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Background: Melioidosis is difficult to diagnose due to its wide range of clinical symptoms. The culture method is time-consuming and less sensitive, emphasizing the importance of rapid and accurate diagnostic tests for melioidosis. Burkholderia invasion protein D (BipD) of Burkholderia pseudomallei is a potential diagnostic biomarker. This study aimed to isolate and characterize single-stranded DNA aptamers that specifically target BipD., Methods: The recombinant BipD protein was produced, followed by isolation of BipD-specific aptamers using Systematic Evolution of Ligands by EXponential enrichment. The binding affinity and specificity of the selected aptamers were evaluated using Enzyme-Linked Oligonucleotide Assay., Results: The fifth SELEX cycle showed a notable enrichment of recombinant BipD protein-specific aptamers. Sequencing analysis identified two clusters with a total of seventeen distinct aptamers. AptBipD1, AptBipD13, and AptBipD50 were chosen based on their frequency. Among them, AptBipD1 exhibited the highest binding affinity with a K d value of 1.0 μM for the recombinant BipD protein. Furthermore, AptBipD1 showed significant specificity for B. pseudomallei compared to other tested bacteria., Conclusion: AptBipD1 is a promising candidate for further development of reliable, affordable, and efficient point-of-care diagnostic tests for melioidosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Aptamers have emerged as prominent ligands in clinical diagnostics because they provide various advantages over antibodies, such as quicker generation time, reduced manufacturing costs, minimal batch-to-batch variability, greater modifiability, and improved thermal stability. In the present study, we isolated and characterized DNA aptamers that can specifically bind to the hemolysin E (HlyE) antigen of Salmonella Typhi for future development of typhoid diagnostic tests. The DNA aptamers against Salmonella Typhi HlyE were isolated using systematic evolution of ligands by exponential enrichment (SELEX), and their binding affinity and specificity were assessed utilizing enzyme-linked oligonucleotide assay (ELONA). A total of 11 distinct aptamers were identified, and the binding affinities and species selectivities of the three most probable aptamers were determined. Kd values were obtained in the nanomolar range, with the highest affinity of 83.6 nM determined for AptHlyE97. In addition, AptHlyE11, AptHlyE45 and AptHlyE97 clearly distinguished S. Typhi HlyE from other tested bacteria, such as Salmonella Paratyphi A, Salmonella Paratyphi B, Shigella flexneri, Klebsiella pneumonia and Escherichia coli, therefore displaying desirable specificity. These novel aptamers could be used as diagnostic ligands for the future development of inexpensive and effective point-of-care tests for typhoid surveillance, especially in developing countries of the tropics and subtropics., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Salinity is a major abiotic stress that affects the agricultural sector and poses a significant threat to sustainable crop production. Nanoparticles (NPs) act as biostimulants and significantly mitigate abiotic stress. In this context, this experiment was designed to assess the effects of foliar application of titanium dioxide (TiO 2 ) nanoparticles at 200 and 400 ppm on the growth of eggplant ( Solanum melongena ) seedlings under moderate (75 mM) and high (150 mM) salinity stress. The TiO 2 -NPs employed were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), and scanning electron microscopy (SEM) analysis. The seedlings were assessed physiologically, growth-wise, and biochemically. The seedlings were significantly affected by their physiological attributes (Fv'/Fm', Fv/Fm, NPQ), growth (root length, shoot length, number of leaves, fresh biomass, dry biomass, leaf greenness), antioxidative enzymes (SOD, POD, CAT, APx, GR), stress indicators (H 2 O 2 , MDA), and toxic ion (Na + ) concentrations. The maximum decrease in physiological and growth attributes in eggplant seedling leaves was observed with no TiO 2 -NP application at 150 mM NaCl. Applying TiO 2 -NPs at 200 ppm showed significantly less decrease in Fv'/Fm', root length, shoot length, number of leaves, fresh biomass, dry biomass, and leaf greenness. In contrast, there were larger increases in SOD, POD, CAT, APx, GR, and TSP. This led to less accumulation of H 2 O 2 , MDA, and Na + . No significant difference was observed in higher concentrations of TiO 2 -NPs compared to the control. Therefore, TiO 2 -NPs at 200 ppm might be used to grow eggplant seedlings at moderate and high salinity.

Chitinases, a glycosyl hydrolase family 18 members, have a wide distribution in both prokaryotes and eukaryotes, including humans. Regardless of the absence of endogenous chitin polymer, various chitinases and chitinase-like proteins (CLPs) have been reported in mammals. However, several other carbohydrate polymers, such as hyaluronic acid and heparan sulfate, show structural similarities with chitin, which could be a potential target of chitinase and CLPs. Heparan sulfate is part of the integral membrane proteins and involves in cell adherence and migration. Hence, to demonstrate the effect of chitinase on cancer cell progression, we selected two chitinases from Serratia marcescens, ChiB and ChiC, which function as exo- and endo-chitinase, respectively. The ChiB and ChiC proteins were produced recombinantly by cloning chiB and chiC genes from Serratia marcescens. The cell viability of the Michigan Cancer Foundation-7 (MCF-7) cells was studied using different concentrations of the purified recombinant proteins. Cell viability assay was performed using 3-(4, 5-dimethyl thiazolyl-2)-2, 5-diphenyltetrazolium bromide and water-soluble tetrazolium salt, and the effect of ChiB and ChiC on cell proliferation was studied by clonogenic assay. The cell migration study was analysed by wound healing, transwell migration, and invasion assays. Cell cycle analysis of propidium iodide-stained cells and cell proliferation markers such as pERK1/2, pAKT, and SMP30 were also done. It was observed that both ChiB and ChiC were able to impede cell viability, cell migration, and invasion significantly. These observations and our in silico molecular docking analysis suggest that ChiC is a potential anticancer agent and is more efficient than ChiB. Since the ChiC is able to inhibit both cancer cell proliferation and migration, it could be a potential candidate for the treatment of metastatic cancer., Competing Interests: Declarations Ethical Approval Not applicable in this study. Consent to Participate Not applicable in this study. Consent for Publication Not applicable in this study as all the figures are original to this study. Competing Interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A highly accurate, rapid, portable, and robust platform for detecting Salmonella enterica serovar Typhi ( S. Typhi) is crucial for early-stage diagnosis of typhoid to avert and control the outbreaks of this pathogen, which threaten global public health. This study presents a proof-of-concept for our developed label-free electrochemical DNA biosensor system for S. Typhi detection, which employs a printed circuit board gold electrode (PCBGE), integrated with a portable potentiostat reader. Initially, the functionalized DNA biosensor and target detection were characterized using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) methods using a benchtop potentiostat. Interestingly, the newly developed DNA biosensor can identify target single-stranded DNA concentrations ranging from 10 nM to 20 μM, achieving a detection limit of 7.6 nM within a brief 5 minute timeframe. Under optimal detection conditions, the DNA biosensor exhibits remarkable selectivity, capable of distinguishing a single mismatch base pair from the target single-stranded DNA sequence. We then evaluated the feasibility of the developed DNA biosensor system as a diagnostic tool by detecting S. Typhi in 50 clinical samples using a portable potentiostat reader based on the DPV technique. Remarkably, the developed biosensor can distinctly distinguish between positive and negative samples, indicating that the miniaturised DNA biosensor system is practical for detecting S. Typhi in real biological samples. The developed DNA biosensor device in this work proves to be a promising point-of-care (POC) device for Salmonella detection due to its swift detection time, uncomplicated design, and streamlined workflow detection system.

