Spider minor ampullate silk protein nanoparticles: an effective protein delivery system capable of enhancing systemic immune responses.

Spider silk proteins (spidroins) are particularly attractive due to their excellent biocompatibility. Spider can produce up to seven different types of spidroins, each with unique properties and functions. Spider minor ampullate silk protein (MiSp) might be particularly interesting for biomedical applications, as the constituent silk is mechanically strong and does not super-contract in water, attributed to its amino acid composition. In this study, we evaluate the potential of recombinant nanoparticles derived from Araneus ventricosus MiSp as a protein delivery carrier. The MiSp-based nanoparticles were able to serve as an effective delivery system, achieving nearly 100% efficiency in loading the model protein lysozyme, and displayed a sustained release profile at physiological pH. These nanoparticles could significantly improve the delivery efficacy of the model proteins through different administration routes. Furthermore, nanoparticles loaded with model protein antigen lysozyme after subcutaneous or intramuscular administration could enhance antigen-specific immune responses in mouse models, through a mechanism involving antigen-depot effects at the injection site, long-term antigen persistence, and efficient uptake by dendritic cells as well as internalization by lymph nodes. These findings highlight the transnational potential of MiSp-based nanoparticle system for protein drug and vaccine delivery.

Predicting rice productivity for ground data-sparse regions: A transferable framework and its application to North Korea.

The use of observation-dependent methods for crop productivity and food security assessment is challenging in data-sparse regions. This study presents a transferable framework and applies it to North Korea (NK) to assess rice productivity based on climate similarity, transferable machine-learning techniques, and extendable multi-source data. We initially divided the primary phenological stages of rice in the study region and extracted dynamic rice distributions based on Moderate Resolution Imaging Spectroradiometer products and phenological observations. We compared the performances of four representative environmentally driven models (Linear Regression, back-propagation Neural Network, Support Vector Machine, and Random Forest) in simulating rice productivity using an extensive dataset that included multi-angle vegetation monitoring, climate variables, and planting distribution information. The framework integrated an optimal environmentally driven model with agricultural management practices for transferability to predict rice productivity in NK over multiple years. Additionally, two crop growth scenarios (whole growth period (WGP) and seeding-heading period (SHP)) were compared to assess pre-harvest forecasting capabilities and identify dominant factors. Finally, independent datasets from the Food and Agriculture Organization, World Food Program, and Global Gridded Crop Models were used to validate the magnitude and spatial distribution of the predicted results. The results showed that phenological identification based on remote sensing can accurately capture rice growth characteristics and map rice distribution. Random Forest outperformed other models in simulating rice productivity variation, with r-squares of 0.87 and 0.83 in the WGP and SHP, respectively. The solar-induced chlorophyll fluorescence, maximum temperature, and evapotranspiration collectively determined approximately 40 % of the variation in yield simulated using Random Forest. Conversely, planting areas contributed over 42 % of the variation in rice production. Compared to Food and Agriculture Organization statistics, the environmentally driven framework explained 78.72 % and 76.89 % of the production variation and 69.42 % and 71.15 % of the yield variation in NK under the WGP and SHP, respectively. Moreover, the environmental management-driven framework captured over 90 % of the yield variation. The predicted spatial pattern of rice productivity exhibited significant concordance with the World Food Program and Global Gridded Crop Model reports. In summary, the proposed transferable framework for crop productivity assessment contributes to early warnings of production reduction and has the potential for scalability across various crops and data-sparse regions.

Building social resilience in North Korea can mitigate the impacts of climate change on food security.

Adaptation based on social resilience is proposed as an effective measure to mitigate hunger and avoid food shocks caused by climate change. But these have not been investigated comprehensively in climate-sensitive regions. North Korea (NK) and its neighbours, South Korea and China, represent three economic levels that provide us with examples for examining climatic risk and quantifying the contribution of social resilience to rice production. Here our data-driven estimates show that climatic factors determined rice biomass changes in NK from 2000 to 2017, and climate extremes triggered reductions in production in 2000 and 2007. If no action is taken, NK will face a higher climatic risk (with continuous high-temperature heatwaves and precipitation extremes) by the 2080s under a high-emissions scenario, when rice biomass and production are expected to decrease by 20.2% and 14.4%, respectively, thereby potentially increasing hunger in NK. Social resilience (agricultural inputs and population development for South Korea; resource use for China) mitigated climate shocks in the past 20 years (2000-2019), even transforming adverse effects into benefits. However, this effect was not significant in NK. Moreover, the contribution of social resilience to food production in the undeveloped region (15.2%) was far below the contribution observed in the developed and developing regions (83.0% and 86.1%, respectively). These findings highlight the importance of social resilience to mitigate the adverse effects of climate change on food security and human hunger and provide necessary quantitative information.

