A total of 38 hybridomas producing monoclonal antibodies (mAbs) was established by immunizing BALB/c mice with extracts of the golden hamster testis. Six mAbs stained the acrosome of developing spermatids by immunofluorescence. Two mAbs (1A11 and 4D8) reacted with spermatid components other than acrosome. The mAbs 1C9 and 4D3 recognized a 103 kilodalton (kDa) protein on immunoblots, and were reactive to spermatocytes and early spermatids, but not to late spermatids and spermatozoa. This finding suggests that the protein functions for meiosis or early spermiogenesis. Four mAbs (3G2, 2E5, 2G3, and 3F10) stained all stages of spermatogenic cells. The remaining 24 mAbs showed a positive reaction to the basement membrane of the seminiferous tubule. Two of them, 3D6 and 3E5, recognized approximately 150 kDa major proteins, indicating that the antigen is an extracellular matrix.

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Estradiol receptors are classified into type I and type II receptors by their affinity and capacity for estradiol binding. The type II receptors are thought to be significant in the suppression of immune response, such as in the pregnancy-associated immunosuppression. The present study was undertaken to show the presence of type II receptors in rat spleen and to examine the change of receptor distribution after estradiol administration and during pregnancy. Scatchard analysis revealed that the type II receptors were present in rat spleen, and dissociation constants were estimated to be 3.23×10^-9 M for cytosol and 4.29×10^-9 M for nuclei. The receptors possessed specificity for estradiol and diethylstilbestrol, but not for promegestone, methyltrienolone and dexamethasone. Administration of estradiol to rats resulted in the increase of nuclear receptor concentrations with concomitant decrease of cytosolic concentrations. During pregnancy, the receptor concentrations were increased in the nuclear fraction, but were not significantly changed in the cytosolic fraction. The dissociation constants of the receptors in pregnant rdt spleen (4.77×10^-9 M for cytosol and 7.20×10^-9 M for nuclei) were similar to those in the non-pregnant control, suggesting the quantitative change of the receptors during pregnancy.

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Pseudorabies virus hemagglutinin was readily adsorbed on mouse erythrocytes at 4, 22, or 37°C, but not on cattle erythrocytes. The adsorbed hemagglutinin could not be eluted from the cells by resuspending in phosphate-buffered saline (PBS), by incubating at 37 or 50°C, or by incubating in the presence of neuraminidase. The receptor on mouse erythrocytes for the hemagglutinin was inactivated by trypsin, but not by neuraminidase, sodium deoxycholate (DOC), potassium periodate (KIO₄), dithiothreitol (DTT), 2-mercaptoethanol(2-ME) and formalin. The hemagglutinin was inactivated by trypsin, α-amylase, pepsin, DOC, KIO₄, and ethylendiamine-tetraacetic acid (EDTA), but not by papain, β-glucosidase, phospholipase C, neuraminidase, DTT, 2-ME, Tween-80, ethylether, chloroform, trichloro-trifluoroethane, β-propiolactone and formalin, suggesting that the hemagglutinin active component involved glycoproteins. The hemagglutinin was stable at 37°C for lower temperatures but not at 60°C or higher. The hemagglutinin activity was resistant to ultraviolet irradiation, while the infectivity was very susceptible. The hemagglutinin and the infectivity were readily sedimented by ultracentrifugation at 48, 000×g for 3 hr. In rate zonal centrifugation of the preparation on a sucrose density gradient, the hemagglutination (HA) activity showed a sharp peak at 1.22 g/ml coinciding with the peak of infectivity. The HA activity in the peak fraction seemed to be structually associated with virus particles. After fractionation of the virus by Nonidet P-40, the HA activity was found only in the fraction of the envelope material, indicating that the hemagglutinin is situated in the viral envelop.

