Immunization of proteins from Toxascaris leonina adult worm inhibits allergic specific Th2 response

Recently, the influence of parasitic infections on the incidence of allergic diseases has become the focus of increased attention. In order to ascertain whether parasite-derived proteins could inhibit the allergic specific Th2 response, we applied excretory-secretory protein (Tl-ES) or total protein (Tl-TP) of the adult worm Toxascaris leonina to asthma model mice prior to or simultaneously with OVA challenge, after which we assessed the OVA-specific Th2 responses. The group subjected to immunization with Tl-ES and Tl-TP (immunized group) evidenced a thinning of the bronchial epithelial and muscle layer, a disruption and shedding of epithelial cells, a reduction in the number of goblet cells, and a reduction in mucus production as compared to the group treated with Tl-ES coupled with OVA challenge (challenge with OVA groups) and the OVA-induced asthma group. The administration of Tl-ES and Tl-TP, regardless of injection time, was shown to inhibit the recruitment of inflammatory cells into the airway, and in particular, macrophages, neutrophils, and lymphocytes were significantly reduced as the result of the parasite proteins. However, the total number of eosinophils was slightly reduced as the result of the administration of parasite proteins. Sensitization and OVA challenge was shown to accelerate the secretion of Th2 cytokines (IL-4 and IL-5) within the lung, but in the immunized groups, those levels were lower. The administration of Tl-TP and OVA challenge group also evidenced a significant reduction in IL-4 levels as compared to the OVA-challenged group. The concentrations of Th2 cytokines in the Tl-ES and OVA challenge group were more similar to those observed in the OVA-challenged group. The concentration of IL-10 and TGF-beta in the lung was decreased substantially in the OVA-only challenge group, but the Tl-TP immunized group exhibited significantly induced IL-10 cytokine. OVA-specific IgG2a, IgG1, and IgE levels in the immunized groups were significantly lower than those detected in the OVA-challenged group. In conclusion, parasite-derived protein is able to inhibit OVA-specific Th2 responses, and in particular, immunization with parasite proteins exerts a more profound protective effect than is seen with the treatment of allergic reactions. The results of our study are encouraging in terms of our further understanding of the molecular basis of immune evasion by nematodes.     

Topical 0.03% tacrolimus for treatment of pemphigus erythematosus in a Korea Jindo dog

Topical 0.03% tacrolimus was used for treatment of a Korea Jindo dog diagnosed with pemphigus erythematosus. The dog was slowly improved following application of tacrolimus but did not achieve complete remission until end of this study. No adverse effects on clinical or laboratory parameters were noted during the topical tacrolimus therapy period.     

Cytochrome c upregulation during capacitation and spontaneous acrosome reaction determines the fate of pig sperm cells: linking proteome analysis

To identify the mechanisms underlying capacitation, we undertook a high-resolution differential proteomic analysis of pig sperm cells. Two-dimensional gel electrophoresis and subsequent MALDI-TOF mass spectrometry analyses led to identification of 56 differentially expressed proteins. After induction of capacitation in vitro, the well-established markers of the capacitation (lactadherin P47, acrosomal protein SP-10 precursor, prohibitin, proteasomes, DJ-1 protein and arylsulfatase-A) and TCA cycle proteins (isocitrate dehydrogenase, malate dehydrogenase and pyruvate dehydrogenase) were identified. During induction, cytochrome c expression via the p53 pathway increased, however apoptotic executors, such as caspase-3, decreased significantly. Therefore, we tested the hypothesis that cytochrome c upregulation in spermatozoa is capable of activating tyrosine phosphorylation for capacitation, rather than apoptosis. Exposure of sperm cells to soluble Na2CrO4 [Cr (VI)], which induces cytochrome c upregulation, caused a dose- and time-dependent increase in tyrosine phosphorylation of sperm proteins in non-capacitating medium. In contrast, supplementation of cyclosporin A, which blocks cytochrome c upregulation, inhibited tyrosine phosphorylation of sperm proteins. Furthermore, spermatozoa in capacitation medium or non-capacitation media supplemented with soluble Cr (VI) showed similar levels of capacitation. These findings indicate that differential expression of many of these proteins has previously been unrecognized in sperm cells incubated in capacitation medium also suggest that a gradual increase of cytochrome c during incubation to induce capacitation determines sperm cell fate, i.e., apoptosis or further development for fertilization.     

