Inhibitor of cardiolipin biosynthesis-related enzyme MoGep4 confers broad-spectrum anti-fungal activity.

Plant pathogens cause devastating diseases, leading to serious losses to agriculture. Mechanistic understanding of pathogenesis of plant pathogens lays the foundation for the development of fungicides for disease control. Mitophagy, a specific form of autophagy, is important for fungal virulence. The role of cardiolipin, mitochondrial signature phospholipid, in mitophagy and pathogenesis is largely unknown in plant pathogenic fungi. The functions of enzymes involved in cardiolipin biosynthesis and relevant inhibitors were assessed using a set of assays, including genetic deletion, plant infection, lipidomics, chemical-protein interaction, chemical inhibition, and field trials. Our results showed that the cardiolipin biosynthesis-related gene MoGEP4 of the rice blast fungus Magnaporthe oryzae regulates growth, conidiation, cardiolipin biosynthesis, and virulence. Mechanistically, MoGep4 regulated mitophagy and Mps1-MAPK phosphorylation, which are required for virulence. Chemical alexidine dihydrochloride (AXD) inhibited the enzyme activity of MoGep4, cardiolipin biosynthesis and mitophagy. Importantly, AXD efficiently inhibited the growth of 10 plant pathogens and controlled rice blast and Fusarium head blight in the field. Our study demonstrated that MoGep4 regulates mitophagy, Mps1 phosphorylation and pathogenesis in M. oryzae. In addition, we found that the MoGep4 inhibitor, AXD, displays broad-spectrum antifungal activity and is a promising candidate for fungicide development.

Repurposing old drugs as antiviral agents for coronaviruses.

BACKGROUND: New therapeutic options to address the ongoing coronavirus disease 2019 (COVID-19) pandemic are urgently needed. One possible strategy is the repurposing of existing drugs approved for other indications as antiviral agents for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Due to the commercial unavailability of SARS-CoV-2 drugs for treating COVID-19, we screened approximately 250 existing drugs or pharmacologically active compounds for their inhibitory activities against feline infectious peritonitis coronavirus (FIPV) and human coronavirus OC43 (HCoV-OC43), a human coronavirus in the same genus (Betacoronavirus) as SARS-CoV-2. METHODS: FIPV was proliferated in feline Fcwf-4 cells and HCoV-OC43 in human HCT-8 cells. Viral proliferation was assayed by visualization of cytopathic effects on the infected Fcwf-4 cells and immunofluorescent assay for detection of the nucleocapsid proteins of HCoV-OC43 in the HCT-8 cells. The concentrations (EC50) of each drug necessary to diminish viral activity to 50% of that for the untreated controls were determined. The viabilities of Fcwf-4 and HCT-8 cells were measured by crystal violet staining and MTS/PMS assay, respectively. RESULTS: Fifteen out of the 252 drugs or pharmacologically active compounds screened were found to be active against both FIPV and HCoV-OC43, with EC50 values ranging from 11 nM to 75 µM. They are all old drugs as follows, anisomycin, antimycin A, atovaquone, chloroquine, conivaptan, emetine, gemcitabine, homoharringtonine, niclosamide, nitazoxanide, oligomycin, salinomycin, tilorone, valinomycin, and vismodegib. CONCLUSION: All of the old drugs identified as having activity against FIPV and HCoV-OC43 have seen clinical use in their respective indications and are associated with known dosing schedules and adverse effect or toxicity profiles in humans. Those, when later confirmed to have an anti-viral effect on SARS-CoV-2, should be considered for immediate uses in COVID-19 patients.

Artificial intelligence approach fighting COVID-19 with repurposing drugs.

