Transcriptome and metabolome analyses reveal the regulatory role of MdPYL9 in drought resistance in apple.

BACKGROUND: The mechanisms by which the apple MdPYL9 gene mediates the response to drought stress remain unclear. Here, transcriptome and metabolome analyses of apple plants under drought were used to investigate the mechanisms by which MdPYL9 regulates the response to drought stress in apple. MdPYL9-overexpressed transgenic and non-transgenic apple histoculture seedlings were rooted, transplanted, and subjected to drought treatments to clarify the mechanisms underlying the responses of apples to drought stress through phenotypic observations, physiological and biochemical index measurements, and transcriptomic and metabolomic analyses. RESULTS: Under drought stress treatment, transgenic plants were less affected by drought stress than non-transgenic plants. Decreases in the net photosynthetic rate, stomatal conductance, and transpiration rate of transgenic apple plants were less pronounced in transgenic plants than in non-transgenic plants, and increases in the intercellular CO2 concentration were less pronounced in transgenic plants than in non-transgenic plants. The relative electrical conductivity and content of malondialdehyde, superoxide anion, and hydrogen peroxide were significantly lower in transgenic plants than in non-transgenic plants, and the chlorophyll content and activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) were significantly higher in transgenic plants than in non-transgenic plants. The number of differentially expressed genes (DEGs) involved in the response to drought stress was lower in transgenic plants than in non-transgenic plants, and the most significant and highly annotated DEGs in the transgenic plants were involved in the flavonoid biosynthesis pathway, and the most significant and highly annotated DEGs in control plants were involved in the phytohormone signal transduction pathway. The number of differentially accumulated metabolites involved in the response to drought stress was lower in transgenic plants than in non-transgenic plants, and up-regulated metabolites were significantly enriched in apigenin-7-O-glucoside in transgenic plants and in abscisic acid in non-transgenic plants. In the flavonoid biosynthetic pathway, the expression of genes encoding chalcone synthase (CHS) and chalcone isomerase (CHI) was more significantly down-regulated in non-transgenic plants than in transgenic plants, and the expression of the gene encoding 4-coumarate-CoA ligase (4CL) was more significantly up-regulated in transgenic plants than in non-transgenic plants, which resulted in the significant up-regulation of apigenin-7-O-glucoside in transgenic plants. CONCLUSIONS: The above results indicated that the over-expression of MdPYL9 increased the drought resistance of plants under drought stress by attenuating the down-regulation of the expression of genes encoding CHS and CHI and enhancing the up-regulated expression of the gene encoding 4CL, which enhanced the content of apigenin-7-O-glucoside.

Maternal exposure to DON during lactation induces testicular toxicity in pubertal and adult offspring mice.

Deoxynivalenol (DON), a type B trichothecene mycotoxin, commonly occurs in cereal grains, and poses significant health risks to humans and animals. Numerous studies reveal its obvious toxic effects on male reproductive performance as well as its ability to transfer from the lactating mother to the suckling offspring through colostrum and milk. The objective of this study was to evaluate the toxic effect of lactational DON exposure on testicular morphology, hormonal levels, inflammation, apoptosis and proliferation of germ cells, tight junction, and sperm quality in male offspring. Sixty-six male offspring mice from lactating dams exposed to DON were euthanized at PND 21 and PND 70 to investigate the reproductive toxicity. Our results indicated that maternal DON exposure had a significant impact on the weight and volume of the testes, caused testicular histopathology, and reduced testosterone levels by downregulating expressions of StAR, CYP11A1, and CYP17A1 in male offspring. We also found that maternal DON exposure led to testicular inflammation in male offspring, which was attributed to increased levels of inflammatory markers, including IL-1ß, IL-6, TNF-α, and IFN-γ. Maternal DON exposure resulted in impaired tight junctions of Sertoli cells in male offspring, as evidenced by decreased expressions of ZO-1, Occludin, and Claudin-3. In addition, maternal DON exposure caused a reduction in the number of Sertoli cells and germ cells, ultimately leading to decreased sperm count and quality in adult male offspring. Collectively, these findings provide compelling evidence that maternal exposure to DON during lactation causes testicular toxicity in both pubertal and adult male offspring.

