PD-L1 mRNA and protein expression in canine mammary carcinomas: Correlation with histopathological grade and molecular markers.

Programmed death ligand 1 (PD-L1) is an immune checkpoint molecule that plays a crucial role in regulating antitumor immune responses. Canine mammary carcinomas (CMCs) are common tumors of dogs. Despite extensive studies on the heterogeneity of CMCs, there is still a lack of effective precision therapies for the treatment of CMCs. In this study, we aimed to investigate the correlation between PD-L1 mRNA and protein expression in CMCs and explore its association with histopathological grade and molecular markers, including the estrogen receptor, epidermal growth factor receptor 2, and cytokeratin 5/6 (CK5/6). Formalin-fixed paraffin-embedded samples were evaluated for PD-L1 mRNA expression using RNA in situ hybridization and PD-L1 protein expression using immunohistochemistry. We observed no substantial correlation between PD-L1 mRNA and protein expression in CMCs; however, PD-L1 mRNA levels were significantly higher in grade 3 than in grade 1 tumors (P = .001). In addition, we observed a positive correlation between PD-L1 protein expression and CK5/6 expression in CMCs (P = .032). These findings suggest that PD-L1 expression in CMCs is heterogeneous and may be regulated post-transcriptionally. Further studies are needed to explore the prognostic and therapeutic implications of PD-L1 expression in different molecular subtypes of CMCs and their potential as predictive biomarkers for immunotherapy.

Analysis of the Prevalence of Canine Splenic Mass Lesions in Republic of Korea via Histopathological Diagnosis with Immunohistochemistry.

The histopathological diagnosis of canine splenic mass lesions is crucial for prognostication. However, thus far, no study has been conducted on the histopathology of canine splenic mass lesions in Republic of Korea. Herein, the prevalence of splenic diseases was analyzed in 137 canine splenic mass lesions via histopathological diagnosis, and the microscopic pattern associated with each disorder was described. Immunohistochemistry was performed for CD31, CD3, PAX5, Iba1, and C-kit for a more accurate diagnosis of splenic tumors. The proportion of non-neoplastic disorders, including nodular hyperplasia (48.2%, n = 66) and hematoma (24.1%, n = 33), was 72.3%. Splenic tumors, including splenic hemangiosarcoma (10.2%, n = 14), splenic lymphoma (nodular and diffuse types, 8.0%, n = 11), splenic stromal sarcoma (7.3%, n = 10), myelolipoma (1.5%, n = 2), and mast cell tumors (0.7%, n = 1), accounted for 27.7% of cases. The results of this study will aid veterinary clinicians in communication with pet owners about prognoses, recommendations for splenectomy, and subsequent histopathological diagnoses. This study will facilitate further investigations with more detailed comparisons of splenic mass lesions between small- and large-breed dogs.

A comparative study of the trends in epidemiological aspects of Lyme disease infections in Korea and Japan, 2011-2016.

Lyme disease (LD) is one of the most prevalent tick-borne emerging infections in North America, Europe, and far Eastern Asia including Korea and Japan. This study was undertaken with the aim of a comparative and quantitative analysis of the epidemiological aspects of LD infections in Korea and Japan from 2011 to 2016. The raw data analyzed in this study were obtained from the websites of the Korea Center for Disease Control and Prevention (KCDC), Korea, and the National Institute of Infectious Diseases (NIID), Japan. In total 65 cases of LD were observed with a cumulative incidence rate (CIR) of 0.22 per million population in Korea from 2011 to 2016. During the same period in Japan, there were 75 cases of LD with a CIR of 0.10 [significantly higher than that in Korea (p <0.01)]. Further, the results showed that in Korea, LD incidence was slightly higher in women than in men; but in Japan, the case was reverse, with incidence much higher in men than women. The proportion of cases differed significantly by age-specific adjusted groups within both countries (p <0.01). In both the countries, the incidence of LD was highest among those aged ≤20 yr (93.8% cases in Korea, and 94.7% cases in Japan). Cases peaked in autumn (46.7% of total cases) in Korea and in summer (60.0%) in Japan (p <0.01). The counties in rural areas had a significantly higher proportion of cases than the capital cities in both the countries (p <0.01). The study indicates that LD in Korea and Japan is an emerging zoonosis, and pose a serious risk to public health. The results underscore the continued emergency of LD and provide a basis for targeting prevention campaigns to a population with increasing incidence. Proper information, education and communication (IEC) activities can be helpful in reducing the spread of infection.

