Fraxinus rhynchophylla (Oleaceae), a deciduous tree, is known to have properties that include anti-inflammatory, convergence, febricide, antiblenophthalmia, antidiarrhea, antileukorrhea, and so forth. In addition, it has been used for traditional herbal medicine in East Asian countries, including Korea. In this study, we investigated the antioxidant and anti-inflammatory effects of Fraxinus rhynchophylla ethanol extract (FRE) in lipopolysaccharide (LPS)-induced murine macrophage Raw 264.7 cells with FRE pretreatment. We performed DPPH-assay, Western blot, and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). FRE showed 85% free radical scavenging activity at concentrations of 80 µg/ml. Results of this study also showed that FRE down-regulates Cox-2 and iNOS expression in mRNA and protein level. In conclusion, crude ethanol extract of Fraxinus rhynchophylla exhibited antioxidant and anti-inflammatory activities, and it may potentially provide a valuable source of natural herbal agent to inhibit inflammation.

The aim of this study was to investigate the morphological development pattern of conical papilla (CP) during prenatal and postnatal periods in native Korean goats by scanning electron microscopy. Tongues were removed from fetuses on days 90, 120, and neonates and from juveniles on days 30, 50, 60, 75, 90, 120, 150, and 180 after birth. The lengths of CP were 194~240 μm in neonates, 335~485 μm in the weaning period of 60-day-old goats, and 526~662 μm in the maturing period of 180-day-old goats. The primordia of CP in 90-day-old fetuses, shaped like mountain berry, were sprouted. The CP of 120-day-old fetuses was a cylinder shape with an apex whose center was slightly concave and the margin was sticking up like a low fence. The CP of neonates was empty inside and the secondary papillae were irregularly arranged fence-like structures on it. In 60-day-old goats, CP was an obliquely sectioned cylinder shape compacted in inner surface; the shapes of CP varied in 120-day-old goats, and had a smooth surface. A low elevation of CP was observed in 150- and 180-day-old goats. The microridges, microplicae, and micropits were well developed on the epithelial surface of lingual papillae in 60- to 120-day-old goats. These findings indicate that CP of goats has a variety of sizes and shapes during development.

Lysyl oxidase (LOX) is an extracellular amine oxidase catalyzing formation of lysine-derived cross-linkages in collagen and elastin in extracellular matrices. Four human paralogs of LOX (LOXL1, LOXL2, LOXL3, and LOXL4) have been identified, each encoding the functional domains required for the amine oxidase activity of LOX. Upregulated expression of LOX and LOXL2 was reported to show significant correlation with absence of lymphovascular invasion in tumor tissues from patients with colorectal adenocarcinoma, suggesting that oxygen tension around tumors may be an important factor in expressional regulation of these genes in colorectal carcinomas. To evaluate the effects of hypoxia on expression of LOX and LOXL2 in colorectal carcinomas, we performed promoter assays and RT-PCR analysis under hypoxia in colorectal carcinoma cell lines, HT-29 and HCT-116. Expression of LOX was upregulated under hypoxia in both HT-29 and HCT-116 cells, whereas LOXL2 expression was not affected by hypoxia in those cells. In the highly metastatic HCT-116 cells, LOX showed a higher level of upregulation than in HT-29 cells, suggesting a possible association of LOX upregulation with increased invasiveness and metastatic potential in colorectal carcinomas.

Human umbilical cord is easy to obtain because it is discarded after birth, so that ethical issues can be avoided. Chondrogenesis studies using MSCs from bone marrow, cord blood, and adipose have indicated that TGFβ3 and BMP6 stimulate chondrogenesis. Therefore, we investigated chondrogenesis of hUC-MSCs on TGFβ3, BMP6, and combination of the two growth factors. We initiated chondrogenesis of cells by application of physical forces to form 3D cell clusters. After initiation, we designated four experimental groups for differentiation of cells, as follows: control, 10 ng/mL TGFβ3, 100 ng/mL BMP6, and the combination of 5 ng/mL TGFβ3 and 50 ng/mL BMP6. For analysis of chondrogenesis, GAG contents, mRNA expression, histological analysis and immunohistochemistry (IHC) were performed. For analysis of GAG contents, GAG assay was performed and RT-PCR was performed for determination of chondrogenic markers. Histological analysis was performed through safranin O, alcian blue, and IHC was performed using collagen type I and II. GAG contents were increased 184% by TGFβ3, 147% by BMP6, and 189% by the combination of TGFβ3 and BMP6, compared to control. The growth factors improved collagen II and aggrecan expression; in particular, TGFβ3 and BMP6 showed a synergistic effect, compared to only TGFβ3 or BMP6 treated. The results of histological and IHC analysis indicated that chondrogenic differentiation in TGFβ3 and the combination of TGFβ3 and BMP6 showed more cartilage deposition. In conclusion, TGFβ3 and BMP6 differentiated hUC-MSCs into chondrogenic clusters of the combination treatment of the two growth factors showed more efficient chondrogenic ability.

