Oat (Avena sativa L.) Sprouts Restore Skin Barrier Function by Modulating the Expression of the Epidermal Differentiation Complex in Models of Skin Irritation.

Oats (Avena sativa L.) are used as therapeutic plants, particularly in dermatology. Despite numerous studies on their skin moisturization, anti-inflammation, and antioxidation effects, the precise molecular mechanisms of these effects are only partially understood. In this study, the efficacy of oat sprouts in the treatment of allergic contact dermatitis (ACD) was investigated, and their specific phytoconstituents and exact mechanisms of action were identified. In the in vivo ACD model, by stimulating the mitogen-activated protein kinase signaling pathway, oat sprouts increased the expression levels of proteins associated with skin barrier formation, which are produced during the differentiation of keratinocytes. In addition, in a lipopolysaccharide-induced skin irritation model using HaCaT, steroidal saponins (avenacoside B and 26-deglucoavenacoside B) and a flavonoid (isovitexin-2-o-arabinoside) of oat sprouts regulated the genetic expression of the same proteins located on the adjacent locus of human chromosomes known as the epidermal differentiation complex (EDC). Furthermore, oat sprouts showed immunomodulatory functions. These findings suggest the potential for expanding the use of oat sprouts as a treatment option for various diseases characterized by skin barrier disruption.

Potential of Aligned Electrospun PLGA/SIS Blended Nanofibrous Membrane for Tendon Tissue Engineering.

Tendons are responsible for transmitting mechanical forces from muscles to bones for body locomotion and joint stability. However, tendons are frequently damaged with high mechanical forces. Various methods have been utilized for repairing damaged tendons, including sutures, soft tissue anchors, and biological grafts. However, tendons experience a higher rate of retear post-surgery due to their low cellularity and vascularity. Surgically sutured tendons are vulnerable to reinjury due to their inferior functionality when compared with native tendons. Surgical treatment using biological grafts also has complications such as joint stiffness, re-rupture, and donor-site morbidity. Therefore, current research is focused on developing novel materials that can facilitate the regeneration of tendons with histological and mechanical characteristics similar to those of intact tendons. With respect to the complications in association with the surgical treatment of tendon injuries, electrospinning may be an alternative for tendon tissue engineering. Electrospinning is an effective method for fabrication of polymeric fibers with diameters ranging from nanometers to micrometers. Thus, this method produces nanofibrous membranes with an extremely high surface area-to-volume ratio, which is similar to the extracellular matrix structure, making them suitable candidates for application in tissue engineering. Moreover, it is possible to fabricate nanofibers with specific orientations that are similar to those of the native tendon tissue using an adequate collector. To increase the hydrophilicity of the electrospun nanofibers, natural polymers in addition to synthetic polymers are used concurrently. Therefore, in this study, aligned nanofibers composed of poly-d,l-lactide-co-glycolide (PLGA) and small intestine submucosa (SIS) were fabricated using electrospinning with rotating mandrel. The diameter of aligned PLGA/SIS nanofibers was 568.44 ± 135.594 nm, which closely resembles that of native collagen fibrils. Compared to the results of the control group, the mechanical strength exhibited by the aligned nanofibers was anisotropic in terms of break strain, ultimate tensile strength, and elastic modulus. Elongated cellular behavior was observed in the aligned PLGA/SIS nanofibers using confocal laser scanning microscopy, indicating that the aligned nanofibers were highly effective with regard to tendon tissue engineering. In conclusion, considering its mechanical properties and cellular behavior, aligned PLGA/SIS is a promising candidate for tendon tissue engineering.

Gastric Hyperplastic Polyp Causing Upper Gastrointestinal Hemorrhage and Severe Anemia in a Dog.