Comparative Study on the Epidemiological Trends and Aspects of Murine Typhus in Korea in the Last Decade (2006-2015).

INTRODUCTION: In this study, we undertook a retrospective analysis of the epidemiological aspects and risk factors of murine typhus (MT) in Korea in the last decade (2006-2015). MATERIALS AND METHODS: MT infections in Korea using a total of 411 cases obtained from the Diseases Web Statistical System of the Korea Center for Disease Control and Prevention. RESULTS: In a total of 411 MT infection cases, the cumulative incidence rate was 0.08/100,000 populations. Men were slightly more infected by MT as compared to women (51.3% vs. 48.7%), and a higher incidence of MT was observed in people aged over 40 years (93.4%; P < 0.01). The seasonal pattern of outbreaks revealed that most infections occurred from October to November (69.1% of the total cases) (P < 0.01). Significantly more outbreaks occurred in the southern part (53.5%) of the Korean peninsula as compared to its northern (33.3%) and central (10.7%) parts, as well as the Jeju Island (1.0%) (P < 0.01). In addition, the number of MT infections was significantly higher in rural and sea-village (87.6%) than in urban areas (12.4%; P < 0.01). CONCLUSION: In conclusion, the rapid reemergence of MT outbreaks can be minimized through health education, and a strong enforcement of control measures against rats and their ectoparasites could markedly reduce the transmission of this infection to humans in high-risk areas.

Effect of Chlorine Dioxide Gas Application to Egg Surface: Microbial Reduction Effect, Quality of Eggs, and Hatchability.

Controlling of microorganisms in the industrial process is important for production and distribution of hatching and table eggs. In the previous study, we reported that chlorine dioxide (ClO2) gas of a proper concentration and humidity can significantly reduce the load of Salmonella spp. on eggshells. In this study, we compared microbial reduction efficacy on egg's surface using hatching eggs and table eggs, internal quality of table eggs, and hatchability after both the conventional method (washing and UV expose, fumigation with formalin) and ClO2 gas disinfection. Application of 40 ppm ClO2 gas to the table and hatching eggs, respectively, reduced the aerobic plate count (APC) with no statistical difference compared with the conventional methods. Additionally, we didn't observed that any significant difference in albumin height, Haugh unit (HU), and yolk color, this result confirms that 40 ppm ClO2 had no effect on the internal quality of the table eggs, when comparing with the UV treatment method. The hatchability of hatching eggs was not statistical different between formaldehyde fumigation and 80 ppm ClO2 gas treatment, though the value was decreased at high concentration of 160 ppm ClO2 gas. From these results, we recommend that ClO2 gas can be used as a safe disinfectant to effectively control egg surface microorganisms without affecting egg quality.

Efficacy evaluation of commercial disinfectants by using Salmonella enterica serovar Typhimurium as a test organism.

The efficacies of six commercial disinfectants were evaluated by using Salmonella enterica serovar Typhimurium under simulated natural conditions such as sub-zero temperature, short disinfecting time, and surface type (uneven or smooth). We used a suspensionmodel test to determine the disinfecting efficacy under varying contact times (1, 5, 10, and 30 min) and temperatures (25°C, 4°C, 0°C, and -10°C). The bactericidal effect according to surface structure was measured by using a carriermodel test at 25°C and -10°C. The effective concentrations of each disinfectant were fixed to give a disinfecting effect within a short time (< 1 min) at 25°C and -10°C. The suspension model results revealed that bactericidal efficacy significantly dropped at low temperature for most of the disinfectants used; a sodium dichloroisocyanurate product showed the strongest efficacy. In the carrier test, bacterial load on a wooden surface was more difficult to remove than that on a stainless-steel surface. The results show that commercial disinfectant products vary in their disinfecting efficacy, which is affected by several field factors including temperature, contact time, and carrier material. Environmental conditions and surface type for disinfection should be considered prior to selecting an optimal disinfectant in the field.

