Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.     

Development of Immune Response to Experimental Bovine Trichophyton vervucosum Infection

Abstract— Cell mediated and humoral immune responses to experimental Trichophyton verrucosum infection were assessed by sequential cutaneous biopsies, antibody assessments and microscopic monitoring of fungal presence. Histopathologic examination showed the accrual of lymphocytes and other inflammatory cells in the dermis of infected sites. Immunoperoxidase staining of frozen sections with monoclonal antibody preparations revealed an influx of macrophages, BoCD4+ and B0CD8+ lymphocytes and γδ T cells from the 5th day to the 33rd day of infection. A moderate influx of B cells was observed. Protein G-colloidal gold staining revealed the presence of immunoglobulins in the dermis and superficial epidermal layers. Trichophyton specific serum antibodies appeared between days 33 and 55. Microscopic assessment of infected tissues revealed an increase in T, verrucosum elements (mycelium and ectothrix spores) from days 19 to 55. Fungal elements in infected areas did not decrease until after both humoral and cell mediated elements of the immune response were established. These responses imply a combination of cell mediated and humoral events were associated with T. verrucosum immunity and clearance in the calf. Résumé— Le réponse immunitaire humorale et cellulaire a une infection expérimentale àT. verrucosum a été appréciée par des biospsies cutanées successives, des dosages d'anticorps et la recherche microscopique de champignons. L'examen histopathologique a montré un afflux de lymphocytes et d'autres cellules inflammatoires dane le derme des sites infectés. Les marquages en immunopéroxydase par un anticorps monoclonal de coupes congelées a montré un influx de macrophages, lymphocytes BoCD4+ et BoCD8+ et des cellules T γδ, du 5e au 33e jour de l'infection. Un marquage par une protéine G—or colloidal a révélé d'immunogolglobulines dans le derme et les couches supéerficielles de l'épiderme. Les anticorps spécifiques de Trichophyton sont apparus entre 33 et 55 jours. L'examen microscopique des tissus infectés a révélé une augmentation du nombre d'éléments de T. verrucosum (mycelium et spores ectothrix) du 19e au 55e jours. Les éléments fongiques dans les zones infectées n'ont pas diminué avant que les réponses humorales et cellulaires ne solent établies. Ces résponses impliquent qu'une coopération des réponses humorales et cellulaires étaient associées dans l'immunité et la défense contre T. verrucosum. Zusammenfassung— Die zellvermittelten und humoralen Immunantworten auf die experimentelle Infektion mit T. verrucosum wurden durch eine Serie von Hautbiopsien, Antikörperuntersuchungen und mikroskopischer Untersuchung auf das Vorhandensein von Pilzen ausgewertet. Die histopathologische Untersuchung zeigte eine Ansammlung von Lymphozyten und anderen Entzündungszellen in der Dermis der infizierten Stellen. Die Immunperoxidasefärbung der Gefrierschnitte mit monoklonalen Antikörperzubereitungen zeigte einen Influx von Makrophagen, BoCd4-und BoCD8-Lymphozyten und gamma-delta-T-Zellen vom 5. bis zum 33. Tag der Infektion. Es wurde auch ein mäßiger Influx von B-Zellen beobachtet. Die Protein-G-kolloidale Goldfärbung zeigte die Anwesenheit von Immunglobulinen in der Dermis und den oberflächlichen epidermalen Schichten. Trichophyton-spezifische Serumantikörper traten zwischen Tag 33 und 55 auf. Die mikroskopische Untersuchung infizierter Gewebe zeigte eine Zunahme von T. verrucosum-Bestandteilen (Myzel und exktothrixe Sporen) vom Tag 19 bis 55. Pilzteile in infizierten Bereichen verminderten sich weder, nachdem humorale, noch nachdem zellvermittelte Elemente der Immunantwort auftraten. Diese Reaktionen deuten an, daß eine Kombination von zellvermittelten und humoralen Vorgängen im Zusammenhang mit T. verrucosum-Immumtät und Abheilung beim Kalb vorliegt. Resumen Por medio de biposias cutáneas secuenciales, medida de anticuerpos y exámen microscópico de presencia de hongos, se estudió la respuesta inmunitaria de tipo humoral y celular producida por la infección experimental con T. verrucosum. El exámen histopatológico reveló la presencia de agregación de linfocitos y otras células inflamatorias procedentes de la dermis de los puntos infectados. La tintura por medio de inmunoperoxidasa de las secciones congeladas, con preparaciones monoclonales de anticuerpos, demostró un aflujo de macrófagos BoCD4 + y BoCD8 + linfocitos y linfocitos, Tαδ, desde el quinto hasta el día 33 la infección. También se observó un aflugo moderado de linfocitos B. La tintura aúrica de proteina coloidal G reveló la presencia de inmunoglobulinas en al dermis y capas superficiales de la epidermis. Los anticuerpos específicos para la especie Trichophyton aparecieron entre los días 33 y 55. El exámen microscópico de los tejidos afectados demostró un incremento, de los elementos füngicos T. verrucosum (micelio y esporas exótricas) desde los días 19 al 55. Los elementos fúngicos en áreas infectadas no disminuyeron hasta después del establecimiento de ambos tipos de respuesta inmunitaria, humoral y celular. Estas respuestas implican que la combinación de ciertos fenómenos de inmunidad celular y humoral, están relacionados con la desaparición y la inmunidad de la infección producia por T. verrucosum en el ternero.     

