Recombinant thymosin β4 (rTβ4) has been reported to migrate and promote vascularization, wound-healing, and hair growth in a mouse hindlimb ischemia model of peripheral vascular disease. C57BL/6 mice (11-weeks-old) were anesthetized and an ischemic model was made by cutting the right aorta femoralis. The ischemic group was intraperitoneally administered with saline (300 μL/mouse) and the muscular administration group received rTβ4 (150 μg in 300 μL of saline) or rTβ4 (150 μg in 300 μL saline) to the abdominal cavity at 3-day intervals for 21 days. Myoatrophy of the ischemic group was observed compared to the normal control group. Generation of adjacent vessels was carried out in the rTβ4 administration group compared to the ischemic group. The biopsy results showed significant fibrosis around the muscular undersurface and perimysium in the musculus quadriceps femoris of the ischemic group, whereas partial fibrosis was observed in the perimysium and endomysium in the rTβ4 administration group. Immunostaining indicated that expression levels of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor-1 (VEGF-1), and endothelial nitric oxide synthase (eNOS) in the rTβ4 group were higher than those of the ischemic group. Western blotting showed that expression levels of HIF-1α, VEGF-1, and eNOS in the rTβ4 group were higher than those of the ischemic group. In conclusion, rTβ4 increases expression levels of HIF-1α, VEGF-1, and eNOS, resulting in angiogenesis.

A 12-year-old spayed female Pomeranian (weighing 2.4 kg) was referred with primary complaints of acute dyspnea, cough, and lethargy. Diagnostic imaging studies found degenerative mitral valve cusps, chordae tendinae rupture, severe mitral regurgitation (5.45 m/s of peak velocity), and marked left atrial and ventricular dilation. The dog was diagnosed as having degenerative mitral valve disease (DMVD) with ISACHC stage IIIa heart failure. Her clinical condition was stabilized after administration of cardiac medication (e.g. diuretics and pimobendan). Ten months later, the dog was referred back to the clinic due to a sudden worsening of clinical signs. Echocardiographic study found pulmonary hypertension in addition to DMVD. After medication was adjusted, clinical signs were stabilized in 2 weeks. The patient was returned after 4 months for cardiac recheck and there was no obvious worsening of clinical signs. Incidental finding of a left-to-right atrial septal defect from rupture of the atrial septum secondary to marked left atrial dilation by DMVD was noted by echocardiography. To diminish left atrial volume overload, the frequencies of both furosemide and pimobendan were increased (i.e. from q 12 hr to q 8 hr) in addition to adding spironolactone (1 mg/kg q 12 hr). Based on diagnostic findings, this case was re-diagnosed as acquired atrial septal defect secondary to rupture of the atrial septum with advanced stage DMVD. The dog was then stabilized and is currently being regularly monitored.

A 2-year-old intact female pomeranian dog presented dyspnea, labored breathing, cough, exercise intolerance, machinery heart murmur, and precordial thrill. A left-to-right patent ductus arteriosus (PDA) was diagnosed based on two-dimensional echocardiography, thoracic radiography, electrocardiography, and blood work. An angiography was performed to accurately evaluate the size and shape of the duct. An interventional approach for transcatheterial occlusion of the PDA was achieved using an Amplatz® Canine Duct Occluder (ACDO), which is a commercially available ductal occluding device. Due to the limited size of the dog’s femoral artery, a device smaller [125% of minimal ductal diameter (MDD); recommended size: 150~200% of MDD] than recommended was mounted. After placement of the ACDO, precordial thrill and continuous heart murmur disappeared, and the patient was discharged the next day after stabilization with O2 supply. Upon follow up examination, dyspnea, labored breathing, cough, exercise intolerance, and cardiomegaly were improved with no complications after the procedure. The ACDO was well maintained in position. This case represents successful clinical application of the Amplatz® Canine Duct Occluder to achieve closure of a PDA using a slightly smaller device than the recommended size.

Guillain-Barre´ syndrome (GBS) is an acute inflammatory demyelinating polyneuropathy most commonly characterized by rapidly progressive, essentially symmetric weakness and areflexia. This study examined clinical symptoms of clinical variants of GBS through a cerebrospinal fluid (CSF) study, nerve conduction (NCV) study, treatment, and prognosis. There were 16 children with GBS who visited our hospital from January 2011 to December 2013. Guillen-Barre´-like syndromes with transient synovitis were noted in three children. Clinical variants of GBS with acute demyelinating encephalomyelitis were observed in one child. Previous infections were noted in 16 children with Guillen-Barre´-like syndrome. There were ascending infections in 16 cases. Fifteen children showed symmetric infections, and one showed asymmetric infection. In NCV, slow waves were noted in two cases. We treated using intravenous immunoglobulin (IVIG) in four cases, IVIG with steroid in two, cases and supportive care in 10 (62.5%) cases. Five children treated with IVIG and 10 with supportive care management were completely improved.Our study suggests that supportive care is effective as a treatment for clinical variants of GBS. Further study is necessary for more patients.

