Dynamics of the thymic transcriptome at stages of acute thymic involution in Japanese Black calves with a poor prognosis.

Transcriptome analysis was performed on the thymus of Japanese Black calves that were necropsied due to poor prognosis, to characterize changes associated with acute thymic involution. Gene expression profiles obtained by DNA microarray analysis of eight calf thymuses were classified into three patterns that correlated with the histopathological stage of acute thymic involution. Using principal component analysis, the first principal component of the global gene expression levels in the calf thymus was associated with the stage of acute thymic involution, suggesting that histopathological changes greatly influence the gene expression profile. Gene ontology enrichment analysis revealed that genes related to cell proliferation, wound healing, and inflammatory responses were the main contributors to the first principal component. Real-time RT-PCR showed that the thymus had lower expression of PCNA, KIFC1, and HES6, and higher expression of SYNPO2, PDGFRB, and TWIST1 during acute thymic involution. Immunohistochemistry demonstrated a decrease in the rate of Ki67-positive cells in the thymic cortex during the late stage of acute thymic involution. The rate of cleaved caspase-1-positive cells increased in the thymic cortex at an earlier stage than the increase in the rate of cleaved caspase-3-positive cells. Vimentin, which was almost absent in the non-involuted thymic cortex, appeared in the thymic cortex during acute thymic involution. These results suggest that in farmed calves with a poor prognosis, inflammatory responses and impaired thymocyte proliferation are primarily involved in acute thymic involution.

Quality of life assessment and cost­utility analysis of initial chemotherapy for patients with non­Hodgkin's lymphoma: A prospective analysis.

Chemotherapy has helped prolong survival in patients with malignant lymphoma, enhancing their quality of life (QOL). Despite the eventual decline in the QOL of patients, the impact of initial chemotherapy remains poorly understood. A prospective patient-reported QOL survey among patients with malignant lymphoma receiving initial chemotherapy was conducted, targeting those treated at Gifu Municipal Hospital (Gifu, Japan) between January 2021 and December 2022. Surveys were conducted pre- and post-chemotherapy based on the EuroQol 5 dimensions. Drug costs were calculated using official prices and analyzed from the cost payer's perspective via cost-utility analysis. Among the 60 patients included in the present study, 28 had diffuse large B-cell lymphoma. Cyclophosphamide, doxorubicin, vincristine, prednisolone ± rituximab therapy was the most common treatment (38 patients) and demonstrated superior cost-effectiveness due to its lowest cost and change in utility value. Initial chemotherapy for patients with malignant lymphoma generally improved the QOL. Clinical trial registration: UMIN000042868 (registered on December 28, 2020).

Acyl modifications in bovine, porcine, and equine ghrelins.

Ghrelin is a peptide hormone with various important physiological functions. The unique feature of ghrelin is its serine 3 acyl-modification, which is essential for ghrelin activity. The major form of ghrelin is modified with n-octanoic acid (C8:0) by ghrelin O-acyltransferase. Various acyl modifications have been reported in different species. However, the underlying mechanism by which ghrelin is modified with various fatty acids remains to be elucidated. Herein, we report the purification of bovine, porcine, and equine ghrelins. The major active form of bovine ghrelin was a 27-amino acid peptide with an n-octanoyl (C8:0) modification at Ser3. The major active form of porcine and equine ghrelin was a 28-amino acid peptide. However, porcine ghrelin was modified with n-octanol (C8:0), whereas equine ghrelin was modified with n-butanol (C4:0) at Ser3. This study indicates the existence of structural divergence in ghrelin and suggests that it is necessary to measure the minor and major forms of ghrelin to fully understand its physiology.

[Establishment of Blinatumomab Home Administration Transition Program Using CADD-Legacy® Pump].

We established a transition program from hospital to home administration of blinatumomab using the CADD-Legacy® pump and evaluated its safety and feasibility in 2 pediatric patients. The program recommended that repeated overnight stays with continued hospitalization would help patients safely transition to home administration of blinatumomab. Pediatric patients who do not attend school, such as preschoolers, or who return to their original school, can be transited to an outpatient setting if people caring for pediatric patients, including teachers in the school, are educated.

Citrin-deficient patient-derived induced pluripotent stem cells as a pathological liver model for congenital urea cycle disorders.