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Perinatal diseases such as weak calf, stillbirth, or abortion were found in 86 calves from about 600 cows in a farm (A Farm) in 1987 and 1988. Such perinatal diseases were decreased to 36 calves in 1989 and 1990 after an additional supply of seaweed was made to the maternal feeds. From these cases, 23 stillborn and weak calves were pathologically examined. The mean weight with standard deviation of the 18 thyroid glands was 36.3±28.6 g before feeding of the seaweed supplement, and 12 (67%) of the glands showed moderate to marked degree of diffuse hyperplastic goiter in histological criteria. On the other hand, 5 glands weighed 12.0±3.4 g and revealed no such histological lesions after the additional supply. These results suggest that the goiter had been associated with birth of weak calves. Further study was performed on 37 newborn calves affected with Akabane disease in 1986 in A Farm and the neighbouring B Farm located. Seven (54%) of 13 thyroid glands from A Farm and 1 (4%) of 24 glands from B Farm showed histologically moderate to marked goitrous lesions. There were dotted "Inugarashi", Rorippa indica, family Cruciferae in both pastures. The mean iodine contents of the orchard grasses were 87 and 121 μg/kg on dry basis in A and B Farms. Marginal deficiency or lower limit of iodine and possible thiocyanate content of "Inugarashi" may have been responsible for the goiter in the farms.

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By in situ hybridization with biotinylated human interleukin 1α(IL-1α) cDNA probe, distribution of feline IL-1α mRNA-expressing cells was examined in the tissues from 49 cases diagnosed as feline infectious peritonitis (FIP) by pathological examination. IL-1α mRNA-expressing cells were found in visceral peritoneum, lymphoid organs, liver, kidney, pancreas, digestive tract, lung, pleura, brain, palpebral conjunctiva, and bone marrow. Hybridization signals for IL-1α mRNA were mostly located in the local inflammatory lesions on the serosal surface of various organs and omentum, which were frequently involved in the lesions of FIP (27.8±5.1 cells/mm²). Morphological examination suggested that they were infiltrated macrophages. However, few IL-1α mRNA-expressing macrophages were in the lesions of other organs. These data suggested that IL-1α produced from macrophages in the local inflammatory sites might participate in the initiation and development of the lesions on the visceral peritoneum in FIP.

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To understand the pathogenesis of equine exercise-induced myopathies and hemolysis, changes of phospholipid peroxidation products in the equine middle gluteal muscle and erythrocytes following the high-speed treadmill exercise were studied. In the skeletal muscle, the peroxidized phosphatidylethanolamine (PE) were increased at 24 hours after the exercise. The malondialdehydes (MDAs) were also increased as the protein-bound MDAs following exercise. In the erythrocytes, the peroxidized PE were significantly decreased at 24 hours after the exercise. The protein-bound MDAs were significantly increased at 5 min after the exercise and returned to the base values at 24 hours after the exercise. These findings indicate that the PE is more susceptible to in vivo oxidative effects than the other phospholipid classes, and the accumulation of the protein-bound MDAs is considered to play some cytotoxic roles in the equine skeletal muscle and erythrocytes following exercise.

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The bone marrow is commonly present in the medullary cavity of long bones in most mammals at birth. In the present study, the persistence of the osseous tissue was examined in the medullary cavities of piglet bones at and after birth. Immediately after birth, medullary cavities of long bones were almost completely filled with the spongy bone. The persisted spongy bone, i.e., medullary bone was prominent in younger pigs but decreased with advance in age. By 90 days of age, the medullary bone almost disappeared to form the extended medullary cavity. During the disappearing process of the medullary bone, osteoclasts showed significant increase in size and number (p<0.05). In irregular and flat bones, osseous trabeculae of the medullary bone became slender with age. The extramedullary hematopoiesis in the liver and spleen was very active till 30 days of age. This may suggest that the persistence of the medullary bone narrows the medullary cavity to cause poor medullary hematopoiesis and is compensated by extramedullary hematopoiesis. The relationship among the medullary bone, extramedullary hematopoiesis and piglet anemia was discussed.

