A quantitative study of the purkinje cells in the cerebellum and the inferior olivary neurons in the chicken

A single olivocerebellar fiber branches off several climbing fibers. One Purkinje cell receives input from only one climbing fiber. A single inferior olivary neuron, therefore, synapses with several Purkinje cells, so that there are more Purkinje cells than the inferior olivary neurons. We aimed to elucidate the numerical ratio of the inferior olivary neurons to Purkinje cells in the chicken. The total numbers were 353,834 +/- 5,274 in the Purkinje cells per the cerebellum and 21,553 +/- 904 in the inferior olivary neurons of both sides. The numerical ratio of inferior olivary neurons to Purkinje cells was 1:16. The ratio of those neurons in mammals is about 1:4-17, so that the ratio in the chicken is within the range of mammals.     

Effects of intramuscular fat deposition on the beef traits of Japanese Black steers (Wagyu)

Effects of intramuscular fat deposition on the chemical composition, tenderness, and free amino acids (FAA) concentration of beef were studied using various classified carcasses of 21 Japanese Black (Wagyu) steers. The Beef Marbling Standard (BMS) No., judged on the market in 1998, and fat content, ranged from 2 to 10 and 4.8 to 39.0% in the M. longissimus thoracis, respectively. Moisture content was negatively correlated with the fat content (r =?0.98, P < 0.01). In contrast, protein content was relatively constant up to approximately 23% fat, corresponding to BMS No.4, and decreased as the fat content increased. Cooking loss was also constant up to approximately 28% fat, corresponding to BMS No.4, and markedly decreased with fat content increase. The beef in these cases were, however, classified into BMS No. around 9 and 11, respectively, according to the BMS model in 1988. A negative correlation was found between the shear‐force value and the fat content (r =?0.83, P < 0.05). Most FAA concentrations on the wet weight of meat were correlated negatively with the fat content, except glutamine, and this negative relationship was still observed when the concentrations were recalculated on the basis of protein. These results indicate that higher marbling Wagyu beef (above approx. 23% fat) would have an extremely lower content of protein, which would partly explain the lower cooking loss and FAA with fat increase. In addition, some other reasons not relating to protein content (e.g. high fat content preventing the breakdown of protein to FAA) seem to explain the negative correlation between fat and FAA.     

Differential cytokine expression in T cell subsets from bovine peripheral blood

Although distinct cytokine expression in T cell subsets is well understood in mice and humans, limited information is available on bovine T cell subsets. In the present study, we analyzed the mRNA expression of 10 kinds of cytokines and CD25 expression in CD4⁺, CD8⁺, WC1⁺ and WC1^-γδ T cell subsets in bovine peripheral blood by Concanavalin A (Con A) stimulation. CD25 expression was significantly increased in CD4⁺, CD8⁺ and WC1⁺γδ T cells, but not in WC1^-γδ T cells by Con A stimulation. In CD4⁺ T cells, the mRNAs of Interleukin (IL)-2, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β and transforming growth factor (TGF)-β were expressed in control cultures, and IL-3, IL-4 and granulocyte-macrophage colony stimulating factor (GM-CSF) were newly expressed when the cells were stimulated with Con A. CD8⁺ T cells expressed the mRNAs of IL-6, TNF-α, TNF-β and TGF-β in control cultures, and newly expressed those of IL-2, IFN-γ and GM-CSF, but did not express those of IL-3, IL-4 or IL-10 after Con A stimulation. The cytokine expression profile of WC1⁺γδ T cells was similar to that of CD8⁺ T cells. However, WC1^-γδ T cells did not express any cytokine mRNA except TGF-â mRNA. These results will contribute to elucidate the participation of T cell subsets in immune responses against infectious disease in cattle.     

Establishment of a BRAF V595E-mutant canine prostate cancer cell line and the antitumor effects of MEK inhibitors against canine prostate cancer

Canine prostate cancer (cPCa) is a malignant neoplasm with no effective therapy. The BRAF V595E mutation, corresponding to the human BRAF V600E mutation, is found frequently in cPCa. Activating BRAF mutations are recognized as oncogenic drivers, and blockade of MAPK/ERK phosphorylation may be an effective therapeutic target against BRAF-mutated tumours. The aim of this study was to establish a novel cPCa cell line and to clarify the antitumor effects of MEK inhibitors on cPCa in vitro and in vivo. We established the novel CHP-2 cPCa cell line that was derived from the prostatic tissue of a cPCa patient. Sequencing of the canine BRAF gene in two cPCa cell lines revealed the presence of the BRAF V595E mutation. MEK inhibitors (trametinib, cobimetinib and mirdametinib) strongly suppressed cell proliferation in vitro, and trametinib showed the highest efficacy against cPCa cells with minimal cytotoxicity to non-cancer COPK cells. Furthermore, we orally administered 0.3 or 1.0 mg/kg trametinib to CHP-2 xenografted mice and examined its antitumor effects in vivo. Trametinib reduced tumour volume, decreased phosphorylated ERK levels, and lowered Ki-67 expression in xenografts in a dose-dependent manner. Although no clear adverse events were observed with administration, trametinib-treated xenografts showed osteogenesis that was independent of dosage. Our results indicate that trametinib induces cell cycle arrest by inhibiting ERK activation, resulting in cPCa tumour regression in a dose-dependent manner. MEK inhibitors, in addition to BRAF inhibitors, may be a targeted agent option for cPCa with the BRAF V595E mutation.     