Cost-effective In Vivo and In Vitro Mouse Models for Evaluating Anticryptosporidial Drug Efficacy: Assessing Vorinostat, Docetaxel, and Baicalein.

BACKGROUND: Cryptosporidiosis is a significant diarrheal disease in humans and animals. Immunodeficient mice are the primary small animal models, but their high costs and specialized breeding/housing requirements limit in vivo drug testing. Numerous anticryptosporidial lead compounds identified in vitro remain untested in vivo. METHODS: Cryptosporidium tyzzeri, a natural mouse parasite closely related to Cryptosporidium parvum and Cryptosporidium hominis, was isolated to establish an infection model in immunocompetent mice. The model was validated using classic anticryptosporidial drugs (paromomycin and nitazoxanide) and then employed to assess the efficacy of 3 new leads (vorinostat, docetaxel, and baicalein). An in vitro culture of C. tyzzeri was also developed to complement the animal model. RESULTS: Chronic C. tyzzeri infection was established in chemically immunosuppressed wild-type mice. Paromomycin (1000 mg/kg/d) and nitazoxanide (100 mg/kg/d) demonstrated efficacy against C. tyzzeri. Vorinostat (30 mg/kg/d), docetaxel (25 mg/kg/d), and baicalein (50 mg/kg/d) were highly effective against C. tyzzeri infection. In vitro, nitazoxanide, vorinostat, docetaxel, and baicalein exhibited low to submicromolar efficacy against C. tyzzeri. CONCLUSIONS: Novel in vivo and in vitro models have been developed for cost-effective anticryptosporidial drug testing. Vorinostat, docetaxel, and baicalein show potential for repurposing and/or optimization for developing new anticryptosporidial drugs.

Lactational exposure to Deoxynivalenol causes mammary gland injury via inducing inflammatory response and impairing blood-milk barrier integrity in mice.

Lactation is a unique physiological process to produce and secrete milk. Deoxynivalenol (DON) exposure during lactation has been demonstrated to affect adversely the growth development of offspring. However, the effects and potential mechanism of DON on maternal mammary glands remain largely unknown. In this study, we found the length and area of mammary glands were significantly reduced after DON exposure on lactation day (LD) 7 and LD 21. RNA-seq analysis results showed that the differentially expressed genes (DEGs) were significantly enriched in acute inflammatory response and HIF-1 signaling pathway, which led to an increase of myeloperoxidase activity and inflammatory cytokines. Furthermore, lactational DON exposure increased blood-milk barrier permeability by reducing the expression of ZO-1 and Occludin, promoted cell apoptosis by upregulating the expression of Bax and cleaved Caspase-3 and downregulating the expression of Bcl-2 and PCNA. Additionally, lactational DON exposure significantly decreased serum concentration of prolactin, estrogen, and progesterone. All these alterations eventually resulted in a decrease of ß-casein expression on LD 7 and LD 21. In summary, our findings indicated that lactational exposure to DON caused lactation-related hormone disorder and mammary gland injury induced by inflammatory response and blood-milk barrier integrity impairment, ultimately resulting in lower production of ß-casein.

The reciprocal relations among self-compassion, and depression among adolescents after the Jiuzhaigou earthquake: A three-wave cross-lagged study.

BACKGROUND: Previous cross-sectional studies have examined the relationship between self-compassion and depression. Although it is often implicitly assumed that self-compassion may increase the vulnerability of an individual to depression, only a few studies have assessed whether self-compassion is a cause or a consequence of depression or both. METHOD: To examine such reciprocal effects, we assessed self-compassion and depression via self-report measures. At the baseline assessment (Time 1, T1), 450 students (M = 13.72, SD = 0.83, 54.2% females) participated 10 months after the Jiuzhaigou earthquake. We reassessed the T1 sample after 6- and 12-month intervals. At Time 2 (T2) assessment, 398 (56.0% female participants) of the Wave 1 participants were retained, and at Time 3 (T3) assessment, 235 (52.5% female participants) of the T1 and T2 participants were retained. RESULTS: Cross-lagged analyses indicated that positive self-compassion could significantly reduce subsequent depression. However, depression did not significantly predict later positive self-compassion. Negative self-compassion at T1 increased depression at T2, but negative self-compassion at T2 did not significantly predict depression at T3. In addition, positive self-compassion significantly reduced subsequent negative self-compassion. CONCLUSIONS: Positive self-compassion appears to protect adolescents against depression and maintain this protection over time, whereas negative self-compassion may worsen depression in adolescents during the initial stages of traumatic events. Additionally, positive self-compassion may decrease the level of negative self-compassion.

