Administration of monosodium aspartate (MSA) to neonatal voles, Microtus arvalis Pallas induced neuronal necrosis in several brain regions including the arcuate and ventromedial hypothalamic nuclei. In the growing stage, 56.7% showed glycosuria. In the mild diabetic voles, blood glucose concentrations rose to 180mg/dl (72mg/dl in normal), and plasma insulin levels were also increased to 112μU/ml (21μU/ml in normal). In the severe diabetic voles, on the other hand, blood glucose concentrations rose over 250mg/dl, and plasma insulin levels decreased under 10μU/ml. Histopathological examination of the pancreatic islets revealed moderately enlarged islets, and nuclear hypertrophy and partial degranulation of B cells in the mild diabetic voles, and moderately to markedly enlarged islets, and marked degranulation, vacuolation and accumulation of glycogen granules of B cells in the severe diabetic voles. These results indicated that the diabetic syndrome in herbivorous voles was induced by administration of MSA and diabetic voles fell into marked insulin deficiency in the severe stage.

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An attempt was made to induce ovulation in 19 cats during the non-breeding season by the administration of a porcine pituitary gland preparation. The dosage was 2mg for the first day and then 1mg for each day until estrus was induced. Studies were made to determine whether fertilization and pregnancy were possible by mating at the time of induced estrus. Estrus was induced in 89.5% (17/19) of the cats, 3-8 days after the first administration. The average number of ovulations induced by mating was 7.3±1.3 (mean ± SE) for 16 cats (excluding one cat that failed to ovulate). The uterine horn was flushed 6-8 days after mating in 9 of the 16 ovulated cats to determine the fertilization rate. An average of 3.6 ova were recovered from 8 out of the 9 cats (including one cat from which no ovum was recovered) which gave an average recovery rate of 38.6%. Expanded blastocysts were recovered from 5 cats, while from the other 3 cats, only unfertilized ova were recovered. Pregnancy was confirmed in the other 7 cats 15-20 days after mating. In two of the 7 cats implantation was observed, but abortion occurred at 24 and 27 days of pregnancy. Plasma progesterone concentrations were lower in the estrus-induced and bred cats as compared to these in the controls, with progesterone values approaching base concentrations, approximately by day 20 in 2 cats and by day 30 in 3 cats. It was concluded that estrus could be induced in cats during the non-breeding season by administration of porcine pituitary gland preparation. Furthermore, mating at this time was observed to cause fertilization and conception. Pregnancy maintenance was difficult because of the failure on the part of the embryos to develop normally or, the regression of the corpora lutea, beginning earlier than on the natural course of Pregnancy.

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The binding specificity of pertussis toxin (PT) was compared with lectins with well-defined specificities by hemagglutination, hemagglutination inhibition and competitive binding assays. Neuraminidase-treated human erythrocytes were much less weakly agglutinated by PT than untreated ones. Hemagglutination of untreated human type A erythrocytes by PT was inhibited by fetuin, haptoglobulin and hog A+H. Mono- and disaccharides, and N-acetylneuraminic acid alone were ineffective at the highest concentrations used. On the other hand, hemagglutination by Ricinus communis agglutinin (RCA-1) was effectively inhibited by these substances. Similar results were also obtained in competitive binding assays with ¹²³I-labeled PT and untreated type A erythrocytes. The binding of ¹²⁵I-labeled PT to type A erythrocytes was not effectively inhibited by any of lectins with well-defined specificities. These results suggest that the combining site of PT may be specific for terminal sialic acid and/or sialic acid-linked carbohydrate portion(s) which can not be recognized by lectins reported previously.

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On day 14 of fetal life in the rat, Mullerian (paramesonephric) duct in both sexes was first observed close to Wolffian (mesonephric) duct. Its caudal end joined Wolffian duct at about the level of the middle of gonadal anlage. Thereafter, Mullerian duct became independent of Wolffian duct through its whole length. Therefore, the main part of Mullerian duct may be formed by budding from Wolffian duct. On day 16, the anogenital distance in male started to increase with thickening of Wolffian duct and disappearance of Mullerian duct. Castration of male fetuses on day 19 stopped later masculinizing development. Therefore, it is clear that the fetal testis is an organ crucial for the masculinization of the urogenital tract as it activates the secretion of masculinizing hormone(s).

