The objective of this experiment was to assess the relationship between electrical resistance of the vaginal mucosa and plasma progesterone for optimal mating time in the bitch. Eight mature beagle bitches were examined, and we observed eight times of estrus. Vaginal electric resistance was recorded weekly using a Draminski ovulation detector in anestrus, and daily in estrus. Plasma progesterone concentration was estimated by radioimmunoassay. In the bitch, incline in vaginal electric resistance (376.20 ± 105.63 units) showed a closely association with the onset of proestrus. Ovulation day was determined as the first day when plasma progesterone concentration increased above 5.0 ng/ml (Day 0). On Day 0, vaginal mucous electric resistance was 438 ± 132 units. Vaginal mucous electric resistance showed a slight decrease or was maintained until Day 0. However, it showed an explosive increase, and peaked on Day 1~3, which was above 600 units. Two of eight cases peaked on Day 1, three of eight cases were revealed on Day 2, and others were revealed on Day 3. After Day 4, resistance showed a rapid drop to below 600 units and reached 200 units on Day 8. The optimal mating time was determined when vaginal mucous electric resistance was above 600 units.

Taste receptors of the anterior tongue are innervated by the chorda tympani (CT) branch of the facial (VIIth) nerve. The CT nerve transmits information on taste to the ipsilateral nucleus of the solitary tract (NST), which is the first taste central nucleus in the medulla. Taste information is known to be transferred ipsilaterally along the taste pathway in the central nervous system. Some patients with unilateral CT damage often retain their ability to sense taste. This phenomenon is not explained by the unilateral taste pathway. We examined whether neurons in the NST receive information on taste from the contralateral side of the tongue by measuring c-Fos-like Immunoreactivity (cFLI) following taste stimulation of the contralateral side of the tongue in the anesthetized rats. We used four basic taste stimuli, 1.0 M sucrose, 0.3 M NaCl, 0.01 M citric acid, 0.03 M QHCl, and distilled water. Stimulation of one side of the tongue with taste stimuli induced cFLI in the NST bilaterally. The mean number of cFLI ranged from 23.28 ± 2.46 by contralateral QHCl to 30.28 ± 2.26 by ipsilateral NaCl stimulation. The difference between the number of cFLI in the ipsilaterl and contralateral NST was not significant. The result of the current study suggests that neurons in the NST receive information on taste not only from the ipsilateral but also the contralateral side of the tongue.

Amoxicillin, a well-known antibiotic, has a broad spectrum against gram-negative and gram-positive bacteria. This experiment was conducted in order to investigate the effect of micronized and non-micronized amoxicillin prepared using different comminution techniques on change in blood concentration of rats. Forty adult male Sprague Dawley rats (6~7 weeks of age, body weight 128.3 ± 10.7 g) were randomly allocated to two treatment groups: micronized amoxicillin (MA) group treated with micronized amoxicillin trihydrate powder (particle size, over 90% of 10 μm), non-micronized amoxicillin (NMA) group treated with non-micronized amoxicillin trihydrate powder (particle size, over 70% of 100 μm), given 480 mg/kg body weight once daily for four days. The results showed a significant increase in serum concentration in the MA group on days 3 and 4, compared to the NMA group (P<0.05). In particular, serum concentration of the MA group on day 4 was increased almost two times that of the NMA group. The results indicate that due to the increase of the drug’s oral bioavailability, higher serum concentration would be achieved with the micronized amoxicillin trihydrate than with the non-micronized drug.

Cancer is the result of damage to the genetic system, i.e., dysfunction of the DNA repair system, resulting in dysregulated expression of various molecules, leading to cancer formation, migration, and invasion. In cancer progression, several proteases play a critical role in metastasis; however, their biological mechanism in cancer metastasis is not clearly understood. Among these proteases, cathepsins are a family of lysosomal proteases found in most animal cells. Cathepsins have an important role in protein turnover of mammalian, and are classified into 15 types based on their structure as serine (cathepsin A and G), aspartic (cathepsin D and E), and cysteine cathepsins (cathepsin B, C, F, H, K, L, O, S, V, X, and W). Cysteine cathepsins appear to accelerate the progression of human and rodent cancers, which can be a biomarker of the potency of malignancy or metastasis in mammalian. Overexpression of cyteine cathepsins causes the activation of angiogenesis promoting factor, whereas their downregulation reduces the angiogenesis of cancer progression. Under physiological conditions, cysteine cathepsins are essential in inflammation, infection, and cancer development. Activity of cysteine proteases, i.e., cathepsin B, is required for cancer progression or metastasis. Elevation of cysteine cathepsin is associated with cancer metastasis, angiogenesis, and immunity. Therefore, in this review, we suggest that cysteine cathepsin may be an anticancer target of strong clinical interest, although the exact mechanism of cathepsins in cancer metastasis is under investigation.

