Immunodominant antigens of zoospores from ovine isolates of Dermatophilus congolensis.

Zoospores of Dermatophilus congolensis were analysed by SDS-PAGE and western blotting. The electrophoretic profiles of zoospores from 13 isolates of D. congolensis were similar but not identical when stained with Coomassie blue or silver. Immunodominant polypeptides with apparent molecular masses of 76 and 31 kDa were identified in western blots of 13 of 13 and 12 of 13 isolates respectively of D. congolensis reacted with hyperimmune, ovine, antizoospore sera. Identical immunodominant polypeptides were observed in western blots reacted with sera obtained from naturally infected sheep. Initial characterisation of the 76 and 31 kDa polypeptides indicated that they were probably surface exposed because (i) antibodies eluted from the surface of live zoospores after adsorption of hyperimmune antizoospore serum, reacted principally against the 76 and 31 kDa subunit polypeptides in western blots, (ii) adsorption of hyperimmune antizoospore serum with live zoospores resulted in significant diminution of reactivity against both the 76 and 31 kDa polypeptides in western blots, (iii) indirect fluorescent immunostaining of zoospores with antiserum prepared against gel-purified 76 kDa polypeptide, resulted in intense staining of the zoospore outer coat. Immuno-gold electron microscopy of negatively stained zoospores with antiserum prepared against gel-purified 31 kDa polypeptide identified this antigen as a flagella subunit.     

Immunohistochemical Localization of Progesterone Receptor Isoforms in the Chick Pre-follicular Ovary

The distribution of progesterone receptor A and B isoforms in different cell types of the chick pre-follicular ovary was studied by immunohistochemistry. Newly hatched chicks were killed and the left ovary was removed, fixed and embedded in paraplast. Sections (5 microns thick) were made for the detection of progesterone receptor isoforms, using a technique of indirect immunoperoxidase. The results indicate that progesterone receptors were localized in the nuclei of germinal epithelium and germ cells of the ovarian cortex and in the interstitial and epithelial cells of the lacunar channels of the ovarian medulla. Undifferentiated cells did not present progesterone receptors. In all cell subpopulations progesterone receptor B was the predominantly expressed isoform. These data suggest that progesterone receptor isoforms are differentially expressed in the chick pre-follicular ovary and that progesterone effects in this tissue are mediated by the progesterone receptor B isoform.     

Identification of the antigenic components of paramyxovirus-3, paramyxovirus-6 and Newcastle disease virus in turkeys

The aim of this study was to use the enzyme-linked immunosorbent assay (ELISA) and the Western immunoblotting as possible tools to differentiate infections in turkeys by different paramyxoviruses. Pooled hyperimmune sera of turkeys infected with either paramyxovirus-3 (PMV-3), paramyxovirus-6 (PMV-6), or Newcastle disease virus (NDV) were assayed for antibodies specific to the three viruses by the ELISA and Western immunoblotting. ELISA results showed cross reactions of turkey antibodies between PMV-3 and PMV-6 antigens, while turkey antibodies to NDV did not cross-react with any of the other paramyxoviruses. The immunoblots of sera from birds infected with PMV-3 (Minnesota turkeys and Iowa chickens) reacted to low molecular weight polypeptides of PMV-3 of 29, 32, and 34 kDa, and to a high molecular weight band of 200 kDa. The same Minnesota turkey sera had a cross reaction to the 200 kDa polypeptide of PMV-6, while the Iowa chicken sera did not. Both sera had no apparent reaction to NDV proteins. Western immunoblotting showed that the turkey PMV-3 sera had a specific reaction to a 220 kDa polypeptide present in PMV-3, but not in PMV-6, while the turkey PMV-6 sera had a specific reaction to a 130 kDa polypeptide present in PMV-6, but not in PMV-3. Immunoblots of pooled sera from turkeys infected with PMV-6 (Minnesota source) reacted to the 200 kDa protein present in both PMV-3 and PMV-6; however, no reaction occurred between this sera and NDV proteins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Carcinogenicity of biphenyl in mice by two years feeding