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Serum osmolalities and osmolar gap were determined in 43 normal healthy beagles (control group) and 40 dogs with dirofilarial hemoglobinuria (hemoglobinuria group). In the control group, the measured and calculated serum osmolality levels were in the means of 296±5 (SD) mOsm/kg and 293±6 mOsm/kg respectively, showing an osmolar gap less than 10 mOsm/kg. In the hemoglobinuria group, the measured serum osmolality ranged from 272 to 370 mOsm/kg. A considerable number of dogs had normal serum osmolalities in spite of severe intravascular hemolysis, suggesting that the changes in serum osmolality would not be the direct cause of intravascular hemolysis. The measured serum osmolality (331±28 mOsm/kg) was significantly higher in 11 dogs which died after a surgical removal heartworms than in 29 dogs which recovered after the removal (302±17 mOsm/kg). The calculated serum osmolality level was 296±16 mOsm/kg in 24 recovered cases, and 304±22 mOsm/kg in 10 fatal cases. The osmolar gap stayed in the normal ranges of 5.4±5.9 mOsm/kg in recovered cases, but it attained a higher level of 22.7±8.9 mOsm/kg in fatal cases, suggesting poor prognosis in cases with large osmolar gaps. There were significant positive correlations between the measured serum osmolality and osmolar gap, serum sodium, potassium, BUN, GOT, GPT, creatinine, bilirubin and plasma hemoglobin values, as well as between the osmolar gap and serum potassium, BUN, GOT, GPT, creatinine and bilirubin values. The plasma hemoglobin concentration fell markedly without significant change in serum osmolality 20 hr after the heartworm removal.

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Some adverse reactions such as shock-like reaction and dirofilarial hemoglobinuria (caval syndrome) occasionally occurred in microfilaremic dogs following milbemycin D (Milbe) administration. This study was carried out to seek the prevention of these adverse reactions. In two groups containing 16 and 9 dogs respectively which were administered either chlorpheniramine maleate (1mg/kg) or indomethacin (2.5mg/kg) simultaneously with Milbe (1mg/kg), the incidence of clinical signs such as the pale color of the visible mucous membranes, respiratory disorders, caval syndrome and shock-like reaction as well as changes in clinical parameters such as RBC and WBC counts, WBC profile and serum total protein, were almost equal to that observed in the group administered Milbe alone. In 41 dogs administered prednisolone (1mg/kg) simulaneously with Milbe (1mg/kg), no shock-like reaction was observed. Changes in clinical parameters were different from those in the group administered Milbe alone, whereas some clinical signs of adverse reactions, including caval syndrome, were observed. These results indicated that prednisolone was effective for prevention of the shock-like reaction in microfilaremic dogs induced by Milbe.

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The microheterogeneity in bovine serum α₁-acid glycoprotein (α₁AGP) was examined by crossed affino-immunoelectrophoresis with concanavalin A(Con A). In healthy cows, serum (α₁AGP revealed 2 fractions which were named fraction 1 and 2 according to the degree of binding affinity to Con A. The mean±SD of serum (α₁AGP concentration was 0.31±0.09 g/l and the relative amounts of fraction 1 and 2 were approximately 67% and 33% respectively on this method. In bovine leukemia cases, 5 out of 8 had a high serum α₁AGP concentration of more than 0.7g/l. On the electrophoretic pattern, the other 2 fractions (Con A weakly and non-reactive fractions, named fraction 3 and 4) were observed in 5 cases and their relative amounts of 4 fractions were 17.8-28.5%, 35.5-46.0%, 17.9-24.4% and 12./17.2%, respectively. In bovie leukemia virus infected cows with no clinical symptoms, however, serum (α₁AGP concentration and the electrophoretic patterns were almost the same as those in healthy cows.

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C-reactive protein (CRP) and serum amyloid P component (SAP), which are known as acute phase reactants in human and many other animals, were purified from cow sera. Affinity chromatography using HE agarose gel was the most effective method to isolate both CRP and SAP from a large volume of bovine serum. Separation of CRP and SAP from the mixed preparation could be performed by DEAE-cellulose column chromatography, gel permeation HPLC using TSK-G3000SW or affinity chromatography using phosphorylcholine derivatives of bovine serum albumin-conjugated Toyopearl HW 65. Bovine CRP and SAP were identified as genuine CRP- and SAP-class proteins by their cross reactivities with anti-human CRP and anti-human SAP, respectively, and by their homology in amino acid compositions compared with those of human CRP and SAP, respectively. Bovine CRP moved slower than β-globulin, and bovine SAP moved in the β-globulin region in agarose gel electrophoresis. Both of them gave single bands in native polyacrylamide gel electrophoresis (PAGE). Bovine CRP and SAP molecular weights were estimated to be 100, 600 and 109, 500 daltons respectively, by sedimentation equilibrium analysis. Bovine CRP showed 23K dalton subunits by sodium laurylsulfate-PAGE and bovine SAP showed 28K and 32K dalton subunits, both of which were glycosylated and had identical amino acid compositions, indicating that both CRP and SAP molecules are pentamers. In fact, they appeared to have pentameric disk-like configurations in electronmicroscopical examination.