Transrectal ultrasonography for the evaluation of stallion accessory sex glands.

This article reviews the capabilities of transrectal ultrasonography for determining the distribution of fluid and tissue within stallion accessory sex glands. Emphasis is placed on describing the normal ultrasonographic appearance of the accessory sex glands, excurrent ducts, and pelvic urethra of stallions during rest, after teasing, and after ejaculation and using this information to detect glandular abnormalities.     

Oxidative damage to the membrane of canine erythrocytes with inherited high Na, K-ATPase activity.

Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions.     

Alkaline phosphatase isoenzymes in feline serum using an agarose gel alkaline phosphatase kit method.

Total serum alkaline phosphatase (ALP) activity is the product of the combined activity of isoenzymes from a number of tissue sources. In this study, a commercially available kit for electrophoretic separation of ALP isoenzymes in an agarose gel was used to separate ALP isoenzymes in feline tissue extracts and serum. Five separate bands of ALP activity were identified. These bands were numbered 1 to 5 with band 1 having the most anodal migration. The tissue of origin corresponding to the migration position of the isoenzymes are as follows: Band 3 was the liver isoenzyme, band 4 was the bone isoenzyme and ALP isoenzymes of both intestine and kidney migrate in the position labelled band 5. Band 1 appears to be related to albumin and does not represent true ALP activity. The tissue source of band 2 (a and b) was not identified. Serum ALP activity of mature, healthy cats is primarily of liver origin. Immature cats (< 1 year of age) have a greater proportion of the bone isoenzyme in the serum.     

Use of autogenous cancellous bone graft for treatment of osteolytic defects in the phalanges of three cattle.

Osteolytic defects were detected radiographically in the distal sesamoid bone of a 16-month-old Bralers heifer, in the middle phalanx of a 14-month-old American Gray Brahman bull, and in the distal phalanx of a 3-year-old American Gray Brahman bull. The articular cartilage was damaged in each animal because of osteolysis or pathologic fracture. After each animal was anesthetized and positioned in lateral recumbency, the lesions were curetted and packed with cancellous bone harvested from the same animal's tuber coxae. Basic postoperative management involved stall rest and immobilization of the graft site with a fiberglass cast (42 to 79 days), after which a support bandage was used for approximately 2 weeks. Recurrence of lameness has not been observed in these animals for 60 months, 58 months, and 21 months, respectively. These cases exemplify the benefit of using an autogenous cancellous bone graft for treatment of severe osteolysis of a digit in cattle.     