The reciprocal relations among self-compassion, posttraumatic stress disorder, and posttraumatic growth among adolescents after the Jiuzhaigou earthquake: a three-wave cross-lagged study.

BACKGROUND: Previous cross-sectional studies have examined the relationship between self-compassion, post-traumatic stress disorder (PTSD), and post-traumatic growth (PTG). But no study has tested whether self-compassion is a cause or a consequence of PTSD, PTG, or both. AIMS: The cross-lag analysis was used to examine the reciprocal effects among self-compassion, PTSD, and PTG. METHOD: We used data from 244 adolescents who had experienced earthquakes. We assessed self-compassion, PTSD, and PTG via self-report measures after the earthquake in Jiuzhaigou, as well as 6 and 12 months later. RESULTS: Cross-lagged analyses indicated that positive self-compassion could significantly predict subsequent PTSD and PTG. Meanwhile, PTSD and PTG also predicted later positive self-compassion. Negative self-compassion at T1 increased PTSD at T2, and neither PTSD nor PTG significantly predicted subsequent negative self-compassion. In addition, negative self-compassion at T1 significantly predicted positive self-compassion at T2, while positive self-compassion at T2 significantly predicted negative self-compassion at T3. CONCLUSIONS: Positive self-compassion is a protective factor of post-traumatic psychological response, and it is maintained for a long time, while negative self-compassion may aggravate the negative psychological outcomes of adolescents in the early stage of experiencing traumatic events. In addition, positive and negative self-compassion can influence each other over time.

Novel functional mutation of the PDIA3 gene affects milk composition traits in Chinese Holstein cattle.

Protein disulfide isomerase family A member 3 (PDIA3) is a multifunctional protein, and it plays a vital role in modulating various cell biological functions under physiological and pathological conditions. Our previous study on Mediterranean buffalo demonstrated that PDIA3 is a potential candidate gene associated with milk yield based on genome-wide association study analysis. However, the genetic effects of the PDIA3 gene on milk performance in dairy cattle and the corresponding mechanism have not been documented. This study aims to explore the genetic effects of PDIA3 polymorphisms on milk production traits in 362 Chinese Holstein cattle. The results showed that 4 SNPs were identified from the 5' untranslated region of the PDIA3 gene in the studied population, of which 2 SNPs (g.-1713 C>T and g.-934 G>A) were confirmed to be significantly associated with milk protein percentage, whereas g.-434 C>T was significantly associated with milk fat percentage. Notably, linkage disequilibrium analysis indicated that 3 SNPs (g.-1713 C>T, g.-934 G>A, and g.-695 A>C) formed one haplotype block, which was found to be significantly associated with milk protein percentage. The luciferase assay demonstrated that allele C of g.-434 C>T exhibited a higher promotor activity compared with allele T, suggesting that g.-434 C>T might be a potential functional mutation affecting PDIA3 expression. Furthermore, overexpression of the PDIA3 gene was found to induce higher levels of triglyceride and BODIPY fluorescence intensity. In addition, PDIA3 overexpression was also found to positively regulate the synthesis and secretion of α-casein, ß-casein, and κ-casein, whereas knockdown of this gene showed the opposite effects. In summary, our findings revealed significant genetic effects of PDIA3 on milk composition traits, and the identified SNP and the haplotype block might be used as genetic markers for dairy cow selected breeding.

Deoxynivalenol exposure inhibits biosynthesis of milk fat and protein by impairing tight junction in bovine mammary epithelial cells.