Background: We investigate the geographical and racial disparities in accessing CAR-T and bispecific antibodies trials for DLBCL., Materials and Methods: ClinicalTrials.gov was searched, and 75 trials with at least 1 open site in the US were included. 2020 US Census Bureau data was used to obtain data on race and ethnicity. SPSS version 26 was used for analysis., Results: There were 62 CAR-T and 13 bispecific antibodies trials with 6221 enrolled or expected to enroll patients. Eighty-five percent of the clinical trials were only open in the US, and the majority 64% were pharmaceutical-funded. There were 126 unique study sites distributed over 31 states with 11 (0-51) mean number of trials per state and 4.5 (1-26) and 4.4 (1-24) mean number of CAR-T and bispecific antibodies trials per site, respectively. Southern states had the most number of trials 31%, followed by Midwestern 25%, Northeastern 24%, and Western 20%. The highest number of study locations were in California 13, New York 9, and Pennsylvania 9, while the highest number of open studies were in California 51, Texas 32, and New York 23. Twenty states had no open CAR-T or bispecific antibodies trials. Only 33% of African Americans (AA) lived in a county with a trial, and 7 out of 10 states with the highest proportion of AA residents (18.6%-41.4%) have no or less than 4 trial sites. Of the 62 counties analyzed, 92% were White predominant, while only 8% were AA predominant (P = .009)., Conclusions: Strategies should be framed to address the observed disparities and to improve access., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This systematic review aimed to evaluate the proportion of primary and secondary endpoints in hematopoietic stem cell transplant (HSCT) phase III randomized clinical trials (RCTs) and analyze their trends in time and study sponsorship status. The Chi-square test and logistic regression analyses were performed using SPSS version 28. A total of 147 HSCT phase III RCTs from 2006 to 2021 reported 197 primary and 600 secondary endpoints. Overall survival (OS, 17 %), progression-free survival (PFS, 15 %), graft versus host disease (GVHD, 8 %), event-free survival (EFS, 8 %), and organ function (8 %) were the most common primary endpoints. GVHD (12.3 %, n = 74), safety/toxicity/adverse events (11.8 %, n = 71), OS (11.5 %, n = 69), PFS (9.3 %, n = 56), and relapse rate (RR; 7.5 %, n = 45) were the most common secondary endpoints during 2006-2021. After 2013, an increase was noted in the use of PFS as a primary endpoint (12 %-18 %, p = 0.196), while the use of OS as a primary endpoint declined (20 %-13 %, p = 0.170). An increase was observed in using the secondary endpoints RR (5 %-10 %, p = 0.047) and NRM (3 %-6 %, p = 0.047). EFS was used more (14 % vs. 4 %, p = 0.012) than ORR (11 % vs. 2 %, p = 0.003) as a primary endpoint in pharmaceutical-compared to non-pharmaceutical-sponsored studies. As secondary endpoints, the use of EFS (4 % vs. 1 %, p = 0.013) and ORR (4 % vs. 1 %, p = 0.028) was higher, whereas that of organ systems/functions (1.5 % vs. 5.5 %, p = 0.022) and GVHD (6.5 % vs. 15 %, p = 0.002) was lower in pharmaceutical-compared to non-pharmaceutical sponsored studies. GVHD-free relapse-free survival was reported as a primary endpoint in 2 % of studies, while only 5 % reported quality of life as a secondary endpoint. We described commonly used endpoints in HSCT phase III RCTs and patterns in their use over time by funding source and study intervention category.

Cadmium, a ubiquitous environmental pollutant, has been implicated in the disruption of various metabolic pathways, contributing to the development of insulin resistance, glucose intolerance, and associated metabolic disorders. This study aimed to investigate the cadmium chloride (CdCl 2 ) exposure on metabolic pathways and to assess the potential therapeutic efficacy of the taxifolin-enriched extract in mitigating these disruptions by modulating biochemical pathways. Taxifolin-enriched extract (TEE) was prepared from Pinus roxburghii bark using a green extraction method. About 60 Wistar albino rats were divided into six groups: the control group ( n = 10), the CdCl 2 group (30 mg/kg) ( n = 10), and four groups (each comprises n = 10) treated with 30 mg/kg CdCl 2 in combination with metformin (100 mg/kg), ascorbic acid, taxifolin (30 mg/kg), and TEE (30 mg/kg), respectively. After the treatment period of 1 month, a comprehensive assessment of metabolic biomarkers and gene expressions that regulate the metabolism of carbohydrates and lipids was conducted to evaluate the impact of CdCl 2 exposure and the potential protective effects of TEE. The results revealed that CdCl 2 exposure significantly increased ( P < 0.001) serum levels of α-glucosidase, α-amylase, insulin, G6PC, hexokinases, TGs, LDL, HMG-CoA reductase, and pro-inflammatory cytokines such as IL-6 and TNF-α. Conversely, CdCl 2 exposure led to a reduction in HDL, antioxidant enzyme levels, phosphofructokinases, and glucose-6-phosphatase dehydrogenase. However, the administration of TEE alongside CdCl 2 substantially mitigated ( P < 0.001) these fluctuations in metabolic and inflammatory biomarker levels induced by CdCl 2 exposure. Both TEE and taxifolin treatment effectively lowered the elevated levels of α-amylase, α-glucosidase, G6PC, insulin, TGs, HMG-CoA reductase, leptin, ALT, AST, blood urea nitrogen, creatinine, and pro-inflammatory cytokines while simultaneously enhancing levels of HDL cholesterol and antioxidant enzymes. Moreover, CdCl 2 exposure suppressed mRNA expression of critical metabolic biomarkers such as glucose transporter 2 (GLUT2), insulin-like growth factor 1 (IGF-1), lactate dehydrogenase, and HMG-CoA lyases while upregulating the mRNA expression of angiotensin receptor 2 and vasopressin, key metabolic biomarkers involved in glucose metabolism and insulin regulation. TEE demonstrated the potential to restore normal metabolic functions and reduce the adverse impacts caused by CdCl 2 exposure by mitigating disturbances in several metabolic pathways and restoring gene expression of critical metabolic biomarkers related to glucose metabolism and insulin regulation. Nevertheless, further investigation is warranted to comprehensively understand the underlying mechanisms and optimize the appropriate dosage and duration of TEE treatment for achieving the most effective therapeutic outcomes., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Background: The study aimed to investigate the effect of urea molasses mineral blocks (UMMB) on nutrient digestibility, productive performance and blood biochemical profile of indigenous yaks under various feeding systems. A total of sixteen yaks were randomly divided into four groups (n = 4 animal per group) and offered the, following feeding systems: (A) stall feeding, (B), urea molasses mineral block (UMMB) + stall feeding, (C) yard feeding and (D) UMMB + yard feeding. Trial lasted for 40 days., Results: Results showed that nutrients intake (g) and nutrient digestibility (%) of dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE) and crude fiber (CF) were significantly higher (p < 0.05) in stall and yard feeding groups with UMMB licking. Blood zinc, cobalt, hemoglobin (Hb), red blood cell (RBC), glucose and serum glutamate private transaminase (SGPT) significantly (p < 0.05) increased in stall and yard feeding with UMMB licking. Milk yield, Ca and monounsaturated fatty acid except milk composition improved significantly (p < 0.05) in stall and yard feeding groups with UMMB licking., Conclusion: It was concluded that feeding of UMMB improved utilization of low-quality roughages and best results were obtained from stall and yard feedings with UMMB licking as compared to other groups., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Citrus fruits are cultivated around the world, and they face drought stress frequently during their growth and development. Previous studies showed that citrus plants biosynthesized flavonoid compounds in response to abiotic stress. In this study, we have quantified 37 flavonoid compounds from the leaves of three distinct citrus species including sour orange (drought-tolerant), pummelo 'Majia you pummelo' (drought-sensitive), and lemon (drought-sensitive). The 37 flavonoids consisted of 12 flavones, 10 flavonols, 6 flavanones, 5 isoflavanones, and 1 each for chalcone, flavanol, flavanonol, and flavone glycoside. Drought stress differentially altered the flavonoid metabolism in drought-tolerant and drought-sensitive citrus species. The kaempferol 3-neohesperidoside was 17-fold higher in sour orange (124.41 nmol/L) after 18 days of drought stress than lemon (7.33 nmol/L). In sour orange, neohesperidin (69.49 nmol/L) was 1,407- and 37-fold higher than pummelo and lemon, respectively. In sour orange, some flavonoids were significantly increased, such as vitexin, neohesperidin, cynaroside, hyperoside, genistin, kaempferol 3-neohesperidoside, eriocitrin, and luteolin, in response to drought stress, whereas in lemon, these flavonoids were significantly decreased or not altered significantly in response to drought stress. Moreover, the total contents of flavonoids and antioxidant activity were increased in sour orange as compared with pummelo and lemon. The genes associated with flavonoid biosynthesis ( PAL , CHI , FLS , GT1 , F3H , F3'M , C4H , 4CL , FLS , FG2 , FG3 , and CYP81E1 ) were more highly expressed in sour orange leaves than in pummelo and lemon after drought stress. These outcomes showed that pummelo and lemon failed to biosynthesize antioxidant flavonoids to cope with the prolonged drought stress, whereas the sour orange biosynthesized fortified flavonoid compounds with increased antioxidant activity to detoxify the harmful effects of reactive oxygen species produced during drought stress., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Rao, Feng, Ahmad, Tahir ul Qamar, Aslam, Khalid, Hussain, Zhong, Ali, Xu, Ma and Wang.)