Although bombesin (BN) regulates colonic motility, the associated mechanism of action remains unclear. The objective of this study was to investigate the effect of BN on colonic motility using isolated rat colon. An isolated rat colon was perfused with Krebs solution via the superior mesenteric artery. Intraluminal pressure was measured via microtip catheter pressure transducers at both proximal and distal portions of the isolated colon. After a control period, BN was administered intraarterially in concentrations of 13, 26, 130, and 260 pM. After pretreatment with hexamethonium (10-3 M), atropine (10-5 M), or tetrodotoxin (10-6 M), BN was administered at a concentration of 260 pM (proximal colon) or 130 pM (distal colon); intraluminal pressure was then monitored. Changes in motility were expressed as the percentage change of motility index (MI) over the basal period. As a result, BN increased colonic motility and a dose-dependent increase in proximal colonic motility was observed. The stimulant effect of BN was almost completely abolished by atropine and tetrodotoxin at both the proximal and distal colon. However, BN was not inhibited by hexamethonium at both the proximal and distal colon. Therefore, the stimulant action of BN may be mediated by local cholinergic muscarinic receptors.

This study was conducted in order to evaluate three different non- invasive blood pressure measurement methods (NIBP) in comparison with direct blood pressure measurements in anesthetized dogs. Ten normal healthy Beagle dogs (mean age; 3.7 ± 1.01 yrs, mean body weight 10.2 ± 1.15 kg) were enrolled in this study. BP was measured using three different methods, i.e., Doppler (DOP), oscillometric, and high density oscillometric (HDO) methods, simultaneously, five times in each subject under anesthesia at three different locations (right, left front legs, and tail). The blood pressure value measured by the Doppler method was the closest to the value measured by the direct method. Although the accuracy and consistency of BP measured by three different non-invasive methods differed according to the method, all methods, except the conventional oscillometric method, were acceptable for monitoring systolic blood pressure in anesthetic dogs. The inter-measurement variations in DOP and HDO were minimal without affecting the consistency of the test results.

Flavonoids have a range of biological activities, including anti-allergic, anti-inflammatory, anti-microbial, and anti-cancer activities, as demonstrated by in vitro studies. In this study, we investigated whether luteolin can be applied to suppression of lipopolysaccharide (LPS)-stimulated inflammatory responses in murine macrophages. Luteolin was found to reduce nitric oxide (NO) production in LPS-stimulated Raw 264.7 cells. In addition, expression of inducible nitric oxide synthetase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokine tumor necrotic factor-α (TNF-α) at the mRNA and protein levels were decreased. These inhibitory effects were found to be caused by the blockade of nuclear factor kappa-light- chain-enhancer of activated B cells (NF-κB) activation and phosphorylation of mitogen-activated protein (MAP) kinase family, extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 MAP kinase. In addition, pre-treatment with luteolin resulted in reduced ganglioside expression levels and inhibited expression of GT1b in Raw 264.7 cells. On the basis of these observations, we suggest that luteolin has potential as an anti-inflammatory drug candidate, and ganglioside GT1b may play a role in the inflammatory process.

The aim of this experiment was to count excreted oocysts and to evaluate gross lesions and histopathological changes in broiler chicks infected with Eimeria; E. acervulina (Ea), E. maxima (Em), and E. tenella (Et). Twelve-day-old broiler chicks without infection with Eimeria spp. were used in this experiment. On day 12, the chicks in group Ea (n=3), Em (n=3), and Et (n=3) received oral inoculation with 5 × 10 sporulated oocysts of Ea, Em, and Et per 1 mL of tap water, while control chicks were injected with tap water of equal amount. The prepatent period (PP) for Ea, Em, and Et was four, five, and six days, respectively. To evaluate the progress of the coccidiosis, post-mortem analysis method and histopathological evaluation were performed two days after PP of Ea and Et, and three days after PP of Em at two-day intervals on one bird from each group. Intestinal segments (Ea, duodenum and upper intestine; Em, mid intestine; and, Et, cecum) of all chicks were obtained from the autopsied birds. Among pathological changes of coccidiosis, target parts of small intestine and caecal pouches showed mucosal petechiation and white plaques streaks in the duodenum. Histopathological evidence showed upper small intestinal villus preserved with different phases of parasite (zygotes, oocysts, macro- and micro-gametocytes) after infection with Ea in broiler chicks. Zygotes in the lamina propria and fusion of middle small intestinal villi in broilers infected with Em and invasion of the caecal crypt of oocysts and granulomatous inflammation of caecal submucosa in broilers infected with Et were found.