Background: The ongoing COVID-19 pandemic has caused more than 193,825 deaths during the past few months. A quick-to-be-identified cure for the disease will be a therapeutic medicine that has prior use experiences in patients in order to resolve the current pandemic situation before it could become worsening. Artificial intelligence (AI) technology is hereby applied to identify the marketed drugs with potential for treating COVID-19. Methods: An AI platform was established to identify potential old drugs with anti-coronavirus activities by using two different learning databases; one consisted of the compounds reported or proven active against SARS-CoV, SARS-CoV-2, human immunodeficiency virus, influenza virus, and the other one containing the known 3C-like protease inhibitors. All AI predicted drugs were then tested for activities against a feline coronavirus in in vitro cell-based assay. These assay results were feedbacks to the AI system for relearning and thus to generate a modified AI model to search for old drugs again. Results: After a few runs of AI learning and prediction processes, the AI system identified 80 marketed drugs with potential. Among them, 8 drugs (bedaquiline, brequinar, celecoxib, clofazimine, conivaptan, gemcitabine, tolcapone, and vismodegib) showed in vitro activities against the proliferation of a feline infectious peritonitis (FIP) virus in Fcwf-4 cells. In addition, 5 other drugs (boceprevir, chloroquine, homoharringtonine, tilorone, and salinomycin) were also found active during the exercises of AI approaches. Conclusion: Having taken advantages of AI, we identified old drugs with activities against FIP coronavirus. Further studies are underway to demonstrate their activities against SARS-CoV-2 in vitro and in vivo at clinically achievable concentrations and doses. With prior use experiences in patients, these old drugs if proven active against SARS-CoV-2 can readily be applied for fighting COVID-19 pandemic.

Inhibition of SARS-CoV-2 by Highly Potent Broad-Spectrum Anti-Coronaviral Tylophorine-Based Derivatives.

Tylophorine-based compounds and natural cardiotonic steroids (cardenolides and bufadienolides) are two classes of transmissible gastroenteritis coronavirus inhibitors, targeting viral RNA and host cell factors, respectively. We tested both types of compounds against two types of coronaviruses, to compare and contrast their antiviral properties, and with view to their further therapeutic development. Examples of both types of compounds potently inhibited the replication of both feline infectious peritonitis virus and human coronavirus OC43 with EC50 values of up to 8 and 16 nM, respectively. Strikingly, the tylophorine-based compounds tested inhibited viral yields of HCoV-OC43 to a much greater extent (7-8 log magnitudes of p.f.u./ml) than the cardiotonic steroids (about 2-3 log magnitudes of p.f.u./ml), as determined by end point assays. Based on these results, three tylophorine-based compounds were further examined for their anti-viral activities on two other human coronaviruses, HCoV-229E and SARS-CoV-2. These three tylophorine-based compounds inhibited HCoV-229E with EC50 values of up to 6.5 nM, inhibited viral yields of HCoV-229E by 6-7 log magnitudes of p.f.u./ml, and were also found to inhibit SARS-CoV-2 with EC50 values of up to 2.5-14 nM. In conclusion, tylophorine-based compounds are potent, broad-spectrum inhibitors of coronaviruses including SARS-CoV-2, and could be used for the treatment of COVID-19.

Assessment of respiratory drive with esophageal diaphragmatic electromyography in patients with acute respiratory distress syndrome treated with prone position ventilation.

BACKGROUND: Prone position ventilation (PPV) is an important strategy for patients with severe acute respiratory distress syndrome (ARDS). This prospective study investigated the use of electromyography of the diaphragm (EMGdi) for monitoring respiratory drive in patients with moderate to severe ARDS during long-term PPV. METHODS: An integrated nostril-gastric feeding tube containing an esophageal electrode and balloon was placed in 14 patients with severe ARDS prior to PPV. EMGdi and trans-pulmonary pressure (∆PL) data were collected before PPV (baseline), every 2 h during PPV, and 2 h after the restoration of supine position ventilation (post-2 h SPV). RESULTS: In ARDS patients, the static compliance of the chest wall was significantly decreased after PPV. EMGdi levels were slightly lower in the early, middle, and late stages of PPV compared with baseline. Patients who received neuromuscular blocker experienced a greater drop in EMGdi from baseline than those who did not. CONCLUSIONS: For ARDS patients, EMGdi was slightly decreased after prolonged PPV. This is contrary to the change in diaphragm electromyography during normal body position changes. Monitoring EMGdi regularly during PPV in ARDS patients is feasible and can be used as a reference for lung protective ventilation strategies.