The morphology of the lingual papillae in a female Bengal tiger (Panthera tigris tigris) was examined by scanning electron microscopy (SEM). The tongue was 22.3 cm in length and 7.1 cm in width. Numerous filiform papillae were distributed over the entire dorsal surface of the tongue. SEM examination of the tongue revealed two types of mechanical papillae, i.e. filiform and conical papilla, and two types of gustatory papillae, i.e. fungiform and vallate papilla, on the dorsal surface of the tongue. Each filiform papilla consisted of one primary papilla and several secondary papillae. The filiform papillae on the anterior part of the tongue were divided into one primary and 6~14 secondary papillae. Unlike other mammalians, however, secondary papillae in the mid-part of the tongue showed pineal-like papillae. In the posterior part of the tongue, secondary papillae were rare or absent. Fungiform papillae were surrounded by filiform papillae and densely distributed on the lingual surface. There were two vallate papillae on the borderline between the lingual body and root of the tongue. A vallate papilla contained two secondary papillae inside the grooves. Conical papillae were located in the area of the vallate papillae and covered the posterior part of the tongue root. No foliate papillae were seen on both margins of the posterior part of the tongue. Our results indicate that the structure on the lingual papillae of the Bengal tiger is somewhat different from that of other mammals.

To investigate the association between maternal iron status at the third trimester and fetal birthweight, maternal serum iron, ferritin, total iron-binding capacity (TIBC), and complete blood count values were measured at 36-weeks gestation. Delivery database on mothers who delivered babies at Chungbuk National University Hospital between January 2008 and March 2013 was extracted. A total of 353 uncomplicated term babies were analyzed using hierarchical regression and ANCOVA. Maternal age (standardized regression coefficient β=0.115, P<0.05), height (β=0.108, P<0.05), BMI (β=0.210, P<0.001), and gestational age (β=0.298, P<0.001) were significantly associated with birthweight. However, birthweight was not associated with maternal iron parameters. After adjusting for maternal age, height, BMI, and gestational age, babies born to mothers with lower mean values of hemoglobin, hematocrit, and serum ferritin were heavier than those born to mothers with higher values. Babies born to lower hemoglobin (11 g/dL) mothers were heavier than those born to higher hemoglobin (12 g/dL) mothers. However, birthweight was not significantly different between mothers with 10 g/dL or 13 g/dL of hemoglobin. Comparing birthweight according to 30 ug/dL of serum iron, 360 ug/dL of TIBC, 15 ng/mL of serum ferritin, and 10% transferrin saturation, babies born to mothers of the lower group were heavier than those born to mothers of the higher group. Therefore, maternal serum iron status at the third trimester seems to not be associated with birthweight.

Pruritus is one of the most important symptoms of allergic inflammatory skin disease. Conjugated linoleic acid (CLA) has been reported to have preventive effects against allergic inflammation. The objective of this study was to determine whether or not oral administration of CLA suppresses pruritus induced by compound 48/80 (composed of N-methyl-p-methoxy phenethylamine with formaldehyde) in mice, and if so, whether or not this effect is associated with serum histamine and prostaglandin (PG) E2 levels. Liquid CLA mixture (36.25% 9c-11t CLA, 36.95% 10t-12c CLA, 1.12% 9c-11c, and 1.94% t9-t11 CLA) was emulsified in 0.5% carboxymethyl cellulose (CMC) sodium salt and orally administered to mice at doses of 200 mg/kg once per day for 3 days. Similarly, disodium chromoglycate (DSCG), an antipruritic substance, was administered orally at the same concentrations as the negative control. Compound 48/80, a pruritus-inducing reagent, was subcutaneously injected 30 minutes after final administration of CLA. Scratching behavior of mice was counted just after compound 48/80 injection. Serum histamine and PGE2 concentrations were evaluated individually. Mice administered with CLA showed reduced frequency of scratching behavior compared to those without CLA. Antipruritic activities in CLA-treated and DSCG-treated groups were 48.5% and 26.8%, respectively. CLA and DSCG also diminished serum concentrations of histamine and PGE2 compared to compound 48/80 alone, respectively. This result suggests that dietary CLA has an antipruritic effect by down-regulating serum histamine and PGE2 levels for relief of compound 48/80-induced scratching behavior in mice, which will be useful in allergic pruritus as a preventive medicine.