An 11-year-old castrated male Shih Tzu was referred for lethargy and melena. The hematocrit level was 18.8% (normal range: 36-56%), indicating severe anemia. Abdominal ultrasound revealed a round-to-oval-shaped mass in the stomach. Computed tomography (CT) revealed an intraluminal mass (17 × 12 × 15 mm) cranial to the pyloric antrum. After obtaining informed consent from the owner, exploratory laparotomy and subsequent gastrostomy were performed, showing an ulcerated mass potentially responsible for the severe anemia. A lump of hair was firmly attached to the ulcerated surface of the mass. After complete removal of the mass, the anemia resolved spontaneously. Histological examination revealed that the mass was a gastric hyperplastic polyp. At the 6-month follow-up, the dog was healthy with a normal hematocrit level. Gastric hyperplastic polyps are tumor-like lesions arising from the mucosal surface of the stomach, and projecting into the lumen. They can appear in any part of the stomach, and are usually found incidentally during gastric endoscopy or necropsy. The clinical signs include chronic occult blood loss, abdominal pain, and gastric tract obstruction. Gastric polyps causing acute blood loss anemia have rarely been reported in human medicine. To our knowledge, this is the first report describing a gastric hyperplastic polyp that caused severe anemia because of acute blood loss in a dog.

Humeral Aneurysmal Bone Cyst in a Cat with Sequential Computed Tomographic Findings.

A 7-year-old spayed female domestic shorthair cat presented with a swollen right forelimb and mild lameness. On physical examination, the mass was palpable in the right humeral region, and the cat exhibited pain on palpation. Radiography revealed an expansile osteolytic lesion at the proximal end of the right humerus. Computed tomography (CT) revealed an expansile bony mass on the proximal end of the right humerus and a mild periosteal reaction around the acromion of the scapula. Amputation of the right forelimb, including the scapula and removal of the axillary lymph node, were strongly recommended to the owner, but were declined. Four months after the initial presentation, the cat presented with a dramatically swollen right forelimb and progressive lameness. CT was performed again. In addition to osteolytic changes in the mass, vascular development had occurred at the cranioproximal region. The right forelimb, including the scapula and ipsilateral lymph nodes, was removed. The cat died during the postoperative recovery period. Based on clinical, diagnostic imaging, and histological findings, the final diagnosis was aneurysmal bone cyst. To the best of our knowledge, this is the first case of an aneurysmal bone cyst in the humerus of a cat.

Immunophenotyping of an Unusual Mixed-Type Extraskeletal Osteosarcoma in a Dog.

A 6-year-old female Maltese dog presented with a cervical mass without pain. The tumor was surrounded by a thick fibrous tissue and consisted of an osteoid matrix with osteoblasts and two distinct areas: a mesenchymal cell-rich lesion with numerous multinucleated giant cells and a chondroid matrix-rich lesion. The tumor cells exhibited heterogeneous protein expression, including a positive expression of vimentin, cytokeratin, RANKL, CRLR, SOX9, and collagen 2, and was diagnosed as extraskeletal osteosarcoma. Despite its malignancy, the dog showed no sign of recurrence or metastasis three months after the resection. Further analysis of the tumor cells revealed a high expression of proliferation- and metastasis-related biomarkers in the absence of angiogenesis-related biomarkers, suggesting that the lack of angiogenesis and the elevated tumor-associated fibrosis resulted in a hypoxic tumor microenvironment and prevented metastasis.

Multiple Undifferentiated Pleomorphic Sarcoma (Malignant Fibrous Histiocytoma) with Extradural Involvement in a 7-Year-Old Labrador Retriever.

A 7-year-old castrated male Labrador retriever was referred for evaluation of progressive hind limb paresis of 4 weeks' duration. On computed tomography and magnetic resonance imaging examination, masses were found in several regions including the lung, right kidney, and peritoneum. Additionally, an extradural mass at the region of T13-L1 was identified, which is assumed to related to the chief complaint, progressive hind limb paresis. With the consent of the owner, a dorsal laminectomy was performed to remove the mass and surrounding tissues for the palliation of the hind limb paresis. Hematoxylin and eosin staining and immunohistochemical examination revealed the mass to be consistent with an undifferentiated (high-grade) pleomorphic sarcoma. The patient presented with recurrence of the hind limb paresis, respiratory discomfort, and urinary incontinence. The owner declined treatment and the dog was euthanized. Systemic metastasis was confirmed on postmortem microscopic examination. To the authors' knowledge, this is the first report describing multiple undifferentiated high-grade pleomorphic sarcoma with extradural involvement developing into the vertebral canal through the intervertebral space, resulting in spinal damage, in a dog.

