Study on Controlling the Surface Structure and Properties of a Cellulose Nanocrystal Film Modified Using Alkoxysilanes in Green Solvents.

Film and sheet products made from naturally derived materials that exhibit high-performance surface functions are important as regards the environment. This study aimed to control the surface structure of a cellulose nanocrystal (CNC) film modified using methyltriethoxysilane and tetraethoxysilane coprecursors with environmentally friendly solvents (water and ethanol) during a spin-coating process. The surface-modified CNC film on the glass substrate was evaluated by microstructure analyses (Fourier transform infrared (FT-IR), nuclear magnetic resonance (NMR), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM)) and water contact angle (hydrophobicity) measurements. Through FT-IR, NMR, and XPS, it was confirmed that the silane compounds were chemically bonded to the surface of the CNC. The AFM images suggested that the local surface structure of the silylation-modified CNC film was formed along with the rod-like shape of the CNC. The water contact angle was approximately 90°, owing to the silylation of the hydroxy group and increased surface roughness of the CNC layer enabled by the sol-gel reaction.

Circulating activin A during equine gestation and immunolocalization of its receptors system in utero-placental tissues and fetal gonads.

Although equine gestation is unique from the standpoint of fetal gonadal enlargement and regression, the activator of this process is still unknown. The present study aimed to show a possible role of activin during equine gestation. In the first experiment, weekly plasma samples from six pregnant mares were used to measure activin A. In the second experiment, eight pregnant mares carrying female (gestational days 110, 140, 180, and 270) and male fetuses (gestational days 120, 180, 225, and 314) were used for immunohistochemistry of activin receptors (IA, IB, IIA, IIB), and their intracellular mediators (Smad2, Smad3, Smad4). Activin A levels in maternal circulation remained low until fourth weeks of gestation, thereafter, started to increase, and peaked first at 11 weeks of gestation. The second significant peak was observed on the day of parturition. Activin receptors type IA, IB, IIA, and IIB were immunostained in interstitial and germ cells of fetal ovaries and testes along with utero-placental tissues. Smad2, Smad3, and Smad4 were also immunolocalized in all these organs. These results demonstrated the activin-producing capacity of utero-placental tissues, and also evidenced the existence of activin receptors and functional signaling molecules in these organs. The first increment in circulating activin A in maternal circulation coinciding with the timing of initiation of fetal gonadal enlargement suggests that activin from the utero-placental tissues may have a stimulatory role in fetal gonad enlargement and utero-placental development in mares, whereas the second peak could be important to follicular development in the maternal ovary for foal heat.

Effects of a custom-made hinged knee brace with knee flexion support for patients with knee osteoarthritis:a preliminary study.

A knee brace for medial knee osteoarthritis (OA) is required to restrict knee adduction moment (KAM), but must not restrict knee flexion during swing phase. There is no report of a knee brace with both functions. The purpose of this study is to investigate the effect of the custom-made hinged knee brace for patients with knee OA compared to the hinged knee brace generally used, and to assess the KAM and knee flexion angle during swing phase. Fifteen patients (average age: 71.6 ± 7.8 years old) with medial knee OA participated. Gait analysis was performed using a 3-D motion analysis system to measure two conditions: hinged knee brace (KB), and a custom-made hinged knee brace with knee-flexion support- equipped rubber tubes at the posterior of the lateral and medial side poles (KBF). The peak KAM with KBF was significantly smaller than those with the KB (P=0.004, the difference between these conditions of KAM: 0.06 Nm/kg). The peak knee flexion angles during swing phase with KBF were also significantly larger than those with the KB (P=0.004, the difference between these conditions of knee flexion angle: 1.5 degrees). The custom-made brace with one type of tube actuator in the present study could decrease KAM and make for a small increase of knee flexion angle as opposed to the hinged knee brace.

Equine major histocompatibility complex class I molecules act as entry receptors that bind to equine herpesvirus-1 glycoprotein D.