Risk factors for dysphagia after anterior cervical spine surgery: A meta-analysis.

BACKGROUND: Dysphagia is a well-known complication following anterior cervical spine surgery. Although risk factors for dysphagia have been reported in the literature, they still remain controversial. This study aims to investigate the risk factors associated with dysphagia following anterior cervical spinal surgery. METHODS: PubMed, EMBASE, and The Cochrane Library were searched up to June 2016 for studies examining dysphagia following anterior cervical spinal surgery. Risk factors associated with dysphagia were extracted. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for outcomes. Data analysis was conducted with RevMan 5.3 and STATA 12.0. RESULTS: The final analysis includes a total of 18 distinct studies. The pooled analysis reveals that there are significant differences in female gender (OR = 2.30, 95% CI: 1.76-2.99, P < 0.001), the use of anterior cervical plate (OR = 1.66, 95% CI: 1.05-2.62, P = 0.03), more than 1 surgical level (OR = 2.07, 95% CI: 1.62-2.66, P < 0.001), the upper surgical level at C3/4 (OR = 3.08, 95% CI: 1.44-6.55, P = 0.004), and the use of bone morphogenetic protein-2 (rhBMP-2) (OR = 5.52, 95% CI: 2.16-14.10, P < 0.001). However, no significant difference is found in revision surgery (OR = 1.67, 95% CI: 0.60-4.68, P = 0.33), the type of fusion (OR = 1.02, 95% CI: 0.62-1.67, P = 0.95), and cervical disc arthroplasty (OR = 1.37, 95% CI: 0.75-2.51, P = 0.30). CONCLUSION: Female gender, the use of anterior cervical plate, more than 1 surgical level, the upper surgical level at C3/4, and the use of rhBMP-2 are the risk factors for dysphagia following anterior cervical spinal surgery. However, revision surgery, the type of fusion, and cervical disc arthroplasty are unassociated with dysphagia. Considering the limited number of studies, this conclusion should be interpreted cautiously, and larger scale studies are required.

Factors predicting venous thromboembolism after spine surgery.

BACKGROUND: A meta-analysis was performed to explore predicted factors of venous thromboembolism (VTE) after surgery in the treatment for spine degeneration diseases. SUMMARY OF BACKGROUND DATA: Many scholars have focused on VTE after spine surgery, but as for the risk factors of VTE have not reached a consensus. METHODS: An extensive search of literature, "spine or spinal," "degeneration," "after surgery or postoperation," and "venous thromboembolism" as key words, was performed in PubMed/MEDLINE, Embase, the Cochrane library, CNKI, and WANFANG databases. The following variables were extracted: wearing elastic stocking, hypertension (HT), heart disease, diabetes, drinking, anticoagulant therapy, walking disability preoperation, smoking, sex, age, surgical duration, fusion versus nonfusion (lumbar fusion vs lumbar discectomy), surgical site (cervical vs lumbar), blood loss, and body mass index. Data analysis was conducted with RevMan 5.3 and STATA 12.0. RESULTS: A total of 12 studies were identified, including 34,597 patients of whom 624 patients had VTE, and the incidence of VTE was 2% in all patients who underwent spine surgery. The incidence of VTE for Asian patients was 7.5%, compared with 1% VTE for Occidental patients; the difference was significant (P < 0.0001). The pooled analysis showed that there were significant differences regarding wearing elastic stocking (odds ratio [OR] = 11.71, 95% confidence interval [CI] [1.46, 94.00], P = 0.02), walking disability preoperation (OR = 4.80, 95% CI [2.53, 9.12], P < 0.00001), surgical site (lumbar surgery) (OR = 0.23, 95% CI [0.20, 0.27], P < 0.00001), HT (OR = 1.59, 95% CI [1.21, 2.10], P = 0.001), and diabetes (OR = 2.12, 95% CI [1.09, 4.10], P = 0.03). However, there were no significant differences in blood loss, heart disease, smoking, sex, surgical duration, body mass index, surgical duration, anticoagulant therapy, wearing elastic stocking, fusion versus nonfusion, drinking, and age (all P > 0.05). CONCLUSIONS: Based on our meta-analysis, Asian patients, patients with walking disability preoperation, patients wearing elastic stocking, patients having undergone lumbar surgery, patients with a history of HT, and patients experiencing diabetes have a higher incidence of VTE after spine surgery.