To evaluate the acute to chronic effects of crude oil exposure on hematological and blood biochemical toxicities, Sprague-Dawley rats were given oral doses of 0, 50, or 100 mg/kg BW/day of Iranian heavy crude oil for a period of four weeks. In the acute phase of exposure (one day after four weeks of oil treatment), decreases in weight of thymus, serum level of interferon gamma (IFN-γ), superoxide dismutase (SOD), and catalase activities in liver or kidney, and increase in weight of adrenal gland occurred after oral administration of crude oil. In body weight, histopathological examination, hematological and blood biochemical analyses in the acute phase of exposure, no significant differences were observed among the experimental groups. In the subchronic and chronic phase of exposure (two months and six months after four weeks of oil treatment), the changes of biomarkers were normalized, except the indicators of oxidative stress. Our findings showed that the bioassay on the indicators of oxidative stress is a sensitive method for determining exposure to crude oil in rats.

Vital pulpotomy is a very useful method for disarming of canine tooth, tooth fracture, periodontitis, and malocclusion in veterinary dentistry. Calcium hydroxide is the material commonly used as a liner during vital pulpotomy. This creates a mineralized barrier by stimulating osteoblastic hard tissue repair, arrests the inflammatory response, and soothes dentin. However, the powder or mix type calcium hydroxide materials have many disadvantages due to complicated procedures for use and are hard to handle when vital pulpotomy is followed under general anesthesia in animals. This study was conducted in order to compare the effect of mix and premixed paste type calcium hydroxide as a liner in vital pulpotomy. Six beagle dogs underwent hemisection on the mesial root of the mandibular first molar and vital pulpotomy on the distal root of the first molar. On the distal root of the left and right mandibular first molar, mix type (DYCAL®, Dentsply, USA) and premixed paste type calcium hydroxide (VITAPEX®, Morita, Japan) were used as liners, respectively. Radiological evaluation was performed at immediate, 4, 12, and 20 weeks after vital pulpotomy. According to the results, all teeth had well-formed dentinal bridges, and there were no periradicular lucency, lamina dura loss, or anomalies of the pulp cavity. According to these results, on vital pulpotomy in animals, premixed paste type calcium hydroxide was easy to handle and decreased the anesthesia period due to a more convenient application procedure. A further study of many clinical cases is needed for evaluation of side effects and other problems.

A 10-year-old intact female Cocker Spaniel weighing 15.3 kg presented with dysbasia. The dog had hyperglycemia and glucosuria at presentation. Neurological examination and magnetic resonance imaging on the following day indicated the onset of cervical disk disease. Four days later, the patient returned due to sudden vomiting and anorexia with severe dehydration. Abnormal laboratory findings included severe hyperglycemia with ketonuria. Plasma progesterone concentration was very high at presentation. She was hospitalized for critical care, and insulin administration with fluid therapy was initiated. Mean blood glucose concentration showed a gradual decline, and hyperglycemia with glucosuria disappeared four days after initiation of insulin therapy. This case demonstrated that high plasma progesterone concentration during diestrus can contribute to development of diabetes mellitus in intact female dogs.

A five-month-old female pomeranian dog (weighing 0.95 kg) with a history of cyanosis, exercise intolerance, and delayed growth presented to Veterinary Teaching Hospital, Gyeongsang National University. Findings on diagnostic imaging studies showed a dextropositioned and over-riding aorta, pulmonic stenosis, ventricular septal defect, right ventricular hypertrophy, and atrial septal defect. Based on the diagnostic findings, the dog was diagnosed as a case of tetralogy of Fallot with atrial septal defect (pentalogy of Fallot).

Competitive ELISA methods were used to measure the level of aflatoxin M1 (AFM1) from urine in 1008 Korean adults. Subjects were selected by random sampling in all areas of Korea, except Cheju-do. The recovery rate of AFM1 using this method was 105% (73-124%). The geometric mean of urinary AFM1 in all subjects was 3.43 pg/mL (3.67 ng/g creatinine). The level of AFM1 in males was statistically higher, compared with female subjects. However, the levels of AFM1 did not differ according to age. Subjects in Chungbuk-do showed the highest urinary AFM1 concentration, whereas subjects in Kyeongnam-do showed the lowest concentration. Assuming an excretion rate of 5%, this AFM1 excretion corresponds to approximately 0.1 microgram/day in Korean adults.

