Hybrid Sequencing-Based Genomic Analysis of Klebsiella pneumoniae from Urinary Tract Infections Among Inpatients at a Tertiary Hospital in Beijing.

Background: Urinary tract infection (UTI) associated with Klebsiella pneumoniae poses a serious threat for inpatients. This study aimed to describe the genomic characteristics of K. pneumoniae causing UTI in a tertiary-care hospital in Beijing, China. Methods: A total of 20 K. pneumoniae strains collected from 2020 to 2021 were performed whole-genome sequencing. The Antibiotic susceptibility of 19 common antimicrobial agents was tested against all strains. The multi-locus sequence types (MLSTs) and serotypes were determined from the WGS data. De novo assemblies were used to identify resistance and virulence genes. The presence and characteristics of the plasmids were detected using hybrid assembly of long and short-read data. Results: These K. pneumoniae strains were clustered into nine sequence types (STs) and twelve K-serotypes. All the carbapenem-resistant K. pneumoniae (CRKP) strains acquired carbapenemase blaKPC-2 (n=7). Two CRKP strains exhibited increased resistance to Polymyxin B with MIC ≥ 4 mg/L due to insertion of an IS5-like sequence in the mgrB gene, and they were also involved in a transmission event in Intensive Care Unit. Long-read assemblies identified many plasmids co-carrying multiple replicons. Acquisition of a new IncM2_1 type blaCTX-M-3 positive plasmid was observed after transfer from ICU to neurovascular surgery by comparing the two strains collected from the same patient. Conclusion: K. pneumoniae is a significant pathogen responsible for urinary tract infections. The ST11-KL47 strain, prevalent at our hospital, exhibits a combination of high drug resistance and hypervirulence. It is imperative to enhance ongoing genomic surveillance of urinary tract infection-causing pathogens.

Spontaneous rupture­induced life­threatening mediastinal mixed germ cell tumor: A case report and therapeutic considerations.

Spontaneous rupture and hemorrhage of mediastinal germ cell tumors is a rare occurrence. In the current report, the case of a 20-year-old male patient who was admitted with chest tightness and dyspnea is presented. An urgent chest CT scan revealed a large tumor in the right anterior mediastinum, measuring ~12 cm in diameter, with associated intratumoral hemorrhage. An emergency thoracotomy was performed to excise the lesion, which revealed that the bleeding was caused by a ruptured tumor. Postoperative pathological findings revealed a mediastinal mixed germ cell tumor consisting of four pathological types: Embryonal carcinoma, seminoma, yolk sac tumor and immature teratoma. Postoperatively, the patient showed marked improvement in the symptoms of dyspnea. However, the follow-up outcome was poor, and the patient succumbed 2 months after surgery. To the best of our knowledge, there are no reports of rupture and hemorrhage involving >4 mixed germ cell tumors. In the present report, the experience of the treatment of the patient is summarized, and literature was reviewed to improve clinicians' awareness of the disease.

A review of mitigation technologies and management strategies for greenhouse gas and air pollutant emissions in livestock production.

One of the key issues in manure management of livestock production is to reduce greenhouse gas (GHG) and air pollutant emissions, which lead to significant environmental footprint and human/animal health threats. This study provides a review of potentially efficacious technologies and management strategies that reduce GHG and air pollutant emissions during the three key stages of manure management in livestock production, i.e., animal housing, manure storage and treatment, and manure application. Several effective mitigation technologies and practices for each manure management stage are identified and analyzed in detail, including feeding formulation adjustment, frequent manure removal and air scrubber during animal housing stage; solid-liquid separation, manure covers for storage, acidification, anaerobic digestion and composting during manure storage and treatment stage; land application techniques at appropriate timing during manure application stage. The results indicated several promising approaches to reduce multiple gas emissions from the entire manure management. Removing manure 2-3 times per week or every day during animal housing stage is an effective and simple way to reduce GHG and air pollutant emissions. Acidification during manure storage and treatment stage can reduce ammonia and methane emissions by 33%-93% and 67%-87%, respectively and proper acid, such as lactic acid can also reduce nitrous oxide emission by about 90%. Shallow injection of manure for field application has the best performance in reducing ammonia emission by 62%-70% but increase nitrous oxide emission. The possible trade-off brings insight to the prioritization of targeted gas emissions for the researchers, stakeholders and policymakers, and also highlights the importance of assessing the mitigation technologies across the entire manure management chain. Implementing a combination of the management strategies needs comprehensive considerations about mitigation efficiency, technical feasibility, local regulations, climate condition, scalability and cost-effectiveness.