[Effects of shenshuning recipe on expression of hepatocyte growth factor in rats with glomerulosclerosis].

OBJECTIVE: To investigate the effects of Shenshuning Recipe (SSNR) on gene expression of hepatocyte growth factor (HGF) in renal tissues in rats with glomerulosclerosis. METHODS: Glomerulosclerosis was induced in 42 rats by unilateral nephrectomy and intravenous injection of doxorubicin. Then these 42 rats were randomly divided into three groups: untreated group, SSNR-treated group and benazepril-treated group. Another eight rats were included into sham-operation group. The rats in the SSNR-treated group and the benazepril-treated group were fed SSNR or benazepril respectively for 8 weeks. The levels of 24 h urine protein (Upr), serum creatinine (Cr) and blood urea nitrogen (BUN) of rats in each group were examined. The renal morphological changes were observed under microscope, and the diameter of glomerular capillary, mesangial matrix and glomerulosclerosis index were analyzed by image analysis software. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the gene expression of HGF in the renal tissues. RESULTS: The levels of 24 h Upr, serum Cr and BUN in the untreated group were remarkably increased than those in the sham-operation group (P<0.01). The pathological morphological changes in the untreated group showed that the glomerulosclerosis was diffused around the renal tissue and the capillaries were shrunk. The expression level of mesangial matrix was up-regulated and the glomerulosclerosis index was 3.32+/-0.35. The expression level of HGF mRNA in the untreated group was obviously lower than that in the sham-operation group (P<0.05). The levels of 24 h Upr, serum Cr and BUN in the SSNR-treated group and the benazepril-treated group were remarkably decreased as compared with those in the untreated group, while the expression levels of HGF mRNA were both obviously higher than that in the untreated group (P<0.01). The pathological morphological changes in the SSNR-treated group and the benazepril-treated group were both alleviated. There was no significant difference in therapeutic effect between the SSNR-treated group and the benazepril-treated group. CONCLUSION: Shenshuning Recipe can up-regulate the expression of HGF mRNA, decrease the mesangial matrix, and improve the renal function, so that it may retard the development of glomerulosclerosis.

Therapeutic effects of clenbuterol in a murine model of amyotrophic lateral sclerosis.

We investigated the effects of clenbuterol, a beta2-adrenoceptor agonist with known anabolic and neuroprotective properties, on G93A-SOD1 mice, a transgenic murine model of familial amyotrophic lateral sclerosis (ALS). Relative to saline-treated vehicle controls (0.2 ml/kg/day; i.p.), early pathologic G93A-SOD1 mice treated with clenbuterol (1.5 mg/kg/day; i.p.) demonstrated a delayed onset of hindlimb signs as measured by rotarod performance, slowed disease progression, as well as trends toward mitigated losses of lumbar motoneurons and body weight. Responses in female G93A-SOD1 mice were favorable to those of males, suggesting synergistic effects between clenbuterol and sex-specific factors. Overall, our data suggest that clenbuterol offers therapeutic effects on ALS-related neuromuscular degeneration.

On the use of biomarkers for environmental health research.

This article discusses the growing interest in the use of biomarker data in environmental health research and considers some of the challenging statistical issues that arise. We specify a modeling framework that links environmental exposure, biomarkers and outcome, and discuss in conceptual terms how such a formulation could be used to inform dose response modeling for the purpose of quantitative risk assessment. We then analyse some biomarker data from a case-control study designed to elucidate the mechanisms of smoking induced lung cancer. Because of sample size limitations, we use a likelihood-based analysis which subsumes both cohort and case-control designs as special cases. Our analysis allows us to 1) investigate the extent to which the markers explain the pathway from exposure to outcome; 2) quantify the degree to which biomarker data can improve on predicting outcome over and above exposure; and 3) estimate the association among multiple markers.