The endotheliotropism of equine herpesvirus-1 (EHV-1) leads to encephalomyelitis secondary to vasculitis and thrombosis in the infected horse central nervous system (CNS). To identify the host factors involved in EHV-1 infection of CNS endothelial cells, we performed functional cloning using an equine brain microvascular endothelial cell cDNA library. Exogenous expression of equine major histocompatibility complex (MHC) class I heavy chain genes conferred susceptibility to EHV-1 infection in mouse NIH3T3 cells, which are not naturally susceptible to EHV-1 infection. Equine MHC class I molecules bound to EHV-1 glycoprotein D (gD), and both anti-gD antibodies and a soluble form of gD blocked viral entry into NIH3T3 cells stably expressing the equine MHC class I heavy chain gene (3T3-A68 cells). Treatment with an anti-equine MHC class I monoclonal antibody blocked EHV-1 entry into 3T3-A68 cells, equine dermis (E. Derm) cells and equine brain microvascular endothelial cells. In addition, inhibition of cell surface expression of MHC class I molecules in E. Derm cells drastically reduced their susceptibility to EHV-1 infection. These results suggest that equine MHC class I is a functional gD receptor that plays a pivotal role in EHV-1 entry into equine cells.

Post-Natal Dynamic Changes in Circulating Follicle-Stimulating Hormone, Luteinizing Hormone, Immunoreactive Inhibin, Progesterone, Testosterone and Estradiol-17ß in Thoroughbred Colts until 6 Months of Age.

The aim of present study was to clarify the post-natal profile of follicle-stimulating hormone (FSH), luteinizing hormone (LH), immunoreactive (ir)-inhibin, progesterone, testosterone, and estradiol-17ß, and their relationships in Thoroughbred colts. Six hundred and thirty-six colts were used for the study. Single plasma samples from each animal were harvested from the blood drawn through jugular venipuncture. The subjects were born with high amounts of progesterone, testosterone, and estradiol-17ß, all of which dropped significantly and remained at lower levels till the end of 6 months. FSH decreased transiently after birth until day 12 and then gradually increased to peak at day 100 which then maintained in lesser levels towards the end of the studied period. LH was highest during birth which decreased until day 26 and then increased slowly to sub-birth levels up to day 90. Animals were born with high amounts of ir-inhibin. It dropped slowly and halved by day 20 and then decreased towards rest of the studied period. The increase in FSH is negatively correlated with the declining ir-inhibin levels. The early increase in FSH can be the indication of early post-natal maturation of the hypothalamic pituitary testicular axis that ultimately might be responsible for priming the testes for future development.

Annual Changes in Day-length, Temperature, and Circulating Reproductive Hormones in Thoroughbred Stallions and Geldings.

Changes in follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, immunoreactive(ir)-inhibin, testosterone, estradiol-17ß, and insulin-like growth factor (IGF)-I in Thoroughbred stallions along with changes in prolactin secretion in geldings were studied. The correlations of day-length with changes in the concentrations of these hormones were also studied. Five stallions and thirteen geldings were employed to draw blood samples in monthly basis and radioimmunoassay was performed to measure these hormones. All hormones showed a seasonal pattern, the levels being highest during the breeding season and lowest during the winter months. Most of the hormones were at their highest concentration during the month of April, the mid of spring in northern hemisphere. The concentration of circulating IGF-I also demonstrated seasonality, the peak lying on the month of April. The plasma concentration of prolactin also increased during the breeding season. This phenomenon was similar both in stallions and geldings although geldings had lower concentration than that of stallions. The changes in concentration of prolactin in stallions and geldings correlated more towards the day-length than towards the temperature. These results clearly indicate the seasonality of pituitary and gonadal hormones of Thoroughbred stallions, the activity being highest during the month of April and May of the breeding season.

Presence of surfactant proteins in the uteri and placentae of pregnant mares.