Candidatus Accumulibacter use fermentation products for enhanced biological phosphorus removal.

Previous research suggested that two major groups of polyphosphate-accumulating organisms (PAOs), i.e., Ca. Accumulibacter and Tetrasphaera, play cooperative roles in enhanced biological phosphorus removal (EBPR). The fermentation of complex organic compounds by Tetrasphaera provides carbon sources for Ca. Accumulibacter. However, the viability of the fermentation products (e.g., lactate, succinate, alanine) as carbon sources for Ca. Accumulibacter and their potential effects on the metabolism of Ca. Accumulibacter were largely unknown. This work for the first time investigated the capability and metabolic details of Ca. Accumulibacter cognatus clade IIC strain SCUT-2 (enriched in a lab-scale reactor with a relative abundance of 42.8%) in using these fermentation products for EBPR. The enrichment culture was able to assimilate lactate and succinate with the anaerobic P release to carbon uptake ratios of 0.28 and 0.36 P mol/C mol, respectively. In the co-presence of acetate, the uptake of lactate was strongly inhibited, since two substrates shared the same transporter as suggested by the carbon uptake bioenergetic analysis. When acetate and succinate were fed at the same time, Ca. Accumulibacter assimilated two carbon sources simultaneously. Proton motive force (PMF) was the key driving force (up to 90%) for the uptake of lactate and succinate by Ca. Accumulibacter. Apart from the efflux of proton in symport with phosphate via the inorganic phosphate transport system, translocation of proton via the activity of fumarate reductase contributed to the generation of PMF, which agreed with the fact that PHV was a major component of PHA when lactate and succinate were used as carbon sources, involving the succinate-propionate pathway. Metabolic models for the usage of lactate and succinate by Ca. Accumulibacter for EBPR were built based on the combined physiological, biochemical, metagenomic, and metatranscriptomic analyses. Alanine was shown as an invalid carbon source for Ca. Accumulibacter. Instead, it significantly and adversely affected Ca. Accumulibacter-mediated EBPR. Phosphate release was observed without alanine uptake. Significant inhibitions on the aerobic phosphate uptake was also evident. Overall, this study suggested that there might not be a simply synergic relationship between Ca. Accumulibacter and Tetrasphaera. Their interactions would largely be determined by the kind of fermentation products released by the latter.

Nodakenin alleviates ovariectomy-induced osteoporosis by modulating osteoblastogenesis and osteoclastogenesis.

Osteoporosis, a systemic bone disease defined by decreased bone mass and deterioration of bone microarchitecture, is becoming a global concern. Nodakenin (NK) is a furanocoumarin-like compound isolated from the traditional Chinese medicine Radix Angelicae biseratae (RAB). NK has been reported to have various pharmacological activities, but osteoporosis has not been reported to be affected by NK. In this study, we used network pharmacology, molecular docking and molecular dynamics simulation techniques to identify potential targets and pathways of NK in osteoporosis. We found that NK treatment significantly promoted osteogenic differentiation of BMSCs while activating the PI3K/AKT/mTOR signalling pathway by measuring alkaline phosphatase activity and the expression of various osteogenic markers. In contrast, LY294002, an inhibitor of PI3K, reversed these changes and inhibited the osteogenic differentiation-enabling effect of NK. Meanwhile, prevent the Akt and NFκB signalling pathways by down-regulating c-Src and TRAF6 thereby effectively inhibiting RANKL-induced osteoclastogenesis. In addition, oral administration of NK to mice significantly elevated bone mass and ameliorated ovariectomized (OVX)-mediated bone microarchitectural disorders. In conclusion, these data suggest that NK attenuates OVX-induced bone loss by enhancing osteogenesis and inhibiting osteoclastogenesis.

