Supplementation of rumen-protected lysine during the close-up period improves vaginal discharge clearance in Holstein dairy cows.

We aimed to evaluate the effects of rumen-protected lysine (RPL) supplementation during the close-up period on uterine involution and the resumption of ovarian function in dairy cows. Fifty-two multiparous Holstein cows were categorized based on parity and expected calving date and randomly assigned to the RPL or control (CON) groups. The RPL group received 80 g of RPL daily from day 21 before the expected calving date until parturition. Blood samples were obtained twice weekly from pre-supplementation to 6 weeks postpartum. The onset of luteal activity postpartum was determined via ultrasonography twice weekly for up to 6 weeks postpartum. Uterine involution was tracked at 3 and 5 weeks postpartum through the vaginal discharge score, percentage of polymorphonuclear cells (PMN) in endometrial cytology samples, presence of intrauterine fluid, and gravid horn diameter via ultrasonography. Before supplementation, the RPL group showed amino acid imbalance, which was improved by RPL supplementation. There were no significant differences in the onset of luteal activity, percentage of PMN, intrauterine fluid, or the diameter of the uterine horn between the two groups. The vaginal discharge score in the RPL group decreased from 3 to 5 weeks postpartum, whereas that in the CON groups did not decrease. The number of cows with clinical endometritis was lower in the RPL group. Overall, RPL supplementation during the close-up period enhanced vaginal discharge clearance, potentially averting clinical endometritis, but did not affect the first ovulation in dairy cows.

Effect of 2% meloxicam injection in Holstein dairy cows on acute clinical mastitis without systemic symptoms.

This study aimed the efficacy of meloxicam (MX) in treating acute clinical mastitis (ACM) without systemic symptoms in Holstein cows by studying improvement in udder pain, changes in prostaglandin E2(PGE2) and bradykinin (BK) levels in the milk, and milk yield (MY) after healing. Forty-two cows with ACM were randomly assigned to the MX treatment group (T group; n=21) and the control group (C group; n=21). At onset of illness (day 0), the T group received a 0.5 mg/kg subcutaneous (SC) injection of MX whereas the C group received 15 mL SC of saline solution as a placebo. Udder tenderness (UT) was measured, and milk samples were collected on days 0-3. There was little change in the MY of the T group before and after healing, whereas MY in the C group was significantly lower than after healing. UT on day 3 in the T group was significantly lower than that in the C group. PGE2 levels significantly decreased from day 0 to day 3 in both groups. A significant negative correlation between PGE2 and linear score was observed on day 1 in the T group, but not in the C group. In ACM without systemic symptoms, the administration MX may be useful for restoring MY and reducing udder pain after healing.

Association between somatic cell count or morbidity of chronic subclinical mastitis and longevity in dairy herds in Eastern Hokkaido, Japan: a cross-sectional study.

An epidemiological analysis was conducted on production records in Hokkaido, Japan, to investigate the potential association between improved milk quality and longevity outcomes. The study found significant variations in herd somatic cell count levels and chronic subclinical mastitis morbidity based on geographical area and herd size. The analysis also revealed a positive correlation between herd somatic cell count and chronic subclinical mastitis morbidity. Although the hypothesis of a causal link between milk quality and longevity was examined, no such association was found. However, intensive assistance for identified high-risk areas and farms is expected to enhance overall milk quality.

Individuals reciprocate negative actions revealing negative upstream reciprocity.

Indirect reciprocity is widely recognized as a mechanism for explaining cooperation and can be divided into two sub-concepts: downstream and upstream reciprocity. Downstream reciprocity is supported by reputation; if someone sees you helping someone else, the person who sees this will think higher of you, and you will be more likely to be helped. Upstream reciprocity is helping someone because you are being helped by somebody else, which often happens in everyday life and experimental games. This paper focuses on the behavior of "take" and examines negative upstream reciprocity using an upstream reciprocity framework. The term "take" is defined as "to steal rather than give resources to others." "If something is taken from you, do you take from others?" is an important extension for indirect reciprocity research; subsequently, this paper discusses experiments conducted on whether negative upstream reciprocity is chained and what causes it. The results demonstrated differences between positive and negative upstream reciprocity. In analyzing the data of nearly 600 participants to determine the extent to which negative upstream reciprocity is observed and the causes of negative upstream reciprocity, the study found that If individual A takes resources from individual B, then B is more likely to take resources from a third-party, individual C. Notably, some causes of positive upstream reciprocity were found to have no effect or the opposite effect on negative upstream reciprocity. The results also demonstrate that the first person to take can cause a chain reaction. This paper demonstrates the importance of the first person not taking from someone else and suggests the need to consider various behavioral options for future research on cooperation.