Influence of age, sex, and production class on liver zinc concentration in calves.

Determination of zinc concentrations in the liver of calves and young stock is commonly requested by practitioners and nutritionists to assess whether they receive an appropriate amount of zinc in their diet. However, interpretation of liver zinc concentrations is currently based on information reported for adult cattle for which the health status was unknown and irrespective of production class, sex, and age. A retrospective study of necropsy reports was undertaken to assess the relationships between liver zinc concentrations and age, sex, and production class for calves that did not have a history compatible with zinc toxicosis or zinc deficiency. Results of a generalized least squares, polynomial regression analysis of 474 records found that zinc concentration was not affected by sex (P = 0.29) or production class (P = 0.50). Zinc concentration was significantly associated with linear (P < 0.00001) and nonlinear (quadratic, P = 0.0039) functions of age (r2 = 0.1503), where the concentration decreased from 93 mg/kg wet weight at 30 days of age to 57 mg/kg wet weight at 9 months of age, after which it began to increase. The age-specific 95% confidence limits of the mean concentration for a group of calves and the 95% prediction limits of a single concentration value for an individual animal estimated in this study suggest reconsideration of the recommended limits for liver zinc concentration in calves. As a consequence of the significant influence of age on liver zinc concentration of calves presumably not experiencing zinc toxicosis or deficiency, diagnosis of zinc imbalances based on liver zinc concentration needs to consider age as a diagnostic covariate.     

Can CT texture analysis parameters be used as imaging biomarkers for prediction of malignancy in canine splenic tumors?

Splenic hemangiosarcoma has morphological similarities to benign nodular hyperplasia. Computed tomography (CT) texture analysis can analyze the texture of images that the naive human eye cannot detect. Recently, there have been attempts to incorporate CT texture analysis with artificial intelligence in human medicine. This retrospective, analytical design study aimed to assess the feasibility of CT texture analysis in splenic masses and investigate predictive biomarkers of splenic hemangiosarcoma in dogs. Parameters for dogs with hemangiosarcoma and nodular hyperplasia were compared, and an independent parameter that could differentiate between them was selected. Discriminant analysis was performed to assess the ability to discriminate the two splenic masses and compare the relative importance of the parameters. A total of 23 dogs were sampled, including 16 splenic nodular hyperplasia and seven hemangiosarcoma. In each dog, total 38 radiomic parameters were extracted from first-, second-, and higher-order matrices. Thirteen parameters had significant differences between hemangiosarcoma and nodular hyperplasia. Skewness in the first-order matrix and GLRLM_LGRE and GLZLM_ZLNU in the second, higher-order matrix were determined as independent parameters. A discriminant equation consisting of skewness, GLZLM_LGZE, and GLZLM_ZLNU was derived, and the cross-validation verification result showed an accuracy of 95.7%. Skewness was the most influential parameter for the discrimination of the two masses. The study results supported using CT texture analysis to help differentiate hemangiosarcoma from nodular hyperplasia in dogs. This new diagnostic approach can be used for developing future machine learning-based texture analysis tools.     

Seroprevalence of leptospirosis in Korean municipal zoo animals

From 2002 to 2005, we collected 118 serum samples from 34 species belonging to 13 families of zoo animals in Korea and determined the prevalence of antibodies for 18 serovars of Leptospira spp. using the microscopic agglutination test (MAT). Twenty-nine (25%) of the serum samples tested were positive for one or more of the serovars. There were no significant differences in relation to genders: 23% and 26% of positives occurring in male and female animals, respectively (P>0.05). However, the seroprevalence for the Leptospira spp. was significantly higher (P<0.05) in herbivores (45%) than in either carnivores (17%) or omnivores (17%). Among the 5 serovars detected in this study, the most common was sejroe (n=27; 87% of all positive reactions). All positive reactions showed low titers (< or = 1:200) and the positives were most frequently detected in 1:25 (58%) and 1:50 (23%) serum dilutions. The highest antibody titer (1:200) was observed for the serovars sejroe (n=1) and bratislava (n=1). We conclude that the exposure of zoo animals to Leptospira spp. is relatively common in Korea and produces low MAT titers, with sejroe being the most commonly encountered serovar.     