Carcinogenicity and chronic toxicity of biphenyl was examined in the male and female BDF1 mice fed a diet containing biphenyl at 667, 2,000 or 6,000 ppm for 2 years. There was no difference in survival rate between any biphenyl-containing diet-fed group of either sex and the respective control. Body weights of the males and females fed 6,000 ppm diet were significantly lower than the respective control. Incidences of hepatocellular carcinomas and hepatocellular adenomas in the females fed diets containing biphenyl were significantly increased in a dose-related manner, and exceeded a range of the historical control data in the Japan Bioassay Research Center. Incidences of basophilic cell foci in the liver were increased in the females fed 2,000 and 6,000 ppm diets. There was no increase in tumor or tumor-related lesion in the males fed diets containing biphenyl. Chronic toxicity of biphenyl was characterized by increased incidences of urothelial desquamation in the renal pelvis in males and females and mineralization in the inner stripe of renal outer medulla in females, as well as changes in serum levels of BUN, ALP and some electrolytes in males and females. In conclusion, the 2-year oral administration of biphenyl-containing diets induced pre-neoplastic and neoplastic lesions in the liver of females and non-neoplastic lesions in the kidney of males and females. Causative factors for the biphenyl-induced hepatocarcinogenicity were discussed in light of our published finding of peroxisome proliferation.     

Neutralizing and Complement-fixing Monoclonal Antibodies as an Aid to the Diagnosis of Equine Herpesvirus-1 Infection

One complement-fixing (C-MAb) and three complement-dependent neutralizing monoclonal antibodies (N-MAbs) were raised against Hisar-90-7 equine herpesvirus-1 (EHV-1) strain. The target antigen of the C-MAb (2A5) and two of the N-MAbs (1H6, 9C4) was identified as a 140 kDa polypeptide in Western blotting. The target antigen of N-MAb (9C6) could not be identified. Purified polypeptides of five EHV-1 strains isolated from different regions and at different times gave intense bands at 140 kDa when reacted with N-MAb (1H6) in Western blots. In sandwich ELISA, all four MAbs captured the viral antigen from clinical materials, giving a reliable and rapid diagnosis of EHV-1 infection in equines.     

Comparative Thickness of the Medulla of Bos taurus and Bos indicus Kidneys

Twenty kidneys of 4 years old Bos taurus cattle and 26 kidneys of 2—4 years old Bos indicus (Zebu cattle) of East African Shorthorn breen were examined lobe by lobe after fixation. The cortex and medulla with subunits were measured and compared in the two sub-species. The width of the cortex was about the same in both. The outer and inner stripes of the outer medullary zone were significantly broader in Bos taurus whereas outer inner zone was significantly broader in Bos indicus. The width of the entire medulla was greater in Bos taurus.     

Distribution of tissue water and electrolytes in normal rhesus macaques.

Techniques for the determination of water and electrolytes in individual tissues of normal rhesus macaques (Macaca mulatta) are described. Base-line values are presented for intracellular and extracellular distributions of water and electrolytes in 14 tissues, including skin, skeletal muscle (gastrocnemius), cardiac muscle (left ventricle), lung, diaphragm, liver, renal cortex, outer medulla and inner medulla, cerebral cortex, cerebellum, thalamus-hypothalamus complex, medulla oblongata, and spinal cord.     

Cyclooxygenase-1 and -2 expression in urothelial carcinomas of the urinary bladder in cows

Bovine urinary bladder tumors occur frequently in animals suffering from chronic enzootic hematuria because of prolonged ingestion of bracken fern (Pteridium spp.). Cyclooxygenase (COX) genes (COX-1 and COX-2) are known to be involved in the carcinogenesis of human and some animal urothelial tumors. The aim of the present study was to investigate COX-1 and COX-2 expression by immunohistochemical methods in 20 bovine urothelial carcinomas collected at public slaughterhouses from cows that had been suffering from chronic enzootic hematuria. COX-1 immunostaining was identified intracytoplasmically in normal urothelium and in 15 of 20 neoplastic specimens. COX-1 immunosignal in the tumor cells was either absent or weak. COX-2 was also expressed intracytoplasmically in 17 of 20 urothelial carcinomas. Moderate to intense COX-2 labeling was detected in both noninvasive and invasive urothelial carcinomas. Coexpression of both enzyme isoforms was also revealed by confocal laser scanning microscopic investigations. This study indicates that COX-2 is overexpressed in naturally occurring urothelial carcinomas of cows.     