Up-regulation of MicroRNAs-21 and -223 in a Sprague-Dawley Rat Model of Traumatic Spinal Cord Injury.

In this experimental animal study, we examined alterations in the degree of transcription of two microRNAs (miRs)-miR-21 and -223-in a Sprague-Dawley (SD) rat model of traumatic spinal cord injury (TSCI). Depending on the volume of the balloon catheter (V), a total of 75 male SD rats were divided into the three experimental groups: the sham group (n = 25; V = 0 µL), the mild group (n = 25; V = 20 µL), and the severe group (n = 25; V = 50 µL). Successful induction of TSCI was confirmed on both locomotor rating scale at 4 h and 1, 3 and 7 days post-lesion and histopathologic examinations. Then, RNA isolation and quantitative polymerase chain reaction (PCR) were performed. No differences in the level of miR-21 expression were found at the first time point studied (4 h post-lesion) between the three experimental groups, whereas such differences were significant at all the other time points (P < 0.05). Moreover, there were significant alterations in the level of miR-223 expression at all time points studied through all the experimental groups (P < 0.05). Furthermore, locomotor rating scale scores had a linear relationship with the level of miR-21 expression (R2 = 0.4363, Y = 1.661X + 3.096) and that of miR-223 one (R2 = 0.9104, Y = 0.8385X + 2.328). Taken together, we conclude that up-regulation of miR-21 and -223 might be closely associated with progression and the early course of TSCI, respectively.

Primary intrapelvic hemangiosarcoma in a dog.

A 12-year-old, spayed female Schnauzer presented with constipation. A mass was observed in the pelvic cavity, and metastasis was not identified. Mass resection was performed through celiotomy with pubic osteotomy, and hemangiosarcoma was diagnosed. At 10 weeks post-operatively, the patient died of multiple metastasis. Primary intrapelvic hemangiosarcoma is rare in dogs.

Changes of the Cytoplasmic Proteome in Response to Alcoholic Hepatotoxicity in Rats.

Proteomic analyses have already been used in a number of hepatological studies and provide important information. However, few reports have focused on changes in the cytoplasmic proteome. The present study therefore aimed to evaluate changes in cytoplasmic proteome of rats in response to alcoholic hepatotoxicity. Rats were fed a Liber-DeCarli liquid diet containing ethanol for four weeks. Cytoplasmic proteins except mitochondrial proteins from the livers of these animals were investigated using two-dimensional gel electrophoresis and mass spectrometry. Alcohol induced a decrease in body weight gain and an increase in alanine transaminase (ALT), cholesterol, and phospholipid levels. Histopathological observations revealed hepatic damage characterized by necrosis and fatty change in alcohol-treated group at week 2, which continues until week 4. Our proteomic analysis revealed that 25 proteins were differentially expressed in the ethanol-fed group. Of these, 12 cytoplasmic proteins are being reported for the first time. Taken together, our results provide further insights into the disease mechanism and therapeutic information of alcoholic liver disease.

Primary lymphoma of the uterine horn in a Lhasa Apso dog.

Primary lymphomas of the canine female genital tract are uncommon tumours. A 9-year-old intact female Lhasa Apso dog presenting with a closed pyometra underwent an ovariohysterectomy (OHE), and the hyperplastic uterine horn along with multiple follicular cysts on the right ovary was examined by histological analysis. Severe infiltration of medium-sized lymphocytes with strong positive immunoreactivity for CD79a and numerous anaplastic features was detected in the unilateral uterine horn, and the dog was diagnosed as having extranodal marginal zone B-cell lymphoma (MZBCL). The present case reports an extremely rare occurrence of primary lymphoma involving the uterine horn in a dog and describes histological characteristics of the tumour for definite diagnosis.

Effect of Rubus coreanus extracts on diabetic osteoporosis by simultaneous regulation of osteoblasts and osteoclasts.