Efficiency of immunocastration with an anti-gonadotropin-releasing hormone vaccine on cryptorchid bulls.

Cryptorchid bulls have low economic value owing to the effects of masculinization. Moreover, surgical removal of an ectopic testis is difficult in certain clinical cases. Recently, immunocastration has garnered popularity as a nonsurgical castration method in pig farming; however, the effects of immunocastration on cryptorchid bulls are yet to be yet. Herein, we investigated endocrine changes due to immunocastration in cryptorchid bulls and studied its effectiveness. This study included 13 Holstein bulls diagnosed with cryptorchidism and classified into two groups based on pubertal period: <8 months of age (pregroup) and ≥8 months of age (postgroup). Antigonadotropin-releasing hormone (GnRH) vaccine was used for immunocastration, and two vaccine doses were administered. Blood testosterone and anti-Müllerian hormone (AMH) levels were measured and analyzed for endocrine evaluation. The testosterone levels significantly decreased following the start of immunocastration in both groups, thereby confirming the efficacy of antiGnRH vaccination in cryptorchid bulls. The AMH levels significantly increased in the pregroup with two antiGnRH vaccination, suggesting a compensatory response via the neutralization of GnRH antibodies. The AMH levels did not significantly change in the postgroup, indicating the partial suppression of AMH secretion in Sertoli cells during sexual maturation and failure of Sertoli cell maturation. Thus, we successfully restrained the serum testosterone levels in cryptorchid bulls using antiGnRH vaccine. The testosterone levels are a useful indicator of the immunocastration effect on cryptorchid bulls. Hereafter, a vaccine program that can sustain the castration effect on cryptorchid bulls is necessary.

Single Cell Analysis of the Fate of Injected Oncogenic RasV12 Cells in Adult Wild Type Drosophila.

We have injected dish-cultured oncogenic RasV12 cells into adult male flies and analyzed by single cell transcriptomics their destiny within the host after 11 days. We identified in the preinjection samples and in the 11-day postinjection samples in all 16 clusters of cells, of which 5 disappeared during the experiment in the host. The other cell clusters expanded and expressed genes involved in the regulation of cell cycle, metabolism, and development. In addition, three clusters expressed genes related to inflammation and defense. Predominant among these were genes coding for phagocytosis and/or characteristic for plasmatocytes (the fly equivalent of macrophages). A pilot experiment indicated that the injection into flies of oncogenic cells, in which two of most strongly expressed genes had been previously silenced by RNA interference, into flies resulted in a dramatic reduction of their proliferation in the host flies as compared to controls. As we have shown earlier, the proliferation of the injected oncogenic cells in the adult flies is a hallmark of the disease and induces a wave of transcriptions in the experimental flies. We hypothesize that this results from a bitter dialogue between the injected cells and the host, while the experiments presented here should contribute to deciphering this dialogue.

The effects of maternal supplementation of rumen-protected lysine during the close-up dry period on newborn metabolism and growth in Holstein calves.

This study aimed to evaluate the effects of rumen-protected lysine (RPL) supplementation during the close-up period on blood metabolites and calf growth. Forty multiparous Holstein dams were selected based on parity, body condition score, and expected calving date, and randomly assigned to a group: with RPL (n = 22) or without (control [CON], n = 18). RPL dams were supplied daily with 80 g of RPL from Day 21 before the expected calving date to parturition. Blood samples were obtained from the dams before the start of supplementation, 1 week before calving, and immediately after calving, and from calves immediately after birth and weekly until 8 weeks of age. Body weight measurements were performed immediately after birth in all calves and at weekly intervals until 8 weeks of age in female calves. No significant difference was observed in serum metabolite levels and plasma amino acid concentrations between the RPL and CON dams before supplementation, whereas plasma lysine concentrations tended to be higher in RPL dams immediately after calving (p = 0.07). Serum total protein levels (p < 0.05) were higher, whereas plasma total amino acid, total essential amino acid, total non-essential amino acid, and other amino acid concentrations were lower in the calves of RPL dams than those of CON dams (p < 0.05). There were no significant differences in calf birth weight between the two groups, although female calves of RPL dams (n = 7) had higher serum total protein (p < 0.05) and tended to have greater body weight (p = 0.09) from 1 to 8 weeks of age than those of CON dams (n = 11). Overall, RPL supplementation during the close-up period may increase placenta-mediated amino acid transfer to the foetus and enhance protein synthesis in the calf, leading to improved weight gain during the suckling period.