Maresin conjugates in tissue regeneration-1 suppresses ferroptosis in septic acute kidney injury.

BACKGROUND: Ferroptosis is unique among different types of regulated cell death and closely related to organ injury. Whether ferroptosis occurs in sepsis-associated acute kidney injury (SA-AKI) is not clear. Nuclear factor-erythroid-2-related factor 2 (Nrf2) is crucial to the regulation of ferroptosis. We and others have shown that Maresin conjugates in tissue regeneration 1 (MCTR1) or other members of specialized pro-resolving mediators (SPMs) can actively regulate inflammation resolution and protect organs against injury in inflammatory diseases by activating the Nrf2 signaling. The aim of this study was to determine whether ferroptosis occurs in SA-AKI. Furthermore, we investigated the potential role and mechanism of MCTR1 in the regulation of ferroptosis in SA-AKI, which mainly focus on the Nrf2 signaling. RESULTS: We demonstrated for the first time that ferroptosis is present in SA-AKI. Moreover, MCTR1 effectively suppressed ferroptosis in SA-AKI. Meanwhile, MCTR1 upregulated the expression of Nrf2 in the kidney of septic mice. Nrf2 inhibitor ML-385 reversed MCTR1-regulated ferroptosis and AKI, implying that Nrf2 is involved in the inhibitory effects of MCTR1 on ferroptosis in SA-AKI. Further, MCTR1 inhibited ferroptosis and elevated the expression of Nrf2 in LPS-induced HK-2 cells. However, Nrf2 siRNA offset the effect of MCTR1 on ferroptosis. Finally, we observed that MCTR1 ameliorates multi-organ injury and improves survival in animal models of sepsis. CONCLUSIONS: These data demonstrate that MCTR1 suppresses ferroptosis in SA-AKI through the Nrf2 signaling. Our study enriches the pathophysiological mechanism of SA-AKI and provides new therapeutic ideas and potential intervention targets for SA-AKI.

γδ T/Interleukin-17A Contributes to the Effect of Maresin Conjugates in Tissue Regeneration 1 on Lipopolysaccharide-Induced Cardiac Injury.

The mechanisms underlying sepsis-induced cardiomyopathy (SIC) remain poorly understood, and there are no specific therapeutics for SIC. We investigated the effects of maresin conjugates in tissue regeneration 1 (MCTR1) on SIC and explored its potential mechanisms. The experiments were conducted using an endotoxemia model induced by lipopolysaccharide (LPS). Mice were given MCTR1 intravenously 6 h after LPS stimulation. Echocardiography was performed to assess cardiac function 12 h after LPS administration. Treatment with MCTR1 significantly enhanced cardiac function and reduced LPS-induced increase of mRNA expression levels of inflammation cytokines. Transcriptomic analysis indicated that MCTR1 inhibited neutrophil chemotaxis via the IL-17 signaling pathway. We confirmed that MCTR1 reduced the expressions of neutrophil chemoattractants and neutrophil infiltration in the LPS-stimulated hearts. MCTR1 also resulted in a considerable reduction in IL-17A production mainly derived from γδ T cells. Moreover, our results provided the first evidence that neutralizing IL-17A or depletion of γδ T cells markedly decreased neutrophil recruitment and enhanced cardiac function in LPS-induced cardiac injury. These results suggest that MCTR1 alleviates neutrophil infiltration thereby improves cardiac function in LPS-induced cardiac injury via the IL-17 signaling pathway. Thus, MCTR1 represented a novel therapeutic strategy for patients with SIC.

Dexmedetomidine promotes inflammation resolving through TGF-ß1 secreted by F4/80+Ly6G+ macrophage.