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Liver lactate dehydrogenase (LDH) isoenzymes in normal and diseased cows were analyzed electrophoretically. This method (histoelectrophoresis) was improved for the direct analysis of tissue LDH isoenzymes. The mean values of LDH1 through LDH5 in the livers of normal cows were 31.7, 24.8, 27.3, 12.8, and 3.3%, respectively. In cases with hydropic degeneration of the liver, the patterns revealed increases of LDH1 and LDH2 as compared to normal cows. The patterns showed a decrease of LDH1 and an increase of LDH2 in fatty change of the liver. Congestion of the liver alone decreased LDH1 and increased LDH3, LDH4 and LDH5. Necrosis of the liver decreased LDH1 and LDH2, and increased LDH3, LDH4 and LDH5. It was suggested that the functional hepatocellular damage due to anoxia might be a important factor of the change of liver LDH isoenzyme patterns. We have attemped to standardize the LDH isoenzyme patterns by using a computer under various conditions. In cases with hydropic lesions, the diagnostic sensitivity (DS) was 53% (28 of 53 cases) and the predictive value of positive tests (PVPT) was 100% (28 cases of 28 cases selected). In cases with congestive or necrotic lesions, DS was 41% (9 of 22 cases) and PVPT was 69% (9 cases of 13 cases selected).

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For the purpose of clarifying the cause of white muscle disease (WMD) in calves, tocopherol and selenium levels and blood glutathione peroxidase (GSH-Px) activity were measured on 10 calves with WMD and nine of their dams. The main clinical symptoms of the 10 calves with WMD were motor disturbances including recumbency and stiffness. Serum enzyme activities (GOT, GPT, CPK, LDH) in calves with WMD increased markedly, and this increase was also observed in some of their dams. Serum tocopherol levels of calves with WMD were low, 70% of which showing deficient levels of less than 70pg/100ml. Serum selenium levels of all the calves were lower than 35 ppb, indicating a deficiency, and were accompanied by low blood GSH-Px activity. α-Tocopherol and selenium concentrations in organs were very low. Dams of calves with WMD showed low serum tocopherol levels, 22% of which indicating deficient levels below 150pg/100ml. Serum selenium levels in dams showed a marked decrease to under 20 ppb, and also low blood GSH-Px activity. Feedstuffs supplied in the farms to affected calves indicated very low α-tocopherol contents (below 3mg/100g DM) and low selenium concentrations below 50 ppb in DM. It was concluded that WMD in calves was attributable to nutritional muscular dystrophy caused by deficiencies in tocopherol and selenium in feedstuffs supplied to their dams.

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Different types of lymphoid malignancy were observed in two sheep inoculated with BLV-containing materials. Sheep 1 showed severe leukemic change in the peripheral blood and splenomegaly but lymphosarcoma in the lymph nodes was absent. Sheep 2 had lymphosarcoma in the lymph nodes and various organs. Neoplastic cells had B-cell marker in both cases and a few neoplastic cells contained intracytoplasmic IgM in sheep 2. It was presumed that B-cells might be transformed into neoplastic cells on the way of their differentiation. Some of neoplastic cells might have ability of immunoglobulin-production in sheep 2.

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An enzyme-linked immunosorbent assay (ELISA) was evaluated for diagnosis of experimental or naturally occurring Fasciola sp. infections in cattle. The positive rate for the ELISA in calves inoculated with Fasciola metacercariae were 21.1% by 2 weeks postinoculation (PI), 94.6% by 4 weeks PI and 100% by 6-21 weeks PI. The positive rate for the immunodiffusion test (Ouchterlony test) reached 91.7% by 2 weeks PI, however, it dropped to 77.8% by 10 weeks PI. The positive rate for the fecal egg examination was 0% by 10 weeks PI, 77.8% by 12 weeks PI and 100% by 14-21 weeks PI. The practical application of ELISA was tested by using 165 cows raised under field condition. All the 24 cows that were positive both in the fecal egg examination and the Ouchterlony test were ELISA positive. Of the 6 cows that were egg positive and Ouchterlony negative, 5 showed ELISA positive reactions. Of the 27 cows that were egg negative and Ouchterlony positive, 24 were ELISA positive. Of the 108 cows that were egg negative and Ouchterlony negative, 90 were ELISA negative. However, the other 18 cows had ELISA positive reactions. Our results suggested that the ELISA using crude adult antigen was superior to the Ouchterlony test and fecal egg examination for diagnosis of experimental or naturally occurring Fasciola sp. infections in cattle.