Effect of salinity on nest building behaviour in the nine-spined stickleback Pungitius sinensis.

The authors evaluated the adaptability of male nine-spined sticklebacks (Pungitius sinensis) at three salinity levels (0, 5 and 10 psu) by comparing nest building success rates with nest structures. Successful nest building decreased as salinity increased. In addition, nests built in fresh water (i.e., 0 psu) were glued together, whereas those built in brackish water (5 and 10 psu) broke easily and fell from the nest site to the gravel bottom. Based on these findings, the authors suggest that P. sinensis adapts to freshwater environments.

Postmortem magnetic resonance imaging findings of tricuspid atresia with ventricular and atrial septal defects and subvalvular pulmonic stenosis in a Japanese native Noma horse.

The necropsy of a 2-day-old Noma horse that died of weakness showed an enlarged cardiac base and a narrow cardiac apex, suggesting cardiac malformation. The excised heart underwent imaging to investigate its luminal structure. On three-dimensional magnetic resonance imaging, the right atrium and right ventricle were discontinuous. The right atrium communicated with the left atrium and the left ventricle communicated with the right ventricle. The lumen narrowed near the pulmonary artery valve. Since the same findings were observed on gross examination, the foal was diagnosed with tricuspid atresia with ventricular and atrial septal defects, along with subvalvular pulmonic stenosis.

Dynamic Regulation of NF-κB Response in Innate Immunity: The Case of the IMD Pathway in Drosophila.

Metazoans have developed strategies to protect themselves from pathogenic attack. These preserved mechanisms constitute the immune system, composed of innate and adaptive responses. Among the two kinds, the innate immune system involves the activation of a fast response. NF-κB signaling pathways are activated during infections and lead to the expression of timely-controlled immune response genes. However, activation of NF-κB pathways can be deleterious when uncontrolled. Their regulation is necessary to prevent the development of inflammatory diseases or cancers. The similarity of the NF-κB pathways mediating immune mechanisms in insects and mammals makes Drosophila melanogaster a suitable model for studying the innate immune response and learning general mechanisms that are also relevant for humans. In this review, we summarize what is known about the dynamic regulation of the central NF-κB-pathways and go into detail on the molecular level of the IMD pathway. We report on the role of the nuclear protein Akirin in the regulation of the NF-κB Relish immune response. The use of the Drosophila model allows the understanding of the fine-tuned regulation of this central NF-κB pathway.

Endoplasmic reticulum-associated protein degradation contributes to Toll innate immune defense in Drosophila melanogaster.

In Drosophila, the endoplasmic reticulum-associated protein degradation (ERAD) is engaged in regulating pleiotropic biological processes, with regard to retinal degeneration, intestinal homeostasis, and organismal development. The extent to which it functions in controlling the fly innate immune defense, however, remains largely unknown. Here, we show that blockade of the ERAD in fat bodies antagonizes the Toll but not the IMD innate immune defense in Drosophila. Genetic approaches further suggest a functional role of Me31B in the ERAD-mediated fly innate immunity. Moreover, we provide evidence that silence of Xbp1 other than PERK or Atf6 partially rescues the immune defects by the dysregulated ERAD in fat bodies. Collectively, our study uncovers an essential function of the ERAD in mediating the Toll innate immune reaction in Drosophila.

Fertility risk factors in transferring Japanese Black embryos into dairy heifers: An epidemiological study.