Dexmedetomidine (DEX) is a highly selective α2-adrenoceptor agonist, which can regulate inflammatory responses. However, whether DEX interferes with the inflammation resolving remains unclear. Here, we reported the effects of DEX on zymosan-induced generalized inflammation in mice during resolution. Mice were administered intraperitoneally with DEX after the initiation of sepsis. The resolution interval (Ri), a vital resolution indice, decreased from twelve hours to eight hours after the administration of DEX. The induction of peritoneal pro-inflammatory interleukin [IL] - 1ß and tumour necrosis factor-α (TNF-α) appeared to be inhibited. Of interest, the anti-inflammatory transforming growth factor-ß1 (TGF-ß1) but not IL-10 levels were up-regulated at twenty-four hours in the DEX group along with 1.0 mg/mice zymosan A (ZyA) treatment. The expression levels of multiple genes related to protective immune processes and clearance functions were detected and revealed the same trends. DEX markedly increased the F4/80+Ly6G+ macrophage population. Additionally, the adequate apoptotic neutrophil clearance from injury after DEX installation could be reverse by opsonization or co-instillation of TGF-ß1 neutralizing antibody in vivo, promoting the inflammation-resolution programs. In conclusion, DEX post-treatment, via the increase of F4/80+Ly6G+ macrophages, provokes further secretion of TGF-ß1, leading to the attenuated cytokine storm and accelerated inflammation resolving.

Monocyte Chemoattractant Protein-1 stimulates the differentiation of rat stem and progenitor Leydig cells during regeneration.

BACKGROUND: Monocyte chemoattractant protein-1(MCP-1) is a chemokine secreted by Leydig cells and peritubular myoid cells in the rat testis. Its role in regulating the development of Leydig cells via autocrine and paracrine is still unclear. The objective of the current study was to investigate the effects of MCP-1 on Leydig cell regeneration from stem cells in vivo and on Leydig cell development in vitro. RESULTS: Intratesticular injection of MCP-1(10 ng/testis) into Leydig cell-depleted rat testis from post-EDS day 14 to 28 significantly increased serum testosterone and luteinizing hormone levels, up-regulated the expression of Leydig cell proteins, LHCGR, SCARB1, CYP11A1, HSD3B1, CYP17A1, and HSD17B3 without affecting progenitor Leydig cell proliferation, as well as increased ERK1/2 phosphorylation. MCP-1 (100 ng/ml) significantly increased medium testosterone levels and up-regulated LHCGR, CYP11A1, and HSD3B1 expression without affecting EdU incorporation into stem cells after in vitro culture for 7 days. RS102895, a CCR2 inhibitor, reversed MCP-1-mediated increase of testosterone level after culture in combination with MCP-1. CONCLUSION: MCP-1 stimulates the differentiation of stem and progenitor Leydig cells without affecting their proliferation.

Material conversion, microbial community composition and metabolic functional succession during green soybean hull composting.

In this study, green soybean hulls and maize straw were used for composting to explore the dynamics of material conversion, bacterial and fungal communities and metabolic functions. The results showed that bacterial and fungal communities had different temporal successions during composting. The bacterium Streptosporangiaceae was a biomarker in the thermophilic stage of composting, and the fungus Chaetomiaceae was a biomarker in the thermophilic stage and cooling stage. In the bacterial network, the germination index (GI) had a time-delayed association with Truepera, Pseudomonas and Methylococcaceae, which represented the key physicochemical characteristics that affect the community. In the fungal community, the GI, pH, fulvic acid (FA) and temperature etc. had a joint effect. Carbohydrate metabolism and amino acid metabolism were the main metabolic pathways, and saprotrophs represented the dominant fungal trophic mode in the composting process. These results provide a reference from screening specific and efficient agents to accelerate natural vegetable composting.

Ranking contributors to traffic crashes on mountainous freeways from an incomplete dataset: A sequential approach of multivariate imputation by chained equations and random forest classifier.