Detection of viable viruses in the air is critical in order to determine the level of risk associated with the airborne diffusion of viruses. Different methods have been developed for the isolation, purification, and detection of viable airborne viruses, but they require an extensive processing time and often present limitations including low physical efficiency (i.e., the amount of collected viruses), low biological efficiency (i.e., the number of viable viruses), or a combination of all. To mitigate such limitations, we have employed an efficient technique based on the magnetic levitation (Maglev) technique with a paramagnetic solution and successfully identified distinct variations in levitation and density characteristics among bacteria ( Escherichia coli ), phages (MS2), and human viruses (SARS-CoV-2 and influenza H1N1). Notably, the Maglev approach enabled a significant enrichment of viable airborne viruses in air samples. Furthermore, the enriched viruses obtained through Maglev exhibited high purity, rendering them suitable for direct utilization in subsequent analyses such as reverse transcription-polymerase chain reaction (RT-PCR) or colorimetric assays. The system is portable, easy to use, and cost-efficient and can potentially provide proactive surveillance data for monitoring future outbreaks of airborne infectious diseases and allow for the induction of various preventative and mitigative measures.

The use of nanoscale fertilizers to boost crop output has increased in recent years. Nanoparticles (NPs) can stimulate the biosynthesis of bioactive compounds in plants. It is the first report on biosynthesized manganese oxide nanoparticles (MnO-NPs) that mediate in-vitro callus induction of Moringa oleifera. To achieve better biocompatibility the leaf extract of Syzygium cumini was used to synthesize MnO-NPs. Scanning electron microscope SEM revealed spherical shaped morphology of MnO-NPs with an average diameter of 36 ± 0.3 nm. Energy-dispersive X-ray spectroscopy (EDX) depicted the formation of pure MnO-NPs. X-ray diffraction (XRD) and Fourier Transform Infrared (FTIR) authenticate the crystalline structure. UV-visible absorption spectroscopy depicted the activity of MnO-NPs under visible light. The biosynthesized MnO-NPs were concentration-dependent and revealed promising results in callus induction of Moringa oleifera. It was found that MnO-NPs enhance callus production of Moringa oleifera and keep the callus infection free by providing an optimum environment for rapid growth and development. Therefore MnO-NPs synthesized through the green process can be utilized in tissue culture studies. This study concludes that MnO is one of the essential plant nutrients that have tailored nutritive properties at a nano scale., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

The lack of preparedness for detecting and responding to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pathogen (i.e., COVID-19) has caused enormous harm to public health and the economy. Testing strategies deployed on a population scale at day zero, i.e., the time of the first reported case, would be of significant value. Next-generation sequencing (NGS) has such capabilities; however, it has limited detection sensitivity for low-copy-number pathogens. Here, we leverage the CRISPR-Cas9 system to effectively remove abundant sequences not contributing to pathogen detection and show that NGS detection sensitivity of SARS-CoV-2 approaches that of RT-qPCR. The resulting sequence data can also be used for variant strain typing, co-infection detection, and individual human host response assessment, all in a single molecular and analysis workflow. This NGS work flow is pathogen agnostic and, therefore, has the potential to transform how large-scale pandemic response and focused clinical infectious disease testing are pursued in the future., Competing Interests: K.B. owns stock in Jumpcode Genomics, Inc. and has filed patents related to this work. Y.H. is a member of the board of directors of Jumpcode Genomics, Inc. N.J.S. is a scientific advisory board member of Jumpcode Genomics, Inc. A.S., Y.D., S.R., K.S.C., M.F.K., J.D., J.B., D.A., Z.G., J.R.A., and Y.H. are employed by Jumpcode Genomics, Inc., (© 2023 The Authors.)

The emergence of coronavirus disease 2019 (COVID-19) motivates continuous efforts to develop robust and accurate diagnostic tests to detect severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Detection of viral nucleic acids provides the highest sensitivity and selectivity for diagnosing early and asymptomatic infection because the human immune system may not be active at this stage. Therefore, this work aims to develop a label-free electrochemical DNA biosensor for SARS-CoV-2 detection using a printed circuit board-based gold substrate (PCBGE). The developed sensor used the nucleocapsid phosphoprotein (N) gene as a biomarker. The DNA sensor-based PCBGE was fabricated by self-assembling a thiolated single-stranded DNA (ssDNA) probe onto an Au surface, which performed as the working electrode (WE). The Au surface was then treated with 6-mercapto-1-hexanol (MCH) before detecting the target N gene to produce a well-oriented arrangement of the immobilized ssDNA chains. The successful fabrication of the biosensor was characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and atomic force microscopy (AFM). The DNA biosensor performances were evaluated using a synthetic SARS-CoV-2 genome and 20 clinical RNA samples from healthy and infected individuals through EIS. The developed DNA biosensor can detect as low as 1 copy per μL of the N gene within 5 minutes with a LOD of 0.50 μM. Interestingly, the proposed DNA sensor could distinguish the expression of SARS-CoV-2 RNA in a patient diagnosed with COVID-19 without any amplification technique. We believe that the proposed DNA sensor platform is a promising point-of-care (POC) device for COVID-19 viral infection since it offers a rapid detection time with a simple design and workflow detection system, as well as an affordable diagnostic assay.

The development of rapid, accurate, and efficient detection methods for Salmonella can significantly control the outbreak of salmonellosis that threatens global public health. Despite the high sensitivity and specificity of the microbiological, nucleic-acid, and immunological-based methods, they are impractical for detecting samples outside of the laboratory due to the requirement for skilled individuals and sophisticated bench-top equipment. Ideally, an electrochemical biosensor could overcome the limitations of these detection methods since it offers simplicity for the detection process, on-site quantitative analysis, rapid detection time, high sensitivity, and portability. The present scoping review aims to assess the current trends in electrochemical aptasensors to detect and quantify Salmonella. This review was conducted according to the latest Preferred Reporting Items for Systematic review and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. A literature search was performed using aptamer and Salmonella keywords in three databases: PubMed, Scopus, and Springer. Studies on electrochemical aptasensors for detecting Salmonella published between January 2014 and January 2022 were retrieved. Of the 787 studies recorded in the search, 29 studies were screened for eligibility, and 15 studies that met the inclusion criteria were retrieved for this review. Information on the Salmonella serovars, targets, samples, sensor specification, platform technologies for fabrication, electrochemical detection methods, limit of detection (LoD), and detection time was discussed to evaluate the effectiveness and limitations of the developed electrochemical aptasensor platform for the detection of Salmonella. The reported electrochemical aptasensors were mainly developed to detect Salmonella enterica Typhimurium in chicken meat samples. Most of the developed electrochemical aptasensors were fabricated using conventional electrodes (13 studies) rather than screen-printed electrodes (SPEs) (two studies). The developed aptasensors showed LoD ranges from 550 CFU/mL to as low as 1 CFU/mL within 5 min to 240 min of detection time. The promising detection performance of the electrochemical aptasensor highlights its potential as an excellent alternative to the existing detection methods. Nonetheless, more research is required to determine the sensitivity and specificity of the electrochemical sensing platform for Salmonella detection, particularly in human clinical samples, to enable their future use in clinical practice.

Background: Idiopathic pulmonary fibrosis (IPF) is a respiratory disorder with a poor prognosis. Our objective is to assess the comparative effectiveness of 22 approved or studied IPF drug treatments., Methods: We searched MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials and clinicaltrials.gov from inception to 2 April 2021. We included randomised controlled trials (RCTs) for adult patients with IPF receiving one or more of 22 drug treatments. Pairs of reviewers independently identified randomised trials that compared one or more of the target medical treatments in patients with IPF. We assessed the certainty of evidence using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach for network meta-analysis. We calculated pooled relative risk (RR) ratios and presented direct or network estimates with 95% credibility intervals (95% CI), within the GRADE framework., Results: We identified 48 (10 326 patients) eligible studies for analysis. Nintedanib [RR 0.69 (0.44 to 1.1), pirfenidone [RR 0.63 (0.37 to 1.09); direct estimate), and sildenafil [RR (0.44 (0.16 to 1.09)] probably reduce mortality (all moderate certainty). Nintedanib (2.92% (1.51 to 4.14)), nintedanib+sildenafil (157 mL (-88.35 to 411.12)), pirfenidone (2.47% (-0.1 to 5)), pamrevlumab (4.3% (0.5 to 8.1)) and pentraxin (2.74% (1 to 4.83)) probably reduce decline of overall forced vital capacity (all moderate certainty). Only sildenafil probably reduces acute exacerbation and hospitalisations (moderate certainty). Corticosteroids+azathioprine+N-acetylcysteine increased risk of serious adverse events versus placebo (high certainty)., Conclusion and Relevance: Future guidelines should consider sildenafil for IPF and further research needs to be done on promising IPF treatments such as pamrevlumab and pentraxin as phase 3 trials are completed., Competing Interests: Competing interests: MK discloses research funding for preclinical work from Boehringer Ingelheim and Pieris. He received research funding for clinical projects from Roche. He has received consulting fees from Boehringer Ingelheim, Roche, Horizon, Cipla, Abbvie, Bellerophon, Algernon and CSL Behring. He has received payments or honoraria for lectures, presentations, speaker bruins, manuscript writings or educational events from Novartis, Boehringer Ingelheim and Roche. He has received payments for expert testimony from Roche. He participates on the data safety monitoring board or advisory board for Covance and United Therapeutics. He is the chief editor for the European Respiratory Journal and received a Chief Editor allowance. No other authors have conflicts of interest to disclose. CS discloses research funding from the Canadian Pulmonary Fibrosis Foundation and Boehringer Ingelheim. He has also received payments for presentations and advisory work from Boehringer Ingelheim., (© Author(s) (or their employer(s)) 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

