited variable degrees of plasmacytic differentiation microscopically. Diagnosis of plasmacytoma was confirmed in paraffin-embedded tissues with a panel of leukocyte differentiation antigen markers that included cross-reactive antibodies for Mb-1 (CD79a), CD3, and vimentin and canine-specific antibodies for CD45RA and CD18. Immunoreactivity for Mb-1 and CD45RA, including staining of multinucleate cells and cells with karyomegaly, confirmed a B-cell origin of neoplasms, while staining for CD3 and CD18 revealed an extensive network of infiltrative T-cells and dendritic cells in tumours suggestive of a directed immune response. These findings (i) demonstrate the value of using a panel of antibodies for leukocyte antigens to differentiate plasmacytomas from other cutaneous and oral round cell tumours, and (ii) suggest that immune recognition and responsiveness within tumours may play a role in the behaviour of plasmacytomas in dogs by affecting tumour cell growth and differentiation.

Antifollicular cell-mediated and humoral immunity in canine alopecia areata.

Abstract A combination of cellular and humoral immunological assaults directed against follicular matrical cells is suspected to cause alopecia areata (AA) in humans. The specific aims of this study were to determine whether cytotoxic T lymphocytes (CTL) were increased in the blood and skin lesions of canine AA. Additionally, we wished to determine if circulating antifollicular antibodies were present in the dog. Finally, we aimed to investigate whether bulbar inflammation was associated with decreased matrical cell proliferation and keratinocyte apoptosis. Canine AA lesions were infiltrated by intrabulbar CTL, perifollicular helper T cells and dendritic cells. A higher percentage of CTL was present in canine AA peripheral blood compared with that of normal dogs. Tissue-fixed and circulating antifollicular IgG antibodies were detected. Reduced matrical proliferation and increased keratinocyte apoptosis was observed in inflamed hair bulbs. This study demonstrates the existence of antifollicular cell-mediated and humoral immunological responses in canine AA. Résumé- L'association d'agressions immunologiques cellulaires et humorales dirigées contre les cellules matricielles folliculaires est suspectée d'être à l'origine de l'Alopécie Areata chez l'homme. Le but de cette étude était de montrer si le nombre de lymphocytes T cytotoxiques est augmenté dans le sang et dans les lésions cutanées lors d'Alopécie Areata chez le chien. En plus, nous souhaitions montrer la présence d'anticorps circulants antifolliculaire chez le chien. Enfin, nous voulions voir si l'inflammation du bulbe pileux était associée à une diminution de la prolifération des cellules matricielles et une apoptose des kératinocytes. Lors d'Alopécie Areata chez le chien, existe une infiltration lésionnelle par des lymphocytes T cytotoxiques intrabulbaires, des lymphocytes T auxiliaires périfolliculaires et des cellules dentritiques. Le pourcentage de lymphocytes T cytotoxiques était plus important dans le sang périphérique provenant de sujets à Alopécie Areata que dans le sang d'animaux sains. Par ailleurs, des anticorps antifolliculaire (IgG) circulants et fixés dans les tissus sont détectés. Une réduction de la prolifération des cellules matricielles et une augmentation de l'apoptose des kératinocytes sont observées dans les bulbes pileux atteints. Cette étude démontre l'existence de réponses immunologiques cellulaires et humorales contre le follicule pileux dans l'Alopécie Areata du chien. [Olivry, T., Moore, P.F., Naydan, D.K., Puget, B.J., Affolter, V.K., Kline, A.E. Antifollicular cell-mediated and humoral immunity in canine alopecia areata (Immunité contre le follicule pileux dans l'Alopécie Areata du chien). Veterinary Dermatology 1996; 7: 67-79.] Resumen Se cree que la alopecia areata (AA) en la especie humana resulta de la combinación de la acción inmunológica celular y humoral contra células de la matriz folicular. Los objetivos especificos de este estudio fueron determinar si los linfocitos T citotóxicos (CTL) se encontraban aumentados en sangre y en las lesiones cutáneas de animales con AA. Además, descamos investigar la posible presencia de anticuerpos antifoliculares circulantes en el perro. Finalmente, nos propusimos investigar si la inflamación bulbar se encontraba asociada a una disminución en la proliferación de células de la matriz y a la apoptosis de queratinocitos. Las lesiones de la AA canina mostraban infiltración por CTL intrabulbares, células T colaboradoras y células dendríticas. Se encontró un porcentaje mayor de CTL en la sangre periférica de perros con AA respecto a perros normales. Se detectaron anticuerpos IgG antifoliculares circulantes y en tejido fijado. Se observó una disminución en la proliferación matrical y un aumento en queratinocitos apoptóticos en los bulbos foliculares inflamados. Este estudio demuestra la existencia de respuestas inmunitarias antifoliculares de tipo celular y humoral en la AA canina. [Olivry, T., Moore, P.F., Naydan, D.K., Puget, B.J., Affolter, V.K., Kline, A.E. Antifollicular cell-mediated and humoral immunity in canine alopecia areata (Immunidad antifolicular en la alopecia areata canina). Veterinary Dermatology 1996; 7: 67-79.] Zusammenfassung- Eine Kombinaton von zellulären und humoralen immunologischen Angriffen, die gegen die follikulären Matrixzellen gerichtet sind, wird als Ursache bei der Alopecia areata (AA) des Menschen vermutet. Die besondere Absicht dieser Studie war, festzustellen, ob die zytotoxischen T-Lymphozyten (CTL) im Blut und in den Hautveränderungen bei kaniner AA erhöht sind. Zusätzlich wollten wir feststellen, ob beim Hund zirkulierende antifollikuläre Antikörper vorkommen. Schließlich beabsichtigten wir, zu untersuchen, ob die bulbäre Entzündung mit verminderter Matrixzellproliferation und Keratinozytenapoptose einhergeht. Die Veränderungen bei kaniner AA wurden von intrabulbären CTL, perifollikulären T-Helferzellen und dendritischen Zellen infiltriert. Verglichen mit dem peripheren Blut gesunder Hunde kam bei kaniner AA eine höherer Prozentsatz an CTL vor. Es wurden gewebsfixierte und zirkulierende antifollikuläre IgG-Antikörper festgestellt. In entzündeten Haarbulbi wurde eine reduzierte Matrixproliferation und eine erhöhte Keratinozytenapoptose beobachtet. Diese Studie zeigt die Existenz von antifollikulären zellvermittelten und humoralen immunologischen Reaktionen bei kaniner AA. [Olivry, T., Moore, P. F., Naydan, D. K., Puget, B. J., Affolter, V. K., Kline, A. E. Antifollicular immunity in canine alopecia areata (Antifollikuläre Immunität bei der Alopecia areata des Hundes). Veterinary Dermatology 1996; 7: 67-79.].