Methionine Supplementation Affects Fecal Bacterial Community and Production Performance in Sika Deer (Cervus nippon).

Amino acid balance is central to improving the efficiency of feed protein utilization and for reducing environmental pollution caused by intensive farming. In previous studies, supplementation with limiting amino acids has been shown to be an effective means of improving animal nutrient utilization and performance. In this experiment, the effects of methionine on the apparent digestibility of nutrients, antler nutrient composition, rumen fluid amino acid composition, fecal volatile fatty acids and intestinal bacteria in antler-growing sika deer were investigated by randomly adding different levels of methionine to the diets of three groups of four deer at 0 g/day (CON), 4 g/day (LMet) and 6 g/day (HMet). Methionine supplementation significantly increased the apparent digestibility of organic matter, neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the LMet group (p < 0.05). The crude protein and collagen protein of antlers were significantly higher in the LMet and HMet groups compared to the CON group and also significantly higher in the HMet group compared to the LMet group, while the calcium content of antlers was significantly lower in the HMet group (p < 0.05). Ruminal fluid free amino acid composition was altered in the three groups of sika deer, with significant changes in aspartic acid, citrulline, valine, cysteine, methionine, histidine and proline. At the phylum level, Firmicutes and Bacteroidetes were highest in the rectal microflora. Unidentified bacterial abundance was significantly decreased in the HMet group compared to the CON group. Based on the results of principal coordinate analysis (PCoA) and Adonis analysis, there was a significant difference in the composition of the intestinal flora between the CON and HMet groups (p < 0.05). At the genus level, compared with the CON group, the abundance of Rikenellaceae_RC9_gut_group and Lachnospiraceae_UCG-010 in the LMet group increased significantly (p < 0.05), the abundance of dgA-11_gut_group in the HMet group decreased significantly (p < 0.05) and the abundance of Lachnospiraceae_UCG-010, Saccharofermentans and Lachnospiraceae_NK3A20_group increased significantly. Taken together, the results showed that methionine supplementation was beneficial in increasing the feed utilization efficiency and improving antler quality in sika deer, while affecting the composition of fecal bacteria.

Altered Respiratory Microbiomes, Plasma Metabolites, and Immune Responses in Influenza A Virus and Methicillin-Resistant Staphylococcus aureus Coinfection.

Influenza A virus (IAV)-methicillin-resistant Staphylococcus aureus (MRSA) coinfection causes severe respiratory infections. The host microbiome plays an important role in respiratory tract infections. However, the relationships among the immune responses, metabolic characteristics, and respiratory microbial characteristics of IAV-MRSA coinfection have not been fully studied. We used specific-pathogen-free (SPF) C57BL/6N mice infected with IAV and MRSA to build a nonlethal model of IAV-MRSA coinfection and characterized the upper respiratory tract (URT) and lower respiratory tract (LRT) microbiomes at 4 and 13 days postinfection by full-length 16S rRNA gene sequencing. Immune response and plasma metabolism profile analyses were performed at 4 days postinfection by flow cytometry and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The relationships among the LRT microbiota, the immune response, and the plasma metabolism profile were analyzed by Spearman's correlation analysis. IAV-MRSA coinfection showed significant weight loss and lung injury and significantly increased loads of IAV and MRSA in bronchoalveolar lavage fluid (BALF). Microbiome data showed that coinfection significantly increased the relative abundances of Enterococcus faecalis, Enterobacter hormaechei, Citrobacter freundii, and Klebsiella pneumoniae and decreased the relative abundances of Lactobacillus reuteri and Lactobacillus murinus. The percentages of CD4+/CD8+ T cells and B cells in the spleen; the levels of interleukin-9 (IL-9), interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), IL-6, and IL-8 in the lung; and the level of mevalonolactone in plasma were increased in IAV-MRSA-coinfected mice. L. murinus was positively correlated with lung macrophages and natural killer (NK) cells, negatively correlated with spleen B cells and CD4+/CD8+ T cells, and correlated with multiple plasma metabolites. Future research is needed to clarify whether L. murinus mediates or alters the severity of IAV-MRSA coinfection. IMPORTANCE The respiratory microbiome plays an important role in respiratory tract infections. In this study, we characterized the URT and LRT microbiota, the host immune response, and plasma metabolic profiles during IAV-MRSA coinfection and evaluated their correlations. We observed that IAV-MRSA coinfection induced severe lung injury and dysregulated host immunity and plasma metabolic profiles, as evidenced by the aggravation of lung pathological damage, the reduction of innate immune cells, the strong adaptation of the immune response, and the upregulation of mevalonolactone in plasma. L. murinus was strongly correlated with immune cells and plasma metabolites. Our findings contribute to a better understanding of the role of the host microbiome in respiratory tract infections and identified a key bacterial species, L. murinus, that may provide important references for the development of probiotic therapies.