Inactivation of Salmonella on Eggshells by Chlorine Dioxide Gas.

Microbiological contamination of eggs should be prevented in the poultry industry, as poultry is one of the major reservoirs of human Salmonella. ClO2 gas has been reported to be an effective disinfectant in various industry fields, particularly the food industry. The aims of this study were to evaluate the antimicrobial effect of chlorine dioxide gas on two strains of Salmonella inoculated onto eggshells under various experimental conditions including concentrations, contact time, humidity, and percentage organic matter. As a result, it was shown that chlorine dioxide gas under wet conditions was more effective in inactivating Salmonella Enteritidis and Salmonella Gallinarum compared to that under dry conditions independently of the presence of organic matter (yeast extract). Under wet conditions, a greater than 4 log reduction in bacterial populations was achieved after 30 min of exposure to ClO2 each at 20 ppm, 40 ppm, and 80 ppm against S. Enteritidis; 40 ppm and 80 ppm against S. Gallinarum. These results suggest that chlorine dioxide gas is an effective agent for controlling Salmonella, the most prevalent contaminant in the egg industry.

Ascorbic acid ameliorates oxidative damage induced by maternal low-level lead exposure in the hippocampus of rat pups during gestation and lactation.

This study was to investigate the effects of ascorbic acid on the hippocampus of suckling rats in the presence of lead (Pb)-induced oxidative stress. Pregnant Sprague-Dawley rats received treatment with drinking water, divided into three groups, as follows: (1) distilled water; (2) 0.2% Pb; (3) 0.2% Pb+ascorbic acid (100mg/kg/day). Rat pups were euthanized at the age of 21days and their brain tissue was examined using light microscopy. Protein levels of Cu/Zn superoxide dismutase (Cu/Zn SOD), manganese superoxide dismutase (Mn SOD), and catalase (CAT) in the hippocampus were determined by Western blotting. We found a significant decrease in levels of Cu/Zn SOD and Mn SOD among Pb-exposed pups. Ascorbic acid supplementation appeared to negate the decrease in protein levels for Cu/Zn SOD and Mn SOD. In the case of CAT, there was no effect from Pb administration alone and Pb plus ascorbic acid appeared to increase the levels. In histopathology, ascorbic acid decreased the number of damaged cells in cornu ammonis areas CA1, CA3, and the dentate gyrus (DG) in hippocampus. Our results showed that administration of ascorbic acid during pregnancy and lactation could ameliorate some of the oxidative damage induced by Pb exposure in the developing rat hippocampus.

Silk fibroin has a protective effect against high glucose induced apoptosis in HIT-T15 cells.

High glucose levels induce cell death in many cell types, including pancreatic ß-cells. Although protective agents against glucotoxicity have been searched for extensively, so far none have been found. In this report, we tested silk fibroin (SF) as a candidate material for antiglucotoxicity in the pancreatic ß-cell (HIT-T15 cell) line. Approximately 50% of cells were killed after treatment with 80 mg/mL glucose. This reduction of cell number was recovered by the addition of SF at 50 mg/mL. SF treatment also decreased cellular reactive oxygen species (ROS) and increased proliferating cellular nuclear antigen (PCNA) immunoreactivity. In addition, TUNEL assays demonstrated that SF protects against glucose-induced apoptosis of HIT-T15 cells, suggesting that SF might protect cells from cell death by lowering cellular ROS levels. SF also induced expression of the insulin-like growth factor-1 (IGF-1) gene, and IGF-1 expression may be the cause of SF-induced protection against glucose toxicity. Taken together, these results suggest that SF could serve as a potential therapeutic agent to treat the hyperglycemia-induced death of pancreatic ß-cells.