A feline case of isolated fourth ventricle with syringomyelia suspected to be related with feline infectious peritonitis

A one-year-old female cat was unable to stand. Magnetic resonance imaging was performed, and an enlargement of the lateral, third, and fourth ventricles and syringomyelia were detected. The cat was diagnosed with an isolated fourth ventricle (IFV) with syringomyelia. The serum isoantibody test for the feline infectious peritonitis (FIP) virus was 1:3,200. After the cat died, a pathological examination revealed nonsuppurative encephalomyelitis. We suspected that the IFV, detected in the cat, was associated with FIP encephalomyelitis. To our knowledge, there has been no report on IFV in veterinary medicine.     

A canine case of necrotizing meningoencephalitis for long-term observation: clinical and MRI findings

A 3-year-old female pug presented with general seizure following a partial seizure. During the remaining 48 months till death, the dog showed various neurological signs such as disturbance of consciousness, myoclonus and various types of partial seizure after each occurrence of the seizure clusters, and the dog eventually exhibited inability to stand and dementia. Magnetic resonance imaging findings revealed atrophy of the brain over the course of the disease. On histopathological analysis, the dog was diagnosed with necrotizing meningoencephalitis. This case of a canine necrotizing meningoencephalitis observed over the long term is valuable.     

Molecular typing of Salmonella enterica serovar 4,[5],12:i:- isolates from humans,animals and river water in Japan by multilocus variable-number tandem repeat analysis and pulsed-field gel electrophoresis

Fifty-one Salmonella enterica serovar 4,[5],12:i:- (S. 4, [5],12:i:-) isolates (14 human strains, 34 animal strains and 3 river water strains) which are assumed to be monophasic variants of S. Typhimurium were analyzed using pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem repeat analysis (MLVA) in order to investigate their genetic diversities and relationships. PFGE, MLVA and combination of them identified 28, 27 and 34 profiles (Simpson’s diversity indices [DI]=0.94, 0.96 and 0.97), respectively. No correlations were detected between MLVA clustering and PFGE clustering or phage typing. These results suggested that S. 4,[5],12:i:- originated from multiple S. Typhimurium ancestors. Two cattle and one pig isolates showing identical phage types as well as PFGE and MLVA profiles to human isolates S. 4,[5],12:i:- suggested the existence of the links between human infections and animal reservoirs.     

Spatial relationships among the cellular tapetum,visual streak and rod density in dogs

The dog visual system is well suited to dim light conditions due to rod-dominated retina and the reflective tapetum. The topographical distributions of rods and thickness of the tapetum of the dog were quantified in retinal whole mounts stained with thionine, and spatial relationships among the tapetum, rod density and visual streak of high ganglion cell density were elucidated. The relationship between the retina and tapetum was analyzed in parasagittal sections stained with thionine or hematoxylin-eosin. The tapetum was thick in its center, and the thickest part consisted of 9 to 12 tapetal cell layers. Rod density ranged from 200,000 to 540,000/mm². Maximum rod density was found in the area dorsal to the visual streak, and the density in that area was significantly higher than the rod density in the visual streak and accorded spatially with the thickest part of the tapetum. The horizontal visual streak was found over the horizontal line through the optic disc in the temporal half and extended slightly into the nasal half. The central area of the highest density of ganglion cells was approximately located midway between the nasal and temporal ends of the visual streak. The visual streak was located within the tapetal area, but ventrally to the thick part of the tapetum.     

Sex differences in stress responses to transportation in goats: Effects of gonadal hormones

The present study examined sex differences and the involvement of gonadal hormones in stress responses caused by road transportation in Shiba goats. In experiment 1, we investigated the stress responses of males and females to transportation. Plasma levels of cortisol (Cor) significantly increased during 1 h of transportation, and those in females were significantly higher than those in males. Plasma glucose (Glu) and free fatty acid (FFA) levels also increased similarly in both females and males by transportation, and there were no sex differences. Food intake following transportation decreased only in males compared with that in the basal session, in which the animals were not transported. Experiment 2 examined the involvement of gonadal hormones in stress responses to transportation using castrated males. Goats were given cholesterol (Cho), 5α‐dihydrotestosterone (DHT) or 17β‐estradiol (Es). The plasma Cor levels increased during transportation regardless of hormone treatment, and those in DHT treated goats were significantly lower than those in Cho or Es treated animals. Plasma Glu and FFA levels also increased during transportation, regardless of hormone treatment, and there were no differences between treatments. Food intake following transportation was significantly lower than that in the basal session only in goats given DHT. In conclusion, gender affects Cor secretion that is increased by transportation and the decrease of food intake following transportation in Shiba goats, and the major cause of these differences is androgen.     

Effects of protozoa on the antioxidant activity in the ruminal fluid and blood plasma of cattle

The objective of this study was to investigate the antioxidant status in ruminal fluid and blood plasma among three faunated and two defaunated (protozoa‐free) cattle (average bodyweight of 225 kg), fed hay plus concentrate. The extra cellular antioxidant activity of the mixed protozoa and bacteria suspensions were also studied in vitro. The antioxidant activity was detected by means of the free radical scavenging ability. The activity (units/microbial nitrogen) of the protozoal suspension increased from 59 (0 h) to 318 (18 h), and decreased to 40 (24 h) during incubation. The activity of the bacterial suspension also increased from 111 (0 h) to 644 (18 h), and decreased to 533 (24 h). The antioxidant activity (units/mL, U/mL) in the ruminal fluid of faunated (ranging from 116 to 254) and defaunated (ranging from 66 to 110) cattle was increased after 2 h and decreased after 5 h of feeding, being significantly higher in the faunated cattle. The antioxidant activity of blood plasma (U/mL) ranged from 248 to 316 in the faunated and 121–170 in the defaunated cattle during 0–5 h after feeding, being significantly higher in the faunated cattle. Therefore, defaunation possibly causes a decrease in the antioxidant level in the ruminal fluid, and may impair the health and performance of ruminants through an oxidant–antioxidant imbalance.