The estimation of the effect of contributors to crash injury severity and the prediction of crash injury severity outcomes suffer often from biases related to missing data in crash datasets that contain incomplete records. As both estimation and prediction would greatly improve if the missing values were recovered, this study proposes a sequential approach to handle incomplete crash datasets and rank contributors to the injury severity of crashes on mountainous freeways in China. The sequential approach consists of two parts: (i) multivariate imputation by chained equations imputes the missing values of independent variables; (ii) a random forest classifier analyses the correlation between the dependent and the independent variables. The first part considers different imputation methods in light of the independent variables being either binary, categorical or continuous, whereas the second part classifies the correlations according to the random forest classifier. The proposed method was applied to the case-study about mountainous freeways in China and compared to the analysis of the raw dataset to evaluate its effectiveness, and the results illustrate that the method improves significantly the classification accuracy when compared with existing methods. Moreover, the classifier ranked the contributors to the injury severity of traffic crashes on mountainous freeways: in order of importance vehicle type, crash type, road longitudinal gradient, crash cause, curve radius, and deflection angles. Interestingly, a lower importance was found for environmental factors.

Automated traffic incident detection with a smaller dataset based on generative adversarial networks.

An imbalanced and small training sample can cause an incident detection model to have a low detection rate and a high false alarm rate. To solve the scarcity of incident samples, a novel incident detection framework is proposed based on generative adversarial networks (GANs). First, spatial and temporal rules are presented to extract variables from traffic data, which is followed by the random forest algorithm to rank the importance of variables. Then, some new incident samples are generated using GANs. Finally, the support vector machine algorithm is applied as the incident detection model. Real traffic data, which were collected from a 69.5-mile section of the I-80 highway, are used to validate the proposed approach. A total of 140 detectors are installed on the section enabling traffic flow to be measured every 30s. During 14 days, 139 incident samples and 946 nonincident samples were extracted from the raw data. Five categories of experiments are designed to evaluate whether the proposed framework can solve the small sample size problem, imbalanced sample problem, and timeliness problem in the current incident detection system. The experimental results show that our proposed framework can considerably improve the detection rate and reduce the false alarm rate of traffic incident detection. The balance of the dataset can improve the detection rate from 87.48% to 90.68% and reduce the false alarm rate from 12.76% to 7.11%. This paper lends support to further studies on combining GANs with the machine learning model to address the imbalance and small sample size problems related to intelligent transportation systems.

In utero cadmium and dibutyl phthalate combination exposure worsens the defects of fetal testis in rats.

Testicular dysgenesis syndrome might be due to the fetal testis defects caused by endocrine disruptors. Here, we report the combined effects of in utero exposure to cadmium (CdCl2, Cd) and di-n-butyl phthalate (DBP) on fetal testis development in rats. Pregnant Sprague-Dawley rats were randomly divided into four groups: control, Cd, DBP (250 mg/kg/day), and Cd + DBP. Cd (0.25 mg/kg/once) was intraperitoneally injected to the dam on gestational day 12 and DBP (250 mg/kg) was daily gavaged to the dam on gestational day 12 for 10 days. Cd, DBP, and Cd + DBP lowered serum testosterone levels in male fetuses. Cd and DBP did not alter fetal Leydig cell (FLC) number, but the combined exposure led to decreased FLC number. Cd did not affect FLC aggregation while DBP caused FLC aggregation and the combined exposure worsened FLC aggregation. Cd lowered FLC mRNA (Lhcgr, Star, Cyp11a1, and Insl3) levels and DBP lowered Lhcgr, Star, Insl3, and Nr5a1 levels. DBP up-regulated Scarb1 expression without affecting Cyp11a1 while the combined exposure antagonized DBP. These two chemicals and its combination did not affect Sertoli cell number and gene (Amh, Fshr, and Sox9) expression at current doses. In conclusion, the combined exposure of Cd and DBP exerts synergically antiandrogenic effects via targeting FLC development.

Triphenyltin chloride reduces the development of rat adrenal cortex during puberty.