Proteinuric conditions demonstrate structural and compositional changes of the foot processes and slit diaphragms between podocytes. β-Catenin in podocytes serves as an adapter protein anchoring P-cadherin of slit diaphragms to actin filaments of the podocyte cytoskeleton. To investigate the effect of ginseng total saponin (GTS) on pathologic changes of podocyte P-cadherin/β-catenin unit induced by diabetic conditions, we cultured mouse podocytes under: 1) normal glucose (5 mM, = control); 2) high glucose (HG, 30 mM); 3) advanced glycosylation end products (AGE)-added; or 4) HG plus AGE-added conditions and treated with GTS. In confocal imaging, β-catenin colocalized with P-cadherin predominantly at intercellular junction area. However, diabetic conditions relocalized and concentrated both molecules at perinuclear cytoplasmic area. In Western blotting, diabetic conditions, especially HG plus AGE-added condition, also decreased cellular β-catenin protein levels at 6, 24, and 48 hours. GTS improved such quantitative and qualitative changes of β-catenin. These findings imply that HG plus AGE have an influence on the redistribution and amount of P-cadherin/ β-catenin unit of podocytes, which can be reversed by GTS.

Expression of epithelial cell adhesion molecule (EpCAM) in the early phase of hepatocarcinogenesis induced by diethylnitrosamine (DEN) was investigated. At 14 days of age, 60 ICR mice were divided into two groups and treated with saline (group 1) or DEN (group 2, 10 mg/kg of body weight, i.p. injection), and were sacrificed at 6 h and 1, 2, 3, 7, and 28 days after treatment with saline or DEN. During necropsy, half of the liver from saline- or DEN-treated mice was processed for histopathological examination and immunohistochemical staining of EpCAM and apoptosis. The remaining liver tissue was snap-frozen in liquid nitrogen for RNA extraction and analysis of EpCAM mRNA expression. Immunohistochemical examination showed that EpCAM expression was detected only in a small number of hepatocytes from saline-treated mice and its expression was detected in bile duct cells and round cells around portal areas, as well as hepatocytes in the livers of DEN-treated mice. In addition, multiple apoptotic cells were found in the livers of mice treated with DEN. EpCAM mRNA expression was significantly higher in DEN-treated mice at 1, 7, and 28 days compared to saline-treated mice at 6 h (P<0.01). Taken together, EpCAM expression and apoptosis were increased in liver by DEN treatment.

Probiotics, enzymes, organic acids, oligosaccharides, antioxidants, and other functional materials are actively being explored as alternatives to antibiotics. Probiotics include live beneficial microorganisms that colonize the intestinal tract and competitively inhibit attachment and growth of harmful microbes. Probiotics also increase feed efficiency by assisting in nutrient absorption and digestion. The current study was conducted in order to evaluate the effect of a new probiotic, CS-A, as a dietary supplement of a fermented product on growth performance, feed intake, and feed conversion efficiency in broiler chickens, and to evaluate its value as an alternative for antibiotics used as a feed additive. Antibacterial and anti-inflammatory effects of CS-A were investigated in vitro and the in vivo effects of a constant concentration of supplemented CS-A on growth rate and feed efficiency were evaluated. In addition, the safety of CS-A was assessed by examination of common symptoms and mortality. Determination of minimal inhibitory concentration revealed an excellent antibacterial effect of CS-A. Cytotoxicity was low and anti-inflammatory effects were achieved at the effective concentration of CS-A. Supplementation with 0.1% CS-A resulted in a feed efficiency score of 1.84 in broilers, compared to 2.00 in the control group. There were no adverse clinical findings, necropsy findings, hematology, and altered serum biochemistry parameters, and no mortality. Thus, it is concluded that CS-A is safe and effective as a feed additive.