This scoping review aims to provide a comprehensive overview of human melioidosis in Southeast Asia as well as to highlight knowledge gaps in the prevalence and risk factors of this life-threatening disease using available evidence-based data for better diagnosis and treatment. Preferred Reporting Items for Systematic Review and Meta-Analyses Extension for Scoping Reviews (PRISMA-ScR) was used as the guideline for this review. The literature search was conducted on 23 March 2022 through two electronic databases (PubMed and Scopus) using lists of keywords referring to the Medical Subject Headings (MeSH) thesaurus. A total of 38 articles related to human melioidosis were included from 645 screened articles. These studies were carried out between 1986 and 2019 in six Southeast Asian countries: Thailand, Cambodia, Malaysia, Myanmar, Singapore, and Vietnam. Melioidosis has been reported with a high disease prevalence among high-risk populations. Studies in Thailand (48.0%) and Cambodia (74.4%) revealed disease prevalence in patients with septic arthritis and children with suppurative parotitis, respectively. Other studies in Thailand (63.5%) and Malaysia (54.4% and 65.7%) showed a high seroprevalence of melioidosis among Tsunami survivors and military personnel, respectively. Additionally, this review documented soil and water exposure, diabetes mellitus, chronic renal failure, thalassemia, and children under the age of 15 as the main risk factors for melioidosis. Human melioidosis is currently under-reported in Southeast Asia and its true prevalence is unknown.

Climate change significantly affects plant growth and productivity by causing different biotic and abiotic stresses to plants. Among the different abiotic stresses, at the top of the list are salinity, drought, temperature extremes, heavy metals and nutrient imbalances, which contribute to large yield losses of crops in various parts of the world, thereby leading to food insecurity issues. In the quest to improve plants' abiotic stress tolerance, many promising techniques are being investigated. These include the use of nanoparticles, which have been shown to have a positive effect on plant performance under stress conditions. Nanoparticles can be used to deliver nutrients to plants, overcome plant diseases and pathogens, and sense and monitor trace elements that are present in soil by absorbing their signals. A better understanding of the mechanisms of nanoparticles that assist plants to cope with abiotic stresses will help towards the development of more long-term strategies against these stresses. However, the intensity of the challenge also warrants more immediate approaches to mitigate these stresses and enhance crop production in the short term. Therefore, this review provides an update of the responses (physiological, biochemical and molecular) of plants affected by nanoparticles under abiotic stress, and potentially effective strategies to enhance production. Taking into consideration all aspects, this review is intended to help researchers from different fields, such as plant science and nanoscience, to better understand possible innovative approaches to deal with abiotic stresses in agriculture.

The Escherichia coli CheY protein belongs to a large bacterial response regulator superfamily. X-ray hydroxy radical foot-printing with mass spectroscopy (XFMS) has shown that allosteric activation of CheY by its motor target triggers a concerted internalization of aromatic sidechains. We reanalyzed the XFMS data to compare polar versus non-polar CheY residue positions. The polar residues around and including the 57D phosphorylated site had an elevated hydroxy radical reactivity. Bioinformatic measures revealed that a water-mediated hydrogen bond network connected this ring of residues with the central 57D. These residues solvated 57D to energetically stabilize the apo-CheY fold. The abundance of these reactive residues was reduced upon activation. This result was supported by the bioinformatics and consistent with the previously reported activation-induced increase in core hydrophobicity. It further illustrated XFMS detection of structural waters. Direct contacts between the ring residues and the phosphorylation site would stabilize the aspartyl phosphate. In addition, we report that the ring residue, 18R, is a constant central node in the 57D solvation network and that 18R non-polar substitutions determine CheY diversity as assessed by its evolutionary trace in bacteria with well-studied chemotaxis. These results showcase the importance of structured water dynamics for phosphorylation-mediated signal transduction.

Infectious diseases are the world's greatest killers, accounting for millions of deaths worldwide annually, especially in low-income countries. As the risk of emerging infectious diseases is increasing, it is critical to rapidly diagnose infections in the early stages and prevent further transmission. However, current detection strategies are time-consuming and have exhibited low sensitivity. Numerous studies revealed the advantages of point-of-care testing, such as those which are rapid, user-friendly and have high sensitivity and specificity, and can be performed at a patient's bedside. The Lateral Flow Immunoassay (LFIA) is the most popular diagnostic assay that fulfills the POCT standards. However, conventional AuNPs-LFIAs are moderately sensitive, meaning that rapid detection remains a challenge. Here, we review quantum dot (QDs)-based LFIA for highly sensitive rapid diagnosis of infectious diseases. We briefly describe the principles of LFIA, strategies for applying QDs to enhance sensitivity, and the published performance of the QD-LFIA tested against several infectious diseases.

Anti-vascular endothelial growth factor (Anti-VEGF) agents are the standard of care for diabetic macular edema (CI-DME) with vision loss. They are commonly administered using several treatment protocols, including fixed, pro re nata (PRN) and treat-and-extend (T&E) regimens. Because of the lack of evidence defining an ideal treatment paradigm, we systematically compared T&E with fixed or PRN regimens. Visual acuity improvement was similar when comparing T&E to fixed or PRN dosing at 12 and 24 months. Regarding anatomic outcomes, no significant difference was found between T&E and fixed regimens for central retinal thickness or central subfoveal thickness at 12 and 24 months. Similarly, no significant difference was found for central retinal thickness at 12 months for T&E versus PRN regimen. Regarding total number of injections, no significant difference existed between T&E versus fixed regimens at 12 months. PRN regimens delivered fewer injections compared to T&E regimens at 12 months. The results of this analysis support that visual acuity and anatomic outcomes at 12 and 24 months are similar between T&E with either fixed or PRN regimens. More head-to-head trials comparing T&E versus fixed and PRN dosing are needed to provide visual and functional outcome data beyond year 2. PROSPERO Registration: CRD42021249362., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Objective: To investigate treat-and-extend (T&E) regimens of anti-vascular endothelial growth factor (anti-VEGF) therapy for the treatment of macular oedema secondary to retinal vein occlusions (RVOs)., Methods: Ovid MEDLINE, Ovid EMBASE and CENTRAL were searched on 25 February 2021. Randomized controlled trials, cohort studies, case-control studies and case series were included. The primary outcome was the change in Early Treatment Diabetic Retinopathy Score (ETDRS) letters from baseline. Conversions from Snellen to ETDRS letters were performed utilizing a published protocol. Secondary outcomes included improvement in retinal thickness from baseline, number of anti-VEGF injections and frequency of adverse events. Outcomes were examined at 12 and 24 months. Certainty of evidence was assessed utilizing GRADE (Grading of Recommendations Assessments, Development and Evaluations) guidelines., Results: Seven hundred eighty-six eyes from 16 studies were included. Meta-analysis demonstrated a mean improvement of 15.7 (95% CI: 13.3-18.0) ETDRS letters at 12 months. Central retinal thickness improved 269.7 μm (95% CI: 233.64-305.90) at 12 months. Injections were performed 8.1 (95% CI: 7.4-8.7) and 13.1 (95% CI: 9.4-16.8) times at 12 and 24 months respectively. Adverse events were infrequent across all studies. Grading of Recommendations Assessments, Development and Evaluations (GRADE) certainty of evidence was very low across all outcomes., Conclusions: The results support the viability of T&E regimens for the treatment of macular oedema secondary to RVOs., (© 2021 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.)