Lactic acid bacteria (LAB) are known to promote activation of macrophages, selectively eliminate harmful intestinal bacteria that enter the body, and increase secretion of cytokines, chemokines, and inflammatory mediators that boost the immune system. In the current study, the immune-enhancing effects of five Leuconostoc stains (Leu. kimchii WK18, Leu. citreum DH33, Leu. mesenteroides DH34, SE34, and GY17) isolated from traditional Korean fermented food, Kimchi, were evaluated. Lactobacillus rhamnosus GG (LGG), which is known for its extraordinary immunopotentiating activity, was used as a reference strain. According to the results, several heat-killed Leuconostoc strains induced a higher level of NO, IL-1β, or TNF-α production than the LGG strain. Collectively, results of the current study suggest that Leuconostoc strains could enhance immune response through activation of macrophages, and could be significant references when verifying the possibility for use of Leuconostoc strains as probiotics, starter, dietary supplement, or immune-enhancing medicine.

To identify subspecies of the sika deer, Cervus nippon, and to select C. n. hortulorum individuals for restoration of this subspecies in Korea, we obtained cytochrome b partial sequences (375 bp) of sika deer (C. nippon) from deer farms in northeastern China, North Korea, mainland Korea, and Jeju Island. Based on noninvasive samples, these cytochrome b sequences were compared with the corresponding haplotypes of sika deer, which were obtained from GenBank. We identified five individuals of sika deer from deer farms in northeastern China and North Korea as C. n. hortulorum, and found that interbreeding between individuals belonging to different subspecies is common on deer farms in far-eastern Asia. We concluded that the five C. n. hortulorum individuals on deer farms from northeastern China and North Korea, detected in this study, cannot be considered as pure descendants of C. n. hortulorum in the wild.

A three-year-old Russian Blue female cat was presented for long-standing infertility and recent development of male behavior. From one to three years of age, the cat had been bred, however, pregnancy never resulted. Abnormal enlargement of the uterus and ovaries was observed by abdominal sonography and radiography. An ovariohystectomy was performed. In the uterus, adenocaricnoma lesions were found and characterized microscopically by replacement of the endometrium with a population of large polyhedral cells with a tubulopapillary or nested arrangement and a supporting fibrous stroma with minimal vascularization. In addition, the pedunculated mass was diagnosed as leiomyoma, which was continuous with the myometrium, composed of closely packed sheets of spindle cells embedded in vascularized stroma, and covered with a discontinuous endometrial epithelium. The ovarian neoplasm was diagnosed as an ovarian interstitial cell tumor. Based on the histopathological characteristics, this case was diagnosed as a simultaneous occurrence of an ovarian interstitial-cell tumor and uterine adenocarcinoma and leiomyoma in a female cat. To the best of our knowledge, this report represents a rare case of multiple tumors in the genital organs of a cat.

To clarify the role of stem cells in hepatocarcinogenesis, octamer-binding transcription factor 4 (Oct4) expression was investigated in mouse liver and embryonic cell lineages. In vivo, at 14 days of age, ten ICR mice were divided into two groups and treated with saline or diethylnitrosamine (DEN), and were sacrificed at 6 h after treatment. Livers were fixed in 10% neutral phosphate buffered formalin, embedded in paraffin, sectioned to a thickness of 5 μm, and immunohistochemical analysis of Oct4 was performed. In vitro, mouse embryonic stem cells, hepatic progenitor cells and hepatocytes, representing 0, 22, and 40 days of differentiation, respectively, were treated with DEN at four doses (0, 1, 5 and 15 mM; G1, G2, G3 and G4, respectively) for 24 h and RNA was isolated; Oct4 and Gadd45a mRNA were investigated. In vivo, Oct4 expression was not detected in saline-treated livers. However, its expression was observed in hepatocytes of mice treated with DEN, showing cytoplasmic staining. In vitro, Oct4 expression differed significantly for G4 on day 0 (P<0.05) and for G2 on day 22 (P<0.01) and G3 and G4 on day 40 (P<0.05 and P<0.01, respectively) compared with G1 at each time point. Gadd45a expression differed significantly in G4 (P<0.01) on day 0 and G4 on day 40 (P<0.01), compared with that of G1 at each time point. Taken together, Oct4 expression was increased by treatment with DEN in hepatocytes, however, not in embroyonic stem cells and hepatic progenitor cells. This finding suggests that Oct4 expression may be modulated in hepatocarcinogenesis induced by DEN.

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).