5-alpha reductase inhibitors and MRI prostates: actively reducing prostate sizes and ambiguity.

BACKGROUND: Magnetic resonance imaging (MRI) scans are increasingly first-line investigations for suspected prostate cancer, and essential in the decision for biopsy. 5-alpha reductase inhibitor (5-ARI) use has been shown to reduce prostate size and prostate cancer risk. However, insufficient data exists on how 5-ARI use affects MRI findings and yield of biopsy. This study explores the differences in imaging and prostate cancer diagnoses between patients receiving and not receiving 5-ARI therapy. METHODS: From 2015 to 2020, we collected retrospective data of consecutive patients undergoing prostate biopsy at one centre. We included patients who were biopsy-naïve, had prior negative biopsies, or on active surveillance for low-grade prostate cancer. Clinical and pathological data was collected, including 5-ARI use, Prostate Imaging Reporting and Data System (PIRADS) classification and biopsy results. RESULTS: 351 men underwent saturation biopsy with or without targeted biopsies. 54 (15.3%) had a history of 5-ARI use. On mpMRI, there was no significant difference between the 5ARI and non-5-ARI groups in PIRADS distribution, number of lesions, and lesion location. Significantly fewer cancers were detected in the 5-ARI group (46.3% vs. 68.0%; p < 0.01). There were no significant differences in PIRADS distribution in 5-ARI patients with positive and negative biopsy. CONCLUSION: Our study found significant differences in biochemical, imaging and biopsy characteristics between 5-ARI and non-5-ARI groups. While both groups had similar PIRADS distribution, 5-ARI patients had a lower rate of positive biopsies across all PIRADS categories, which may suggest that the use of 5ARI may confound MRI findings. Further studies on how 5-ARI therapy affects the imaging characteristics of prostate cancer should be performed.

Effects of Film Thickness of ALD-Deposited Al2O3, ZrO2 and HfO2 Nano-Layers on the Corrosion Resistance of Ti(N,O)-Coated Stainless Steel.

The goal of this stydy was to explore the potential of the enhanced corrosion resistance of Ti(N,O) cathodic arc evaporation-coated 304L stainless steel using oxide nano-layers deposited by atomic layer deposition (ALD). In this study, we deposited Al2O3, ZrO2, and HfO2 nanolayers of two different thicknesses by ALD onto Ti(N,O)-coated 304L stainless steel surfaces. XRD, EDS, SEM, surface profilometry, and voltammetry investigations of the anticorrosion properties of the coated samples are reported. The amorphous oxide nanolayers homogeneously deposited on the sample surfaces exhibited lower roughness after corrosion attack compared to the Ti(N,O)-coated stainless steel. The best corrosion resistance was obtained for the thickest oxide layers. All samples coated with thicker oxide nanolayers augmented the corrosion resistance of the Ti(N,O)-coated stainless steel in a saline, acidic, and oxidising environment (0.9% NaCl + 6% H2O2, pH = 4), which is of interest for building corrosion-resistant housings for advanced oxidation systems such as cavitation and plasma-related electrochemical dielectric barrier discharge for breaking down persistent organic pollutants in water.