OBJECTIVE: Diabetes mellitus has been known to be associated with a high risk of osteoporosis. Rubus coreanus Miquel, a traditional Asian herbal medicine, has various uses, such as antiobesity and antiosteoporosis treatment, among others. We investigated the effect of R. coreanus extracts on diabetic osteoporosis. METHODS: Rats were not treated, or treated with streptozotocin or R. coreanus, or ovariectomized, in various combinations. After 6 weeks of treatment, the rats were killed, and serum biochemistry, histopathology, immunohistochemistry, and semiquantitative reverse transcription polymerase chain reaction were performed. In addition, in vitro studies were performed in MC3T3-E1 and RAW 264.7 cells. RESULTS: Rats treated using R. coreanus showed significant improvement in trabecular bone histopathology. Increased expression of osteocalcin was observed in rats treated with streptozotocin and R. coreanus, whether ovariectomized or not. In addition, the expression levels of cannabinoid receptors 1 and 2 and receptor activator for nuclear factor κß ligand were increased in rats that were ovariectomized and treated with streptozotocin and R. coreanus but decreased in those treated with streptozotocin and R. coreanus alone. These results indicate that the antiosteoporotic effect of R. coreanus in postmenopausal diabetic osteoporosis is attributable to the cannabinoid receptor-dependent maximal up-regulation of osteoblastogenesis. CONCLUSIONS: The present study shows that R. coreanus may rescue diabetic osteoporotic bone loss by simultaneous alteration of activation in osteoblasts and osteoclasts. Furthermore, these effects may be partially influenced by the up-regulation of the endocannabinoid system. In conclusion, dietary R. coreanus may be of use in improving the conditions of diabetic osteoporosis.

Tracheal replacement with fresh and cryopreserved aortic allograft in adult dog.

BACKGROUND: Many reports have described tracheal replacement using aortic allografts, with varying results and minimal understanding of the mechanism of tracheal regeneration. The present study attempted tracheal regeneration in adult dogs using fresh aortic allografts (FAA) and cryopreserved aortic allografts (CAA). MATERIALS AND METHODS: Twelve adult beagles underwent tracheal resection and were transplanted with FAA (n = 5) or CAA (n = 7). Animals were followed-up with serial radiography and magnetic resonance imaging, and were euthanized at predetermined times up to 16 mo post-surgery. RESULTS: There were no procedural deaths, but two animals died due to stent migration. Stent migration occurred in seven of the 12 animals. Evidence of regeneration of tracheal epithelium was observed in the surviving animals, with the transformation of squamous metaplasia to mucociliary epithelium being time-dependent. Islet of cartilage were observed in animals after 6 mo, but ring-like cartilage structures were absent, even after 16 mo. During autopsy, axial graft contractions up to 68% were observed. Serial radiographs show that most of the contraction occurred within 1 mo. The results of the MRI showed that the graft area was strongly enhanced for up to 2 mo, but was clearly reduced after 3 mo. CONCLUSIONS: Tracheal replacement in adult dogs using FAA or CAA is feasible. However, immaturity of the neotracheal cartilage did not allow the tissue to function as native tracheal tissue. Prolonged stenting should be considered in adult if the procedure is to be clinically contemplated.

Activation of MAP kinases during progression of radiation-induced pneumonitis in rats.

Radiation-induced pneumonitis is closely associated with the interplay of various stress-activated signals and immune responses related to the progression of lung injury. Mitogen-activated protein (MAP) kinase pathways play critical roles in the progression of inflammation via a cellular damage. Here, we examined the regional distribution of phosphorylated MAP kinases (p-JNK, p-ERK, and p-p38) in the progression of pneumonitis after exposure of a single dose irradiation with 10 Gy for 0, 4, and 8 weeks in rats. Also, we identified positive cells for these kinases using specific cell-type markers related to inflammation and type II pneumocyte. p-JNK was present abundantly in activated macrophages, CD8(+)T-cells, peribronchiolar smooth muscle cells, and weakly type II pneumocytes at 4 weeks or 8 weeks after irradiation. p-p38 and p-ERK was predominantly expressed in macrophages, CD4(+) T-cells, fibrotic cells as well as present in various lung parenchymal cells including alveolar epithelial cells and type II pneumocytes. In conclusion, it is considered that MAP kinase pathways play a pivotal role in early damage of residual cells as well as in the long-term regulation of distinct inflammatory cells during the progression of radiation-induced pneumonitis.

The protective effect of ENA Actimineral resource A on CCl4-induced liver injury in rats.