Citrin deficiency is a congenital secondary urea cycle disorder lacking useful disease models for effective treatment development. In this study, human induced pluripotent stem cells (iPSCs) were generated from two patients with citrin deficiency and differentiated into hepatocyte-like cells (HLCs). Citrin-deficient HLCs produced albumin and liver-specific markers but completely lacked citrin protein and expressed argininosuccinate synthase only weakly. In addition, ammonia concentrations in a medium cultured with citrin-deficient HLCs were higher than with control HLCs. Sodium pyruvate administration significantly reduced ammonia concentrations in the medium of citrin-deficient HLCs and slightly reduced ammonia in HLCs differentiated from control iPSCs, though this change was not significant. Our results suggest that sodium pyruvate may be an efficient treatment for patients with citrin deficiency. Citrin-deficient iPSCs are a pathological liver model for congenital urea cycle disorders to clarify pathogenesis and develop novel therapies.

Impact of Polypharmacy and Risk Factors for Exacerbation of Lower Urinary Tract Symptoms in Patients with Urological Conditions: A Retrospective Study in a Japanese Municipal Hospital.

Polypharmacy exacerbates lower urinary tract symptoms (LUTS). Japan exhibits a higher prevalence of concomitant medication use in drug therapy than other countries. Previous age- and sex-specific reports exist; however, none include patients of all ages. Therefore, this retrospective study determined the impact of polypharmacy and its associated risk factors on LUTS exacerbation in outpatients with urological conditions. We included patients receiving medication who visited the Department of Urology at the Gifu Municipal Hospital (Gifu, Japan) between January, 2018 and December, 2018. The association between LUTS and polypharmacy and the risk factors for LUTS exacerbation were investigated. Patients were categorized into two groups according to their polypharmacy status. We performed propensity score matching and compared the International Prostate Symptom Score (IPSS) between the groups using the unpaired t-test. Multiple logistic regression analysis was performed to examine the risk factors, including "polypharmacy" and "taking multiple anticholinergic medications" for LUTS exacerbation. When comparing the IPSS between the groups, the polypharmacy group was found to have significantly higher scores than the non-polypharmacy group in six items, including "total score" and "storage score." Multiple logistic regression analysis results showed high significance in three items, including "polypharmacy" (odds ratio (OR) = 1.67, 95% confidence interval (CI): 1.03-2.71) and "taking multiple anticholinergic medications" (OR = 8.68, 95% CI: 1.05-71.7). In conclusion, this study revealed that "polypharmacy" and "taking multiple anticholinergic medications" were risk factors for LUTS. Particularly, "polypharmacy" is associated with storage symptom exacerbation. Therefore, eliminating "polypharmacy" and "taking multiple anticholinergic medications" is expected to improve LUTS.

Anatomical View of the Internal Carotid Artery Occlusion in Japanese Black Cattle.

The internal carotid artery (ICA) is a branch of the common carotid artery (CCA), along with the external carotid artery (ECA), which together provide the blood supply for the brain. The description of the ICA in cattle is vague, including denial of its existence or degeneration at an early stage after birth. This anatomical study investigated the internal carotid artery in Japanese black cattle. Sixty-five heads of Japanese black cattle aged from newborn to 13 years were dissected and injected with colored latex from the CCA after separating the head and body. Diameter measurements of the artery branches from the CCA on its bifurcation were conducted. Furthermore, a histological examination of the ICA wall's structures, which consist of the tunica intima, tunica media, and tunica externa, was performed. The ICA of Japanese black cattle is closed on the left side after age 3 years, except for a small lumen at 13 years, whereas the right ICA remains open at all ages. The location of occlusion of the left internal carotid artery (LICA) shows thickness of the tunica intima and an increased connective tissue layer area. The diameter of the ICA does not differ between the left and right sides, and there is no correlation with age. Therefore, further studies are needed, especially of ICA occlusion related to Japanese black cattle's physiology or cerebrospinal disease.

Comparative Analysis of Maternal Colostrum and Colostrum Replacer Effects on Immunity, Growth, and Health of Japanese Black Calves.