Deoxynivalenol (DON) is one of the most common feed contaminants, and it poses a serious threat to the health of dairy cows. The existing studies of biological toxicity of DON mainly focus on the proliferation, oxidative stress, and inflammation in bovine mammary epithelial cells, while its toxicity on the biosynthesis of milk components has not been well documented. Hence, we investigated the toxic effects and the underlying mechanism of DON on the bovine mammary alveolar cells (MAC-T). Our results showed that exposure to various concentrations of DON significantly inhibited cell proliferation, induced apoptosis, and altered the cell morphology which was manifested by cell distortion and shrinkage. Moreover, the transepithelial electrical resistance (TEER) values of MAC-T cells exposed to DON were gradually decreased in a time- and concentration- dependent manner, but lactate dehydrogenase (LDH) leakage was significantly increased with the maximum increase of 2.4-fold, indicating the cell membrane and tight junctions were damaged by DON. Importantly, DON significantly reduced the synthesis of ß-casein and lipid droplets, along with the significantly decreases of phospho-mTOR, phospho-4EBP1, phospho-JAK2, and phospho-STAT5. Gene expression profiles showed that the expressions of several genes related to lipid synthesis and metabolism were changed, including acyl-CoA synthetase short-chain family member 2 (ACSS2), fatty acid binding protein 3 (FABP3), 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1), and insulin-induced gene 1 (INSIG1). GO and KEGG enrichment analyses revealed that the differentially expressed genes (DEGs) were significantly enriched in ribosome, glutathione metabolism, and lipid biosynthetic process, which play important roles in the toxicological process induced by DON. Taken together, DON affects the proliferation and functional differentiation of MAC-T cells, which might be related to the cell junction disruption and morphological alteration. Our data provide new insights into functional differentiation and transcriptomic alterations of MAC-T cells after DON exposure, which contributes to a comprehensive understanding of DON-induced toxicity mechanism.

Host cells with transient overexpression of MDR1 as a novel in vitro model for evaluating on-target effect for activity against the epicellular Cryptosporidium parasite.

OBJECTIVES: To rapidly generate host cells with resistance to multiple compounds for differentiating drug action on parasite target or the host cell target (i.e. on-target or off-target effect) against the zoonotic enteric parasite Cryptosporidium parvum. METHODS: Transient overexpression of a multidrug resistance protein 1 (MDR1) gene in host cells (HCT-8 cell line) was explored to increase drug tolerance of the host cells to selected anti-cryptosporidial leads. In vitro cytotoxicity and anti-cryptosporidial efficacy of selected compounds were evaluated on the parasite grown in WT parental and transiently transfected HCT-8 cells. The approach was based on the theory that, for an epicellular parasite receiving consistent exposure to compounds in culture medium, overexpressing MDR1 in HCT-8 cells would increase drug tolerance of host cells to selected compounds but would not affect the anti-cryptosporidial efficacy if the compounds acted solely on the parasite target and the drug action on host cell target played no role on the antiparasitic efficacy. RESULTS: Six known anti-cryptosporidial leads were tested. Transient overexpression of MDR1 increased drug tolerance of HCT-8 cells on paclitaxel, doxorubicin HCl and vincristine sulphate (2.11- to 2.27-fold increase), but not on cyclosporin A, daunorubicin HCl and nitazoxanide. Increased drug tolerance in host cells had no effect on antiparasitic efficacy of paclitaxel, but affected that of doxorubicin HCl. CONCLUSIONS: Data confirmed that, at efficacious concentrations, paclitaxel acted mainly on the parasite target, while doxorubicin might act on both parasite and host cell targets. This model can be employed for studying the action of additional anti-cryptosporidial leads, and adapted to studying drug action in other epicellular pathogens. The limitation of the model is that the anti-cryptosporidial leads/hits need to be MDR1 substrates.

The Association between Obesity and Severity in Patients with Coronavirus Disease 2019: a Retrospective, Single-center Study, Wuhan.