The aims of this study were 1) to summarize the current status of Japanese Black (JB) embryo transfer into Holstein heifers, which is carried out on a commercial basis in Japan, and 2) to reveal fertility risk factors, including those from the environment (year and season of transfer), recipient (age, number of transfers, clinical status of the ovaries) and embryo (quality, stage, state, genetic background). We used data from 4467 JB fresh or frozen embryo transfers into Holstein heifers conducted by Zen-noh Embryo Transfer Center during 2016-2018, and the differences in fertility risk due to factors related to the environment, recipient, and embryo were statistically evaluated. Differences in fertility risk due to each variable were observed, leading to significant differences in fertility with respect to year of transfer, embryo quality, embryo state, and embryo breed. These results suggest that the fertility of JB embryos might depend on differences in genetic background. There have been no previous reports of differences in embryo fertility due to the differences among JB's bloodline combinations. In the future, overall reproductive efficiency must be monitored, including the effects of different bloodline combinations on the success of embryo recovery and transfer.

Effects of the formation and regression of accessory corpus lutea during pregnancy on plasma progesterone concentration and pregnancy status in cross-bred beef heifers.

There was examination of effects of accessory corpus lutea (CLs) formation and regression during pregnancy on circulating progesterone (P4) concentrations and pregnancy maintenance in beef heifers. Heifers (Experiment 1, n = 75; Experiment 2, n = 496) were randomly assigned to either a human chorionic gonadotropin (hCG) treatment or untreated group 5 days post-estrus, followed by embryo transfer (ET) on Days 6-8 (Day 0 = Estrus). In Experiment 1, blood samples were collected from pregnant heifers on Days 33, 40, and 47 for conducting P4 assays. Plasma P4 concentrations were greater in hCG-treated heifers than in untreated heifers on Day 33. In hCG-treated heifers with accessory CL regression between Days 33 and 47, plasma P4 decreased to concentrations similar to those of untreated heifers after Day 40. In hCG-treated pregnant heifers in Experiment 2, CL regression by Day 50 of gestation was more frequent when CLs were contralateral (49.3 %) rather than ipsilateral (4.4 %, P < 0.001) to the original CL. The hCG treatment resulted in a greater pregnancy percentages on Days 30 (80.5 % and 68.6 %, P = 0.002) and 50 (76.2 % and 65.3 %, P = 0.007) compared with untreated heifers. There, however, were no differences in either pregnancy percentages on Days 30 and 50 or pregnancy losses between hCG-treated heifers with ipsilateral and contralateral accessory CLs. These results indicate accessory CL formation improves pregnancy percentages resulting from ET. Furthermore, plasma P4 decreases associated with accessory CL regression does not affect pregnancy loss in beef heifers.

Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila.

Drosophila immune deficiency (IMD) pathway is similar to the human tumor necrosis factor receptor (TNFR) signaling pathway and is preferentially activated by Gram-negative bacterial infection. Recent studies highlighted the importance of IMD pathway regulation as it is tightly controlled by numbers of negative regulators at multiple levels. Here, we report a new negative regulator of the IMD pathway, Verloren (Velo). Silencing of Velo led to constitutive expression of the IMD pathway dependent antimicrobial peptides (AMPs), and Escherichia coli stimulation further enhanced the AMP expression. Epistatic analysis indicated that Velo knock-down mediated AMP upregulation is dependent on the canonical members of the IMD pathway. The immune fluorescent study using overexpression constructs revealed that Velo resides both in the nucleus and cytoplasm, but the majority (~ 75%) is localized in the nucleus. We also observed from in vivo analysis that Velo knock-down flies exhibit significant upregulation of the AMP expression and reduced bacterial load. Survival experiments showed that Velo knock-down flies have a short lifespan and are susceptible to the infection of pathogenic Gram-negative bacteria, P. aeruginosa. Taken together, these data suggest that Velo is an additional new negative regulator of the IMD pathway, possibly acting in both the nucleus and cytoplasm.

A temporal Ca2+ desensitization of myosin light chain kinase in phasic smooth muscles induced by CaMKKß/PP2A pathways.

