Comparative mapping of crawling-cell morphodynamics in deep learning-based feature space.

Navigation of fast migrating cells such as amoeba Dictyostelium and immune cells are tightly associated with their morphologies that range from steady polarized forms that support high directionality to those more complex and variable when making frequent turns. Model simulations are essential for quantitative understanding of these features and their origins, however systematic comparisons with real data are underdeveloped. Here, by employing deep-learning-based feature extraction combined with phase-field modeling framework, we show that a low dimensional feature space for 2D migrating cell morphologies obtained from the shape stereotype of keratocytes, Dictyostelium and neutrophils can be fully mapped by an interlinked signaling network of cell-polarization and protrusion dynamics. Our analysis links the data-driven shape analysis to the underlying causalities by identifying key parameters critical for migratory morphologies both normal and aberrant under genetic and pharmacological perturbations. The results underscore the importance of deciphering self-organizing states and their interplay when characterizing morphological phenotypes.

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.

Optogenetic relaxation of actomyosin contractility uncovers mechanistic roles of cortical tension during cytokinesis.

Actomyosin contractility generated cooperatively by nonmuscle myosin II and actin filaments plays essential roles in a wide range of biological processes, such as cell motility, cytokinesis, and tissue morphogenesis. However, subcellular dynamics of actomyosin contractility underlying such processes remains elusive. Here, we demonstrate an optogenetic method to induce relaxation of actomyosin contractility at the subcellular level. The system, named OptoMYPT, combines a protein phosphatase 1c (PP1c)-binding domain of MYPT1 with an optogenetic dimerizer, so that it allows light-dependent recruitment of endogenous PP1c to the plasma membrane. Blue-light illumination is sufficient to induce dephosphorylation of myosin regulatory light chains and a decrease in actomyosin contractile force in mammalian cells and Xenopus embryos. The OptoMYPT system is further employed to understand the mechanics of actomyosin-based cortical tension and contractile ring tension during cytokinesis. We find that the relaxation of cortical tension at both poles by OptoMYPT accelerated the furrow ingression rate, revealing that the cortical tension substantially antagonizes constriction of the cleavage furrow. Based on these results, the OptoMYPT system provides opportunities to understand cellular and tissue mechanics.

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.

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.

Collateral uterine blood flow in Holstein cows during the third trimester of pregnancy.

Blood flow to the gravid and nongravid uterine horns of four multiparous Holstein cows (mean +/- SD, BW=641.8 +/- 95.4 kg; age=4.8 +/- 1.2 years; parity=3.0 +/- 1.2) was measured on days 225, 248, and 266 of gestation. Surgery was conducted on day 214.5 +/- 4.0 of gestation through the flank of the standing cows. Transit-time ultrasonic flow probes (diameter 12 or 14 mm) were fitted surgically around the uterine arteries of each cow. Surgery was completed within two hours of anesthesia, and the animals recovered rapidly following surgery. Uterine blood flow (UBF, l/min) was recorded at 10 sec intervals for approximately 23.5 hours; these values were averaged to determine UBF. The mean gravid UBF was significantly (P<0.05) greater than the nongravid UBF in this study. The range of the gravid and nongravid UBFs varied from 3.61 to 14.05 and 0.72 to 6.54 l/min, respectively. There were no changes (P>0.1) in the mean gravid and nongravid UBFs from day 225 to 266 of gestation.