Transcription factor MafB in podocytes protects against the development of focal segmental glomerulosclerosis.

Focal segmental glomerulosclerosis (FSGS) is a common cause of steroid-resistant nephrotic syndrome. Spontaneous remission of FSGS is rare and steroid-resistant FSGS frequently progresses to renal failure. Many inheritable forms of FSGS have been described, caused by mutations in proteins that are important for podocyte function. Here, we show that a basic leucine zipper transcription factor, MafB, protects against FSGS. MAFB expression was found to be decreased in the podocytes of patients with FSGS. Moreover, conditional podocyte-specific MafB-knockout mice developed FSGS with massive proteinuria accompanied by depletion of the slit diaphragm-related proteins (Nphs1 and Magi2), and the podocyte-specific transcription factor Tcf21. These findings indicate that MafB plays a crucial role in the pathogenesis of FSGS. Consistent with this, adriamycin-induced FSGS and attendant proteinuria were ameliorated by MafB overexpression in the podocytes of MafB podocyte-specific transgenic mice. Thus, MafB could be a new therapeutic target for FSGS.

Improving quality of common reed (Phragmites communis Trin.) silage with additives.

OBJECTIVE: Common reed (Phragmites communis Trin.) could potentially provide an alternative resource for silage; however, its silage quality is poor. The aim of this study was to investigate the factors in reed that contribute to poor quality and determine how the use of additives at ensiling could improve fermentation quality. METHODS: In Experiment 1, we determined the chemical composition and the presence of indigenous lactic acid bacteria (LAB) in reed. We further examined fermentation quality of reed silage under conditions without additives (NA) and treated glucose (G), lactic acid bacteria (L), and their combination (G+L). In Experiment 2, silage of NA, and with an addition of cellulase and lactic acid bacteria (CL) were prepared from harvested reed. The harvested reeds were fertilized at nitrogen concentrations of 0, 4, 8, and 12 g N/m2 and were harvested thrice within one year. RESULTS: The indigenous LAB and fermentable carbohydrates are at extremely low concentrations in reed. Reed silage, to which we added G+L, provided the highest quality silage among treatments in Experiment 1. In Experiment 2, N fertilization had no negative effect on silage quality of reed. The harvest times decreased fermentable carbohydrate content in reed. The CL treatment provided a higher lactic acid content compared to the NA treatment. However, the quality of CL treated silage at the second and third harvests was significantly lower than at the first harvest, due to a reduction in carbohydrates caused by frequent harvesting. CONCLUSION: The causes of poor quality in reed silage are its lack of indigenous LAB and fermentable carbohydrates and its high moisture content. In addition, reed managed by frequent harvesting reduces carbohydrate content. Although the silage quality could be improved by adding CL, higher-quality silage could be prepared by adding fermentable carbohydrates, such as glucose (rather than adding cellulases).

Identification of further diversity among posaviruses.

Recently, there have been reports of new members of posavirus-like viruses in the order Picornavirales. In this study, using a metagenomics approach, 11 posavirus-like sequences (>7,000 nucleotides) were detected in 155 porcine fecal samples. Phylogenetic analysis revealed that the newly identified virus sequences, together with other posavirus-like viruses, form distinct clusters within the order Picornavirales, composed of eight genogroups and unassigned sequences based on amino acid sequences of the helicase and RNA-dependent RNA polymerase regions, with <40 % and <50 % sequence identity, respectively. We propose further classifications of highly diverse posavirus populations based on newly identified sequences from Japanese pig feces.

Non-randomized controlled trial examining the effects of livestock on motivation and anxiety in patients with chronic psychiatric disorders.

Objectives: Patients with chronic schizophrenia exhibit negative symptoms, including decreased work motivation. Animal-assisted therapy programs have been reported to benefit such patients; hence, there is a possibility that sheep-rearing, rather than conventional employment training, may motivate these patients. Therefore, we investigated the effects of a one-day experiential learning program of sheep-rearing on the work motivation and anxiety of patients with chronic schizophrenia. Methods: Fourteen patients were included in a non-randomized controlled trial conducted between August 2018 and October 2018. The patients' participation in the sheep-rearing experiential learning (one day; intervention day) and normal day care (one day; control day) programs were compared. The salivary cortisol and testosterone levels and State-Trait Anxiety Inventory (STAI) scores of the patients were analyzed. Results: The patients' salivary testosterone was significantly higher on the intervention day (p = 0.04) than on the control day (p = 0.02). Their salivary cortisol was lower on the control day than on the intervention day, although the difference was not significant. Regression analysis was performed based on the change in salivary cortisol levels and STAI-Trait scores (p = 0.006), and a regression equation was established. Conclusions: The study revealed that participation in sheep-rearing may have promoted the testosterone production but did not increase anxiety in patients with schizophrenia. Additionally, regression equations for salivary cortisol levels in such patients may provide information on individual differences in anxiety levels.