Aim: In other respiratory infectious diseases, obesity may be associated with a poor outcome. For coronavirus disease 2019 (COVID-19), the association between obesity and severity or prognosis requires further analysis. Methods: This was a retrospective, single-center study. Hospitalized patients were recruited in Renmin Hospital of Wuhan University from January 2, 2020 to February 20, 2020. The data of body mass index (BMI) was obtained from follow-up of surviving patients. According to BMI, normal weight was defined as 18.5-23.9 kg/m2, overweight as 24.0-27.9 kg/m2 and obesity as > 28.0 kg/m2. Results: A total of 463 patients were enrolled, of which 242 (52.3%) patients were in the normal weight group; 179 (38.7%) were in the overweight group; and 42 (9.1%) were in the obesity group. Compared to the normal group, obese patients were more likely to have a higher heart rate; lower finger oxygen saturation; higher levels of white blood cells, neutrophil counts, basophil counts, intravenous glucose, triacylglycerol, uric acid, alanine aminotransferase, creatine kinase-MB, CD19+ cell counts and percentage; and lower levels of monocyte percentage, high density lipoprotein and CD3+ cell percentage. In addition, the proportions of hypertension (21.5% vs. 42.6%) and severe+critical illness (47.8 vs. 81.0 %) were significantly higher in the obesity group than those in normal group. However, no significant differences were observed between the normal and obesity groups in critical illness, organ damage and defined endpoint (mechanical ventilation or intensive care unit). Multiple logistic regression showed that obesity increased the risk of developing severe+critical illness (Odd ratio 3.586, 95% CI 1.550-8.298, P=0.003) in patients with COVID-19, and did not affect the risk of critical illness, organ damage and endpoints. Overweight did not affect the risk of severity, organ damage or endpoint in patients with COVID-19. Conclusion: Obesity may be a risk factor for developing severity in patients with COVID-19.

Training with Agency-Inspired Feedback from an Instrumented Glove to Improve Functional Grasp Performance.

Sensory feedback from wearables can be effective to learn better movement through enhanced information and engagement. Facilitating greater user cognition during movement practice is critical to accelerate gains in motor function during rehabilitation following brain or spinal cord trauma. This preliminary study presents an approach using an instrumented glove to leverage sense of agency, or perception of control, to provide training feedback for functional grasp. Seventeen able-bodied subjects underwent training and testing with a custom-built sensor glove prototype from our laboratory. The glove utilizes onboard force and flex sensors to provide inputs to an artificial neural network that predicts achievement of "secure" grasp. Onboard visual and audio feedback was provided during training with progressively shorter time delay to induce greater agency by intentional binding, or perceived compression in time between an action (grasp) and sensory consequence (feedback). After training, subjects demonstrated a significant reduction (p < 0.05) in movement pathlength and completion time for a functional task involving grasp-move-place of a small object. Future work will include a model-based algorithm to compute secure grasp, virtual reality immersion, and testing with clinical populations.

Concurrent Continuous Versus Bandwidth Visual Feedback With Varying Body Representation for the 2-Legged Squat Exercise.

CONTEXT: Continuous visual feedback (VF) can improve abilities to achieve desired movements and maximize rehabilitation outcomes by displaying actual versus target body positions in real time. Bandwidth VF reduces the reliance on feedback by displaying movement cues only when performance errors exceed specified thresholds. As such, bandwidth VF may better train independent movement abilities through greater development of intrinsic body control. In this study, continuous and bandwidth VF were investigated across modes of display (abstract and representative) that differed in body-discernibility. OBJECTIVE: To compare the performance of the 2-legged squat during training with concurrent feedback (real-time VF) and short-term retention (immediately after training, VF removed). DESIGN: Cross-sectional. SETTING: University research laboratory. PARTICIPANTS: Eighteen healthy individuals. METHODS: Marker-based motion capture displayed real-time position. MAIN OUTCOME MEASURES: Four VF cases (continuous-abstract, bandwidth-abstract, continuous-representative, and bandwidth-representative) were evaluated for accuracy and consistency to a target trajectory and target depth. RESULTS: During training, both continuous VF cases showed significantly (P < .05) higher accuracy and consistency to the target trajectory compared with both bandwidth VF cases. Bandwidth VF resulted in greater potential learning (retention performance relative to a training baseline) compared with continuous-abstract. CONCLUSIONS: Continuous-representative may offer unique performance benefits in both training and retention of multisegment movement tasks. Bandwidth VF showed greater potential for learning. For long-term learning, an optimal VF paradigm should consider continuous-representative with bandwidth features.

The effects of CRISPR-Cas9 knockout of the TGF-ß1 gene on antler cartilage cells in vitro.

