Stroke is a major cause of death and long-term disability. Chlorogenic acid is a phenolic compound with a potent neuroprotective effect. γ-enolase is a phosphopyruvate hydratase found in mature neurons and plays an important role in neuronal survival. This study investigated whether chlorogenic acid regulates the expression of γ-enolase during cerebral ischemia. Middle cerebral artery occlusion (MCAO) was performed to induce cerebral ischemia. Adult male rats were used and chlorogenic acid (30 mg/kg) or phosphate buffered saline (PBS) was injected intraperitoneally 2 hours after MCAO surgery. Cerebral cortical tissues were collected 24 hours after MCAO surgery. Our proteomic approach identified the reduction of γ-enolase caused by MCAO damage and the mitigation of this reduction by chlorogenic acid treatment. Results of reverse transcription-polymerase chain reaction and Western blot analyses showed a decrease in γ-enolase expression in the PBS-treated MCAO group. However, chlorogenic acid treatment attenuated this decrease. Results of immunofluorescence staining showed the change of γ-enolase by chlorogenic acid treatment. These results demonstrated that chlorogenic acid regulates the γ-enolase expression during MCAO-induced ischemia. Therefore, we suggest that chlorogenic acid mediates the neuroprotective function by regulating the γ-enolase expression in cerebral ischemia and may be used as a therapeutic agent for brain diseases including stroke.

This study investigated dexamethasone (DXM) residues in the milk from intramuscularly dosed dairy cows and established the withdrawal time (WT) of DXM in milk. Eighteen healthy Holstein cows were injected with 20 (DXM-1) or 40 mL (DXM-2) of a drug containing 1 mg/mL of DXM. After administering DXM, milk samples were collected from all cows at 12-hour intervals for five days. The DXM residue concentrations in milk were determined by liquid chromatography-tandem mass spectrometry. The correlation coefficient of the calibration curve was 0.9966, and the limits of detection and quantification (LOQ) were 0.03 and 0.1 μg/kg, respectively. The recoveries were 97.0% to 104.0%, and the coefficient of variations was less than 7.22%. After treatment, DXM in DXM-1 was detected above the LOQ in two milk samples at 36 hours and below the LOQ in all milk samples of DXM-2 at 48 hours. Using the WT calculation program WT 1.4, the withdrawal periods of DXM-1 and DXM-2 in milk were established to be two days. In conclusion, the developed analytical method is sensitive and reliable for detecting DXM in milk. The estimated WT of DXM in bovine milk is shorter than the current milk WT recommendation of three days for DXM in lactating dairy cows.

5-Fluorouracil, doxorubicin, and vincristine are chemotherapy agents used to treat various cancers, such as breast cancer and lymphoma for decades, and their effects on cancer have been proven. On the other hand, these anti-cancer drugs cause fatal side effects, including immunosuppression. This study investigated whether sonicated Bordetella bronchiseptica bacterin (B. bronchiseptica) can attenuate the immunosuppression of spleen cells induced by these chemotherapy agents and which subsets of spleen cells were affected. B. bronchiseptica increased the metabolic activity of spleen cells treated with 3 anti-cancer drugs. Cell death analysis using Annexin V/propidium iodide showed that B. bronchiseptica markedly decreased the death of spleen cells. The subsets of spleen cells were analyzed by flow cytometry using a surface marker-specific antibody. B. bronchiseptica increased nitric oxide production in the spleen cells treated with anti-cancer drugs (p less than 0.0001). Despite the pharmacological effects of anti-cancer drugs, many patients suffer from the fatal side effects of immunosuppression. This study provides valuable information on how to overcome chemotherapy-induced immunosuppression.

With the increasing population of companion dogs, the social cost derived from their behavioral problems is increasing. Therefore, it is important to understand the environment to provide experience for dogs via interactions with their owners to prevent and solve these problems. The parenting behavior of dog owners as an environmental factor has a profound impact on the behavior development of dogs, as does the parenting behavior on children. Therefore, this study aimed to develop a scale to assess the dog owner’s parenting behavior. Exploratory factor analysis involving 300 participants resulted in a scale with four subscales, Positive Education, Involvement of Socialization, Intimacy, and Stable Responses, and 19 items. Confirmatory factor analysis was then performed to verify its reliability and validity. The result of parenting behaviors assessed by this scale was significantly different between a group with dogs with problematic behavior (n = 141) and those without (n = 159). Overall, a dog owner’s parenting behavior involves affection and control aspects, but the role required specifically in the control aspect is distinguished from the parenting behavior with children. The findings in the present study will provide dog owners with effective education that will help prevent dogs from developing behavioral problems.

