Effects of Multi-Component Exercise on Sleep Quality in Middle-Aged Adults.

Sleep is a crucial factor in healthy aging. However, most middle-aged adults experience high levels of sleep disorders. While previous findings have suggested exercise training could benefit the quality of sleep, the effects of multi-component exercise on sleep quality are less examined. Accordingly, the current study aimed to assess the effectiveness of a multi-component exercise program on the quality of sleep among middle-aged adults. Twenty-four middle-aged adults were randomly assigned either to a multi-component exercise (MCE) group or a control group. The participants in the MCE group attended a 90-min session per week for 12 weeks. The control group was instructed to maintain their daily routine for 12 weeks. The primary outcome was the sleep quality evaluated by the Pittsburgh Sleep Quality Index (PSQI). The secondary outcome was physical fitness, including muscular strength and endurance, balance, and flexibility. Regarding sleep quality, the global mean score (p = 028), sleep disturbances (p = 011), and sleep efficiency (p = 035) of the PSQI scores were significantly reduced in the MCE group after the 12-week intervention. Regarding physical fitness, the flexibility of the MCE group improved significantly after the intervention (p = 028), yet, no significant change was observed in the control group. Additionally, the muscular strength of the control group declined significantly after the 12-week period (p = 034). Our results revealed the effectiveness of the MCE intervention in improving sleep quality and physical fitness in middle-aged adults. Further studies using larger sample sizes, objective measures of sleep quality, different types of exercise training, as well as different populations, are warranted to extend our current findings.

Neospora caninum infection activated autophagy of caprine endometrial epithelial cells via mTOR signaling.

Neosporosis, caused by infection with the protozoan parasite Neospora caninum, is one of the main causes of abortion in cattle and small ruminants (e.g., goats), negatively influencing animal health and production costs. The uterus is an adhesion organ of placenta that is important for pregnancy and embryonic development. However, the underlying molecular pathogenic mechanisms of N. caninum in the uterus are still unclear. Autophagy regulates innate and adaptive immunity for eliminating pathogens by xenophagy, while pathogens can manipulate autophagy to facilitate their propagation. To study the role of host cell autophagy during N. caninum infection, a N. caninum infection model in caprine endometrial epithelial cells (EECs) was successfully established. In this in vitro model, N. caninum infection increased the expression of LC3-II (a standard marker for autophagosomes) from 6 h post infection (pi) to 48 h pi and the number of autophagosomes in caprine EECs at 48 h pi. Expression of p62 protein (a classical receptor of autophagy) levels were significantly decreased (P < 0.05) in caprine EECs infected with N. caninum tachyzoites at both 24 h pi and 48 h pi. Enhanced autophagic flux was also detected at 48 h pi in caprine EECs infected with N. caninum tachyzoites by transfecting Ad-mCherry-GFP-LC3B recombinant adenovirus. Treatments using a mechanistic target of rapamycin (mTOR)-specific inhibitor (rapamycin) and an autophagy inhibitor (chloroquine) indicated that cell autophagy induced by N. caninum infection promoted the intracellular propagation of parasite tachyzoites. Further studies showed that N. caninum infection induced autophagy through inhibition of mTOR phosphorylation. To the best of our current knowledge, this is the first study to reveal the role of autophagy during N. caninum infection in caprine EECs, and the findings provided significant information for uncovering mechanisms of abortion and pathogenicity caused by N. caninum infection.

Diagnostic Value of Circulating Antigens in the Serum of Piglets with Experimental Acute Toxoplasmosis.