Effect of Methionine Supplementation on Serum Metabolism and the Rumen Bacterial Community of Sika Deer (Cervus nippon).

Methionine is the first or second limiting amino acid for ruminants, such as sika deer, and has a variety of biological functions such as antioxidant activity, immune response, and protein synthesis. This study aimed to investigate the effects of methionine supplementation on antler growth, serum biochemistry, rumen fermentation, and the bacterial community of sika deer during the antler-growing period. Twelve 4-year-old male sika deer were randomly assigned to three dietary groups supplemented with 0 g/day (n = 4, CON), 4.0 g/day (n = 4, LMet), and 6.0 g/day (n = 4, HMet) methionine. No significant difference (p > 0.05) was found in the production performance between the three groups, but antler weight was higher in both the LMet and HMet groups than in the CON group. Methionine supplementation significantly increased the serum glutathione peroxidase activity (p < 0.05). The serum immunoglobulin G level was significantly higher in the HMet group than in the other two groups (p < 0.05). No significant effect was found on the apparent amino acid digestibility of the three groups, but cysteine and methionine digestibility were higher in the LMet group. The serum hydroxylysine level was significantly lower in the LMet and HMet groups, whereas the serum lysine level was significantly lower in the HMet group compared with the CON group (p < 0.05). The LMet group had the highest but a nonsignificant total volatile fatty acid content and significantly higher microbial protein content in the rumen than the CON group (p < 0.05). The phyla Bacteroidetes, Firmicutes, and Proteobacteria were dominant in the rumen of the sika deer. The principal coordinate analysis (PCoA) and analysis of similarities (ANOSIM) results showed a significant change in the bacterial composition of the three groups (p < 0.05). The relative abundance of Prevotella and Rikenellaceae-RC9 was significantly higher in the LMet group compared with the CON group and CON and HMet groups, respectively. These results revealed that methionine supplementation improved the antioxidant activity and immune status, affecting amino acid metabolism and rumen microbial composition of the sika deer.

Rapid PCR-Based Nanopore Adaptive Sequencing Improves Sensitivity and Timeliness of Viral Clinical Detection and Genome Surveillance.

Nanopore sequencing has been widely used for the real-time detection and surveillance of pathogens with portable MinION. Nanopore adaptive sequencing can enrich on-target sequences without additional pretreatment. In this study, the performance of adaptive sequencing was evaluated for viral genome enrichment of clinical respiratory samples. Ligation-based nanopore adaptive sequencing (LNAS) and rapid PCR-based nanopore adaptive sequencing (RPNAS) workflows were performed to assess the effects of enrichment on nasopharyngeal swab samples from human adenovirus (HAdV) outbreaks. RPNAS was further applied for the enrichment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from nasopharyngeal swab samples to evaluate sensitivity and timeliness. The RPNAS increased both the relative abundance (7.87-12.86-fold) and data yield (1.27-2.15-fold) of HAdV samples, whereas the LNAS increased only the relative abundance but had no obvious enrichment on the data yield. Compared with standard nanopore sequencing, RPNAS detected the SARS-CoV-2 reads from two low-abundance samples, increased the coverage of SARS-CoV-2 by 36.68-98.92%, and reduced the time to achieve the same coverage. Our study highlights the utility of RPNAS for virus enrichment directly from clinical samples, with more on-target data and a shorter sequencing time to recover viral genomes. These findings promise to improve the sensitivity and timeliness of rapid identification and genomic surveillance of infectious diseases.

Facile Synthesis of 4,4'-biphenyl Dicarboxylic Acid-Based Nickel Metal Organic Frameworks with a Tunable Pore Size towards High-Performance Supercapacitors.