Immunohistochemical investigations of the expression of surfactant protein A (SP-A) and surfactant protein D (SP-D) in the uterine and placental tissues of 13 pregnant mares were performed using anti-horse monoclonal primary antibodies. Strong positive reactions for both SP-A and SP-D were observed in the trophoblasts in the microcotyledons of the placentae at 182 to 314 days of gestation; in uterine glandular epithelial cells, faint-to-weak reactions were observed during gestation. This study describes, for the first time, the changes in the SP-A and SP-D expression levels in the endometrium of mares during gestation; the SP-A and SP-D expression levels increased after the second trimester of gestation.

Jugular thrombophlebitis developed from buccal ulcer in a thoroughbred horse.

A 22-year-old Thoroughbred stallion had severe left jugular thrombophlebitis. Macroscopic and microscopic examinations revealed extension of the lesions from a penetrating ulcer on the left buccal mucosa to the underlying muscle and local vein, and sequentially to the left jugular vein. This was a rare case of equine jugular thrombophlebitis caused by direct extension of infection from a traumatic oral lesion.

Hepatocellular carcinoma in a Hokkaido brown bear (Ursus arctos yesoensis).

Hepatocellular carcinoma with metastases to lymph nodes and adrenal glands was found in a 26-year-old male Hokkaido brown bear (Ursus arctos yesoensis). Left hepatic lobe was largely replaced by well-differentiated neoplastic cells, whereas poorly differentiated tumor cells had proliferated in part of the left hepatic lobe, in scattered nodules in the remaining liver tissue, and at the metastatic sites. Immunoreactivity for hepatocyte antigen (hepatocyte paraffin 1 antibody) and alpha-fetoprotein was observed in both well- and poorly differentiated neoplastic cells in the liver and metastatic foci. To our knowledge, this case is the first report of hepatocellular carcinoma in brown bears.

Non-cytopathic bovine viral diarrhea virus infection inhibits differentiation of bovine neural stem/progenitor cells into astrocytes in vitro.

Bovine viral diarrhea virus (BVDV) causes fetal brain malformations in ruminants when the fetuses are infected transplacentally in mid-pregnancy. In both cytopathic and non-cytopatic virus infections, viral lytic infection in actively replicating cells and interruption of vascular integrity have been suggested as the pathogenesis, but functional disturbance of infected neural developing cells has been unclear. In this study, we examined the effect of infection with non-cytopathic BVDV2 on the differentiation of neural stem/precursor cells isolated from the bovine fetus. In the process of differentiation to three types of neural cells, neurons, astrocytes and oligodendrocytes, virus infection significantly and selectively inhibited the differentiation of neural stem/precursor cells into the astrocytic lineage. This inhibition is possibly important for the pathogenesis of congenital brain malformations associated with non-cytopathic BVDV infection.

Systemic candidiasis and mesenteric mast cell tumor with multiple metastases in a dog.

A 5-year-old female miniature dachshund presenting with persistent vomiting and diarrhea had two concurrent rare pathological conditions: systemic candidiasis and mesenteric mast cell tumor with multiorgan metastases. Neoplastic mast cells formed mass in the mesentery of the cecal-colonic region and were also found in the liver, spleen, kidneys, lungs, adrenal grands, ovaries, bone marrow and other tissues. The cells had intracytoplasmic granules with metachromasia and were immunohistochemically positive for c-kit and histamine. Granulomatous lesions with fungal organisms were present in the heart, lungs, kidneys, pancreas, subserosal and surrounding adipose tissue of the duodenum, thyroid glands and mesenteric mass, and phagocytosed organisms were detected in the liver and bone marrow. Bacteriologically and immunohistochemically, the fungi were consistent with Candida albicans.

Two cases of bovine sarcoma in clinically long-standing lesions.