This study was performed to investigate the anti-obesity and anti-lipidemic ability of linalool (LL) in ApoE deficient mice fed a high-fat diet (HFD). Mice were divided into four experimental groups of eight each. Mice in the control group received a basic diet and oral repeated dose of the vehicle only for 12 weeks; mice in the HFD group received a HFD and oral repeated dose of the vehicle only for 12 weeks; and the HFD&LL25 and HFD&LL50 groups received a HFD and oral repeated dose of LL 25 and 50 mg/kg/day for 12 weeks, respectively. Mice in the HFD group showed a significant increase in body weight, spleen weight, and adipose tissue weight, compared with the control group. An increase in the serum levels of aspartate aminotransferase and alanine aminotransferase activities, total cholesterol (T-CHO), triglyceride (TG), and low density lipoprotein cholesterol was also observed in the HFD group. Histopathological examinations showed severe liver injuries, characterized by extensive fatty changes and hepatocyte degeneration/necrosis. On the contrary, oral administration with LL resulted in significantly improved HFD-induced obesity and hyperlipidemia, indicated by a decrease in adipose tissue weight, T-CHO, TG, and histopathological lesions. The results indicate that LL suppressed HFD-induced obesity, hyperlipidemia, hypercholesterolemia, and hepatic steatosis, suggesting that LL might be a promising adjuvant therapy for treatment of these metabolic disorders related to corpulence.

The aim of this study was to investigate the morphological development of fungiform papillae in native Korean goats by scanning electron microscopy (SEM). Tongues were removed from fetuses on days 120 and 150 of gestation and from juveniles on days 30, 60, 90, 120, 150, and 180 after birth. The sizes of fungiform papillae in the newborn were 225.0-250.0 μm, the weaning period of 60-day-old goats was 271-294 μm and the maturing period of 180-day-old goats was 385-398 μm, respectively. The primordia of fungiform papillae had the typical feature of a mushroom shape in a 120-day-old fetus. In a 150-day-old fetus (neonate), fungiform papillae had features similar to those of the adult form and showed a gradual increase in size with slight morphological changes until 150-day-old goats. Microridges, microplicae, and micropits were well developed on the epithelial surface of lingual papillae from 60 to 150-day-old goats. The results obtained in this study might provide some basic data on the mechanisms that control the morphogenesis and development of the lingual papillae in ruminants.

Inter-breed and individual variations in thoracic conformation often resulted in incorrect diagnosis during interpretation of canine thoracic radiographs. Therefore, it may be helpful to build a collection of normal thoracic radiographs of different breeds for useful reference. The aim of this study is to establish a normal canine thoracic radiograph database according to breed, age, and body weight. Medical records and thoracic radiographs of Veterinary Medical Center, Chungbuk National University were reviewed retrospectively. Normal thoracic radiographs of 170 dogs out of 640 patients who had thoracic radiographs were obtained. In 170 normal dogs, Maltese, Shih-tzu, Yorkshire Terriers, and Miniature Poodle were the most prevalent breeds, with 90 dogs. In this study, the normal canine thoracic radiograph database was established in the four breeds; it can be used as normal references for interpretation of canine thoracic radiographs.

The previous studies of p16^INK4a, pRb, p53, and Ki-67 expression suggested that these markers may be preferentially expressed in cervical neoplasms. The purpose of this study was to assess the expression and the clinical significance of p16^INK4a, pRb, p53, and Ki-67 proteins in cervical lesions. We obtained 106 cases with various categories of cervical squamous mucosa, including squamous cell carcinoma (n=35), cervical intraepithelial neoplasia (CIN) II/III (n=26), CIN I (n=10), squamous metaplasia (n=15), and normal squamous mucosa (n=20). Immunohistochemical staining was performed for p16INK4a, pRb, p53, and Ki-67 proteins in formalin-fixed and paraffin-embedded tissue sections of the uterine cervix. Evaluation of immunohistochemical staining was based on the frequencies of expression and the mean immunoreactivity scores (IS) in each diagnostic category. p16INK4a positive sotaining was observed in 26 of 35 cases (74.3%) of squamous cell carcinoma, in 16 of 26 cases (61.5%) of CIN II/III, in six of 10 cases (60%) of CIN I, in nine of 15 cases (60%) of squamous metaplasia, and negative in normal squamous mucosa. pRb expression was detected in all diagnostic categories; however, the proportion of pRb positive cells was relatively decreased in CIN II/III (38.5%) and squamous cell carcinomas (51.4%), compared to normal squamous epithelium (90%) and squamous metaplasia (73.3%). No significant differences in expression of p53 were observed in any diagnostic categories. Ki-67 expression was increased in squamous cell carcinoma (37.1%), CIN II/ III (42.3%), and CIN I (40%), but negative in squamous metaplasia and normal mucosa. In 35 cases of squamous cell carcinomas, multivariate analysis revealed no differences in p^INK4a, pRb, p53, and Ki-67 expression according to the age of the patient, lymph node metastasis and clinical stage. In conclusion, the combined use of p16^INK4a and Ki-67 immunoreactivity could improve the diagnostic specificity of squamous cell carcinoma of the uterine cervix.