BACKGROUND: Deer antler is the only mammalian organ that can be completely regenerated every year. Its periodic regeneration is regulated by multiple factors, including transforming growth factor ß (TGF-ß). This widely distributed multi-functional growth factor can control the proliferation and differentiation of many types of cell, and it may play a crucial regulatory role in antler regeneration. This study explored the role of TGF-ß1 during the rapid growth of sika deer antler. METHODS: Three CRISPR-Cas9 knockout vectors targeting the TGF-ß1 gene of sika deer were constructed and packaged with a lentiviral system. The expression level of TGF-ß1 protein in the knockout cell line was determined using western blot, the proliferation and migration of cartilage cells in vitro were respectively determined using EdU and the cell scratch test, and the expression levels of TGF-ß pathway-related genes were determined using a PCR array. RESULTS: Of the three gRNAs designed, pBOBI-gRNA2 had the best knockout effect. Knockout of TGF-ß1 gene inhibits the proliferation of cartilage cells and enhances their migration in vitro. TGF-ß signaling pathway-related genes undergo significant changes, so we speculate that when the TGF-ß pathway is blocked, the BMP signaling pathway mediated by BMP4 may play a key role. CONCLUSIONS: TGF-ß1 is a newly identified regulatory factor of rapid growth in sika deer antler.

Posttraumatic stress disorder symptoms and posttraumatic growth among college students with a history of childhood maltreatment in China: A latent profile analysis.

BACKGROUND: Childhood maltreatment is prevalent in China, and research indicates that trauma-exposed individuals may exhibit concurrent negative and positive posttraumatic psychological responses. OBJECTIVE: To examine the co-occurring patterns of posttraumatic stress disorder (PTSD) symptoms and posttraumatic growth (PTG) among college students in China having a history of childhood maltreatment. METHOD: Participants (N = 2,968) were college students from China who had experienced childhood maltreatment and had completed the PTSD Checklist, a PTG inventory, and a childhood maltreatment questionnaire. RESULTS: The results revealed four heterogeneous profiles characterized by low symptoms (n = 666, 22.4%), growth (n = 960, 32.3%), struggle (n = 1,285, 43.3%), and distress (n = 57, 1.9%). The three-step approach revealed emotional abuse and sexual abuse as crucial risk factors for the distress profile. In addition, age, sex, parental education, and forms of childhood maltreatment play distinctive roles in the four profiles of posttraumatic reactions. CONCLUSION: Posttraumatic reactions exhibit heterogeneous characteristics among Chinese college students exposed to childhood maltreatment, and professionals provide the service targeting their needs. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Study on the Effect of Additives on the Performance of Cement-Based Composite Anti-Corrosion Coatings for Steel Bars in Prefabricated Construction.

The influence of polymer emulsion, pigment filler, and dispersant on the corrosion resistance of polymer cement-based composite anti-corrosion coatings were investigated in this study. Adhesion loss rate tests and electrochemical tests were conducted on samples. The research results show that optimal corrosion resistance can be achieved with a 45 wt% dosage of emulsion, a 6 wt% dosage of pigment filler, and a 0.30 wt% dosage of dispersant. The bonding properties of bare steel bars, epoxy-coated steel bars, and polymer cement-based composite anti-corrosion coated steel bars with grout were compared. The results show that the polymer cement-based composite anti-corrosion coating can enhance the bonding properties of the samples. Furthermore, the microscopic analysis was conducted on the samples. The results demonstrate that the appropriate addition of emulsion can fill internal pores of the coating, tightly bonding hydration products with unhydrated cement particles. Moreover, incorporating a suitable dosage of functional additives enhances the stability of the coating system and leads to a denser microstructure.

Lower micromolar activity of the antifungal imidazoles on the bacterial-type bifunctional aldehyde/alcohol dehydrogenase (AdhE) in Cryptosporidium parvum and in vitro efficacy against the zoonotic parasite.