The objective of this study was to investigate the antimicrobial effects of carvacrol (CV) against Staphylococcus aureus (S. aureus) and Escherichia coli O157:H7 (E. coli O157:H7) strains in milk. The minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of CV against S. aureus and E. coli O157:H7 were determined. In addition, bactericidal kinetics and antimicrobial activity of CV against the aforementioned pathogens in milk over a period of 2 weeks were investigated. CV exhibited antibacterial activity against both foodborne pathogens tested. The MIC and MBC of CV against S. aureus were 15.0 and 20 mg/mL, respectively, whereas those against E. coli O157:H7 were 16.0 and 32 mg/mL, respectively. In time-kill assays, CV at MBC reduced the number of S. aureus and E. coli O157:H7 in milk to undetectable levels within 24 hr. The antibacterial effects of CV persisted for 14 days without any loss of activity. Results of this study suggest that CV has a potential antibacterial activity against foodborne pathogens such as S. aureus and E. coli O157:H7 in milk.

Cytokeratin (CK) comprises the intermediate filament cytoskeleton of epithelial cells. Patterns of CK expression can be regarded as a specific marker for epithelial differentiation status. The aim of this study was to identify CK expression on tongues of Korean native goats ranging from 60-day-old fetuses to newborns during prenatal development using immunohistochemistry. The tongues of fetuses were removed from 2- to 4-year-old female Korean native goats by caesarean section performed under general anesthesia. Immunohistochemistry was performed to assess CK expression patterns on developing goat tongues using serial paraffin-embedded sections. Light zones signifying CK immunoreactivity in dorsal lingual epithelia were weakly positive in 60-day-old fetuses. In 90-day-old fetuses, deep areas in dorsal lingual epithelia were strongly positive for CK expression and superficial areas were moderately positive. In 120-day-old fetuses, light zones of lingual epithelia in the vallate papilla were strongly positive for CK expression, whereas ducts of von Ebner’s glands were moderately positive. In neonates, taste buds were positive for CK expression, whereas non-taste epithelial cells and von Ebner’s glands were negative. These findings indicate that goat tongues have different patterns of CK expression during development and provide a morphological basis for studies on the biological mechanism of epithelial differentiation.

Ischemic stroke is the most common type of stroke in humans. The purpose of this study was to evaluate the diagnostic value of magnetic resonance imaging (MRI) in a canine model of stroke. Ischemic stroke was induced by using prepared autologous thrombus. The dogs were placed in lateral recumbency on the operation table and the cervical area of each dog was sterilized by using alcohol. After making a cervical incision, the common carotid artery and internal carotid artery (a branch of the common carotid artery that supplies an anterior part of the brain) were exposed. A 200 μL injection of the autologous thrombus prepared 24 hr prior to surgery was delivered with a 20 gauge venous catheter through an internal carotid artery. After successful delivery of the autologous thrombus, the venous catheter was removed, and the cervical incision was sutured. Neurologic signs including generalized seizures, tetraparesis, and altered mental status, were observed in all 3 dogs after induction of ischemic stroke and the signs manifested immediately after awakening from anesthesia. T1- and T2-weighted images and fluid-attenuated inversion recovery (FLAIR) images of the brain were acquired 1 day before and 1 day after surgery. On the day following ischemic stroke induction, MRI revealed multifocal lesions in the cerebral cortex and subcortex such as T1 hypointensity, T2 hyperintensity, FLAIR hyperintensity, and diffusion-weighted hyperintensity in all 3 dogs. Upon postmortem examination, ischemic lesions were found to be consistent with the MRI findings and they were unstained with 2% triphenyltetrazolium chloride. Histologic features of the earliest neuronal changes such as cytoplasmic eosinophilia with pyknotic nuclei were identified. Neuropil spongiosis and perivascular cuffing were also prominently observed at the infarcted area. The present study demonstrated the features of MRI and histopathologic findings in canine ischemic stroke models.