Triphenyltin has been classified as an endocrine disruptor. However, whether triphenyltin interferes with the adrenal glands during puberty remains unknown. Here, we reported the effects of triphenyltin on the adrenal glands in rats. Male Sprague Dawley rats (age of 35 days) were orally administered with 0, 0.5, 1, or 2 mg/kg/day triphenyltin for 18 days. Triphenyltin significantly lowered corticosterone levels at 1 and 2 mg/kg and adrenocorticotropic hormone at 2 mg/kg. The RNA-Seq analysis detected multiple differentially expressed genes. Four down-regulated genes were transcription factor genes (Nr4a1, Nr4a2, Nr4a3, and Ppard), which might be associated with the suppression of the adrenal cortex function. RNA-seq and qPCR showed that triphenyltin dose-dependently down-regulated the expression of the genes for cholesterol transport and biosynthesis, including Scarb1, Ldlr, Hmgcs1, Hmgcr, and Hsd17b7. Further Western blotting revealed that it lowered NR4A1, PPRAD, LDLR, and HMGCS1 protein levels. We treated H295R adrenal cells with 1-100 nM triphenyltin for 72 h. Triphenyltin induced significant higher ROS production at 100 nM and did not induce apoptosis at 10 and 100 nM. In conclusion, triphenyltin inhibits production of corticosterone via blocking the expression of cholesterol uptake transporters and cholesterol biosynthesis.

Maresin conjugates in tissue regeneration 1 prevents lipopolysaccharide-induced cardiac dysfunction through improvement of mitochondrial biogenesis and function.

Mitochondrial dysfunction is increasingly considered as the center of pathophysiology in sepsis-induced cardiac dysfunction. Maresin conjugates in tissue regeneration 1 (MCTR1) is a newly identified specialized pro-resolving mediator (SPM) and has been shown to accelerate tissue regeneration and exert positive inotropic effects. Our present study aims to investigate the effect of MCTR1 on lipopolysaccharide (LPS)-induced cardiac dysfunction and explore its potential mechanisms. Mice were treated with LPS to generate LPS-induced cardiac dysfunction. H9C2 cells were used to verify the effect of MCTR1 in vitro. LPS injection triggered cardiac dysfunction and increased mRNA expression of inflammation cytokines, which were significantly attenuated by post-treatment of MCTR1. Mechanistically, we found that MCTR1 ameliorated LPS-mediated reduction of protein expression of mitochondrial biogenesis factors and silent information regulator 1 (Sirt1), accompanied by enhancement of mitochondrial biogenesis and function. Besides, Sirt1 inhibitor EX527 inhibited effects of MCTR1 on mitochondrial biogenesis and function, blunted the protective effect of MCTR1 on cardiac function, and prevented enhancement of survival rate. MCTR1 protected against LPS-induced cardiac dysfunction through improvement of mitochondrial biogenesis and function in a Sirt1-dependent manner. Our studies showed that MCTR1 might represent a novel therapeutic strategy for cardiac dysfunction caused by sepsis.

Projections of drought characteristics in China based on a standardized precipitation and evapotranspiration index and multiple GCMs.

Droughts have destructive impacts on agricultural production; thus, drought projections are vital for the development of future drought mitigation strategies. This work aimed to project a standardized precipitation and evapotranspiration index (SPEI) at 3-, 6- and 12-month timescales for the period 2011-2100 under two representative concentration pathway (RCP) scenarios - RCP 4.5 and RCP 8.5 in mainland China and to assess the changes in various drought indices over a baseline period of 1961-2000. The spatiotemporal variations in drought characteristics (e.g., the drought occurrence time, duration, severity, peak, and frequency and the percentage of stations suffering from drought (PSSD) were estimated by the projected SPEI for the periods 2011-2040, 2041-2070 and 2071-2100. The results showed that mainland China would experience more frequent and severe droughts in the future than in the baseline period, as denoted by SPEI and the generated drought variables. In particular, drier areas of northwestern China were likely to suffer from worse drought conditions than those in other areas, with PSSD values of 60% and 81% by 2100 under the RCP4.5 and RCP 8.5 scenarios, respectively. Although the annual precipitation was projected to increase in most regions, drought conditions would still worsen because of increased the minimum and maximum air temperatures. However, the GCMs contributed more uncertainties to the projection of the SPEI than the stations or the RCPs, because the GCMs made a larger contribution to the variance (>40%). The SPEI performed better than the other indices that only accounted for the influence of a single variable. The relationship between crop yields and the three drought indices varied by month, crop (maize and cotton), and timescale (3- and 6-month). The drought projections from our study can provide invaluable information for stakeholders in developing regionally specific drought adaptation strategies in the face of climate change.