Histopathological examination of clinically long-standing lesions with durations of one year or more in the extremities of two cattle revealed the presence of sarcomas with distant metastases. In case 1, neoplastic cells were fusiform to pleomorphic, stained for no specific differentiation markers, and diagnosed as undifferentiated sarcoma. Neoplastic growth in case 2 was composed of spindle to histiocytoid cells and a significant number of multinucleated giant cells, both of which were immunoreactive to histiocyte markers, and diagnosed as giant cell malignant fibrous histiocytoma. Neoplastic cells of both cases were immunohistochemically positive for nitric oxide-related antigens, which were recognized as markers of inflammation-induced carcinogenesis in human and laboratory animals.

Histologic evaluation of the diversity of epidermal laminae in hooves of horses without clinical signs of laminitis.

OBJECTIVE: To evaluate the histologic diversity of epidermal laminae in hooves from horses without clinical signs of laminitis. SAMPLE POPULATION: Formalin-fixed samples of stratum internum obtained from the mid region of the dorsal aspect of the hoof wall from the forelimbs of 35Thoroughbred cadavers (including foals [n = 9], yearlings [5], 2 year olds [6], racing horses [5], and mares [10]). PROCEDURES: Paraffin-embedded laminar tissues were stained with H&E for the evaluation of architectural variety of primary epidermal laminae (PEL) and secondary epidermal laminae (SEL). For detection of cytokeratin (CK) expression in epidermal laminae, immunohisto-chemical staining was performed by use of anti-CK14 and anti-CK8.12 antibodies. RESULTS: The morphology of the PEL, SEL, and tips of PEL was classified into 3, 5, and 3 patterns, respectively. Differences in the predominant type of SEL depended on their location with respect to the laminar interface. In SEL attached to the sides of PEL, the basal cells were immunoreactive to CK14 and CK8.12, which was interpreted as a normal pattern. In some SEL at the tips of PEL, the suprabasal cells expressed CK14, CK8.12, or both, which constituted a hyperplastic pattern. CONCLUSIONS AND CLINICAL RELEVANCE: The histologic diversity of epidermal laminae from hooves of Thoroughbreds was attributable to the combined morphology of PEL and SEL. Detection of hyperplastic changes in the laminar interface does not justify a diagnosis of laminitis because such changes can develop independent of clinical disease. The classification system used here should aid investigators in making a more accurate histologic evaluation of laminae.

Borna disease virus RNA detected in Japanese macaques (Macaca fuscata).

We have examined the seroprevalence of BDV in wild Japanese macaques (Macaca fuscata) in the peninsula (Chiba prefecture), Japan. Serum samples from macaques were examined by the ELISA, Western blot and immunofluorescence assays to detect the presence of serum antibodies that react specifically to BDV antigens. Among 49 investigated individuals, 6 (12.2%) showed positive reaction to BDV antigens. RT-PCR studies detected BDV sequences in brain tissue of one case among four seropositive cases examined. Sequence analysis revealed a high degree of genetic conservation between BDV sequences derived from Japanese macaques and those documented for other animal species. Nevertheless, phylogenetic analysis revealed unique differences between macaque and other species derived BDV sequences.

Retrospective serological analysis of spontaneous CDV infection in 192 dogs.

Spontaneous cases of canine distemper virus (CDV) infection were serologically evaluated. The 192 dogs in which CDV antigen was confirmed from tonsil by immunohistological examination were 2- to 4-months old, of various breeds, and unvaccinated. The prognoses were good in 74 dogs with significantly high levels of anti-CDV passive hemolytic aggregation (PHA) titer. In the other 118 dogs with poor prognoses, anti-CDV PHA titer was not detected. Anti-CDV PHA titer had the most significant association with the prognoses of CDV infection, and could be the most reliable and useful indicator for evaluating such prognoses.

Tubulopapillary carcinoma with spindle cell metaplasia of the mammary gland in a cat.