Canine herpesvirus (CHV) is a member of the alphaherpesvirus subfamily, which can cause severe hemorrhagic diseases in neonatal pups as well as mild or subclinical respiratory infections in adult dogs. We examined the effects of cold stress on disease progression of CHV, an alphaherpesvirus, in neonatal puppies. Eight puppies were challenged intranasally with CHV suspension and divided into a cold stress treatment group and a hyperthermal group. Four pups were left uninoculated as controls and divided into cold and hyperthermal groups. In the challenged cold treatment group, all pups showed CHV-related disease within 5 days; pathological changes were observed in organs of puppies showing clinical symptoms. Grossly, numerous petechial red foci were scattered throughout lungs, kidneys, livers, and intestines of all CHV-infected puppies exposed to cold stress. Most puppies showed typical clinical signs and macroscopic lesions, and CHV infection was confirmed by isolation of the virus. However, in the challenged hyperthermal group, only one of the pups showed mild symptoms of CHV-induced disease. None of the puppies in the uninoculated group showed abnormal signs, although they were exposed to cold stress. These findings indicate that cold stress can cause rapid disease progression of CHV, an alphaherpesvirus.

A 7-year-old, spayed female, domestic short hair cat showed signs of a 2-week history of chronic anorexia, depression, and severe weight loss. Upon physical examination, pyrexia, mild gingivitis, and pale mucus membranes were noted. Laboratory analysis revealed normocytic normochromic non-regenerative anemia, severe thrombocytopenia, and hypergammaglobulinemia. Serum protein electrophoresis revealed the presence of elevated alpha-2 fraction within the globulin concentration. Based on history, clinical signs, and laboratory results, systemic viral infection was strongly suspected. Reverse transcriptase polymerase chain reaction identified the presence of feline immunodeficiency virus (FIV) in the serum. Furthermore, gene sequencing revealed the virus as FIV subtype A. Treatment with anti-retroviral agents, including azidothymidine (AZT) and recombinant human interferon-alpha, was continued for 4 weeks. However, the patient’s clinical condition deteriorated, resulting in death 1 month after initiation of treatment due to progressive renal failure. Necropsy and histopathology revealed hepatic and renal necrosis with hyper-cellular bone marrow mainly comprised of myeloid precursor cells. This case report is the first to describe phylogenetic subtyping, anti-retroviral combination treatment, and clinical outcomes in an FIV-infected cat in Korea. In addition, this report suggests that treatment should be initiated during the early phase of infection that could be effective for the virus.

Renal dysplasia is a developmental disorder of the renal parenchyma involving anomalous differentiation. It is characterized by persistent metanephric ducts surrounded by primitive mesenchyme, fetal or immature glomeruli, fetal or immature tubules, interstitial fibrosis, and dysontogenic metaplasia involving tissues such as cartilage. Renal dysplasia has been rarely reported in rats. Here, we observed a small left kidney in a rat used in a short-term repeat toxicity study. The rat showed no clinical signs throughout the study. All parameters, including those reflecting kidney functions, were normal upon hematological examination and urinalysis. Grossly, the kidney was small (5 × 8 mm) and its surface appeared normal. Histological examination revealed that the cortex and medulla were poorly demarcated and contained immature/atrophic glomeruli, immature renal tubules, and mesenchymal cells. The cortex contained immature glomeruli, atrophic glomeruli with cystic dilatation of Bowman’s capsular space, and some atypical tubules. Primitive metanephric tubules in the medulla were larger in diameter than normal collecting ducts, lined by a tall columnar epithelium with pale cytoplasm and basal nucleus, and surrounded by loose mesenchymal cells. Occasional tubules contained pale eosinophilic homogenous material in the lumen. Thus, this was diagnosed as a case of renal dysplasia on the basis of histologic features and is the first reported case of renal dysplasia in Sprague Dawley rats.

The current study was conducted to evaluate the biocompatibility of α-1,3 galactosyltransferase knockout pig bone graft in a rat calvarial defect model. Porcine cancellous bones were harvested from general and alpha-gal KO pigs and washed with 70% ethanol solution and normal saline. Bone pieces of the alpha-gal KO pig underwent a chemical treatment process to delipidize and deproteinize the bone. Bone graft particles were freeze-dried and stored at −70°C until use. Each bone graft was implanted into the rat calvarial defect in a fresh general pig, fresh transgenic pig, and chemical-treated pig bone group. There was no systemic adverse effect on hematology or necropsy findings in all groups at 1 week and 4 weeks. In the microcomputed tomography analysis, bone volume increased significantly in the chemical-treated transgenic pig bone group, whereas bone mineral density decreased significantly in the fresh general pig bone group compared with other groups. Histological evaluation showed cellular infiltration located at the margin of the bone graft particles, especially in the fresh general pig bone group. These results indicate that fresh general pig bone can elicit a greater local inflammatory response than fresh transgenic pig bone. Further, chemical-treated transgenic pig bone graft was less immunogenic than fresh bone graft. In conclusion, transgenic pig bone is a more biocompatible graft material. In addition, chemical treatment can reduce bone graft immunogenicity by delipidizing and deproteinizing bone.