Background: There is no consensus on the most effective treatments of pulmonary arterial hypertension (PAH). Our objective was to compare effects of medications for PAH., Methods: We searched MEDLINE, Embase, the Cochrane Central Register of Controlled Trials and Clinicaltrials.gov from inception to December 2021. We performed a frequentist random-effects network meta-analysis on all included trials. We rated the certainty of the evidence using the Grades of Recommendation, Assessment, Development, and Evaluation approach., Results: We included 53 randomised controlled trials with 10 670 patients. Combination therapy with endothelin receptor antagonist (ERA) plus phosphodiesterase-5 inhibitors (PDE5i) reduced clinical worsening (120.7 fewer events per 1000, 95% CI 136.8-93.4 fewer; high certainty) and was superior to either ERA or PDE5i alone, both of which reduced clinical worsening, as did riociguat monotherapy (all high certainty). PDE5i (24.9 fewer deaths per 1000, 95% CI 35.2 fewer to 2.1 more); intravenous/subcutaneous prostanoids (18.3 fewer deaths per 1000, 95% CI 28.6 fewer deaths to 0) and riociguat (29.1 fewer deaths per 1000, 95% CI 38.6 fewer to 8.7 more) probably reduce mortality as compared to placebo (all moderate certainty). Combination therapy with ERA+PDE5i (49.9 m, 95% CI 25.9-73.8 m) and riociguat (49.5 m, 95% CI 17.3-81.7 m) probably increase 6-min walk distance as compared to placebo (moderate certainty)., Conclusion: Current PAH treatments improve clinically important outcomes, although the degree and certainty of benefit vary between treatments., Competing Interests: Conflict of interest: L. Mielniczuk discloses speaker fees/advisory boards/consulting fees from Janssen. She is a clinician scientist supported by Heart and Stroke Foundation. S. Mehta discloses speaker fees and consulting fees from Acceleron Pharmaceuticals and Janssen Pharmaceuticals, as well as institutional research support from Acceleron, Bellerophon, Gossamer Bio, Janssen, and United Therapeutics. The remaining authors have nothing to disclose., (Copyright ©The authors 2022.)

The development of precise and efficient diagnostic tools enables early treatment and proper isolation of infected individuals, hence limiting the spread of coronavirus disease 2019 (COVID-19). The standard diagnostic tests used by healthcare workers to diagnose severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection have some limitations, including longer detection time, the need for qualified individuals, and the use of sophisticated bench-top equipment, which limit their use for rapid SARS-CoV-2 assessment. Advances in sensor technology have renewed the interest in electrochemical biosensors miniaturization, which provide improved diagnostic qualities such as rapid response, simplicity of operation, portability, and readiness for on-site screening of infection. This review gives a condensed overview of the current electrochemical sensing platform strategies for SARS-CoV-2 detection in clinical samples. The fundamentals of fabricating electrochemical biosensors, such as the chosen electrode materials, electrochemical transducing techniques, and sensitive biorecognition molecules, are thoroughly discussed in this paper. Furthermore, we summarised electrochemical biosensors detection strategies and their analytical performance on diverse clinical samples, including saliva, blood, and nasopharyngeal swab. Finally, we address the employment of miniaturized electrochemical biosensors integrated with microfluidic technology in viral electrochemical biosensors, emphasizing its potential for on-site diagnostics applications.

Background: Patients with idiopathic pulmonary fibrosis have a poor overall prognosis and there are few evidence-based drug therapies that reduce mortality., Objective: We aimed to perform a systematic review and meta-analysis to assess whether sildenafil reduces mortality, disease progression and adverse side effects., Methods: We reviewed randomized controlled studies (RCTs) from MEDLINE, Cochrane registry of clinical trials, and EMBASE. Our outcomes of interest included mortality, change in FVC, acute exacerbations and hospitalizations and adverse drug effects leading to discontinuation. We used an inverse variance fixed effects meta-analysis method to calculate pooled relative risk (RR) and mean difference (MD)., Results: A total of 4 studies were included in the systematic review. Sildenafil probably reduces mortality when compared to placebo or to standard care, [RR 0.73 (95% CI 0.51 to 1.04); moderate certainty]. Pooled estimates showed sildenafil may not alter the rate of change of FVC [MD 0.61% (95% CI -0.29 to 1.52)], or DLCO [MD 0.97% (95% CI 0.04 to 1.90)] (both low certainty). Pooled estimated showed sildenafil may not reduce the number of hospitalizations or acute exacerbations, [RR 1.10 (95% CI 0.61 to 1.98); low certainty]. There is probably no difference in drug discontinuation due to adverse effects when comparing sildenafil to the control group, [RR 0.79 (95% CI 0.56, 1.10); moderate certainty]., Conclusion: Sildenafil probably reduces all-cause mortality in IPF patients. More studies need to be done to confirm the magnitude and reliability of the point estimate., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Recently, CRISPR-Cas system-based assays for bacterial detection have been developed. The aim of this scoping review is to map existing evidence on the utilization of CRISPR-Cas systems in the development of bacterial detection assays. A literature search was conducted using three databases (PubMed, Scopus, and Cochrane Library) and manual searches through the references of identified full texts based on a PROSPERO-registered protocol (CRD42021289140). Studies on bacterial detection using CRISPR-Cas systems that were published before October 2021 were retrieved. The Critical Appraisal Skills Programme (CASP) qualitative checklist was used to assess the risk of bias for all the included studies. Of the 420 studies identified throughout the search, 46 studies that met the inclusion criteria were included in the final analysis. Bacteria from 17 genera were identified utilising CRISPR-Cas systems. Most of the bacteria came from genera such as Staphylococcus , Escherichia , Salmonella , Listeria , Mycobacterium and Streptococcus . Cas12a (64%) is the most often used Cas enzyme in bacterial detection, followed by Cas13a (13%), and Cas9 (11%). To improve the signal of detection, 83% of the research exploited Cas enzymes' trans-cleavage capabilities to cut tagged reporter probes non-specifically. Most studies used the extraction procedure, whereas only 17% did not. In terms of amplification methods, isothermal reactions were employed in 66% of the studies, followed by PCR (23%). Fluorescence detection (67%) was discovered to be the most commonly used method, while lateral flow biosensors (13%), electrochemical biosensors (11%), and others (9%) were found to be less commonly used. Most of the studies (39) used specific bacterial nucleic acid sequences as a target, while seven used non-nucleic acid targets, including aptamers and antibodies particular to the bacteria under investigation. The turnaround time of the 46 studies was 30 min to 4 h. The limit of detection (LoD) was evaluated in three types of concentration, which include copies per mL, CFU per mL and molarity. Most of the studies used spiked samples (78%) rather than clinical samples (22%) to determine LoD. This review identified the gap in clinical accuracy evaluation of the CRISPR-Cas system in bacterial detection. More research is needed to assess the diagnostic sensitivity and specificity of amplification-free CRISPR-Cas systems in bacterial detection for nucleic acid-based tests.

The aim of current study was to investigate the inhibitory activities of resveratrol and taxifolin against α-amylase, α-glucosidase, and DPP-IV enzymes via in vitro analysis which was further validated by in silico studies. The analysis of molecular docking was also done to determine the binding capabilities of resveratrol and taxifolin with α-amylase, α-glucosidase, and DPP-IV enzymes. Resveratrol and taxifolin having IC 50 values, 47.93 ± 5.21 μ M and 45.86 ± 3.78 μ M , respectively, showed weaker effect than acarbose (4.6 ± 1.26 μ M ) on α-amylase but showed significant effect to inhibit α-glucosidase (32.23 ± .556 μ M and 31.26 ± .556 μ M , respectively). IC 50 value of resveratrol and taxifolin (5.638 ± .0016 μ M and 6.691 ± .004 μ M ) in comparison to diprotin A (IC 50 : 7.21 ± .021 μ M ) showed that they have significant inhibitory effect on DPP-IV enzyme. Our results illustrated that resveratrol and taxifolin have potential to prevent the metabolism of carbohydrates via inhibition of α-amylase and α-glucosidase, and prolongs metabolic function of incretin by inhibiting the enzymatic activity of DPP-IV. The results of molecular docking have also revealed that resveratrol and taxifolin have significant affinity to bind with α-amylase, α-glucosidase, and DPP-IV in comparison with standard drugs such as acarbose, miglitol, and diprotin., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2022.)