In situ radiometric mapping of soil erosion and field-moist bulk density on cultivated fields

Abstract. Recent developments in in situγ ray spectrometry offer a new approach to measuring the activity of radionuclides such as ¹³⁷Cs and ⁴⁰K in soils, and thus estimating erosion or deposition rates and field moist bulk density (ρ_m). Such estimates would be rapid and involve minimal site disturbance, especially important where archaeological remains are present. This paper presents the results of a pilot investigation of an eroded field in Scotland in which a portable hyper pure germanium (HPGe) detector was used to measure γ ray spectra in situ. The gamma (γ) photon flux observed at the soil surface is a function of the ¹³⁷Cs inventory, its depth distribution characteristics and ρ_m. A coefficient, Q_Cs, derived from the forward scattering of ¹³⁷Cs γ ray photons within the soil profile relative to the ¹³⁷Cs full energy peak (662 keV), was used to correct the in situ calibration for changes in the ¹³⁷Cs vertical distribution in the ploughed field, a function of tillage, soil accumulation and ρ_m. Based on only 8 measurements, the agreement between in situγ ray spectrometry and soil sample measurements of ¹³⁷Cs inventories improved from a non significant r²=0.05 to a significant r²=0.62 (P<0.05). Erosion and deposition rates calculated from the corrected in situ¹³⁷Cs measurements had a similarly good agreement with those calculated from soil cores. Mean soil bulk density was also calculated using a separate coefficient, Q_K, derived from the forward scattering γ photons from ⁴⁰K within the soil relative to the ⁴⁰K full energy peak (1460 keV). Again there was good agreement with soil core measurements (r²=0.64; P<0.05). The precision of the in situ¹³⁷Cs measurement was limited by the precision with which Q_Cs can be estimated, a function of the low ¹³⁷Cs deposition levels associated with the weapons testing fallout and relatively low detector efficiency (35%). In contrast, the precision of the in situ ρ_m determination was only limited by the spatial variability associated with soil sampling.     

Control of Johne's disease in four commercial dairy herds in Iowa

A 6-year study was conducted in 4 dairy herds in Iowa in which Johne's disease was diagnosed previously. Fecal specimens were collected at 6-month intervals from animals 2 years of age and over for mycobacteriologic examination. Serum samples were obtained at 3-month intervals and tested by enzyme-linked immunosorbent assay (ELISA). The antigen used in the ELISA was a potassium chloride extract of a field strain of Mycobacterium paratuberculosis. The ELISA reactions were observed in 87% of the cows from which M. paratuberculosis was isolated. Dairy producers that participated in the Johne's control program reported reduced economic losses. Increased income was attributed to improved milk production, increased value of vaccinated animals sold as replacements to other dairy herds in which Johne's disease had been diagnosed, and the increased market value of slaughter animals removed from the herd.     

Squamous cell carcinoma of the corneoscleral limbus in a dog.

Squamous cell carcinoma is an uncommon tumor of the cornea in dogs. A 12-year-old Shih Tzu with a history of chronic keratoconjunctivitis sicca was diagnosed as having squamous cell carcinoma of the corneoscleral limbus. The lesion was treated by surgical resection and cryotherapy. One year after surgery, multiple tumors, apparently unrelated to the first tumor, appeared on the same cornea. Keratoconjunctivitis sicca may have predisposed the dog to corneal squamous cell carcinoma development.     

Persistence and mobility of chlorsulfuron and metsulfuron under different soil and climatic conditions