Cell signaling pathways regulating myosin regulatory light chain (LC20) phosphorylation contribute to determining contractile responses in smooth muscles. Following excitation and contraction, phasic smooth muscles, such as the digestive tract and urinary bladder, undergo relaxation due to a decline of cellular Ca2+ concentration and decreased Ca2+ sensitivity of LC20 phosphorylation, named Ca2+ desensitization. Here, we determined the mechanisms underlying the temporal Ca2+ desensitization of LC20 phosphorylation in phasic smooth muscles using permeabilized strips of the mouse ileum and urinary bladder. Upon stimulation with pCa6.0 at 20°C, contraction and LC20 phosphorylation peaked within 30 s and then declined to about 50% of the peak force at 2 min after stimulation. During the relaxation phase after the contraction, LC20 kinase [myosin light chain kinase (MLCK)] was inactivated, but no fluctuation in LC20 phosphatase activity occurred, suggesting that MLCK inactivation is a cause of the Ca2+-induced Ca2+ desensitization of LC20 phosphorylation. MLCK inactivation was associated with phosphorylation at the calmodulin-binding domain of the kinase. Treatment with STO-609 and TIM-063 antagonists for Ca2+/calmodulin (CaM)-dependent protein kinase kinase-ß (CaMKKß) attenuated both the phasic response of the contraction and MLCK phosphorylation, whereas neither CaM kinase II, AMP-activated protein kinase, nor p21-activated kinase induced MLCK inactivation in phasic smooth muscles. Conversely, protein phosphatase 2A inhibition amplified the phasic response. Signaling pathways through CaMKKß and protein phosphatase 2A may contribute to regulating the phasic response of smooth muscle contraction.

Asymmetric effects of social and economic incentives on cooperation in real effort based public goods games.

Many practitioners as well as researchers explore promoting environmentally conscious behavior in the context of public goods systems. Numerous experimental studies revealed various types of incentives to increase cooperation on public goods. There is ample evidence that monetary and non-monetary incentives, such as donations, have a positive effect on cooperation in public goods games that exceeds fully rational and optimal economic decision making. Despite an accumulation of these studies, in the typical setting of these experiments participants decide on an allocation of resources to a public pool, but they never exert actual effort. However, in reality, we often observe that players' real effort is required in these public goods game situations. Therefore, more analysis is needed to draw conclusions for a wider set of incentive possibilities in situations similar to yet deviating from resource allocation games. Here we construct a real effort public goods game in an online experiment and statistically analyze the effect different types of incentives have on cooperation. In our experiment, we examine combinations of monetary and social incentives in a setting aimed closer to practical realities, such as financial costs and real effort forming part of the decision to cooperate on a public good. In our real effort public goods game participants cooperate and defect on image-scoring tasks. We find that in our setting economic and social incentives produce an asymmetric effect. Interestingly economic incentives decreased the share of highly uncooperative participants, while social incentives raised the share of highly cooperative participants.

A time course transcriptomic analysis of host and injected oncogenic cells reveals new aspects of Drosophila immune defenses.

Oncogenic RasV12 cells [A. Simcox et al., PLoS Genet 4, e1000142 (2008)] injected into adult males proliferated massively after a lag period of several days, and led to the demise of the flies after 2 to 3 wk. The injection induced an early massive transcriptomic response that, unexpectedly, included more than 100 genes encoding chemoreceptors of various families. The kinetics of induction and the identities of the induced genes differed markedly from the responses generated by injections of microbes. Subsequently, hundreds of genes were up-regulated, attesting to intense catabolic activities in the flies, active tracheogenesis, and cuticulogenesis, as well as stress and inflammation-type responses. At 11 d after the injections, GFP-positive oncogenic cells isolated from the host flies exhibited a markedly different transcriptomic profile from that of the host and distinct from that at the time of their injection, including in particular up-regulated expression of genes typical for cells engaged in the classical antimicrobial response of Drosophila.

cGMP signaling pathway that modulates NF-κB activation in innate immune responses.

The nuclear factor-kappa B (NF-κB) pathway is an evolutionarily conserved signaling pathway that plays a central role in immune responses and inflammation. Here, we show that Drosophila NF-κB signaling is activated via a pathway in parallel with the Toll receptor by receptor-type guanylate cyclase, Gyc76C. Gyc76C produces cyclic guanosine monophosphate (cGMP) and modulates NF-κB signaling through the downstream Tollreceptor components dMyd88, Pelle, Tube, and Dif/Dorsal (NF-κB). The cGMP signaling pathway comprises a membrane-localized cGMP-dependent protein kinase (cGK) called DG2 and protein phosphatase 2A (PP2A) and is crucial for host survival against Gram-positive bacterial infections in Drosophila. A membrane-bound cGK, PRKG2, also modulates NF-κB activation via PP2A in human cells, indicating that modulation of NF-κB activation in innate immunity by the cGMP signaling pathway is evolutionarily conserved.