Self-reports of workloads and aberrant driving behaviors as predictors of crash rate among taxi drivers: A cross-sectional study in China.

Objective: Taxis provide an important mode of public transport in China, but there has been very little in-depth research on the crash involvement propensity of taxi drivers. Thus, this study was conducted to develop a quantitative model for predicting the crash rate of taxi drivers. Methods: A total of 2,391 taxi drivers from 29 companies in 4 Chinese cities completed a structured and anonymous face-to-face questionnaire reporting their demographic information, workload conditions, aberrant driving behaviors, and crash history within the 2 years prior to the survey. Using the self-reported variables, a negative binomial model was implemented to predict taxi drivers' property damage only (PDO) and personal injury (PI) crash rates and identify the factors contributing to this risk. Results: Descriptive analysis of the survey data revealed that the workload of taxi drivers in China is relatively heavy. Seven significant predictors of PDO and PI crash rates were identified, including crossing red lights, dangerous overtaking, honking at a slow driver, failure to use an indicator lamp, driving while fatigued, stopping in forbidden areas to pick up or drop off a passenger, and driving with one hand. Taxi drivers' average off-duty days per week, aggressive driving behaviors, and preventing another driver from merging had significant effects only on PDO crash rate, and sleep problems were found to be significantly correlated with PI crash rate. Conclusions: To improve the safety of taxi drivers, considerable measures should be strictly implemented, ranging from periodic driver training and safety education to workload reduction, with the cooperation of government agencies and taxi companies. The findings of this study contribute to the design of potentially useful policy initiatives as well as targeted safety promotion programs.

Fibroblast Growth Factor 1 Promotes Rat Stem Leydig Cell Development.

Fibroblast growth factor 1 (FGF1) is reported to be expressed in the testis. How FGF1 affects stem Leydig cell development remains unclear. Here, we report the effects of FGF1 on rat stem Leydig cell development in an ethane dimethane sulfonate (EDS)-treated model. FGF1 (100 ng/testis) significantly increased serum testosterone level, increased PCNA-positive Leydig cell percentage and Leydig cell number, but down-regulated the expression of Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1, and Hsd11b1 in Leydig cells per se, after its daily intratesticular injection from post-EDS day 14 for 14 days. Primary culture of the seminiferous tubules showed that FGF1 stimulated EdU incorporation to stem Leydig cells but blocked the differentiation into the Leydig cell lineage, possibly via FGFR1-mediated mechanism. In conclusion, FGF1 promotes stem Leydig cell proliferation but blocks its differentiation.

Amycolatopsis panacis sp. nov., isolated from Panax notoginseng rhizospheric soil.

A novel Gram-positive bacterium, designated strain YIM PH21725T, was isolated from a sample of rhizospheric soil of Panaxnotoginseng cultivated in Anning, Yunnan. The strain contained meso-diaminopimelic acid in the cell-wall peptidoglycan. The main fatty acids identified were C17 : 0, iso-C15 : 0, iso-C16 : 0 and C16 : 0. The main menaquinone was MK-9 (H4). The polar lipids included phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, phospholipids and phospholipids of an unidentified structure containing glucosamine. The G+C content of genomic DNA was 69.43 mol%. On the basis of its 16S rRNA gene sequence, strain YIM PH21725T should belong to the genus Amycolatopsis, and was closely related to Amycolatopsis sulphurea DSM 46092T (98.57 %), Amycolatopsis jejuensis JCM13280T (97.27 %), Amycolatopsis jiangsuensis KCTC 19885T (96.88 %) and Amycolatopsis ultiminotia JCM 16989T (96.8 %). The phenotypic, chemotaxonomic, phylogenetic and digital DNA-DNA hybridization results clearly indicated that strain YIM PH21725T represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsispanacis sp. nov. is proposed. The type strain is YIM PH21725T (=CCTCC AA 2017044T=KCTC 49031T=DSM 105902T).

The relation between working conditions, aberrant driving behaviour and crash propensity among taxi drivers in China.