Metal-organic frameworks (MOFs) have attracted significant research interest for supercapacitor applications due to their high-tunable conductivity and their structure's pore size. In this work, we report a facile one-step hydrothermal method to synthesize nickel-based metal-organic frameworks (MOF) using organic linker 4,4'-biphenyl dicarboxylic acid (BPDC) for high-performance supercapacitors. The pore size of the Ni-BPDC-MOF nanostructure is tuned through different synthesization temperatures. Among them, the sample synthesized at 180 °C exhibits a nanoplate morphology with a specific surface area of 311.99 m2·g-1, a pore size distribution of 1-40 nm and an average diameter of ~29.2 nm. A high specific capacitance of 488 F·g-1 has been obtained at a current density of 1.0 A·g-1 in a 3 M KOH aqueous electrolyte. The electrode shows reliable cycling stability, with 85% retention after 2000 cycles. The hydrothermal process Ni-BPDC-MOF may provide a simple and efficient method to synthesize high-performance hybrid MOF composites for future electrochemical energy storage applications.

Nitrogen-Doped Nanoporous Anodic Stainless Steel Foils towards Flexible Supercapacitors.

In this work, we report the fabrication and enhanced supercapacitive performance of nitrogen-doped nanoporous stainless steel foils, which have been prepared by electrochemical anodization and subsequent thermal annealing in ammonia atmosphere. The nanoporous oxide layers are grown on type-304 stainless steel foil with optimal thickness ~11.9 µm. The N-doped sample exhibits high average areal capacitance of 321.3 mF·cm-2 at a current density of 1.0 mA·cm-2, 3.6 times of increment compared with untreated one. Structural and electrochemical characterizations indicate that the significant enhancement is correlated to the high charge transfer efficiency from nitriding nanosheet products Fe3N. Our report here may provide new insight on the development of high-performance, low-cost and binder-free supercapacitor electrodes for flexible and portable electronic device applications with multiple anions.

aBVA Procedure by Uniportal Video-Assisted Thoracoscopic Surgery for Right Upper Peripheral Lung Cancer: A Randomized Trial.

OBJECTIVE: This study aims to determine the optimal dividing order of anatomic pulmonary resection under uniportal video-assisted thoracoscopic surgery (uni-VATS) for patients with right upper peripheral lung cancer. METHODS: Patients who met the eligibility criteria were randomly allocated into the aBVA and VAB groups. In the aBVA group, the surgical procedure proceeded from the posterior to the anterior region (from the deeper to the superficial site). In the VAB group, the dissection orders were vein first followed by arterial branches, followed by the bronchus. Clinical data were collected and analyzed. RESULTS: Sixty patients were randomly allocated to the aBVA group (n = 30) and the VAB group (n = 30). The operation time in the aBVA group (230.500 ± 68.360 min) was significantly shorter than that in the VAB group (305.600 ± 107.821 min) (p = 0.01). The blood loss in the aBVA group (104.000 ± 70.935 ml) was significantly lower than that in the VAB group (391.000 ± 625.175 ml) (p = 0.01). Two patients in the VAB group underwent conversion to 2-portal VATS. The number of lymph nodes (13.367 ± 5.436 vs. 10.333 ± 7.279, p = 0.072) and lymph node stations (5.067 ± 1.574 vs. 4.467 ± 2.345, p = 0.567) were comparable between the two groups. The differences in the postoperative drainage tube time (5.033 ± 3.113 vs. 6.467 ± 4.447 days, p = 0.278) and hospital stay (8.233 ± 3.390 vs. 9.433 ± 4.523 days, p = 0.361) were not significantly different between the two groups. CONCLUSION: Compared with the VBA procedure, aBVA is easier for patients with right upper peripheral lung cancer who undergo uni-VATS lobectomy.

Combination of ultrasonic and acidic pretreatments for enhancing biohythane production from tofu processing residue via one-stage anaerobic digestion.