Biology of epidermal and hair pigmentation in cattle: a mini-review

Coat colours in cattle have been of interest to both breeders and researchers as genes regulating pigmentation not only affect the phenotype but also have economic implications in the event of genetic mutations. The genes controlling pigmentation act as a complex and interact with each other to cause phenotypic and genotypic variations. Pigmentation of coat broadly depends on the ratio of eumelanin and pheomelanin, the two components of melanin. Increase in eumelanin imparts a black coat colour while raise in pheomelanin is responsible for a yellowish or reddish colour. The main enzymes responsible for melanogenesis are regulated by the genes of the tyrosinase family. It is speculated that the wild-type gene present in the ancestral breeds of the present day cattle have more pheomelanin content and that, over time, mutations have introduced more variations leading to many shades. This could have occurred either because of interactions or because of deletions in the responsible genes. The environmental conditions have also contributed to mutations in these genes, helping in the adaptability of the animals to different geographical regions. The switching between the syntheses of melanin components depends on several genes like melanocortin-1receptor gene (MC1r) - also known as melanocyte-stimulating hormone receptor gene (MSHr)-, agouti (A), attractin (Atrn) and mahogunin (Mgrn1). The purpose of this review is to summarize the recent advances in the field of pigment biology and to highlight possible areas of research that may benefit a breeder or a farmer in the selection of animals on the basis of phenotype.     

Cellular characterization of multidrug resistance P-glycoprotein, alpha fetoprotein, and neovascular endothelium-associated antigens in canine hepatocellular carcinoma and cirrhotic liver

P-glycoprotein (P-gp), which is encoded by the multidrug resistance gene (MDR-1); alpha fetoprotein (AFP); and vascular endothelium-associated antigens are well-known markers for human and canine hepatic diseases. We obtained liver tissues from 5 dogs with hepatocellular carcinoma (HCC) and 12 dogs with cirrhosis, and we performed histopathologic and immunohistochemical evaluations using anti-P-gp, anti-AFP, anti-CD31, and anti-CD34 antibodies. P-gp was expressed at higher levels in HCC than in cirrhotic livers ( P < .01), and was most commonly localized in biliary canaliculi and small ductuli. AFP was localized mainly in the cytoplasm in HCC ( P < .01) and in a few cases of cirrhosis. In both HCC and cirrhosis, the AFP-positive cells were morphologically similar to normal hepatocytes and showed an even cytoplasmic distribution of AFP. The endothelial markers CD31 and CD34 were used to investigate vascular distribution. CD31 was expressed strongly in the portal area and parenchyma in HCC, but it was rarely observed in the parenchyma in cirrhosis. CD34 expression could not be detected in both HCC and cirrhosis. This study constitutes the first comprehensive study of P-gp, AFP, and endothelial markers in canine HCC and cirrhosis. The importance of these markers in HCC and cirrhosis in dogs was demonstrated and provides a more accurate basis for a definitive diagnosis of HCC and cirrhosis in dogs.     

Molecular characterization of cDNA encoding porcine IP-10 and induction of porcine endothelial IP-10 in response to human TNF-alpha

Donor-derived chemokines may play an important role in xenograft rejection, as well as allograft rejection. We have cloned the cDNA encoding porcine IP-10 (interferon-gamma-inducible protein 10), and evaluated its induction in miniature porcine endothelial cells in response to various human cytokines. The cloned sequence is 764 nucleotides long, and the open reading frame consists of 312 nucleotides. The deduced protein sequence includes a predicted mature protein of approximately 83 residues. The comparison of porcine IP-10, and its human, mouse, rat, goat, and sheep counterparts exhibited high similarity among different mammalian species. The sequences of important regulatory elements such as the interferon-stimulated response element (ISRE), and two NF-kappaB binding elements are conserved in the proximal promoter region of porcine IP-10, like other mammalian IP-10s. Human TNF-alpha induced the expression of IP-10 mRNA in porcine endothelial cells, while both human IFN-gamma and human IL-1beta failed. Together, the data of this study suggest that porcine IP-10 may interact with human leukocytes across the species barrier.