Telemedicine modalities for patient care have seen significant global uptake during the COVID-19 pandemic. This study aimed to bibliometrically evaluate the evolution and current landscape of telemedicine literature in Canada. The Scopus database was searched to identify telemedicine publications for which the first or last author had a Canadian institutional affiliation. Study selection and data abstraction were conducted by two pairs of independent reviewers. Between 1976 and January 2021, 810 of 3,620 retrieved citations were telemedicine publications originating from Canada, including 29 randomized controlled trials and 6 systematic reviews. The annual publication output increased substantially from 1/year in 1976 to 80/year in 2020. Based on author keyword analysis, the most frequently investigated disciplines or disease entities were primary care, COVID-19, telepsychiatry, heart failure, and mental health. The insights this study provides will aid scientists, policy makers, and other stakeholders in identifying opportunities for future investigation and clinical application.

Dengue is one of the most dominant arthropod-borne viral diseases, infecting at least 390 million people every year throughout the world. Despite this, there is no effective treatment against dengue, and the only available vaccine has already been withdrawn owing to the significant adverse effects. Therefore, passive immunotherapy using monoclonal antibodies is now being sought as a therapeutic option. To date, many dengue monoclonal antibodies have been identified, most of which are serotype-specific, and only a few of which are cross-reactive. Furthermore, antibodies that cross-react within serotypes are weakly neutralizing and frequently induce antibody-dependent enhancement, which promotes viral entry and replication. Therefore, broadly neutralizing antibodies with no risk of antibody-dependent enhancement are required for the treatment of dengue. Here, we developed a single-chain variable fragment (scFv) antibody from an anti-fusion loop E53 antibody (PDB: 2IGF). We introduced previously predicted favorable complementarity-determining region (CDR) mutations into the gene encoding the scFv antibody for affinity maturation, and the resultant variants were tested in vitro against the highly conserved fusion and bc epitope of the dengue virus envelope protein. We show some of these scFv variants with two to three substitution mutations in three different CDRs possess affinity constants (K D ) ranging from 20 to 200 nM. The scFv-mutant15, containing D31L, Y105W, and S227W substitutions, showed the lowest affinity constant, (K D  = 24 ± 7 nM), approximately 100-fold lower than its parental construct. We propose that the scFv-derivative antibody may be a good candidate for the development of an effective and safe immunotherapy., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The identification of viral RNA using reverse transcription quantitative polymerase chain reaction (RT-qPCR) is the gold standard for identifying an infection caused by SARS-CoV-2. The limitations of RT-qPCR such as requirement of expensive instruments, trained staff and laboratory facilities led to development of rapid antigen tests (RATs). The performance of RATs has been widely evaluated and found to be varied in different settings. The present systematic review aims to evaluate the pooled sensitivity and specificity of the commercially available RATs. This review was registered on PROSPERO (registration number: CRD42021278105). Literature search was performed through PubMed, Embase and Cochrane COVID-19 Study Register to search studies published up to 26 August 2021. The overall pooled sensitivity and specificity of RATs and subgroup analyses were calculated. Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) was used to assess the risk of bias in each study. The overall pooled sensitivity and specificity of RATs were 70% (95% CI: 69-71) and 98% (95% CI: 98-98), respectively. In subgroup analyses, nasal swabs showed the highest sensitivity of 83% (95% CI: 80-86) followed by nasopharyngeal swabs 71% (95% CI: 70-72), throat swabs 69% (95% CI: 63-75) and saliva 68% (95% CI: 59-77). Samples from symptomatic patients showed a higher sensitivity of 82% (95% CI: 82-82) as compared to asymptomatic patients at 68% (95% CI: 65-71), while a cycle threshold (Ct) value ≤25 showed a higher sensitivity of 96% (95% CI: 95-97) as compared to higher Ct value. Although the sensitivity of RATs needs to be enhanced, it may still be a viable option in places where laboratory facilities are lacking for diagnostic purposes in the early phase of disease.

Consequences of warming and postwarming events on photosynthetic thermotolerance (P T ) and photoprotective responses in tropical evergreen species remain elusive. We chose Citrus to answer some of the emerging questions related to tropical evergreen species' P T behaviour including (i) how wide is the genotypic variation in P T ? (ii) how does P T respond to short-term warming and (iii) how do photosynthesis and photoprotective functions respond over short-term warming and postwarming events? A study on 21 genotypes revealed significant genotypic differences in P T , though these were not large. We selected five genotypes with divergent P T and simulated warming events: T max 26/20°C (day-time highest maximum/night-time lowest maximum) (Week 1) < T max 33/30°C (Week 2) < T max 36/32°C (Week 3) followed by T max 26/16°C (Week 4, recovery). The P T of all genotypes remained unaltered despite strong leaf megathermy (leaf temperature > air temperature) during warming events. Though moderate warming showed genotype-specific stimulation in photosynthesis, higher warming unequivocally led to severe loss in net photosynthesis and induced higher nonphotochemical quenching. Even after a week of postwarming, photoprotective mechanisms strongly persisted. Our study points towards a conservative P T in evergreen citrus genotypes and their need for sustaining higher photoprotection during warming as well as postwarming recovery conditions., (© 2021 John Wiley & Sons Ltd.)

Water scarcity restricts citrus growth and productivity worldwide. In pot conditions, tetraploid plants tolerate water deficit more than their corresponding diploids. However, their tolerance mechanisms remain elusive. In this study, we focused on which mechanisms (i.e., hydraulic, osmotic or antioxidative) confer water-deficit tolerance to tetraploids. We exposed diploid and tetraploid Volkamer lemon rootstock (Citrus volkameriana Tan. and Pasq.) to quickly (fast) and slowly (slow) developing water-deficit conditions. We evaluated their physiological, antioxidative defense and osmotic adjustment responses, and mineral distribution to leaves and roots. Water-deficit conditions decreased the photosynthetic variables of both diploid and tetraploid plants. Moreover, the corresponding decrease was greater in diploids than tetraploids. Higher concentrations of antioxidant enzymes, osmoprotectants and antioxidant capacity were found in the leaves and roots of tetraploids than diploids under water deficit. Diploid plants showed fast response in slow water-deficit condition, but that response did not persist as the deficit intensified. Meanwhile, tetraploids had lower water loss, which slowed the onset of slow water deficit relative to diploids. This response allowed stronger photosynthesis, while antioxidant and osmoprotectant production allowed for further tolerance once desiccation began. Overall, our results concluded that Volkamer lemon tetraploid plants tolerate rapid and slow water deficit by maintaining their photosynthesis due to low conductance (stem or roots), which helps to avoid desiccation, and stronger biochemical defense machinery than their corresponding diploids., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Since its first detection in December 2019, more than 232 million cases of COVID-19, including 4.7 million deaths, have been reported by the WHO. The SARS-CoV-2 viral genomes have evolved rapidly worldwide, causing the emergence of new variants. This systematic review and meta-analysis was conducted to provide a global mutational profile of SARS-CoV-2 from December 2019 to October 2020. The review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA), and a study protocol was lodged with PROSPERO. Data from 62 eligible studies involving 368,316 SARS-CoV-2 genomes were analyzed. The mutational data analyzed showed most studies detected mutations in the Spike protein ( n = 50), Nucleocapsid phosphoprotein ( n = 34), ORF1ab gene ( n = 29), 5'-UTR ( n = 28) and ORF3a ( n = 25). Under the random-effects model, pooled prevalence of SARS-CoV-2 variants was estimated at 95.1% (95% CI; 93.3-96.4%; I 2 = 98.952%; p = 0.000) while subgroup meta-analysis by country showed majority of the studies were conducted 'Worldwide' ( n = 10), followed by 'Multiple countries' ( n = 6) and the USA ( n = 5). The estimated prevalence indicated a need to continuously monitor the prevalence of new mutations due to their potential influence on disease severity, transmissibility and vaccine effectiveness.