One of the major problems in the modern pig industry is infectious disease. Susceptibility to infectious diseases is influenced by both constitutional (e.g. genotype, age, gender, and reproductive status) and environmental factors (e.g. nutrition, management, infections, and other forms of stress). Genetic variations within individual animals or herds can cause differences in the execution of immune functions against infectious agents in domestic pigs. The objectives of the present study were to identify genetic factor(s) responsible for piglet survival and mortality under commercial field conditions with infectious diseases as well as determine QTL regions for immune capacity in a pig reference family. Sex ratio was examined between normal grown and presumed dead groups of F2 animals generated by Korean native pigs (KNP) and Yorkshire (YS) breeds. The ratio of males was significantly higher in the presumed dead group than in the normal grown group. In order to study genetic factors associated with presumed mortality under disease outbreak, allelic frequencies of 239 single nucleotide polymorphisms (SNPs) were compared between normal grown and presumed dead groups. Exactly 22 SNPs showed significant differences in their allelic frequencies between the two groups, and four of them were validated with another commercial population divided by normal and emaciated pigs. Furthermore, the relationships between 239 SNPs and immune-related traits were studied. These results demonstrate that identification of genetic components of animal immune systems and susceptibility to infectious diseases is possible and will be useful to improve disease resistance in individuals as well as in breeding programs.

Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-β1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-β1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.

Background and Purpose: The Ankle-Brachial Index (ABI) is the ratio of blood pressure in the lower legs to that in the arms. The intima-media thickness (IMT) of extracranial carotid arteries determined by B-mode ultrasound is a measurable index of the presence of atherosclerosis. A low ABI and a high carotid IMT are independently related to increased risk of cardiovascular events. This study examined the association between carotid IMT and ABI in patients with ischemic stroke. Materials and Methods: Retrospectively, 116 hospitalized patients with ischemic stroke were recruited. Using a pulse wave velocity ABI device along with carotid duplex sonography, we measured carotid IMT and ABI and investigated the correlation between average values. Results: There was a significant difference in carotid IMT between the normal and abnormal ABI groups (P=0.0262). The group with an abnormal ABI was more than five times as likely to have increased carotid IMT as the group with a normal ABI (age, sex-adjusted OR 5.67 (95% CI 1.85~17.38)). The ABI and carotid IMT showed a weak inverse linear correlation in patients with ischemic stroke (correlation coefficient −0.378 after adjusting for age and sex). Conclusion: Our study suggests that an abnormal ABI is associated with a high carotid IMT in patients with ischemic stroke.

An evidence suggests that even low-dose irradiation can lead to progressive cognitive decline as well as memory deficits in both humans and experimental animals in part due to hippocampal dysfunction. To determine whether or not green tea (GT) and epigallocatechin gallate (EGCG) could attenuate memory impairment as well as suppress hippocampal neurogenesis, passive avoidance and object recognition memory test as well as TUNEL assay and immunohistochemical detection with markers of neurogenesis (Ki-67 and doublecortin (DCX)) were performed using adult mice treated with relatively low-dose gamma irradiation (2.0 Gy). GT was administered intraperitonially at a dosage of 50 mg/kg of body weight at 36 and 12 hr pre-irradiation and at 30 minutes post-irradiation, or orally at a dosage of 250 mg/kg of body weight/day for 7 days before autopsy. EGCG (25 mg/kg of body weight) was administered intraperitonially at 36 and 12 hr pre-irradiation and at 30 minutes post-irradiation. In the passive avoidance and object recognition memory test, mice trained for 1 day after acute irradiation (2 Gy) showed significant memory deficits compared with sham controls. The number of TUNEL-positive apoptotic nuclei in the dentate gyrus increased by 12 h after irradiation. In addition, the numbers of Ki-67- and DCX-positive cells significantly decreased. GT treatment prior to irradiation attenuated memory defects, blocked apoptotic death, as well as reduced the number of DCX-positive cells. Therefore, GT may attenuate memory defects in adult mice exposed to a relatively low dose of radiation possibly by inhibiting the detrimental effects of irradiation on hippocampal neurogenesis.