ENA Actimineral Resource A (ENA-A) is alkaline water that is composed of refined edible cuttlefish bone and two different species of seaweed, Phymatolithon calcareum and Lithothamnion corallioides. In the present study, ENA-A was investigated as an antioxidant to protect against CCl(4)-induced oxidative stress and hepatotoxicity in rats. Liver injury was induced by either subacute or chronic CCl(4) administration, and the rats had free access to tap water mixed with 0% (control group) or 10% (v/v) ENA-A for 5 or 8 weeks. The results of histological examination and measurement of antioxidant activity showed that the reactive oxygen species production, lipid peroxidation, induction of CYP2E1 were decreased and the antioxidant activity, including glutathione and catalase production, was increased in the ENA-A groups as compared with the control group. On 2-DE gel analysis of the proteomes, 13 differentially expressed proteins were obtained in the ENA-A groups as compared with the control group. Antioxidant proteins, including glutathione S-transferase, kelch-like ECH-associated protein 1, and peroxiredoxin 1, were increased with hepatocyte nuclear factor 3-beta and serum albumin precursor, and kininogen precursor decreased more in the ENA-A groups than compared to the control group. In conclusion, our results suggest that ENA-A does indeed have some protective capabilities against CCl(4)-induced liver injury through its antioxidant function.

Isolation and characterization of avian metapneumovirus from chickens in Korea.

Avian metapneumovirus (aMPV) causes upper respiratory tract infections in chickens and turkeys. Although the swollen head syndrome (SHS) associated with aMPV in chickens has been reported in Korea since 1992, this is the study isolating aMPV from chickens in this country. We examined 780 oropharyngeal swab or nasal turbinate samples collected from 130 chicken flocks to investigate the prevalence of aMPV and to isolate aMPV from chickens from 2004-2008. Twelve aMPV subtype A and 13 subtype B strains were detected from clinical samples by the aMPV subtype A and B multiplex real-time reverse transcription polymerase chain reaction (RRT-PCR). Partial sequence analysis of the G glycoprotein gene confirmed that the detected aMPVs belonged to subtypes A and B. Two aMPVs subtype A out of the 25 detected aMPVs were isolated by Vero cell passage. In animal experiments with an aMPV isolate, viral RNA was detected in nasal discharge, although no clinical signs of SHS were observed in chickens. In contrast to chickens, turkeys showed severe nasal discharge and a relatively higher titer of viral excretion than chickens. Here, we reveal the co-circulation of aMPV subtypes A and B, and isolate aMPVs from chicken flocks in Korea.

Multiple perianal infundibular follicular cysts in a dog.

This case report describes a 7-year-old male cocker spaniel dog with multiple perianal infundibular follicular cysts. Clinically the dog had moderate anal sacculitis, peri-anal pruritus causing it to 'scoot' and lick the area. On examination of the perianal area, there were over 100 firm, well circumscribed papules, ranged from 0.2 to 0.5 cm in diameter with a central pore, and were found in the perianal region. Alopecia was present in the perianal region. The skin tissue in the perianal region resected surgically was submitted for histological examination. Microscopically, the tissue revealed multiple dilated cysts filled with keratins and the papules corresponded to infundibular follicular cysts. The affected dog showed moderate anal sacculitis. Anal sacculitis commonly causes repeated scooting or licking the area around the anus. Therefore, the multiple follicular cysts in the present case appear to be primarily a sequela to chronic external trauma to the perianal area, probably in response to anal sacculitis. To the best of the authors' knowledge, the present report is the first documented case of multiple perianal infundibular follicular cysts in a dog.

Helicobacter pylori promotes hepatic fibrosis in the animal model.