The effect of crash experience on changes in risk taking among urban and rural young people.

A 20-month prospective study was conducted to investigate the effect of motorcycle crash experience on changes in risk taking among 2514 urban and 2304 rural students in Taiwan. Risk taking was assessed using a 14-item self-administered questionnaire at the beginning and end of the study. A risk-taking score for each student at the initial and the last follow-up assessments was generated from adding up points across all 14 items. For exposure variables, the study documented past motorcycle crash history at the initial assessment and collected detailed information about any motorcycle crash involvement that occurred during the study period. A general linear mixed model was applied to assess the effects of prior and recent crash involvements on the path of risk-taking behavior. The results show that at the initial assessment, students with crash experience had higher risk-taking levels than those without crash experience. However, crash experience, irregardless of whether it was measured in terms of crash history prior to the study, crash frequency, time elapsed since the last crash, or crash severity, did not significantly change the risk-taking path among students, even though its effect differed between urban and rural areas.

Factors associated with severity of motorcycle injuries among young adult riders.

STUDY OBJECTIVE: A cohort of 4,729 junior college students in Taiwan was studied to determine risk factors for increased severity of motorcycle injuries. METHODS: Crash characteristics were collected by using self-administered questionnaires soon after a student was involved in a motorcycle crash. The proportional odds model with generalized estimating equations, with correlated ordinal responses for collisions categorized into not injured, mildly injured, and severely injured categories, was used to determine the odds of injury severity. RESULTS: A total of 1,889 motorcycle crashes involving 1,284 persons occurred over a 20-month period from November 1994 to June 1996. There were 1,339 noninjuries, 474 mild injuries, and 76 severe injuries. The adjusted odds ratio (OR) of rural to urban roads having a greater level of injury severity was 1.64. Compared with noncollisions, collisions with a moving car (adjusted OR=1.76), a parked car (adjusted OR 1.90), or another stationary object (adjusted OR=2.31) increased the odds for a greater level of injury severity. Riders using Sanyang (adjusted OR=1.63) and Yamaha (adjusted OR=1.39) motorcycles had greater odds of being involved in a crash with a greater level of injury compared with those riding Kymco motorcycles. Darkness (adjusted OR=1.65) and greater speeds (adjusted OR=1.63 to 4.69) also increased the odds of greater injury severity. CONCLUSION: At the time of motorcycle crashes, factors such as being on rural roads, collisions with a heavier object, some motorcycle makes, darkness, and greater speeds increased the severity of motorcycle injuries among these young adult riders.

The impact of the Chi-Chi earthquake on quality of life among elderly survivors in Taiwan--a before and after study.

This paper examines the impact of the Chi-Chi earthquake, which hit central Taiwan on September 21, 1999, on the quality of life among the elderly survivors. The 28-item Taiwanese-adapted brief version of the World Health Organization's quality of life questionnaire (WHOQOL-BREF) was used to measure quality of life in four domains: physical capacity, psychological well-being, social relationships, and environment. These measures were coincidently collected in a separate study from 368 subjects aged 65 and older in the affected area shortly before the earthquake. Of these subjects, 268 were interviewed in a follow-up assessment 12 months after the earthquake. Linear mixed models were applied to investigate how quality of life in each of the four domains changed from the pre-earthquake assessment to 12 months after the earthquake, and how these changes depended on the level of damage to residences. In conclusion, elderly survivors tended to report lower quality of life in physical capacity, psychological well-being, and environment 12 months after the earthquake than at the assessment prior to the earthquake, regardless of the level of damage to their residences during the earthquake. However, those whose residences completely collapsed during the earthquake reported a higher quality of life in social relationships while others reported the opposite.