The helicase genes and hypervariable regions (HVRs) of three avian hepatitis E viruses (HEVs) detected at three different farms were sequenced and characterized. Two isolates (DW-L and GI-B2) were classified as genotype 2 and one isolate (GR-B) was classified as genotype 1. A phylogenetic tree, based on the helicase gene and HVR nucleotide sequences, revealed the newly detected viruses and other avian HEVs were classified similarly. Unlike previously reported avian HEVs, the DW-L isolate detected in broiler breeders with characteristic lesions of avian HEV had no prolinerich motif in its HVR, suggesting that the proline-rich motif is non-essential for viral replication and infection.

Eight rabbits exhibited head tilt and subsequently died. At necropsy, three rabbits had crusty deposits in ears and four had reddish lungs. The main histopathological features were severe diffuse suppurative meningoencephalitis (75.0% of rabbits), fibrinopurulent pneumonia (37.5%), and otitis externa (37.5%). Pasteurella multocida (P. multocida) was isolated from brains, ears, and lungs. The capsular serogroups of the isolates were untypable. Based on histopathological features and bacterial analysis results, the rabbits were diagnosed as P. multocida infection. P. multocida infections might result in considerable economic loss in commercial rabbit production facilities in Korea.

Blood group determination in dogs is an important factor in transfusion medicine to minimize immediate or delayed adverse reactions after red blood cells transfusion in small animal clinics. Dog erythrocyte antigen (DEA) 1 is the most important blood type due to its high degree of antigenicity causing acute transfusion adverse reactions. The aim of this study was to investigate the prevalence of DEA 1 in various dog breeds in Korea. As a result of testing 592 blood samples from more than 35 dog breeds, DEA 1 blood typing for each breed showed that 57.8% of Malteses, 63.3% of Poodles, 76.2% of Mastiff-like dogs, 72.5% of Pomeranians, 47.7% of Shih Tzus, 70.3% of mixed breeds, 60.0% of Yorkshire Terriers, and 71.4% of Beagles were DEA 1-positive. Miniature Schnauzers and Jindo breeds had a significantly high prevalence (100%) of DEA 1-positive dogs compared to that in other small breed dogs. This is the first report of immunochromatography-detected DEA 1 prevalence in various domestic dog breeds. Although additional studies need clarifying the potential blood transfusion risks in domestic breed dogs with DEA 1, the results of this study may be useful when selecting a blood donor.

This study assessed the effects of Weissella cibaria (W. cibaria) CMU on oral health in male and female beagles (n = 18) by measuring oral malodor and periodontal disease-related parameters (calculus, plaque, and gingivitis indices). Oral malodor and indicators of periodontal disease were assessed in five treatment groups: negative control (scaling and 0.24 mg of maltodextrin, n = 3), positive control (0.24 mg of maltodextrin, n = 3), and W. cibaria CMU groups (each n = 4) at low (CMU-L, 2 × 107 colony forming unit [CFU]), medium (CMU-M, 2 × 108 CFU), and high (CMU-H, 2 × 109 CFU) concentrations. After feeding with W. cibaria CMU for 6 weeks, total volatile sulfur compound concentrations in the CMU-L (2.0 plus-minus 1.04 ng/10 mL), CMU-M (2.4 plus-minus 1.05 ng/10 mL), and CMU-H (2.6 plus-minus 1.33 ng/10 mL) groups were significantly lower than in the positive control group (3.2 plus-minus 1.65 ng/10 mL). Also, CMU-L (1.4 plus-minus 0.83 ng/ 10 mL) and CMU-H (1.9 plus-minus 1.14 ng/10 mL) groups had methyl mercaptan levels lower than that in the positive control group (2.4 plus-minus 1.21 ng/10 mL) at week 2. The plaque index was significantly lower in the CMU-H group (4.5 plus-minus 0.28) than in the positive control group (5.9 plus-minus 1.08) at week 6. W. cibaria CMU could be useful as a novel oral hygiene probiotics for reducing volatile sulfur compounds production and inhibiting plaque growth in companion animals.

Canine mammary tumors are among the most frequently observed cutaneous tumors in female dogs. Cancer stem cells (CSCs), referred to as tumor-initiating cells, are thought to have properties similar to normal stem cells such as the ability to self-renewal and to differentiate into various cell types. Biological understanding of CSCs and the critical pathways involved in their maintenance are important in research and therapy for mammary tumors. We conducted the present study on sphere formation from REM134 cells by using methylcellulose to produce tumorspheres on a large scale and compared the specific markers of the spheres-formed and plating-cultured REM134 cells. The results revealed that the tumorspheres cultured in methylcellulose had higher seeding density and improved morphology compared to those produced in normal sphere formation medium. Expression levels of stemness markers and CSCrelated markers were higher in tumorsphere-forming cells than in plating-cultured cells. Subsequently, we transplanted the tumorsphere-forming and plating-cultured cells into female nude mice to examine their tumorigenic potential. Tumor volume increased rapidly in mice transplanted with tumorsphere-derived cells compared to plating-cultured cells. We observed a novel sphere-forming condition for REM134 cells and showed that REM134 cell tumorspheres can exhibit improved CSC properties.

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-β1 and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3DTN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.