Hemagglutinin glycosylation modulates the pathogenicity and antigenicity of the H5N1 avian influenza virus

The location and number of glycosylation in HA proteins exhibit large variations among H5 subtype avian influenza viruses (AIVs). To investigate the effect of glycosylation in the globular head of HA on the pathogenicity and antigenicity of H5N1 AIVs, seven rescued AIVs differing in their glycosylation patterns (144N, 158N and 169N) within the HA globular head of A/Mallard/Huadong/S/2005 were generated using site directed mutagenesis. Results showed that loss of glycosylation 158N was the prerequisite for H5 AIV binding to the α2,6-linked receptor. Only in conjunction with the removal of the 158N glycosylation, the H5 AIVs harboring both 144N and 169N glycosylations obtained an optimal binding preference to the α2,6-linked receptor. Compared with the wild-type virus, growth of viruses lacking glycosylation at either 158N or 169N was significantly reduced both in MDCK and A549 cells, while replication of viruses with additional glycosylation 144N was significantly promoted. Mutant viruses with loss of 158N or 169N glycosylation sites showed increased pathogenicity, systemic spread and pulmonary inflammation in mice compared to the wild-type H5N1 virus. In addition, chicken studies demonstrated that inactivated de-glycosylation 169N mutant induced cross-reaction HI and neutralization antibody against various clades of H5N1 AIVs. Moreover, this type of glycan pattern vaccine virus provided better cross-protection in chickens compared to wild-type vaccine virus. Thus, the glycosylation alteration of HA should be considered in the global surveillance and vaccine design of H5 subtype AIVs.     

RENAL MEDULLARY RIM SIGN: ULTRASONOGRAPHIC EVIDENCE OF RENAL DISEASE

An echogenic line in the outer zone of the renal medulla, paralleling the corticomedullary junction is described as the renal medullary rim sign. This renal ultrasonographic change is demonstrated in 4 dogs and 2 cats with a range of renal diseases. The renal medullary rim sign provides additional ultrasonographic criteria indicating primary renal disease in some patients. However the renal medullary rim sign may prove to be a poor correlate for prognosis across the range of differentials present in these clinical patients.     

Expression of transforming growth factor-beta isoforms in the skin, kidney, pancreas and bladder in a German shepherd dog affected by renal cystadenocarcinoma and nodular dermatofibrosis

The present study was performed to assess the expression of isoforms 1, 2 and 3 of transforming growth factor (TGF)-beta in skin nodular dermatofibrosis lesions, kidney, bladder and pancreas from a 10-year-old female German shepherd dog (GSD) affected by renal cystadenocarcinoma and nodular dermatofibrosis (RCND) compared with normal GSDs (n = 2). Formalin-fixed, paraffin-embedded tissues obtained from the dog affected by RCND, diagnosed by renal ultrasonography and histopathological examination were analysed by immunohistochemistry using polyclonal antibodies to TGF-beta1, 2 and 3, and evaluated semiquantitatively using an immunoreactivity score. Similar expression of TGF-beta2 and TGF-beta3 was observed in all tissue specimens in both the RCND-affected animal and normal dogs. In contrast, TGF-beta1 immunoreactivity was increased in the derma of the RCND canine. Comparable TGF-beta1 serum levels were found between the diseased and normal animals. The increased local cutaneous production of TGF-beta1 in the RCND dog, compared with the normal animals, suggests that this cytokine may play an important role in the induction of nodular dermatofibrosis associated with renal cystadenocarcinoma.     

鸟类飞羽羽轴髓质结构的定量分析

鸟类飞羽羽轴机械性能一部分来自髓质,但是髓质泡沫结构如何影响羽轴机械性能的问题仍未被研究。本文对5种鸟类初级飞羽羽轴的髓质形态结构进行研究,测定了腔室的大小、壁厚度等形态计量学参数,分析了这些结构参数与体重的关系以及腔室在抵抗形变的作用过程。结果显示,髓质腔室断面的平均周长(l,表示腔室的大小)介于74.26±11.27～153.02±59.01μm之间,在一些种类(如丹顶鹤和毛脚鵟),外周髓质的l值显著大于中央髓质(P<0.05),而在另一些种类(鸿雁、银鸥和短耳鸮)择二者差异不显著(P>0.05);腔室壁厚度(t)介于0.51±0.15～2.14±1.11μm之间,在所有种类外周髓质的t值均显著(P<0.05)或极显著(P<0.01)大于中央髓质。l和t之间没有显著的相关性(P>0.05),但是外周髓质的t值与翈面面积、体重均呈显著的线性相关(P<0.05)。上述结果表明,空球形腔室可以把某一点上的压力有效地传递和分配到整个羽轴内部,并在每个形变的腔室壁上积累势能;外周髓质对羽轴机械性能的贡献大于中央髓质,这种贡献主要缘于腔室壁的厚度而不是腔室的大小。但是,腔室大小在不同鸟类显出与扑翼强度相一致的关系。     

Clinical aspects of experimentally induced chloride deficiency in Holstein calves

A severe total body chloride deficit was induced in Holstein calves by feeding a low-chloride ration (0.063% Cl) and removing digesta daily from the abomasum through a surgically implanted cannula. Clinical signs of the deficit observed included polydipsia, polyuria, dehydration, anorexia, scleral injection, decreased respiratory rate, and blood and mucus in the feces. Necropsy findings included dehydration, blood in the lumen of the small intestine, and renal lesions. The most extensive histopathologic changes occurred in the renal tubular epithelium of the outer medulla where mineralization of the tubular epithelium and basement membranes was frequently seen.     