Cryptosporidium parvum is a waterborne and foodborne zoonotic protozoan parasite, a causative agent of moderate to severe diarrheal diseases in humans and animals. However, fully effective treatments are unavailable for medical and veterinary uses. There is a need to explore new drug targets for potential development of new therapeutics. Because C. parvum relies on anaerobic metabolism to produce ATP, fermentative enzymes in this parasite are attractive targets for exploration. In this study, we investigated the ethanol-fermentation in the parasite and characterized the basic biochemical features of a bacterial-type bifunctional aldehyde/alcohol dehydrogenase, namely CpAdhE. We also screened 3892 chemical entries from three libraries and identified 14 compounds showing >50% inhibition on the enzyme activity of CpAdhE. Intriguingly, antifungal imidazoles and unsaturated fatty acids are the two major chemical groups among the top hits. We further characterized the inhibitory kinetics of selected imidazoles and unsaturated fatty acids on CpAdhE. These compounds displayed lower micromolar activities on CpAdhE (i.e., IC50 values ranging from 0.88 to 11.02 µM for imidazoles and 8.93 to 35.33 µM for unsaturated fatty acids). Finally, we evaluated the in vitro anti-cryptosporidial efficacies and cytotoxicity of three imidazoles (i.e., tioconazole, miconazole and isoconazole). The three antifungal imidazoles exhibited lower micromolar efficacies against the growth of C. parvum in vitro (EC50 values ranging from 4.85 to 10.41 µM and selectivity indices ranging from 5.19 to 10.95). The results provide a proof-of-concept data to support that imidazoles are worth being further investigated for potential development of anti-cryptosporidial therapeutics.

Neddylation inhibition affects early embryonic development by disrupting maternal-to-zygotic transition and mitochondrial function in mice.

Post-translational modifications (PTMs) are critical for early development in mice because early cleavage-stage embryos are characterized by transcriptional inactivity. Neddylation is an important ubiquitin-like PTM that regulates multiple biophysical processes. However, the exact roles of neddylation in regulating early embryonic development remain largely unknown. In the present study, we found that inhibition of neddylation by specific inhibitor MLN4924 led to severe arrest of early embryonic development. Transcriptomic analysis showed that neddylation inhibition changed the expression of 3959 genes at the 2-cell stage. Importantly, neddylation inhibition blocked zygotic genome activation and maternal mRNA degradation, thus disrupting the maternal-to-zygotic transition. Moreover, inhibition of neddylation induced mitochondrial dysfunction including aberrant mitochondrial distribution, decreased mitochondrial membrane potential, and reduced ATP content. Further analysis showed that inhibition of neddylation resulted in the accumulation of reactive oxygen species and superoxide anion, thereby resulting in oxidative stress and severe DNA damage at the 2-cell stage. Overall, this study demonstrates that neddylation is vital for early embryonic development in mice. Our findings suggest that proper neddylation regulation is essential for the timely inter-stage transition during early embryonic development.

Requirement of microtubules for secretion of a micronemal protein CpTSP4 in the invasive stage of the apicomplexan Cryptosporidium parvum.

The zoonotic Cryptosporidium parvum is a global contributor to infantile diarrheal diseases and opportunistic infections in immunocompromised or weakened individuals. Like other apicomplexans, it possesses several specialized secretory organelles, including micronemes, rhoptry, and dense granules. However, the understanding of cryptosporidial micronemal composition and secretory pathway remains limited. Here, we report a new micronemal protein in C. parvum, namely, thrombospondin (TSP)-repeat domain-containing protein-4 (CpTSP4), providing insights into these ambiguities. Immunostaining and enzyme-linked assays show that CpTSP4 is prestored in the micronemes of unexcysted sporozoites but secreted during sporozoite excystation, gliding, and invasion. In excysted sporozoites, CpTSP4 is also distributed on the two central microtubules unique to Cryptosporidium. The secretion and microtubular distribution could be completely blocked by the selective kinesin-5 inhibitors SB-743921 and SB-715992, resulting in the accumulation of CpTSP4 in micronemes. These support the kinesin-dependent microtubular trafficking of CpTSP4 for secretion. We also localize γ-tubulin, consistent with kinesin-dependent anterograde trafficking. Additionally, recombinant CpTSP4 displays nanomolar binding affinity to the host cell surface, for which heparin acts as one of the host ligands. A novel heparin-binding motif is identified and validated biochemically for its contribution to the adhesive property of CpTSP4 by peptide competition assays and site-directed mutagenesis. These findings shed light on the mechanisms of intracellular trafficking and secretion of a cryptosporidial micronemal protein and the interaction of a TSP-family protein with host cells.IMPORTANCECryptosporidium parvum is a globally distributed apicomplexan parasite infecting humans and/or animals. Like other apicomplexans, it possesses specialized secretory organelles in the zoites, in which micronemes discharge molecules to facilitate the movement and invasion of zoites. Although past and recent studies have identified several proteins in cryptosporidial micronemes, our understanding of the composition, secretory pathways, and domain-ligand interactions of micronemal proteins remains limited. This study identifies a new micronemal protein, namely, CpTSP4, that is discharged during excystation, gliding, and invasion of C. parvum sporozoites. The CpTSP4 secretion depends on the intracellular trafficking on the two Cryptosporidium-unique microtubes that could be blocked by kinesin-5/Eg5 inhibitors. Additionally, a novel heparin-binding motif is identified and biochemically validated, which contributes to the nanomolar binding affinity of CpTSP4 to host cells. These findings indicate that kinesin-dependent microtubular trafficking is critical to CpTSP4 secretion, and heparin/heparan sulfate is one of the ligands for this micronemal protein.