Tofu processing residues (TPR) have received more attention as a source of bioenergy. However, their low solubility has hindered biohythane generation. Consequently, the ultrasonic and H2SO4 pretreatments were combined and compared for the first time to improve the hydrolysis of organic matter and carbohydrate and increase free amino nitrogen generation from TPR. Besides, the impact of pretreatments on biohythane generation was investigated. Under the optimal conditions of 7.54% substrate level, 8% H2SO4 concentration, 80 °C and 50 min, the coincident ultrasonic-H2SO4 pretreatment enriched the contents of soluble chemical oxygen demand, reducing sugar, and free amino nitrogen to 49675 mg/L, 26 g/L, and 1721 mg/L, respectively, greater than individual pretreatments. Also, Biohythane yield increased by 4.24-13.61% over control (389.42 ± 23.7 ml/g-VSfed). Furthermore, hydrogen yield at 42.5 ± 2.08 and 28.1 ± 1.07 ml/g-VSfed and sulfate removal efficiency at 93 and 92% were significantly improved with ultrasonic-H2SO4 and H2SO4 pretreatments, respectively, indicating acidogenic and sulfidogenic activity enhancement.

Therapeutic Exon Skipping Through a CRISPR-Guided Cytidine Deaminase Rescues Dystrophic Cardiomyopathy in Vivo.

BACKGROUND: Loss of dystrophin protein causes Duchenne muscular dystrophy (DMD), characterized by progressive degeneration of cardiac and skeletal muscles, and mortality in adolescence or young adulthood. Although cardiac failure has risen as the leading cause of mortality in patients with DMD, effective therapeutic interventions remain underdeveloped, in part, because of the lack of a suitable preclinical model. METHODS: We analyzed a novel murine model of DMD created by introducing a 4-bp deletion into exon 4, one of the exons encoding the actin-binding domain 1 of dystrophin (referred to as DmdE4* mice). Echocardiography, microcomputed tomography, muscle force measurement, and histological analysis were performed to determine cardiac and skeletal muscle defects in these mice. Using this model, we examined the feasibility of using a cytidine base editor to install exon skipping and rescue dystrophic cardiomyopathy in vivo. AAV9-based CRISPR/Cas9-AID (eTAM) together with AAV9-sgRNA was injected into neonatal DmdE4* mice, which were analyzed 2 or 12 months after treatment to evaluate the extent of exon skipping, dystrophin restoration, and phenotypic improvements of cardiac and skeletal muscles. RESULTS: DmdE4* mice recapitulated many aspects of human DMD, including shortened life span (by ≈50%), progressive cardiomyopathy, kyphosis, profound loss of muscle strength, and myocyte degeneration. A single-dose administration of AAV9-eTAM instituted >50% targeted exon skipping in the Dmd transcripts and restored up to 90% dystrophin in the heart. As a result, early ventricular remodeling was prevented and cardiac and skeletal muscle functions were improved, leading to an increased life span of the DmdE4* mice. Despite gradual decline of AAV vector and base editor expression, dystrophin restoration and pathophysiological rescue of muscular dystrophy were long lasted for at least 1 year. CONCLUSIONS: Our study demonstrates the feasibility and efficacy to institute exon skipping through an enhanced TAM (eTAM) for therapeutic application(s).

Rapid identification and metagenomics analysis of the adenovirus type 55 outbreak in Hubei using real-time and high-throughput sequencing platforms.

The rise in human adenovirus (HAdV) infections poses a serious challenge to public health in China. Real-time (RT) sequencing provides solutions for achieving rapid pathogen identification during outbreaks, whereas high-throughput sequencing yields higher sequence accuracy. In the present study, we report the outcomes of applying nanopore and BGI platforms in the identification and genomic analysis of an HAdV outbreak in Hubei province, China in May of 2019. A mixed sample of nine nasopharyngeal swabs and one single sample were submitted to direct nanopore sequencing (MinION device), generating their first HAdV-55 reads within 13 and 20 min, respectively. The sequences were confirmed by RT-polymerase chain reaction (PCR). Ten HAdV-positive samples were further sequenced using next-generation high-throughput sequencing (BGISEQ-500 device). Phylogenetic analysis revealed that the outbreak strain had a close genetic relation to strains isolated in Sichuan province. Metagenomic analysis showed that HAdV-55 was not a dominant species in samples from which the whole HAdV-55 genome could not be assembled. The present results highlight the value of combining sequencing platforms and using mixed samples for nucleic acid enrichment in pathogen detection of infectious disease outbreaks.