Characteristics of glycoprotein B of equine herpesvirus 1 expressed by a recombinant baculovirus.

A recombinant baculovirus (Bac-EgB) containing the complete open reading frame of equine herpesvirus 1 glycoprotein B (EHV-1 gB) expressed recombinant products of 107-133 kDa, 58-75 kDa and 53-57 kDa, corresponding to EHV-1 gB precursor, large and small subunits respectively. High molecular mass products (>200 kDa) in the Bac-EgB infected insect cells were consistent with oligomerisation of the recombinant EHV-1 gB products, and analysis with tunicamycin and endoglycosidases indicated that the baculovirus-expressed gB contained N-linked sugars with high mannose and hybrid chains. N-terminal amino acid sequence analysis of the gB forms revealed identical signal and endoproteolytic cleavage sites to those of gB in EHV-1 infected mammalian cells, and authenticity of processing and transport was supported by the presence of EHV-1 gB antigen at the surface of infected insect cells. Immunogold labelling and electron microscopy of recombinant baculovirus particles indicated that the recombinant gB was also present in baculovirus envelopes. Bac-EgB infected insect cells were able to induce low levels of complement dependent virus neutralising antibody, and have been shown to evoke protective immune responses in murine models of respiratory disease and abortion.     

Definition of the specificity of monoclonal antibodies against porcine CD45 and CD45R: report from the CD45/CD45R and CD44 subgroup of the Second International Swine CD Workshop

Swine cell binding analyses of a set of 48 monoclonal antibodies (mAbs), including eleven standards, assigned to the CD44 and CD45 subset group of the Second International Swine CD Workshop yielded 13 clusters. Although none of these corresponded to CD44, seven mAbs formed a cluster which was identified as being specific for restricted epitopes of CD45 (CD45R). In addition, a T-cell subset specific cluster comprised of four mAbs was also identified. Two mAbs (STH106 and SwNL 554.1) reacted exclusively with CD8 bright lymphocytes, the other two (2B11 and F01G9) with a subset of CD4 lymphocytes. The other 10 clusters were either specific for MHC-class I like molecules or overlapped with clusters identified by the adhesion molecule subgroup and are therefore just briefly discussed in this report. The specificity of all the mAbs in the CD45R cluster was verified by their ability to immunoprecipitate distinct proteins and to react with CHO cells expressing individual isoforms of CD45. Three CD45R mAbs (3a56, MIL5, −a2) did react with a 210 kDa isoform(s), while another three (STH267, FG2F9, 6E3/7) only recognized a 226 kDa isoform(s). The remaining one (MAC326) precipitated both a 210 and 226 kDa protein. The specificity of all the mAbs in the CD45R cluster, and of the CD45 common mAbs, was confirmed by their reactivity with CHO cells transfected with cDNAs encoding the extracellular and transmembrane portions of distinct CD45R isoforms. Those mAbs recognizing a 210 kDa protein reacted with CHO cells expressing the CD45RC isoform, while those capable of precipitating a 226 kDa, but not the 210 kDa, polypeptide recognized CHO cells expressing either the CD45RAC and the relatively rare CD45RA isoform. MAC326 was unique in its inability to react with CHO cells engineered to produce the CD45RC and CD45RAC isoforms. Thus, three mAbs (6E3/7, STH267, and FG2F9) appear to be specific for an epitope(s) encoded by the A exon, while one (MAC326) recognizes a determinant encoded by the C exon. The remaining three mAbs (3a56, −a2, MIL5) are apparently specific for an epitope(s) which results from the fusion of the C exon to the invariant leader sequence and is destroyed by inclusion of the A exon. All three CD45 common mAbs, K252.1E4, MAC323 and 74.9.3, did react with the CHO cells lines expressing either the CD45RA, CD45RC, CD45RAC or CD45RO isoforms, but not with untransfected CHO cells. When the natural expression of CD45 isoforms was examined by reacting lymphocytes with CD45R mAbs, a high level expression of isoforms containing the A exon-generated domain was detected in all B cells while the majority of CD4⁺ T cells had undetectable or lower expression density of this protein than B cells. In contrast, the density of expression of the CD45 isoform(s) containing the C exon-generated domain ranged from undetectable to high in CD4⁺ T cells whereas the amounts were approximately ten-fold lower in B cells. Overall this panel of CD45 mAbs will be very useful in analyzing the maturation and differentiation of swine lymphoid cells subsets.     