Molecular identification of duck and quail common cytokine receptor γ chain genes.

Common cytokine receptor γ chain (γ(c)) family cytokines play crucial roles in the regulation of innate and adaptive immune responses. Unlike mammals, chickens possess two different γ(c) transcripts. To determine if this difference is present in other avian species, γ(c) cDNA and genomic clones from ducks and quails were investigated. Two different γ(c) transcripts were identified in both species and designated as duck γ(c)-a (duγ(c)-a), duγ(c)-b, quail γ(c)-a (quγ(c)-a), and quγ(c)-b. Comparisons between the duck and quail γ(c) cDNA and genomic sequences indicated that the two transcripts were produced by alternative splicing. Unexpectedly, the duγ(c)-b contained the fifth intron, a frame-switching 88-bp insertion, resulting in a receptor molecule lacking a transmembrane region. These findings indicate a possibility that avian species, unlike mammals, express two different γ(c) transcripts due to alternative splicing. This study is the first demonstration of an alternatively spliced γ(c) isoform that lacks a transmembrane domain.

Monoclonal antibodies reactive with chicken interleukin-17.

Chicken interleukin-17 (chIL-17) gene was previously characterized through cloning from a chicken intestinal expressed sequence tag (EST) cDNA library. To further investigate the biological properties of chIL-17, six monoclonal antibodies (mAbs) against a bacterially expressed chIL-17 recombinant protein were produced and their binding specificities characterized. Antibodies which were initially selected on the basis of their specific binding reactivity with recombinant chIL-17 in ELISA were further characterized by Western blot analysis. Monoclonal antibodies specific for chIL-17 identified 20 and 21kDa protein bands in the culture supernatant and cell lysate of CU205 cells. These mAbs also recognized specific bands for chIL-17 in the cell lysate from conconavalin A (Con A)-activated, but not from normal splenic lymphocytes. Furthermore, these mAbs detected a 16kDa protein in the lysate of CU205 cells treated with tunicamycin and stained an intracellular protein in CU205 cells in flow cytometric analysis. Together, these results indicate that these new mAbs are specific for chIL-17 and will be a useful tool for structural and immunological studies of IL-17 in poultry.

Protective effects of ascorbic acid against lead-induced apoptotic neurodegeneration in the developing rat hippocampus in vivo.

Lead is a neurotoxin that affects the developing central nervous system and may potentially induce apoptotic cell death. We investigated the effect of ascorbic acid against lead-induced neurotoxicity in the developing rat hippocampus. Female Sprague-Dawley rats were divided into three groups: control group, lead-treated group and lead plus ascorbic acid-treated group. Lead (0.2% lead acetate) was administered to female rats during pregnancy and lactation, in their drinking water. During this period, rats in the lead plus ascorbic acid-treated group received 100 mg/kg/day ascorbic acid, orally. At the end of the treatment, neuronal damage, apoptosis and blood lead levels were determined and the levels of Bax and Bcl-2 were immunodetected in the hippocampus of 21-day-old male pups. Histopathological evaluation demonstrated that ascorbic acid significantly attenuates apoptosis in the developing hippocampus and also spares hippocampal CA1, CA3 and dentate gyrus (DG) neurons. Simultaneous administration of ascorbic acid and lead lowered the level of Bax protein and increased Bcl-2 in pup hippocampus and reduced lead level in blood of dams compared with lead-treated only. Based on these results, it seems that ascorbic acid may potentially be beneficial in treating lead-induced brain injury in the developing rat brain.

In vitro anti-oxidative and anti-inflammatory effects of solvent-extracted fractions from Suaeda asparagoides.