Helicobacter pylori infection has been reported to be very common in patients with chronic liver diseases, including cirrhosis. To elucidate the pathological effect of H. pylori infection on the progression of hepatic fibrosis, C57BL/6 mice and Sprague-Dawley rats were orally inoculated with H. pylori, and hepatic fibrosis was induced with carbon tetrachloride (CCl(4)) administration. We observed the histopathological changes and the presence of H. pylori genes by PCR in the liver. Significant increase in the fibrotic score as well as in serum alanine aminotransferase and aspartate aminotransferase levels was shown in the CCl(4)+H. pylori group compared with that in the CCl(4)-treated group. Compared with the CCl(4)-treated group, alpha-smooth muscle actin and transforming growth factor-beta1 were enhanced; however, senescence marker protein-30, a multifunctional protein protecting hepatocytes against oxidative stress and apoptosis, was suppressed in the CCl(4)+H. pylori group. The 16S rRNA (400 bp) was demonstrated by PCR for H. pylori genes from genomic DNA extracted from the liver, and H. pylori-infected mice showed 93.8% (15 of 16) seropositivity by contrast with seronegativity in all H. pylori-noninfected mice. In addition, immunohistochemical study against H. pylori showed positive antigen fragments in the liver of the infected groups. Consequently, our data suggest that H. pylori infection could be an important contributing infectious factor to the development of liver cirrhosis.

Inhibition of radiation-induced apoptosis via overexpression of SMP30 in Smad3-knockout mice liver.

Apoptosis occurs early after irradiation and may be a good indicator of radiation damages. Since elevated levels of TGF-beta are associated with radiation-induced inflammation, the null mice of Smad3, a key downstream mediator of TGF-beta, show accelerated healing of irradiated injury. In order to evaluate resistance to radiation-induced liver injuries in Smad3-null mice, we determined the occurrence of apoptosis and the expression of senescence marker protein-30 (SMP30), as an anti-apoptotic marker, after irradiation to the liver. The livers of Smad3-mutant mice were exposed to local irradiation of 15 gray, from a (60)Co-gamma radiation. One week after irradiation, in Smad3-KO mice, radiation-induced apoptosis was at lower levels compared to those of irradiated WT mice. These findings were well matched with the expression of CYP2E1, which plays a role in hepatic injuries produced by oxidative stress. In addition, antioxidant related protein, the SMP30 levels were reduced by gamma irradiation in both groups. Interestingly, the increased expression of SMP30 expression in Smad3-KO mice liver was preserved at a higher level than that of the WT mice after irradiation. Therefore, these results suggest that the interruption of TGF-beta signaling by deletion of Smad3 brings about inhibition of hepatic apoptosis after ionizing irradiation. Moreover, the protective effect to ionizing radiation might be in correlation with the overexpression of SMP30 in the Smad3-null mice, which may act as an anti-apoptotic signaling molecule. The alteration of SMP30 by interruption of Smad3 might be a useful therapeutic target and diagnostic marker for radiation-induced liver damages.

Primary biliary cirrhosis, similar to that in human beings, in a male C57BL/6 mouse infected with Helicobacter pylori.

We report a case of primary biliary cirrhosis (PBC) that occurred in a 24-month-old male C57BL/6 mouse infected with Helicobacter pylori (H. pylori). Microscopically, the portal tract in the liver showed nonsuppurative destructive cholangitis with variable cytologic distortion of the epithelial cells and peribiliary lymphoplasmacytic infiltration. Immunohistochemistry using alpha-smooth muscle actin demonstrated fibrous bands associating with the wall of vasculature. The level of serum antivacuolating toxin IgG in this mouse showed the highest value (optical density=2.1470) of the H. pylori-infected group (n=13) (optical density=1.7168+/-0.1759, mean+/-SD). Spontaneously developed PBC-like lesions in C57BL/6 mice have been reported by several authors. However, this case strikingly resembles human PBC with its characterized histological features. Therefore, we propose that the increase in vacuolating toxin caused by H. pylori infection may be related to the development of PBC by molecular mimicry.

Macrophages, myofibroblasts and mast cells in a rat liver infected with Capillaria hepatica.

We trapped a rat (Rattus norvegicus) infected with Capillaria hepatica. At necropsy, grossly yellowish-white nodules (2-3 mm in diameter) were noted to be scattered on the liver's surface. Microscopically, granulomatous and fibrotic nodules that contained the eggs and/or adult worms of Capillaria hepatica were detected in the liver. Septal fibrosis was diffusely formed throughout the liver. There were a number of ED1-positive macrophages located in the sinusoids of the pseudolobules. On the double staining, myofibroblasts and mast cells were generally observed within the fibrous septa with the mast cells in close proximity to the myofibroblasts. We suggest that the interactions between macrophages, myofibroblasts and mast cells play a role in the septal fibrosis observed in rats infected by Capillaria hepatica.