Maternal colostrum (MC) is an important source of nutrients and immune factors for newborn calves. However, when colostrum is unavailable or of poor quality, a colostrum replacer (CR) may be a suitable alternative to MC. As stock-raising farmers must make informed decisions about colostrum feeding management, this study was conducted to determine the effect of feeding MC versus CR on the promotion of immunological status, growth, and health in pre-weaned Japanese black (JB) calves. Sixteen newborn JB calves were fed MC after birth, and 16 JB calves were fed CR. For the MC group, the numbers of γδ T cells, CD4+ cells, CD8+ cells, CD4+CD8+ cells, B cells, and MHC class II+ cells were significantly higher compared with the CR group. Furthermore, the expression levels of interleukin (IL)-1ß-, IL-2-, and interferon-γ (IFN-γ)-encoding mRNAs were significantly higher in the MC group compared with the CR group. A lower incidence of disease in 1-month-old calves and higher carcass weight in the MC group were observed compared with the CR group. These results suggest that CR activates the immune system delayed in calves compared with MC. MC increases populations of various immunocompetent cells, which can reduce infection rates and improve body weight gain.

Probable effects of polypharmacy and equivalent doses of psychotropic drugs on prevalence of adverse drug events among psychiatric inpatients in a general hospital in Japan.

OBJECTIVE: In psychiatry, polypharmacy or high psychotropic drug doses increase adverse drug event (ADE) prevalence. However, the full relationship between polypharmacy and ADEs is unclear, and few studies have evaluated dose equivalents for psychotropic drugs for ADEs. Thus, we conducted a retrospective analysis to clarify the effects of polypharmacy and chlorpromazine (CP)-, diazepam (DAP)-, and imipramine- equivalent doses on all ADEs in inpatients. METHODS: Psychiatric inpatients in a Japanese hospital from April 1, 2016 to March 31, 2018, were enrolled. ADE severity and causality were assessed. Multiple logistic regression analyses were performed to evaluate ADE risk factors. RESULTS: Among 462 patients analyzed, out of 471 patients enrolled, 145 (31.4%) experienced ADEs. The causality assessment determined that "possible" was 96.5%. The most common ADEs were nervous system disorders (35%). Multiple logistic regression analyses indicated an increase in ADE prevalence with the number of drugs used (≥5; p = 0.026); CP-equivalent dose (p = 0.048); and endocrine, nutritional, and metabolic disorders (p = 0.045). DAP-equivalent dose; infectious and parasitic diseases; and injury, poisoning, and consequences of other external causes decreased ADE prevalence (p = 0.047, 0.022, and 0.021, respectively). CONCLUSIONS: Avoiding polypharmacy in psychiatric inpatients and adjusting drug regimens to safe equivalent doses could reduce ADEs during hospitalization.

Effects of Maternal Supplementation with Organic Trace Minerals including Zinc, Manganese, Copper, and Cobalt during the Late and Post-Partum Periods on the Health and Immune Status of Japanese Black Calves.

In this study, we evaluated the effects of supplementation of the maternal diet with organic trace minerals including Zn (zinc), Mn (manganese), Cu (copper), and Co (cobalt) on the health and immune status of beef calves. We examined 19 pregnant cows, which were divided into a group of 9 cows fed a basal diet (control) and 10 cows fed a diet with organic trace minerals (treated). Cows were fed for a period of 45 days before the predicted calving date until 45 days after calving. The number of treatments needed for respiratory and digestive diseases within 14 days of birth was significantly lower in the treated group (p < 0.05) than the control group. In addition, the concentration of serum zinc in the treated group on day 1 was significantly higher (p < 0.05) than that in the control group. The numbers of CD4+ and CD8+ cells in the treated group on days 30 and 60 were significantly increased (p < 0.01) compared with those in the control group, as was the number of γδ T cells on days 1 and 30 (p < 0.05). The number of IgM+ cells in the treated group on days 30 and 60 was significantly increased (p < 0.01) compared with that in the control group, as was the number of MHC class II+ cells on day 60 (p < 0.01). The number of NK cells in the treated group on day 60 was also significantly increased (p < 0.05) compared with that in the control group. The expression levels of mRNAs encoding interlukin-2 (IL-2), interlukin-4 (IL-4), interlukin-12 (IL-12), and interferon-γ (IFN-γ) in the treated group were significantly higher than those in the control group (p < 0.05) on days 1 and 60. The results indicate that maternal supplementation with trace minerals is a promising approach for producing highly disease-resistant calves and enhancing calf immunity.