Exploring gender differences in the relationships among childhood maltreatment, PTSD, and depression in young adults through network analysis.

BACKGROUND AND OBJECTIVES: College students who have experienced childhood maltreatment commonly exhibit adverse psychological consequences such as post-traumatic stress disorder (PTSD) and depression. The aim of this study is to use network analysis to investigate the gender differences in complex connections among various types of childhood maltreatment, PTSD, and depression. METHODS: This study involved 481 participants (M = 19.25 years, 54.5 % female) who were selected from a larger sample of 5231 college students. These participants had experienced childhood maltreatment and exhibited significant clinical symptoms of PTSD and depression. The participants completed validated measures that assessed childhood trauma, PTSD, and depression. RESULTS: The network analysis performed on the entire sample revealed robust connections among various types of childhood maltreatment, symptom clusters of PTSD, and depression. The most central symptom that emerged was negative alterations in cognitions and mood (NACM), with emotional abuse displaying the highest centrality measure among the various types of childhood maltreatment. Upon comparing the subnetworks, the centrality analysis identified significant gender differences in nodes such as sexual abuse, physical neglect, emotional neglect, and avoidance. CONCLUSIONS: The study's findings confirm that various forms of childhood abuse have intricate and multifaceted connections with depression and PTSD symptoms in adulthood. The study suggests that NACM could be the most significant symptom, and emotional abuse may play a vital role in adverse psychological outcomes. Furthermore, notable gender discrepancies were identified in the relationship between various forms of maltreatment and psychopathological symptoms.

Augmented feedback modes during functional grasp training with an intelligent glove and virtual reality for persons with traumatic brain injury.

Introduction: Physical therapy is crucial to rehabilitating hand function needed for activities of daily living after neurological traumas such as traumatic brain injury (TBI). Virtual reality (VR) can motivate participation in motor rehabilitation therapies. This study examines how multimodal feedback in VR to train grasp-and-place function will impact the neurological and motor responses in TBI participants (n = 7) compared to neurotypicals (n = 13). Methods: We newly incorporated VR with our existing intelligent glove system to seamlessly enhance the augmented visual and audio feedback to inform participants about grasp security. We then assessed how multimodal feedback (audio plus visual cues) impacted electroencephalography (EEG) power, grasp-and-place task performance (motion pathlength, completion time), and electromyography (EMG) measures. Results: After training with multimodal feedback, electroencephalography (EEG) alpha power significantly increased for TBI and neurotypical groups. However, only the TBI group demonstrated significantly improved performance or significant shifts in EMG activity. Discussion: These results suggest that the effectiveness of motor training with augmented sensory feedback will depend on the nature of the feedback and the presence of neurological dysfunction. Specifically, adding sensory cues may better consolidate early motor learning when neurological dysfunction is present. Computerized interfaces such as virtual reality offer a powerful platform to personalize rehabilitative training and improve functional outcomes based on neuropathology.