Effect of climatic factors on urinary cortisol and peripheral blood leukocytes in lambs grazing on a semi-natural grassland in the Hokuriku District of Japan.

The effects of climatic factors on stress and immune functions of grazing lambs in summer and autumn in the Hokuriku District of Japan were evaluated by determining urinary cortisol (U-COR) levels and peripheral blood leukocyte populations and comparing those with lambs kept indoors. Two groups of five lambs, consisting of those grazed on a semi-natural grassland (GRL) and those housed indoors in a domestic animal shelter (INL), were maintained at from July to October. The temperature-humidity index at each location was indicative of heat stress during summer; however, the U-COR elevation was not observed in both groups. The elevation was observed in GRL in autumn and was higher than INL in October. Climatic conditions in autumn were characterized by high humidity and a sudden drop in temperature. U-COR was positively correlated with the relative humidity. The GRL was exposed to low-nutrient conditions for a relatively long time. The CD4+ /CD8+ T cell ratio in GRL decreased in October. Subsequently, the total leucocyte, including granulocytes, monocytes, and lymphocytes, sharply increased. The responses indicated an immune deficiency caused by immunosuppression because of a low nutrition caused by grazing and high-stress conditions in autumn.

Generation of reconstituted hemato-lymphoid murine embryos by placental transplantation into embryos lacking HSCs.

In order to increase the contribution of donor HSC cells, irradiation and DNA alkylating agents have been commonly used as experimental methods to eliminate HSCs for adult mice. But a technique of HSC deletion for mouse embryo for increase contribution of donor cells has not been published. Here, we established for the first time a procedure for placental HSC transplantation into E11.5 Runx1-deficient mice mated with G1-HRD-Runx1 transgenic mice (Runx1-/-::Tg mice) that have no HSCs in the fetal liver. Following the transplantation of fetal liver cells from mice (allogeneic) or rats (xenogeneic), high donor cell chimerism was observed in Runx1-/-::Tg embryos. Furthermore, chimerism analysis and colony assay data showed that donor fetal liver hematopoietic cells contributed to both white blood cells and red blood cells. Moreover, secondary transplantation into adult recipient mice indicated that the HSCs in rescued Runx1-/-::Tg embryos had normal abilities. These results suggest that mice lacking fetal liver HSCs are a powerful tool for hematopoiesis reconstruction during the embryonic stage and can potentially be used in basic research on HSCs or xenograft models.

Digestibility of common reed (Pharagmites communis Trin.) silage as ruminant feed and effects of inclusion levels in the diet of breeding cows on feed intake, ruminal fermentation and blood metabolites.

To examine the digestibility of common reed (Pharagmites communis Trin.) silage and effects of inclusion levels in the diet of breeding cows on nutrient intakes, ruminal fermentation and nutrient status, two digestion trials using four sheep and cows, respectively, at the maintenance stage were conducted. In the trial using sheep, common reed silage contained 20% crude protein (CP) and 50% total digestible nutrients (TDN) on a dry matter basis. In the trial using cows, treatments were the control (92.4% Sudangrass hay and 7.6% soybean meal) and CS25, CS50 and CS80 (replacing 25%, 50%, and 80% of the control with silage, respectively). The intake and sufficiency rate of CP increased by increasing the silage level in the diet (P < 0.05), and the CP requirements of cows at maintenance was satisfied in CS80. In contrast, the sufficiency rate of TDN decreased in CS80, although it was 99.8%. Ruminal pH and NH3 -N concentration changed among the treatments (P < 0.05); however, the blood metabolites were not affected by silage. The results indicated that including common reed silage of up to 80% in the diet of breeding cows was possible and did not have adverse effects on the blood metabolites.

Effects of inclusion levels of pelleted silvergrass (Miscanthus sinensis Andress.) in the diet on digestibility, chewing activity, ruminal fermentation and blood metabolites in breeding Japanese Black cows.