[Effectiveness of Pharmaceutical Intervention While Administering 5-Aminolevulinic Acid Hydrochloride through a Collaboration of Physicians and Pharmacists].

5-aminolevulinic acid hydrochloride is a highly effective drug in reducing tumor residuals in transurethral resection of the bladder tumors; however, hypotension is a serious side effect that causes clinical problems. To avoid serious side effects, a pharmacist, in consultation with a physician, decided to discontinue the antihypertensive medication, and the effect of this pharmaceutical intervention was examined retrospectively. This study included patients who received 5-aminolevulinic acid hydrochloride at Gifu Municipal Hospital and were instructed to continue receiving their usual antihypertensive medication on the day of surgery. The control group comprised 17 patients before the pharmaceutical intervention, and the intervention group comprised 18 patients after the pharmaceutical intervention. The difference in systolic blood pressure before and after 5-aminolevulinic acid hydrochloride administration was -19.4±22.5 mmHg in the control group and -2.8±16.0 mmHg in the intervention group. The intervention group showed a significantly lower decrease in blood pressure(p=0.019). Intervention to avoid hypotension through the collaboration between physicians and pharmacists may be effective in improving the safety of 5-aminolevulinic acid hydrochloride.

Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR.

The mechanistic/mammalian target of rapamycin (mTOR) is involved in a wide range of cellular processes. However, the role of mTOR in podocytes remains unclear. In this study, we aimed to clarify the role of mTOR in podocyte differentiation from human induced pluripotent stem cells (hiPSCs) and to establish an efficient differentiation protocol for human podocytes. We generated podocytes from hiPSCs by modifying protocol. The expression of the podocyte-specific slit membrane components nephrin and podocin was measured using PCR, western blotting, flow cytometry, and immunostaining; and the role of mTOR was evaluated using inhibitors of the mTOR pathway. Nephrin and podocin were found to be expressed in cells differentiated from hiPSCs, and their expression was increased by mTOR inhibitor treatment. S6, a downstream component of the mTOR pathway, was also found to be involved in podocyte differentiation. we evaluated its permeability to albumin, urea, and electrolytes. The induced podocytes were permeable to the small molecules, but only poorly permeable to albumin. We have shown that the mTOR pathway is involved in podocyte differentiation. Our monolayer podocyte differential protocol, using an mTOR inhibitor, provides a novel in vitro model for studies of kidney physiology and pathology.

Melphalan Febrile Neutropenia Risk Factors.

This study aimed to identify the risk factors of febrile neutropenia(FN)onset associated with melphalan(L-PAM)therapy. Thirty-nine patients(21 men, 18 women)were administered L-PAM intravenously for multiple myeloma(MM)from April 2011 to February 2022 at the Department of Hematology of Gifu Municipal Hospital. Patients were classified into those with and without FN(Grade 3 or higher), complete blood count and liver function tests were performed immediately before starting therapy. Univariate analysis with Fisher's exact probability test was performed. Factors with p<0.2 were considered as independent variables for multivariate analysis in the multiple logistic regression analysis. A multivariate analysis with 2 independent variables, lactate dehydrogenase(LD)level>222 U/L(upper limit of the facility reference value)and white <3.3×103/µL(lower limit of the facility reference value)from the univariate analysis, and FN onset(Grade 3 or higher)as the dependent variable showed that LD level>222 U/L(odds ratio: 6.33, 95% confidence interval: 1.12-35.8, p=0.037)was a significant factor. In conclusion, patients with LD levels >222 U/L immediately before starting therapy require adequate monitoring for FN onset following L-PAM administration.

The molecular signals that regulate activity-dependent synapse refinement in the brain.

The formation of appropriate synaptic connections is critical for the proper functioning of the brain. Early in development, neurons form a surplus of immature synapses. To establish efficient, functional neural networks, neurons selectively stabilize active synapses and eliminate less active ones. This process is known as activity-dependent synapse refinement. Defects in this process have been implicated in neuropsychiatric disorders such as schizophrenia and autism. Here we review the manner and mechanisms by which synapse elimination is regulated through activity-dependent competition. We propose a theoretical framework for the molecular mechanisms of synapse refinement, in which three types of signals regulate the refinement. We then describe the identity of these signals and discuss how multiple molecular signals interact to achieve appropriate synapse refinement in the brain.