Sarcomatous proliferation of spindle cells was present in the mammary gland and many metastatic sites in a 10-year-old female domestic cat with tubulopapillary carcinoma in the mammary gland. Transition from neoplastic tubular structures to spindle cells in the primary site and fascicular proliferation of the spindle cells with or without coexistance of tubulopapillary carcinoma in the primary and metastatic sites were observed. Most of spindle cells were positive for cytokeratin CAM5.2 as well as the normal luminal epithelium but not the myoepithelium. From these results, this case was diagnosed as tubulopapillary carcinoma with spindle cell metaplasia and it was clarified that neoplastic luminal epithelial cells can transform to sarcomatous appearence.

Suprascapular nerve paralysis due to streptococcal meningoradiculitis in a cow.

A cow, presenting with lameness with atrophy of the right supraspinatus and infraspinatus muscles, was clinically diagnosed with suprascapular nerve paralysis. Histological examination revealed necrosuppurative lesions with Gram-positive cocci arranged in chains in multiple organs, including the cardiac valves, lungs, muscles, joints, brain, cerebral and spinal meninges. Spinal meningitis progressed into the roots of the right 6th to 8th cervical nerves. The suprascapular nerve showed partial loss of nerve fibers in the periphery, and innervated muscle fibers were atrophied. Bacterial culture of the cerebrospinal fluid revealed the presence of Streptococcus pyogenes. From these findings, the suprascapular nerve paralysis in this case was considered to result from meningoradiculitis associated with systemic streptococcal infection. This is a rare bovine case of suprascapular nerve paralysis due to central nerve damage of infectious cause.

Aldosterone-, corticosterone- and cortisol-secreting adrenocortical carcinoma in a dog: case report.

A 12-year-old, intact female beagle exhibited symptoms of polyuria-polydipsia and hyperorexia for two months. Blood tests showed elevated asparate aminotransferase, alanine aminotransferase, alkaline phosphatase and creatine kinase levels, as well as marked hypokalemia. The results of adrenocorticotropic hormone stimulation test showed elevated cortisol, aldosterone and corticosterone concentrations. Abdominal ultrasonography confirmed a mass in the left adrenal gland. Masses were also seen in the liver and caudal vena cava. Diagnosis was a tumor of the adrenal cortex with metastases. Trilostane administration was initiated. The dog initially showed improved demeanor as a result of regulating hormone secretion. However, after 88 days, the dog weakened rapidly, before dying on the 117th day. Pathological findings confirmed a diagnosis of adrenocortical carcinoma.

Characterization of a Borna disease virus field isolate which shows efficient viral propagation and transmissibility.

To investigate the biological characteristics of field isolates of Borna disease virus (BDV), as well as to understand BDV infections outside endemic countries, we isolated the virus from brain samples of a heifer with Borna disease in Japan. We demonstrate that the brain lysate contained replication products of BDV and induced viral propagation in rat glioma cells, suggesting that a replication-competent BDV existed in the bovine brain. This field strain of BDV, named Bo/04w, showed efficient viral release and transmissibility and also displayed a distinct pattern of expression of viral phosphoprotein (P) during infection, as compared with laboratory-adapted BDV strains. Interestingly, we found the level of P to be significantly low in cells infected with Bo/04w, and the transcription of this isolate to be more efficient than that of laboratory strain of BDV. These results indicated that the field isolate may regulate the expression of P at an optimal level in infected cells. We also confirmed that Bo/04w maintains biological significance in neonatal gerbil brain. Sequencing revealed that despite the biological differences, the field isolate is closely related genetically to the laboratory strains of BDV. We discuss here the sequence similarities between BDV isolates from endemic and nonendemic countries.

Distribution of inclusion bodies in tissues from 100 dogs infected with canine distemper virus.

One hundred dogs that were positive for canine distemper virus antigen and inclusion bodies in the tonsils were examined for the distribution of inclusion bodies in various tissues. Inclusion bodies were found in the lungs (70 dogs), brains (20 dogs), urinary bladders (73 dogs), stomachs (78 dogs), spleens (77 dogs), and lymph nodes (81 dogs) of the dogs. Based on these results, the tonsils may be the most suitable tissue for detection of inclusion bodies in canine distemper.