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The central nervous system (CNS) of DBA/2 mice inoculated i.p. with the D variant of encephalomyocarditis virus (EMC-D) (10³ PFU/head) was examined up to 28 days postinoculation (28 DPI). The virus titer of CNS reached a maximum level at 4 DPI, and infectious viruses became undetectable by 28 DPI. Histopathologically, degeneration of neurons with virus antigens was observed in St. pyramidale hippocampi, Nuc. amigdaloideus corticalis and St. granulosum cerebelli of the brain and in Cornu ventrale of the thoracic to lumbar spinal cord at 6 DPI. In addition, in the spinal cord, demyelination was found in Funiculus ventralis and Funiculus lateralis at 6 DPI and it progresscd to form spongiosis at 10 DPI. In correspondence with these virological and histopathological findings, hind limb paralysis developed in some mice at 6 DPI, and its incidence increased markedly to about 60% at 10 DPI.

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The serum concentration of C-reactive protein (CRP) in horses was measured by single radial immunodiffusion (SRID) using rabbit anti-equine CRP serum to evaluate the significance of CRP as an acute-phase reactive protein. In serum samples of clinically normal newborn thoroughbred foals before being given colostrum, serum CRP was not detectable (≤1 μg/ml). The serum CRP concentration was found to increase quickly and reach a peak level of approximately 14.1 μg/ml in 12-month-old horses. Then, it decreased gradually to reach a low value of 5.4 μg/ml in the 4-year-old. In horses over 5-year-old, serum CRP concentration was stable at values of 7 to 8 μg/ml. In mares during the peri-natal period, serum CRP concentration decreased at 2 months before delivery, showed moderate changes just before and after delivery, and then increased to relatively high level by 2 months after delivery. The concentration of serum CRP was found to be increased at 24 hours after experimentally inflammatory stimuli in adult horses, and reached peak values of 3 to 6 times as high as the base line values at 3 to 5 days. Serum CRP concentration returned to base line values by 2 to 3 weeks after treatments. It is clear from these data that CRP is one of the acute-phase reactive proteins in horses.

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We investigated the source of immunoreactive relaxin (IR) in the dog during pregnancy using the following:(1)controls, (2)dogs ovariectomized during pregnancy and maintained on progesterone, (3)dogs hysterectomized during pregnancy or immediately postpartum, (4)corpora lutea, uteri, and placentas collected at various times during pregnancy for determination of IR, and (5)maternal (overian and uterine vein) and fetal (cardiac) blood and amniotic and allantoic fluid. Plasma IR patterns remained unchanged in animals subjected to ovariectomy, whereas in the controls IR was first detected at about Day 20 of pregnancy, peaked on Day 35, and was then remained at the same level until parturition. Hysterectomy, on the other hand, resulted in IR values becoming undetectable within 2 to 3 days. The highest tissue concentrations of IR were found in the placenta. These findings indicate that the source of relaxin in the pregnant dog is the placenta.

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The present study describes histological alterations and immunohistochemical distributions of extracellular matrices (ECMs) and the carbonic anhydrase isozyme-III (CA-III) during the period of bovine palatine ridge formation. Morphogenesis of bovine palatine ridges was preceded by epidermal placodes and the mesenchymal condensation (MC). During the early stages of less than 44 cm crown rump length (CRL), fibronectin (FN) was distributed densely in the MC. Strong reactions against type I collagen (C-1) were detected outer to the FN positive site. In the stages of more than 44 cm CRL, FN and C-1 weie distributed diffusely in subepithelial mesenchyme. Laminin (LN) and type IV collagen were distributed in the epithelial and endothelial basement membranes (BMs) in dll of the stages examined, except in the stage of 7 cm CRL, where LN was not detected only in the BM just beneath the epidermal placode. CA-III was detected in basal epithelial cells except for palatine ridge rudiments in the stages of more than 21 cm CRL. It is suggested that the expressions of LN and CA-III might play a role in the spatial determination of rudiments of bovine fetal palatine ridges.