Although the taxi industry is playing an important role in Chinese everyday life, little attention has been posed towards occupational health issues concerning the taxi drivers' working conditions, driving behaviour and road safety. A cross-sectional survey was administered to 1021 taxi drivers from 21 companies in four Chinese cities and collected information about (i) sociodemographic characteristics, (ii) working conditions, (iii) frequency of daily aberrant driving behaviour, and (iv) involvement in property-damage-only (PDO) and personal injury (PI) crashes over the past two years. A hybrid bivariate model of crash involvement was specified: (i) the hybrid part concerned a latent variable model capturing unobserved traits of the taxi drivers; (ii) the bivariate part modelled jointly both types of crashes while capturing unobserved correlation between error terms. The survey answers paint a gloomy picture in terms of workload, as taxi drivers reported averages of 9.4 working hours per day and 6.7 working days per week that amount on average to about 63.0 working hours per week. Moreover, the estimates of the hybrid bivariate model reveal that increasing levels of fatigue, reckless behaviour and aggressive behaviour are positively related to a higher propensity of crash involvement. Lastly, the heavy workload is also positively correlated with the higher propensity of crashing, not only directly as a predictor of crash involvement, but also indirectly as a covariate of fatigue and aberrant driving behaviour. The findings from this study provide insights into potential strategies for preventive education and taxi industry management to improve the working conditions and hence reduce fatigue and road risk for the taxi drivers.

Oncostatin M inhibits differentiation of rat stem Leydig cells in vivo and in vitro.

Oncostatin M (OSM) is a pleiotropic cytokine within the interleukin six family of cytokines, which regulate cell growth and differentiation in a wide variety of biological systems. However, its action and underlying mechanisms on stem Leydig cell development are unclear. The objective of the present study was to investigate whether OSM affects the proliferation and differentiation of rat stem Leydig cells. We used a Leydig cell regeneration model in rat testis and a unique seminiferous tubule culture system after ethane dimethane sulfonate (EDS) treatment to assess the ability of OSM in the regulation of proliferation and differentiation of rat stem Leydig cells. Intratesticular injection of OSM (10 and 100 ng/testis) from post-EDS day 14 to 28 blocked the regeneration of Leydig cells by reducing serum testosterone levels without affecting serum luteinizing hormone and follicle-stimulating hormone levels. It also decreased the levels of Leydig cell-specific mRNAs (Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1 and Hsd11b1) and their proteins by the RNA-Seq and Western blotting analysis. OSM had no effect on the proliferative capacity of Leydig cells in vivo. In the seminiferous tubule culture system, OSM (0.1, 1, 10 and 100 ng/mL) inhibited the differentiation of stem Leydig cells by reducing medium testosterone levels and downregulating the expression of Leydig cell-specific genes (Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1 and Hsd11b1) and their proteins. OSM-mediated action was reversed by S3I-201 (a STAT3 antagonist) or filgotinib (a JAK1 inhibitor). These data suggest that OSM is an inhibitory factor of rat stem Leydig cell development.

Delayed Puberty by Ziram Is Associated with Down Regulation of Testicular Phosphorylated AKT1 and SIRT1/PGC-1α Signaling.

Ziram is a dimethyldithiocarbamate fungicide, which may influence the male reproductive system as a potential endocrine disruptor. We interrogated the disruption of ziram on rat progenitor Leydig cell development. Prepubertal male Sprague-Dawley rats were orally treated with 0, 2, 4, or 8 mg/kg ziram for 2 weeks. We investigated the effects of ziram on serum testosterone levels, Leydig cell number, and Leydig and Sertoli cell gene and protein expression, SIRT1/PGC-1α levels, and phosphorylation of AKT1, ERK1/2, and AMPK in vivo. We also interrogated the effects of ziram on reactive oxidative species (ROS) level, apoptosis rate, and mitochondrial membrane potential of progenitor Leydig cells in vitro. Ziram decreased serum testosterone and follicle-stimulating hormone levels, the down-regulated Leydig cell-specific gene ( Lhcgr, Scarb1, Star, Cyp17a1, and Hsd17b3), and their protein expression. However, ziram stimulated anti-Müllerian hormone production. Ziram lowered SIRT1/PGC-1α and phosphorylated protein levels of AKT1. Ziram induced ROS and apoptosis and lowered the mitochondrial membrane potential of progenitor Leydig cells in vitro. In conclusion, ziram disrupts Leydig cell development during the prepubertal period potentially through the SIRT1/PGC-1α and phosphorylated AKT1 signaling.