Introduction: Virtually all hospitalized coronavirus disease-2019 (COVID-19) outcome data come from urban environments. The extent to which these findings are generalizable to other settings is unknown. Coronavirus disease-2019 data from large, urban settings may be particularly difficult to apply in military medicine, where practice environments are often semi-urban, rural, or austere. The purpose of this study is compare presenting characteristics and outcomes of U.S. patients with COVID-19 in a nonurban setting to similar patients in an urban setting., Materials and Methods: This is a retrospective case series of adults with laboratory-confirmed COVID-19 infection who were admitted to Hershey Medical Center (HMC), a 548-bed tertiary academic medical center in central Pennsylvania serving semi-urban and rural populations, from March 23, 2020, to April 20, 2020 (the first month of COVID-19 admissions at HMC). Patients and outcomes of this cohort were compared to published data on a cohort of similar patients from the New York City (NYC) area., Results: The cohorts had similar age, gender, comorbidities, need for intensive care or mechanical ventilation, and most vital sign and laboratory studies. The NYC's cohort had shorter hospital stays (4.1 versus 7.2 days, P < .001) but more African American patients (23% versus 12%, P = .02) and higher prevalence of abnormal alanine (>60U/L; 39.0% versus 5.9%, P < .001) and aspartate (>40U/L; 58.4% versus 42.4%, P = .012) aminotransferase, oxygen saturation <90% (20.4% versus 7.2%, P = .004), and mortality (21% versus 1.4%, P < .001)., Conclusions: Hospitalists in nonurban environments would be prudent to use caution when considering the generalizability of results from dissimilar regions. Further investigation is needed to explore the possibility of reproducible causative systemic elements that may help improve COVID-19-related outcomes. Broader reports of these relationships across many settings will offer military medical planners greater ability to consider outcomes most relevant to their unique settings when considering COVID-19 planning., (© The Association of Military Surgeons of the United States 2021. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: The optimal approach to deal with severe coronary artery calcification (CAC) during percutaneous coronary intervention (PCI) remains ill-defined., Methods: We conducted an electronic database search of all published studies comparing Orbital versus Rotational Atherectomy in patients undergoing PCI., Results: Eight observational studies were included in the analysis. Overall, there were no significant differences in Major-adverse-cardiac-events/MACE (OR: 0.81, CI: 0.63-1.05, p = .11), myocardial-infarction/MI (OR: 0.75, CI: 0.56-1.00, p = .05), all-cause mortality (OR: 0.82, CI: 0.25-2.64, p = .73) or Target-vessel-revascularization/TVR (OR: 0.72, CI: 0.38-1.36, p = .31). However, OA was associated with lower long-term MACE (1-year), (OR: 0.66, CI: 0.44-0.99, p = .04), long-term TVR (OR: 0.40, CI: 0.18-0.89, p = .03), and short-term MI (in-hospital and 30-day) (OR: 0.64, CI: 0.44-0.94, p = .02). OA was associated with more coronary artery dissections (OR: 2.61, CI: 1.38-4.92, p = .003) and device-related coronary perforations (OR: 2.79, CI: 1.08-7.19, p = .03). There were no differences in cardiac tamponade (OR: 1.78, CI: 0.37-8.69, p = .47). OA was noted to have significantly lower fluoroscopy time (MD: -3.96 min, CI: -7.67, -0.25; p = .04) compared to RA. No significant difference was noted in terms of contrast volume between the two groups (OR: -4.35 ml, CI: -14.52, 23.22; p = .65)., Conclusion: Although there was no difference in overall MACE, MI, all-cause mortality and TVR, OA was associated with lower long-term MACE and short-term MI. OA is associated with lower fluoroscopy time but higher rates of coronary artery dissection and coronary perforation., (© 2020 Wiley Periodicals LLC.)

Large-scale food-borne outbreaks caused by Salmonella are rarely seen nowadays, thanks to the advanced nature of the medical system. However, small, localised outbreaks in certain regions still exist and could possess a huge threat to the public health if eradication measure is not initiated. This review discusses the progress of Salmonella detection approaches covering their basic principles, characteristics, applications, and performances. Conventional Salmonella detection is usually performed using a culture-based method, which is time-consuming, labour intensive, and unsuitable for on-site testing and high-throughput analysis. To date, there are many detection methods with a unique detection system available for Salmonella detection utilising immunological-based techniques, molecular-based techniques, mass spectrometry, spectroscopy, optical phenotyping, and biosensor methods. The electrochemical biosensor has growing interest in Salmonella detection mainly due to its excellent sensitivity, rapidity, and portability. The use of a highly specific bioreceptor, such as aptamers, and the application of nanomaterials are contributing factors to these excellent characteristics. Furthermore, insight on the types of biorecognition elements, the principles of electrochemical transduction elements, and the miniaturisation potential of electrochemical biosensors are discussed.

Typhoid fever, also known as typhoid, is a life-threatening bacterial infection that remains a global health concern. The infection is associated with a significant morbidity and mortality rate, resulting in an urgent need for specific and rapid detection tests to aid prevention and management of the disease. The present review aims to assess the specificity and sensitivity of the available literature on the immunodiagnostics of typhoid fever. A literature search was conducted using three databases (PubMed, ProQuest and Scopus) and manual searches through the references of identified full texts to retrieve relevant literature published between 1 January 2011 and 31 December 2020. Of the 577 studies identified in our search, 12 were included in further analysis. Lipopolysaccharides (LPS) and hemolysin E (HlyE) were the most frequently studied antigens. The specimens examined in these studies included serum and saliva. Using blood culture as the gold standard, anti-LPS IgA gave the highest sensitivity of 96% (95% CI: 93-99) and specificity of 96% (95% CI: 93-99) for distinguishing between typhoid cases and healthy controls, whereas the combination of anti-LPS and anti-flagellin total IgGAM gave the highest sensitivity of 93% (95% CI: 86-99) and specificity of 95% (95% CI: 89-100) for distinguishing typhoid cases and other febrile infections. A comparably high sensitivity of 92% (95% CI: 86-98) and specificity of 89% (95% CI: 78-100) were shown in testing based on detection of the combination of anti-LPS (IgA and IgM) and anti-HlyE IgG as well as a slightly lower sensitivity of 91% (95% CI: 74-100) in the case of anti-50kDa IgA. Anti-50kDa IgM had the lowest sensitivity of 36% (95% CI: 6-65) against both healthy and febrile controls. The development of a rapid diagnostic test targeting antibodies against lipopolysaccharides combined with flagellin appeared to be a suitable approach for the rapid detection test of typhoid fever. Saliva is added benefit for rapid typhoid diagnosis since it is less invasive. As a result, further studies could be done to develop additional approaches for adopting such samples.

Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has attracted public attention. The gold standard for diagnosing COVID-19 is reverse transcription-quantitative polymerase chain reaction (RT-qPCR). However, RT-qPCR can only be performed in centralized laboratories due to the requirement for advanced laboratory equipment and qualified workers. In the last decade, clustered regularly interspaced short palindromic repeats (CRISPR) technology has shown considerable promise in the development of rapid, highly sensitive, and specific molecular diagnostic methods that do not require complicated instrumentation. During the current COVID-19 pandemic, there has been growing interest in using CRISPR-based diagnostic techniques to develop rapid and accurate assays for detecting SARS-CoV-2. In this work, we review and summarize reverse-transcription loop-mediated isothermal amplification (RT-LAMP) CRISPR-based diagnostic techniques for detecting SARS-CoV-2.

Competing Interests: Competing interests: None declared.

PERCEPTRON is a next-generation freely available web-based proteoform identification and characterization platform for top-down proteomics (TDP). PERCEPTRON search pipeline brings together algorithms for (i) intact protein mass tuning, (ii) de novo sequence tags-based filtering, (iii) characterization of terminal as well as post-translational modifications, (iv) identification of truncated proteoforms, (v) in silico spectral comparison, and (vi) weight-based candidate protein scoring. High-throughput performance is achieved through the execution of optimized code via multiple threads in parallel, on graphics processing units (GPUs) using NVidia Compute Unified Device Architecture (CUDA) framework. An intuitive graphical web interface allows for setting up of search parameters as well as for visualization of results. The accuracy and performance of the tool have been validated on several TDP datasets and against available TDP software. Specifically, results obtained from searching two published TDP datasets demonstrate that PERCEPTRON outperforms all other tools by up to 135% in terms of reported proteins and 10-fold in terms of runtime. In conclusion, the proposed tool significantly enhances the state-of-the-art in TDP search software and is publicly available at https://perceptron.lums.edu.pk. Users can also create in-house deployments of the tool by building code available on the GitHub repository (http://github.com/BIRL/Perceptron)., (© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Hydroxyapatite (HA) has been widely used as a scaffold in tissue engineering. HA possesses high mechanical stress and exhibits particularly excellent biocompatibility owing to its similarity to natural bone. Nonetheless, this ceramic scaffold has limited applications due to its apparent brittleness. Therefore, this had presented some difficulties when shaping implants out of HA and for sustaining a high mechanical load. Fortunately, these drawbacks can be improved by combining HA with other biomaterials. Starch was heavily considered for biomedical device applications in favor of its low cost, wide availability, and biocompatibility properties that complement HA. This review provides an insight into starch/HA composites used in the fabrication of bone tissue scaffolds and numerous factors that influence the scaffold properties. Moreover, an alternative characterization of scaffolds via dielectric and free space measurement as a potential contactless and nondestructive measurement method is also highlighted.