The persistence and mobility of chlorsulfuron and metsulfuron were studied in Italy at four field locations. Chlorsulfuron was applied pre-emer-gence to wheat at 15 and 30 g a.i. ha^?1 in the autumn of 1986 and metsulfuron early post-emergence at 8 g a.i. ha^?1 in the spring of 1987. Soil samples were taken periodically at each location for 1 year. Trials were conducted until 1989 at three locations. After the wheat harvest in 1989, chlorsulfuron and metsulfuron residues in the soil were measured by bioassay in plots that had been treated annually for 3 years and also in plots that had received single treatments. Chlorsulfuron at both application rates dissipated to levels below 0.3 g a.i. ha^?1 over 12 months at three locations while, at the fourth, which received only 250 mm rain during the study, significant levels of residues were still present after 15 months. The degradation of chlorsulfuron applied at 30 g a.i. ha^?1 followed first-order kinetics, with half-lives of 149, 70, 59 and 51 days at the four locations. Chlorsulfuron mobility was high at three locations while, in the silty clay soil of the fourth, the herbicide was confined to the upper 30 cm, despite the high rainfall. Metsulfuron applied in spring was detected only in the first soil sample collection. It was present in all sampled layers at one location but, in the others, was confined to the surface layer. There was no evidence of herbicide accumulation in the soil with repeated applications. Persistance et mobilité du chlorsulfuron et du met-sulfuron dans différents sols et sous différentes conditions climatiques La persistance et la mobilité du chorsulfuron et du metsulfuron au champ ont étéétudiées en Italie sur quatre sites. Le chlorsulfuron était appliqué en pré-levée sur le blé aux doses de matière active (m.a.) 15 et 30 g ha^?1à l'automne 1986 et le metsulfuron en post-levée précoce à la dose 8 g m.a. ha^?1 au printemps 1987. Des échantillons de sol étaient prélevés périodiquement sur chaque site pendant 1 an. Les essais ont été conduits jusqu'en 1989 sur trois des sites. En 1989, après la récolte du blé. les résidus de chlorsulfuron et de metsulfuron dans le sol ont été mesurés par des essais biologiques dans des parcelles qui avaient été traitées chaque année pendant 3 ans, ainsi que dans des parcelles qui n'avaient reçu qu'un seul traitement. Le chlorsulfuron aux deux doses disparaissait jusqu'à des niveaux inférieurs à 0.3 g m.a. ha^?1 en 12 mois sur trois des sites, alors que sur le quatrième, qui n'a reçu que 250 mm de précipitations pendant l'étude, des résidus étaient présents en quantité significative après 15 mois. La dégradation du chlorsulfuron appliquéà la dose 30 g m.a. ha^?1 suivait une cinétique du premier ordre, avec des demi-vies 149, 70, 59 et 51 j sur les quatre sites. La mobilité du chlorsulfuron était élevée sur trois des sites alors que sur le quatrième oú le sol était une argile limoneuse, l'herbicide était confiné aux 30 cm superficiels en dépit de précipitations élevées. Le metsulfuron appliqué au printemps n'était détecté que dans le premier prélèvement. Sur un des sites, il était présent dans toutes les couches, mais était confinéà la couche superficielle sur les autres sites. Après des applications répétées, aucune accumulation d'herbicide n'était observée dans le sol. Persistenz und Mobilität von Chlorsulfuron und Metsulfuron in verschiedenen Böden und bei unterschiedlicher Witterung Die Persistenz und Mobilität von Chlorsulfuron und Metsulfuron wurden in Italien an 4 Standorten unter Freilandbedingungen untersucht. Chlorsulfuron wurde im Herbst 1986 bei Weizen im Vorauflauf mit 15 und 30 g AS ha^?1 und Metsulfuron im Frühjahr 1987 im frühen Nachauflauf mit 8 g AS ha^?1 angewandt. Von jeder Versuchsfläche wurden über einen Zeitraum von 1 Jahr periodisch Bodenproben entnommen. Die Versuche wurden an 3 Standorten bis 1989 durchgeführt. Nach der Weizenernte 1989 wurden mit Biotests die Rückstände von Chlorsulfuron und Metsulfuron in den Böden mit jährlicher (über 3 Jahre) sowie mit einmaliger Behandlung bestimmt. Die Chlorsulfuron-Rückstände nahmen an 3 Standorten bei beiden Aufwandmengen nach 12 Monaten auf 0,3 g AS ha^?1 ab. Am 4. Standort, wo während der Untersuchung nur 250 mm Niederschlag gefallen waren, wurden nach 15 Monaten noch signifikante Rückstände gefunden. Der Abbau von Chlorsulfuron folgte an allen Standorten bei einer Aufwandmenge von 30 g AS ha^?1 einer Kinetik erster Ordnung mit Halbwertszeiten von 149, 70, 59 und 51 Tagen. Die Mobilität von Chlorsulfuron war an 3 Standorten hoch, während es in dem schluffigen Tonboden des 4. Standortes trotz hohen Niederschlags in den oberen 30 cm verblieb. Das im Frühjahr ausgebrachte Metsulfuron wurde nur in der 1. Bodenprobenserie gefunden. An einem Standort lagen in allen beprobten Bodenschichten Rückstände vor, doch sonst waren sie auf die oberste Bodenschicht beschränkt. Bei wiederholter Anwendung konnte keine Herbizidakkumulation im Boden festgestellt werden.