Interaction between chromaffin and sustentacular cells in adrenal medulla of viscacha (Lagostomus maximus maximus)

New evidence provides valuable information about the participation of sustentacular cells in chromaffin cell catecholamine secretion. In this process, calcium ions play an important role. It has been shown that there is an intense ionic traffic between both types of cells. Moreover, sustentacular cells take an active part in calcium metabolism, regulating levels of the ion and indirectly, the synthesis and release of catecholamines. This background information encouraged us to study the sustentacular population of Lagostomus adrenal medulla and its morphologic relationship with the chromaffin population. The animals were captured, transported to the animal facilities, anaesthetized and killed. The adrenal gland was processed by immunohistochemistry using antiserum against S-100 (subunit alpha and beta), a specific marker. Through the morphological and immunohistochemical study, it was found that there are sustentacular cells in deferent regions of adrenal medulla, mainly in the basal zone of chromaffin cells, which constitute the glomerular structure around blood capillaries. Cytoplasmic extentions of sustentacular cells penetrate into chromaffin cells and make contact with the basal membrane of the capillary endothelium. The relationship among chromaffin cells, capillaries and sustentacular cells suggests that they may intervene actively in the adrenal medulla metabolism.     

Renal Tubular Cyst Formation in Newborn Rats Treated with p-Cumylphenol

In this study, we investigated the sequential changes in the development of renal tubular cysts in newborn rats treated with p-cumylphenol (PCP). Fifteen animals per sex were treated orally with 300 mg/kg/day of PCP for up to 18 days from postnatal day (PND) 4 and were sacrificed on PNDs 8, 12, 19 and 22 and after a 7 day recovery period. On PNDs 8 and 12, slight dilatation of the collecting ducts was frequently observed in the medulla and slight papillary necrosis was also noted in some cases. These dilated collecting ducts were lined with slightly hyperplastic epithelial cells. On PNDs 19 and 22, multiple large cystic changes arising from the collecting ducts in the outer medulla were seen. These cystically dilated ducts were also lined with hyperplastic epithelial cells. During the dosing period, the labeling index of proliferating cell nuclear antigen in the collecting duct epithelium was higher in the PCP-treated group than in the control group at all time points. After a 7 day recovery period, the cystic change still remained, although the cell density was decreased and the epithelial cells became flattened. On the other hand, basophilic tubules with peritubular lymphoid cell infiltration were multifocally observed in the cortex. In conclusion, PCP induced multiple renal cysts that developed in the collecting ducts of the outer medulla in neonatal rats, and it is suggested that epithelial cell proliferation may play some roles in the induction of cystic lesions.     

Pharmacokinetics of phenylbutazone and oxyphenabutazone in the pig.

The kinetics of phenylbutazone (pbz) and its main metabolite oxyphenbutazone (oxpbz) were investigated in seven pigs in order to determine if this animal might serve as a model for human investigations. The study was performed in two stages, one as single dose experiments using intravenous injection, the second as infusion experiments. The rate of elimination for both compounds was demonstrated to be much faster than in man. Similar results have been observed in several other animal species. The degree of protein binding (pbz 98%, ox-pbz 97%), the apparent specific volume of distribution (pbz 0.18 1/kg, ox-pbz 0.28 1/kg) and the renal clearance were found to be similar to results obtained in man and rat. Ox-pbz had a higher renal clearance (0.13 ml/min/kg) than pbz (0.003 ml/min/kg) but still the renal excretion constituted only a small fraction of the total elimination (pbz 0.5%, ox-pbz 5%). Provided real steady state conditions were obtained during the infusion experiments calculations based on the results from the single dose experiments in the same animal revealed that between 20 and 60% of the injected pbz was metabolized to the ring-hydroxylation product (ox-pbz). It is concluded that the pig model is not superior to other animal models for studies of pbz/ox-pbz. The rapid elimination pattern is the main problem in relation to human investigations.