A descriptive analysis of concurrent alcohol and substance use among patients living with HIV/HCV co-infection.

The objectives of this study were to estimate the prevalence of concurrent alcohol and substance use among patients living with HIV/HCV co-infection and to compare demographic and clinical characteristics of those with concurrent alcohol and substance to those with alcohol or substance use, and to those who were abstinent. We conducted an analysis of patient reported outcomes data of patients living with HIV/HCV co-infection (n = 327) who transitioned from primary care to sub-specialty care for evaluation of candidacy for HCV treatment at a university-affiliated HIV Clinic. The prevalence of self-reported concurrent alcohol and substance use was 33%. A higher proportion of those with concurrent alcohol and substance use were currently smoking tobacco, and those who were abstinent had higher ratings of health-related quality of life compared to those with alcohol or substance use. To reduce patients' risk for progression to advanced stages of HIV, HCV, and liver-related disease due to continued alcohol and substance and tobacco use, social workers and other health care professionals are encouraged to develop and implement intervention strategies to assist patients living with HIV/HCV co-infection in efforts to achieve behavioral change.

Physical Activity Maintenance Following Home-Based, Individually Tailored Print Interventions for African American Women.

African American women report low participation in physical activity and are disproportionately burdened by related conditions (obesity, breast, and colon cancer). Physical activity interventions have shown promising results among African American women, but most studies in this area have focused on short-term increases. More enduring changes in health behavior will be needed to eliminate existing health disparities. Thus, the current study examined 12-month physical activity and psychosocial outcomes from a pilot randomized controlled trial (N = 84) of a Home-based Individually tailored Physical activity Print (HIPP) intervention for African American women in the Deep South. Retention was 77.4% at 12 months. HIPP participants increased self-reported moderate-to-vigorous physical activity from 35.1 minutes/week (standard deviation [SD] = 47.8) at baseline to 124 minutes/week (SD = 95.5) at 12 months, compared with the wellness contact control participants who reported increases from 48.2 minutes/week (SD = 51.3) to 102.5 minutes/week (SD = 94.5) over 12 months (between-group p > .05). Results indicate that modest improvements in moderate-to-vigorous physical activity and related psychosocial variables occurred during the active intervention phase (months 0-6) and were sustained during the tapered maintenance period (months 6-12). Low-cost, high-reach, home-based strategies have great potential for supporting sustained participation in physical activity and achieving long-term health benefits among African American women in the Deep South.

Dichloroacetic acid (DCA) synergizes with the SIRT2 inhibitor Sirtinol and AGK2 to enhance anti-tumor efficacy in non-small cell lung cancer.

Combination chemotherapy is a potentially promising approach to enhance anticancer activity, overcome drug resistance, and improve disease-free and overall survival. The current study investigates the antitumor activity of sodium dichloroacetic acid (DCA) in combination with SIRT2 inhibitor Sirtinol and AGK2. We found that combining DCA with Sirtinol produced a synergistic therapeutic benefit in A549 and H1299 NSCLC cells in vitro and in a mouse A549 xenograft model. Synergistic potentiation of oxidative phosphorylation (OXPHOS) was observed, including decreased glucose consumption, decreased lactate production, increased OCR and increased ROS generation, possibly via co-targeting pyruvate dehydrogenase alpha 1(PDHA1). Mechanically, AGK2 and Sirtinol were found to increase the lysine-acetylation and decrease the serine-phosphorylation of PDHA1, which enabled the two inhibitors to synergize with DCA to further activate PDHA1. Besides, a AMPKα-ROS feed-forward loop was notably activated after the combined treatments compared with mono-therapy. Our results indicate that the combination of DCA and SIRT2 inhibitor may provide a promising therapeutic strategy to effectively kill cancer cells.