Suaeda asparagoides Miq. (Chenopodiaceae: S. asparagoides) is a salt-marsh plant that has long been prescribed in traditional Oriental medicine for the treatment of hypertension and hepatitis. In order to elucidate the pharmacological mechanisms of the herb, we conducted an examination of the anti-oxidative and anti-inflammatory properties of solvent-extracts of S. asparagoides. All of the solvent fractions showed potent anti-oxidative effects, as assessed using a radical generation assay system (xanthine oxidase assay) and an electron-donating activity system (DPPH [2,2-diphenyl-l-picrylhydrazyl radical] assay), with IC50 values ranging from 9 to 42 microg/ml. In agreement with this pattern, the total phenolic contents were widely distributed in the various solvent fractions, and ranged from 36.5 to 50.3 mg/g of dry weight. All of the solvent fractions significantly suppressed NO production in RAW264.7 cells induced by lipopolysaccharide (LPS, 0.1 microg/ml) and of the fractions, only the chloroform (CHC) fraction completely blocked the expression of inducible NO synthase (iNOS). Additionally, the hexane (HEX) and CHC fractions suppressed the mRNA expression of granulocyte/macrophage colony-stimulating factor (GM-CSF) and monocyte chemoattractant protein 1 (MCP-1), respectively, in the LPS-stimulated RAW264.7 cells. Therefore, these results suggest that the pharmacological action of S. asparagoides is due to its potent anti-oxidative effects and anti-inflammatory effects, and that therefore it can be applied to other diseases caused by oxidative stress and inflammation, such as cardiovascular diseases.

Stable transcriptional status in the apoptotic erythroid genome.

When a cell is destined for apoptosis, will its genome reprogram its transcriptional machinery to overcome the life-threatening challenge? To address this issue, we performed a genome-wide transcriptome analysis in EPO (erythropoietin) deprivation-induced apoptotic erythroid cells using the SAGE method. The results show that the transcript contents for the majority of the genes remain unchanged in the apoptotic cells, including the apoptotic genes and the heat shock genes. Of the small number of genes with an altered expression, they are mainly associated with cellular structure. Our study reveals that there is no genetic reprogramming for the transcriptional machinery in the apoptotic genome. Apoptosis, as defined by programmed cell death, is not a crisis but a peaceful physiological process.

Induction of heme oxygenase-1 mediates the anti-inflammatory effects of the ethanol extract of Rubus coreanus in murine macrophages.

Foods of plant origin, especially fruits and vegetables, draw increased attention because of their potential benefits to human health. The aim of the present study was to determine in vitro anti-inflammatory activity of four different extracts obtained from the fruits of Rubus coreanus (aqueous and ethanol extracts of unripe and ripe fruits). Among the four extracts, the ethanol extract of unripe fruits of R. coreanus (URCE) suppressed nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. We also demonstrated that URCE by itself is a potent inducer of heme oxygenase-1 (HO-1). Inhibition of HO-1 activity by tin protoporphyrin, a specific HO-1 inhibitor, suppressed the URCE-induced reductions in the production of NO and PGE(2) as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2). Our data suggest that URCE exerts anti-inflammatory effects in macrophages via activation of the HO-1 pathway and helps to elucidate the mechanism underlying the potential therapeutic value of R. coreanus extracts.

Expression of PSD95 in the rat sciatic nerve.

This study was designed to elucidate the existence of PSD95 in the rat sciatic nerve. Immunohistochemical stains of cryosection and teased fiber of sciatic nerves were performed with goat polyclonal antibody against PSD95. Western blot analysis was also accomplished with the same antibody. We got an interesting result that the rat sciatic nerve obviously showed PSD95 immunoreactivity especially in the nodal and paranodal regions, and we also identified a distinct band of PSD95 by western blot. These results suggest PSD95 exists in the sciatic nerve as well as it does in the central nervous system. We suppose PSD95 may have some important roles in ion channel clustering, junctional plasticity and signal transduction in the peripheral nerves as well.