Toxoplasmosis, caused by Toxoplasma gondii, an apicomplexan parasite, infects all warm-blooded animals, including a third of the human population. Laboratory diagnosis of acute toxoplasmosis is based on the detection of anti-T. gondii IgM and IgG and T. gondii nucleic acid; however, these assays have certain limitations. Circulating Ags (CAgs) are reliable diagnostic indicators of acute infection. In this study, we established a model of acute T. gondii infection in Large White pigs. CAg levels peaked between 3 and 5 d after inoculation, and 28 CAgs were identified using an immunoprecipitation-shotgun approach, among which dolichol-phosphate-mannose synthase family protein (TgDPM), C3HC zinc finger-like protein (TgZFLP3), and ribosomal protein RPL7 (TgRPL7) were selected to further investigate their value in the diagnosis of acute toxoplasmosis. Immunofluorescence assays revealed that TgDPM and TgRPL7 were localized in the membrane surface, while TgZFLP3 was localized in the apical end. Western blotting revealed the presence of the three proteins in the serum during acute infection. Indirect ELISA results indicate that TgZFLP3 is likely to be a novel candidate for the diagnosis of acute toxoplasmosis. However, these three proteins may not be useful as candidate vaccines against toxoplasmosis owing to their low protective ability. In addition, deletion of the zflp3 gene partially attenuated virulence in Kunming mice. Collectively, we identified 28 CAgs in the serum of piglets with experimental acute toxoplasmosis and confirmed that TgZFLP3 is a potential biomarker for acute T. gondii infection. The results of this study provide data to improve the detection efficiency of acute toxoplasmosis.

Genome-Wide CRISPR/Cas9 Screen Identifies New Genes Critical for Defense Against Oxidant Stress in Toxoplasma gondii.

Toxoplasma gondii is one of the most widespread apicomplexans and can cause serious infections in humans and animals. Its antioxidant system plays an important role in defending against oxidant stress imposed by the host. Some genes encoding the antioxidant enzymes of T. gondii have been identified; however, critical genes that function in response to reactive oxygen species (ROS) stress are still poorly understood. Here, we performed genome-wide CRISPR/Cas9 loss-of-function screening in the T. gondii RH strain to identify potential genes contributing to the ROS stress response. Under hydrogen peroxide treatment, 30 single guide RNAs targeting high-confidence genes were identified, including some known important antioxidant genes such as catalase and peroxiredoxin PRX3. In addition, several previously uncharacterized genes were identified, among which five hypothetical protein-coding genes, namely, HP1-HP5, were selected for further functional characterization. Targeted deletion of HP1 in T. gondii RH led to significant sensitivity to H2O2, suggesting that HP1 is critical for oxidative stress management. Furthermore, loss of HP1 led to decreased antioxidant capacity, invasion efficiency, and proliferation in vitro. In vivo results also revealed that the survival time of mice infected with the HP1-KO strain was significantly prolonged relative to that of mice infected with the wild-type strain. Altogether, these findings demonstrate that the CRISPR/Cas9 system can be used to identify potential genes critical for oxidative stress management. Furthermore, HP1 may confer protection against oxidative damage and contributes to T. gondii virulence in mice.

Salidroside-Mitigated Inflammatory Injury of Hepatocytes with Non-Alcoholic Fatty Liver Disease via Inhibition TRPM2 Ion Channel Activation.

PURPOSE: Oxidative stress plays an important role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). TRPM2 ion channel functions as a molecular sensor for oxidative stress. The aim of this study was to examine the protective effects of Salidroside, a powerful antioxidative plant, on TRPM2 in an established in vitro model of NAFLD. METHODS: NAFLD model was established by palmitic acid (PA) in hepatic L02 cell lines and was added to the media at a final concentration of 400 µM. Cells were used as normal group, PA group and PA receiving varied concentrations of Salidroside (75µg/mL, 150µg/mL, 300µg/mL). After treating 24 hrs, MTT assay was used to detect cell viability, and ALT level was measured using an appropriate kit assay. Intracellular lipid accumulation was observed by Oil red O staining. Cytosolic Ca2+ concentrations were evaluated by flow cytometer with Fluo-3/AM. Quantitative RT-PCR was used to measure the mRNA expression of TRPM2, IL-1ß and IL-6, and the protein expressions of TRPM2, p-CaMKII and autophagy (LC3B, p62) were determined using Western blot. RESULTS: Treatment with Salidroside effectively restored liver injury and alleviated lipid droplet deposition in a dose-dependent manner, which was associated with inhibition of TRPM2/Ca2+/CaMKII pathway. Additionally, autophagic clearance was enhanced by intervention with Salidroside in a dose-dependent manner. Further investigation indicated that Salidroside down-regulated the mRNA expression of IL-1ß and IL-6-pro-inflammatory cytokines. CONCLUSION: These results suggest that Salidroside could alleviate inflammatory injury and steatosis via autophagy activation mediated by downregulation of the TRPM2/Ca2+/CaMKII pathway. Targeting the TRPM2 ion channel is a novel treatment strategy for oxidative stress-induced liver in NAFLD.