Prevalence of anatomic variants in coronary arteries of Japanese Black cattle.

Coronary artery anomalies that can cause sudden cardiac death have been described in some mammals. However, few studies have investigated coronary anomalies and coronary artery branching patterns in cattle. Therefore, understanding of bovine coronary arteries is incomplete. The aim of this study was to clarify anatomic variants in coronary arteries of Japanese Black cattle. The gross anatomy of the coronary arteries was examined in the hearts of 100 cattle. A total of 14 of 100 hearts exhibited confirmed coronary anomalies in the left main trunk. These 14 hearts were classified into 5 types potentially associated with cardiac ischemia. Regarding the coronary branches, the subsinuosal interventricular branch originated from the left circumflex branch (LCX) in 98 hearts and from the right coronary artery in 2 hearts. The origin of the sinoatrial nodal branch was classified into 3 types. In 99 hearts, the atrioventricular nodal branch originated from the LCX at the heart crux. This study characterized the prevalence of coronary anomalies, branching patterns of coronary arteries, and coronary artery anatomic variants in Japanese Black cattle.

Protective effects of probiotics against tannin-induced immunosuppression in broiler chickens.

Tannins (TAs) are an anti-nutritional substance commonly used as a natural feed additive for livestock. However, our previous study described the dose-dependent adverse effects of TA on immune responses and growth in chickens. In this study, we evaluated the protective effects of a probiotic preparation (BT) consisting of three different bacteria (Bacillus mesenteric, Clostridium butyricum, and Streptococcus faecalis) against TA-induced immunosuppression in chickens. Forty chicks were divided into 4 groups as follows: the CON group (basal diet), BT group supplemented with 3 g BT/kg diet, tannic acid (TA) group supplemented with 30 g TA/kg diet, and BT+TA group supplemented with 3 g BT/kg diet + 30 g TA/kg diet. The feeding trial lasted for 35 days. Lymphocyte subset, macrophage phagocytosis, cytokine mRNA expression, and primary and secondary IgY immune responses were evaluated. BT supplementation significantly improved TA-induced reductions in final body weight, body weight gain, feed intake, and relative weights of lymphoid organs compared with the TA group. Furthermore, in the spleen and cecal tonsil (CT), the relative populations of CD4+, CD8+, and CD4+CD8+ cells in the BT+TA group were significantly ameliorated compared with the TA group. Additionally, comparison with the TA group showed that the chickens in the BT+TA group had an improved relative population of B cells in the CT and that macrophage phagocytosis in the spleen was significantly increased. Chickens in the BT+TA group showed significant increases in IFN-γ and IL-4 mRNA expression in the spleen compared with the TA group. The primary and secondary IgY responses were significantly improved. These results revealed that supplementation with BT protects against TA-induced immunosuppression in chickens.

Feeding ecology of Swinhoe's tree lizard (Diploderma swinhonis (Günther, 1864)) in Hyuga City, Miyazaki Prefecture, Japan.

Swinhoe's tree lizard (Diploderma swinhonis: D. swinhonis) is an arboreal agamid that is native to Taiwan. In Taiwan, the lizard is considered to be a generalist that feeds primarily on ants and a diversity of small insect prey by employing an opportunistic sit-and-wait foraging strategy. In Japan, D. swinhonis is considered as an invasive alien species that was discovered in Hyuga city, Miyazaki Prefecture, in 2016. Despite concerns about the impact of D. swinhonis on native fauna, little information about the diet of this alien species has been published to date. This study, therefore, investigated the feeding ecology of D. swinhonis in Hyuga city to evaluate their potential impact on the ecosystem. Specifically, prey preference was investigated by examining the stomach contents of males, females, and juveniles captured from April to December 2020 and in March 2021. The results showed that the lizards in Hyuga preyed upon a wide variety of invertebrates as in Taiwan, while ants accounted for the largest proportion of the prey items consumed regardless of sex, age or changes in season. These findings indicated that D. swinhonis might cause a decrease in the abundance of the native insect fauna of Hyuga city or competition with native lizards for foods in Hyuga city. Since its impact is not currently apparent, it's necessary to monitor its effect in the future.