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The relationship between cerebral lesions such as amyloid angiopathy or senile plaques and amyloid deposition in the visceral organs were studied in 90 autopsy cases of dogs, 0 to 19-year-old. Cerebral amyloid angiopathy was detected in 28 aged dogs (mean age: 13.7-year-old) and was found mostly in or around the wall of cerebral meningeal arterioles and capillaries of the neocortex. That condition was often accompanied by cerebral hemorrhage in dogs more than 9 years of age. Senile plaques were detected in the neocortex of the brain of 12 dogs (mean age: 13.2-year-old) and classified into 3 subtyped, i.e., "diffuse plaque", "primitive plaque" and "classical plaque". Among those 3 subtypes of senile plaques, amyloid containing plaques were small in number. In the visceral organs of dogs with cerebral amyloid angiopathy, amyloid deposition was found in the vascular walls or cornnective tissues of small intestines at a high frequency and sometimes in the vascular walls of the heart, lung, liver and thyroid gland as well as in atrioventricular valves. Amyloid in both cerebral and visceral organs was congophilic and showed green birefringence under poralized light even after potassium permanganate oxidation.

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Five inbred strains of mice were tested for their susceptibility to Streptococcus suis type 2 including the type strain, two isolates from meningitis in pigs and two isolates from tonsils of clinically healthy pigs. C57BL/6, ICR and ddY strain mice showed lower susceptibility to all strains of S. suis type 2 than BALB/c and SS strain mice. The type strain and the isolates from diseased pigs produced septicaemia and meningitis in BALB/c and SS mice inoculated with 10⁸ colony forming unit of the bacteria and 6O to 1OO% of these infected mice died. On the other hand, mice inoculated with the isolates from healthy pigs showed mild clinical signs but none of them died. In BALB/c mice which died or developed nervous signs, the purulent meningo-encephalitis, myocarditis, ophthalmitis, labyrinthitis and otitis media were observed. S. suis type 2 antigen was demonstrated in these lesions by immunoperoxidase staining using rabbit S. suis type 2 antiserum. These results were similar to those in the experimentally infected pigs with these virulent and avirulent strains against mice. These results indicate that BALB/c and SS strains of mice are useful as an experimental model of S. suis type 2 infections in pigs, and that there are virulent and avirulent strains against mice and pigs among the strains of S. suis type 2.

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Systemic mycoses were found in 19 (4.7%) of 406 calves less than 6 months old which were autopsied during the past 10 years. Alimentary mycosis occurred in 12 (63.2%) of 19 cases. In alimentary mycosis, mucormycosis showed the highest rate of occurrence (91.7%, 11/12 calves) followed by aspergillosis 41.7% and candidiasis was 9.3%. Mucormycosis and aspergillosis were characterized by focal hemorrhagic necroses with hyphal proliferation and thrombi in the mucosa and muscular layers of the forestomach, abomasum, and small intestine. Candidiasis was characterized by hyperkeratosis with pseudohyphae and microconidia in the mucosa of the omasum. Four of 12 calves (33.3%) had mixed infections of the alimentary tract consisting of Mucorales and Aspergillus species. Pulmonary aspergillosis was found in 10 (52.6%) of 19 calves. There were micro-abscesses with hyphal proliferation or asteroid bodies in the lungs. Infections involving both the alimentary tract and respiratory organs were noted in 3 (10.5%) of 19 calves. Disseminated mycosis was found only in one calf. In alimentary mycosis, administration of antibiotics for the treatment of diarrhea and early weaning were thought to be an important predisposing factor.

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Seventeen field isolates of sialodacryoadenitis (SDA) virus had been isolated from the lung of rats with clinical SDA during epizootics of SDA from 1976 to 1978 in the research laboratory. Based on their neutralization patterns against antisera to strains 681, 930-10, Lu-3, Lu-4 and Lu-7, these isolates were divided into 3 antigenic groups. The first group consisted of 8 isolates which were neutralized by 4 out of 5 antisera at high serum dilution. The second group consisted of 6 isolates which were neutralized by only 2 antisera at high serum dilution. The third group exhibited intermediate neutralization pattern and 3 isolates belonged to it. Considering the time course of virus isolation, it was concluded that three antigenically different SDA viruses had been spread irregularly and ocassionally two had been spread simultaneously in an animal house of rats during the several epizootics of SDA.

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