A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify Salmonella Typhi and Salmonella Paratyphi A from suspected carriers. The lower detection limit for S . Typhi and S . Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 10 2 CFU/mL, respectively. Lateral flow biosensor was used for visual detection of mPCR amplicons (stgA, SPAint, ompC, internal amplification control) by labeling forward primers with fluorescein-isothiocyanate (FITC), Texas Red, dinitrophenol (DNP) and digoxigenin (DIG) and reverse primers with biotin. Binding of streptavidin-colloidal gold conjugate with the amplicons resulted in formation of a red color dots on the strip after 15-20 min of sample exposure. The nucleic acid lateral flow analysis of the mPCR-LFB was better in sensitivity and more rapid than the conventional agarose gel electrophoresis. Moreover, the mPCR-LFB showed 100% sensitivity and specificity when evaluated with stools spiked with 100 isolates of Salmonella genus and other bacteria. A prospective cohort study on stool samples of 1176 food handlers in outbreak areas (suspected carriers) resulted in 23 (2%) positive for S . Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program.

Water shortage is among the major abiotic stresses that restrict growth and productivity of citrus. The existing literature indicates that tetraploid rootstocks had better water-deficit tolerance than corresponding diploids. However, the associated tolerance mechanisms such as antioxidant defence and nutrient uptake are less explored. Therefore, we evaluated physiological and biochemical responses (antioxidant defence, osmotic adjustments and nutrient uptake) of diploid (2x) and tetraploid (4x) volkamer lemon (VM) rootstocks grafted with kinnow mandarin (KM) under two water-deficit regimes. The KM/4xVM (VM4) and KM/2xVM (VM2) observed decrease in photosynthetic variables, i.e., photosynthetic rate (Pn), stomatal conductance (gs), transpiration rate (E), leaf greenness (SPAD), dark adopted chlorophyll fluorescence (Fv/Fm), dark adopted chlorophyll fluorescence (Fv´/Fm´), relative water contents (RWC) and leaf surface area (LSA), and increase in non-photochemical quenching (NPQ) under both water-deficit regimes. Moreover, oxidative stress indicators, i.e., malondialdehyde (MDA) and hydrogen peroxide, and activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APx), glutathione reductase (GR) were increased under both water-deficit regimes. Nonetheless, increase was noted in osmoprotectants such as proline (PRO) and glycine betaine (GB) and other biochemical compounds, including antioxidant capacity (AC), total phenolic content (TPC) and total soluble protein (TSP) in VM2 and VM4 under both water-deficit regimes. Dry biomass (DB) of both rootstocks was decreased under each water-deficit condition. Interestingly, VM4 showed higher and significant increase in antioxidant enzymes, osmoprotectants and other biochemical compounds, while VM2 exhibited higher values for oxidative stress indicators. Overall, results indicated that VM4 better tolerated water-deficit stress by maintaining photosynthetic variables associated with strong antioxidant defence machinery as compared to VM2. However, nutrient uptake was not differed among tested water-deficit conditions and rootstocks. The results conclude that VM4 can better tolerate water-deficit than VM2. Therefore, VM4 can be used as rootstock in areas of high-water deficiency for better citrus productivity., Competing Interests: The authors have declared that no competing interests exist.

Hepatitis C is an inflammatory liver disease caused by the single-stranded RNA (ssRNA) hepatitis C virus (HCV). The genetic diversity of the virus and quasispecies produced during replication have resulted in viral resistance to direct-acting antivirals (DAAs) as well as impediments in vaccine development. The recent adaptation of CRISPR-Cas as an alternative antiviral approach has demonstrated degradation of viral nucleic acids in eukaryotes. In particular, the CRISPR-effector Cas13 enzyme has been shown to target ssRNA viruses effectively. In this work, we have employed Cas13a to knockdown HCV in mammalian cells. Using a computational screen, we identified several potential Cas13a target sites within highly conserved regions of the HCV internal ribosomal entry site (IRES). Our results demonstrate significant inhibition of HCV replication as well as translation in huh-7.5 cells with minimal effects on cell viability. These findings were validated using a multi-modality approach involving qRT-PCR, luciferase assay, and MTT cell viability assay. In conclusion, the CRISPR-Cas13a system efficiently targets HCV in vitro, suggesting its potential as a programmable therapeutic antiviral strategy., (Copyright © 2021 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)

The unfolded protein response (UPR) is activated in pancreatic pathologies and suggested as a target for therapeutic intervention. In this study, we examined activating transcription factor 3 (ATF3), a mediator of the UPR that promotes acinar-to-ductal metaplasia (ADM) in response to pancreatic injury. Since ADM is an initial step in the progression to pancreatic ductal adenocarcinoma (PDAC), we hypothesized that ATF3 is required for initiation and progression of PDAC. We generated mice carrying a germline mutation of Atf3 (Atf3 -/- ) combined with acinar-specific induction of oncogenic KRAS (Ptf1a creERT/+ Kras G12D/+ ). Atf3 -/- mice with (termed APK) and without KRAS G12D were exposed to cerulein-induced pancreatitis. In response to recurrent pancreatitis, Atf3 -/- mice showed decreased ADM and enhanced regeneration based on morphological and biochemical analysis. Similarly, an absence of ATF3 reduced spontaneous pancreatic intraepithelial neoplasia (PanIN) formation and PDAC in Ptf1a creERT/+ Kras G12D/+ mice. In response to injury, KRAS G12D bypassed the requirement for ATF3 with a dramatic loss in acinar tissue and PanIN formation observed regardless of ATF3 status. Compared to Ptf1a creERT/+ Kras G12D/+ mice, APK mice exhibited a significant decrease in pancreatic and total body weight, did not progress through to PDAC, and showed altered pancreatic fibrosis and immune cell infiltration. These findings suggest a complex, multifaceted role for ATF3 in pancreatic cancer pathology.

Melioidosis is a severe disease caused by Burkholderia pseudomallei ( B. pseudomallei ), a Gram-negative environmental bacterium. It is endemic in Southeast Asia and Northern Australia, but it is underreported in many other countries. The principal routes of entry for B. pseudomallei are skin penetration, inhalation, and ingestion. It mainly affects immunocompromised populations, especially patients with type 2 diabetes mellitus. The laboratory diagnosis of melioidosis is challenging due to its non-specific clinical manifestations, which mimic other severe infections. The culture method is considered an imperfect gold standard for the diagnosis of melioidosis due to its low sensitivity. Antibody detection has low sensitivity and specificity due to the high seropositivity among healthy people in endemic regions. Antigen detection using various proteins has been tested for the rapid determination of B. pseudomallei ; however, it presents certain limitations in terms of its sensitivity and specificity. Therefore, this review aims to frame the present knowledge of a potential target known as the Burkholderia invasion protein D (BipD), including future directions for its detection using an aptamer-based sensor (aptasensor).

The presence of concurrent chronic total occlusion (CTO) is a strong predictor for both short-term and long-term mortality. Successful percutaneous coronary intervention (PCI) of CTO has been associated with clinical benefit. We sought to perform a meta-analysis comparing CTO-PCI versus optimal medical therapy. PubMed, ClinicalTrials.gov, Google scholar and the Cochrane Central Register of Controlled Trials were searched for studies published from 2006 to 2019. A total of 16 studies, with 11,314 patients were included. We analyzed data on mortality, cardiac deaths, myocardial re-infarction, major adverse cardiac events, stroke, and repeat CTO-PCI using random-effects models. The odds ratios (OR) with 95% confidence interval (CI) were computed and P < 0.05 was considered as a level of significance. Compared with medical therapy alone, CTO-PCI was associated with lower mortality (OR: 0.45, CI: 0.32-0.63, P < 0.00001) and cardiac deaths (OR: 0.58, CI: 0.38-0.89, P = 0.01). These results were primarily driven by observational studies with no difference observed in randomized controlled trials. There was no significant difference in the incidence of major adverse cardiac events (OR: 0.71, CI: 0.48-1.05, P = 0.54), myocardial re-infarction (OR: 0.71, CI: 0.48-1.05, P = 0.54), stroke (OR: 0.61, CI: 0.32-1.17, P = 0.14, and repeat PCI (OR: 1.28, CI: 0.91-1.78, P = 0.16). This meta-analysis shows lower long-term mortality and cardiac deaths in CTO-PCI group as compared to OMT driven by observational studies with no difference observed in randomized controlled trials. Further randomized trials are needed to confirm these findings and evaluate long term results., (Copyright © 2020 Elsevier Inc. All rights reserved.)