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The vascular architecture of the dog penis was studied by resin casts and histological preparations. Morphological evidence was found for penile engorgement. The crus and the bulb were compressed by the ischiocavernosus and the bulbospongiosus muscles, respectively, indicating that these muscles act as a booster pump. The distal part of the bulb was also strangulated by the external anal sphincter, indicating that this muscle prevents blood in the corpus spongiosum penis from going back into the bulb. The common trunk of the dorsal vein of the penis was significantly occluded by the ischiourethralis muscle. The internal pudendal vein was also throttled by the levator ani, the coccygeus, and the internal obturator muscles. The extremely narrow diameter of each vein draining the erectile body was insufficient to drain the increased blood flow and, therefore, formed a sluice channel. It is inferred that these muscle activities and sluice channels facilitate the erection of the penis.

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In order to obtain three-dimensional information on the fine architecture of the red pulp of the mink spleen, especially the circulation pattern in the red pulp, perfusion-fixed and freeze-cracked specimens were examined with a scanning electron microscope. A latticework was formed by elongated endothelial cells (rod cells) with side processes and the spongy reticular tissue. The outer surface of the sinus was covered with fine processes of reticular cells. Numerous sheathed arteries were found in the splenic cord. The sheath was composed of a few layers of flat reticular cells. The arterial capillaries of the red pulp opened directly into the cordal space. No evidence could be detected to prove or suggest any direct continuity between arterial capillaries and splenic sinuses. These results strongly support the concept of "open circulation", at least in the red pulp of the mink spleen, with the possibility of a "functionally closed circulation" under some physiological conditions.

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The erythropoietic effects of erythropoietin (EPO) have been investigated in the different administration schedule and injection routes. EPO was intravenously, intramuscularly or subcutaneously injected to partially nephrectomized anemic rats in 3 types of prescriptions (300 units of EPO per kg of body weight was respectively given in a dose at the first day, in 4 divided doses every 4 days, and in 7 divided doses every 2 days for 2 weeks). Repeated injections of EPO in divided doses caused stronger erythropoiesis than single injection. Especially, seven repeated injections promoted the strongest erythropoiesis. The serum iron concentration and reticulocyte counts suggested that erythropoiesis was continued less than 2 weeks after single injection but erythropoiesis in repeated injection groups had been accelerated for 2 weeks. Intravenous injections were less effective than either intramuscular or subcutaneous injections at 2 weeks after 7 repeated injections of 300 units of EPO per kg of body weight in 7 divided doses. Erythropoietic effects of EPO on the same total dose are dependent on the frequency of EPO injection and the durability of serum EPO concentration. On EPO usage, one bolus intravenous injection of excessive dose is considered to be wasteful and the repeating injections to maintain plasma EPO concentration is expected for the rational treatment of uremic anemia.

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O-sec-butylphenyl methylcarbamate (BPMC), an anticholinesterase carbamate, was injected once (100mg/kg, s.c.) or repeatedly (50mg/kg/day for 10 days) into mice. Animals were examined for their behavior and for parameters of cholinergic activity in the forebrain. Mice that received only a single injection exhibited reduced ambulation, hypothermia, and impairment of rotarod performance for up to 3 hr after a single injection. BPMC increased levels of acetylcholine (ACh) in the forebrain for up to 6 hr, and decreased acetylcholinesterase (AChE) activity for up to 24 hr. Both high-affinity choline uptake (HACU) and binding of [³H] quinuclidinyl benzilate (QNB) were reduced 20 min after a single injection without any effect on choline acetyltransferase (ChAT) activity. In behavioral tests conducted 10 min prior to the daily injections, rotarod performance and ambulation were slightly impaired for a few days before and after cessation of injection. Repeated treatment decreased HACU and binding of [³H] QNB for 24 hr after the final injection without any changes in levels of ACh content, AChE activity and ChAT activity. BPMC may reversibly impair cholinergic functions through effects not only on AChE activity but also on HACU and binding of [³H] QNB.

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