To examine the effects of inclusion levels of pelleted silvergrass (PS) in the diet on digestibility, ruminal fermentation and nutrient status of breeding Japanese Black cows, four cows were allotted to a 4 × 4 Latin square design experiment. Treatments were control fed a diet consisting of 89.4% Sudangrass hay and 10.6% soybean meal on a dry matter (DM) basis, and PS18, PS27 and PS45 fed the diet replaced with 18%, 27% and 45% of control with PS, respectively. The total digestible nutrients (TDN) content of PS was 45.6% on a DM basis. The TDN intakes were significantly decreased by increasing PS level in the diet (P < 0.05), but were higher than the TDN requirement of maintenance cows in all treatments. The total chewing time was decreased significantly by increasing PS level in the diets (P < 0.05). However, the pH and concentration of volatile fatty acid in the ruminal fluid and serum metabolite concentrations were not significantly different among the treatments. The results suggested that including PS up to 45% in the diet did not have adverse effects on the ruminal fermentation and nutrient status in breeding Japanese Black cows at the maintenance stage.

MafB is a critical regulator of complement component C1q.

The transcription factor MafB is expressed by monocytes and macrophages. Efferocytosis (apoptotic cell uptake) by macrophages is important for inhibiting the development of autoimmune diseases, and is greatly reduced in Mafb-deficient macrophages. Here, we show the expression of the first protein in the classical complement pathway C1q is important for mediating efferocytosis and is reduced in Mafb-deficient macrophages. The efferocytosis defect in Mafb-deficient macrophages can be rescued by adding serum from wild-type mice, but not by adding serum from C1q-deficient mice. By hemolysis assay we also show that activation of the classical complement pathway is decreased in Mafb-deficient mice. In addition, MafB overexpression induces C1q-dependent gene expression and signals that induce C1q genes are less effective in the absence of MafB. We also show that Mafb-deficiency can increase glomerular autoimmunity, including anti-nuclear antibody deposition. These results show that MafB is an important regulator of C1q.

Diversity in VP3, NSP3, and NSP4 of rotavirus B detected from Japanese cattle.

Bovine rotavirus B (RVB) is an etiological agent of diarrhea mostly in adult cattle. Currently, a few sequences of viral protein (VP)1, 2, 4, 6, and 7 and nonstructural protein (NSP)1, 2, and 5 of bovine RVB are available in the DDBJ/EMBL/GenBank databases, and none have been reported for VP3, NSP3, and NSP4. In order to fill this gap in the genetic characterization of bovine RVB strains, we used a metagenomics approach and sequenced and analyzed the complete coding sequences (CDS) of VP3, NSP3, and NSP4 genes, as well as the partial or complete CDS of other genes of RVBs detected from Japanese cattle. VP3, NSP3, and NSP4 of bovine RVBs shared low nucleotide sequence identities (63.3-64.9% for VP3, 65.9-68.2% for NSP3, and 52.6-56.2% for NSP4) with those of murine, human, and porcine RVBs, suggesting that bovine RVBs belong to a novel genotype. Furthermore, significantly low amino acid sequence identities were observed for NSP4 (36.1-39.3%) between bovine RVBs and the RVBs of other species. In contrast, hydrophobic plot analysis of NSP4 revealed profiles similar to those of RVBs of other species and rotavirus A (RVA) strains. Phylogenetic analyses of all gene segments revealed that bovine RVB strains formed a cluster that branched distantly from other RVBs. These results suggest that bovine RVBs have evolved independently from other RVBs but in a similar manner to other rotaviruses. These findings provide insights into the evolution and diversity of RVB strains.

Ground-based assessment of JAXA mouse habitat cage unit by mouse phenotypic studies.

The Japan Aerospace Exploration Agency developed the mouse Habitat Cage Unit (HCU) for installation in the Cell Biology Experiment Facility (CBEF) onboard the Japanese Experimental Module ("Kibo") on the International Space Station. The CBEF provides "space-based controls" by generating artificial gravity in the HCU through a centrifuge, enabling a comparison of the biological consequences of microgravity and artificial gravity of 1 g on mice housed in space. Therefore, prior to the space experiment, a ground-based study to validate the habitability of the HCU is necessary to conduct space experiments using the HCU in the CBEF. Here, we investigated the ground-based effect of a 32-day housing period in the HCU breadboard model on male mice in comparison with the control cage mice. Morphology of skeletal muscle, the thymus, heart, and kidney, and the sperm function showed no critical abnormalities between the control mice and HCU mice. Slight but significant changes caused by the HCU itself were observed, including decreased body weight, increased weights of the thymus and gastrocnemius, reduced thickness of cortical bone of the femur, and several gene expressions from 11 tissues. Results suggest that the HCU provides acceptable conditions for mouse phenotypic analysis using CBEF in space, as long as its characteristic features are considered. Thus, the HCU is a feasible device for future space experiments.