Morphology and morphometry of the inner ear of the dromedary camel and their influence on the efficiency of hearing and equilibrium.

BACKGROUND: The inner ear morphology and size are linked to hearing and balance ability. The goal of this study was to determine the morphology and morphometrics of the dromedary camel's inner ear and how it influences hearing accommodation and equilibrium in the desert environment. MATERIALS AND METHODS: Gross morphology, computed tomography images, and the endocast were used to show the inner ear morphology. A caliper and ImageJ software were used to take measurements on a plastic endocast. RESULTS: The presence of the subarcuate fossa, flat cochlea, radii curvature of the semicircular canals, particularly the lateral semicircular canal, orthogonality, and the union between the semicircular canals, along with slightly increased saccule and utricle size, maintains camel balance on sandy ground, even during heavy sandstorms. The cochlear basilar membrane length and cochlea radii ratio aided low-frequency hearing and perception over a wide octave range. CONCLUSION: The camel's cochlear characteristics revealed a lengthy basilar membrane, a high radii ratio, 3.0 cochlear canal turns, and a very broad cochlea. The orthogonality of the semicircular canals, the high curvature of the lateral semicircular canal, the presence of the subarcuate fossa, and the confluence between the lateral and posterior semicircular canal were particular specifications that allowed the inner ear of the camel to adapt to desert living.

Toltrazuril and diclazuril: comparative evaluation of anti-coccidial drugs using a murine model.

Intestinal coccidiosis caused by Eimeria protozoan species is an economically important disease, especially in poultry and cattle. Anti-coccidial drugs commonly used for controlling coccidiosis are toltrazuril (TTZ) and diclazuril (DCZ). In this study, the efficacies of TTZ and DCZ were compared using a murine model, and the effect of these treatments on the induction of acquired resistance was evaluated. Male C57BL/6J mice were inoculated with 1,000 sporulated E. vermiformis oocytes and treated with TTZ or DCZ. The recommended TTZ dose for cattle (15 mg/kg) completely prevented oocyte excretion. But, mice required 5 mg/kg of DCZ, which is five times the recommended dose for cattle, to reduce oocyte excretion. In E. vermiformis re-infection, TTZ (15 mg/kg) and DCZ (5 mg/kg) treatments did not interfere with the development of acquired resistance. Bodyweight gain was significantly higher in the TTZ-treated group than in the control (untreated/infected) group and the DCZ-treated group, and no significant difference in bodyweight gain was observed between the TTZ-treated group and the healthy (uninfected/untreated) group. Analysis of T lymphocyte subsets in the spleen and mesenteric lymph nodes indicated that the relative populations of CD4+ and CD8+ T cells were reduced in the DCZ-treated and control (untreated/infected) groups, suggesting there was immunosuppression during the infection. However, no reductions in T cell populations were observed in the TTZ-treated group. The results indicated that an optimal anti-coccidial drug is one that can completely break the parasite life cycle in the host animal.

Effects of Probiotics on Growth and Immunity of Piglets.

Growth promoting antibiotics are used in modern animal husbandry to promote growth and avoid infections. Negative effects of these antibiotics on human health are a big concern and they need to be replaced. Probiotics are expected to be a good replacement for growth promoting antibiotics. In this study, we evaluated effects of probiotics on growth and immune status of liver and secondary lymphoid organs of piglets. Body weights of probiotic-fed piglets were heavier than those of control piglets (p < 0.05) at days 30 and 45 of the experiment. Relative populations of CD4+ and IgM+ cells isolated from the liver were significantly increased (p < 0.05 and p < 0.01, respectively) in probiotic-fed piglets compared to control piglets. CD4+CD8+ T cells were significantly decreased (p < 0.05) in jejunal Peyer's patches of treated piglets. Phagocytosis of MHC class II+ cells isolated from the liver of probiotic-fed piglets was significantly higher (p < 0.05) than that of control piglets. Phagocytosis of granulocytes isolated from the liver and peripheral blood of probiotic-fed piglets were also higher than those of control piglets. These results indicate excellent effects on growth and immune status of piglets. In conclusion, probiotics have beneficial effects on the growth and health of piglets and could be good replacement for growth promoting antibiotics.