Mycobacterium avium complex (MAC) infections are a significant clinical challenge. Determining drug-susceptibility profiles and the genetic basis of drug resistance is crucial for guiding effective treatment strategies. This study aimed to determine the drug-susceptibility profiles of MAC clinical isolates and to investigate the genetic basis conferring drug resistance using whole-genome sequencing (WGS) analysis. Drug-susceptibility profiles based on minimum inhibitory concentration (MIC) assays were determined for 38 MAC clinical isolates (12 Mycobacterium avium and 26 Mycobacterium intracellulare). Mutations associated with drug resistance were identified through genome analysis of these isolates, and their phylogenetic relationships were also examined. Drug resistance, based on MIC values, was most commonly observed for moxifloxacin (81.6%), followed by linezolid (78.9%), clarithromycin (44.7%) and amikacin (36.8%). We identified specific mutations associated with resistance to amikacin. These include the rrs mutation at C464T in amikacin intermediate-resistance M. avium, and two mutations at T250A and G1453T in amikacin non-susceptible M. intracellulare. Mutations in rrl at A2058G, A2059C and A2059G were potentially linked to clarithromycin resistance. MAC clinical isolates not susceptible to linezolid exhibited mutations in rplC at G237C and C459T, as well as two rplD mutations at G443A and A489G. GyrB substitution Thr521Ala (T521A) was identified in moxifloxacin non-susceptible isolates, which may contribute to this resistance. A phylogeny of our MAC isolates revealed high levels of genetic diversity. Our findings suggest that the standard treatment regimen for MAC infections using moxifloxacin, linezolid, clarithromycin and amikacin may be driving development of resistance, potentially due to specific mutations. The combination of phenotypic and genotypic susceptibility testing can be valuable in guiding the clinical use of drugs for the treatment of MAC infections. Mycobacterium avium complex (MAC), which mainly consists of Mycobacterium avium and Mycobacterium intracellulare, are common pathogens found in patients [Nontuberculous mycobacteria (NTM) are ubiquitous organisms that can cause chronic disease which is increasing in incidence and prevalence globally . Among Npatients  and in vpatients . MAC canpatients . Due to Mycobacterium avium complex is difficult to treat. Treatment failure can be observed in approximately one-third of cases [in vitro (phenotypic) drug-susceptibility tests and observed in vivo resistance of NTM [rrs gene confer aminoglycoside resistance [rrl confer macrolide resistance [gyrA or gyrB genes confer fluoroquinolone resistance [rplC or rplD genes confer linezolid resistance [of cases . The extof cases , 7. Drugof cases . Also, te of NTM , 10. Thesistance . Mutatiosistance , those isistance  and chansistance . TherefoRecent advances in whole-genome sequencing (WGS) and bioinformatics have provided powerful tools for analyzing genetics in various microorganisms . This teM. avium, M. intracellulare and other species. However, few have reported relevant drug-susceptibility profiles based on MIC tests and genome analysis. Therefore, we aimed to determine the drug-susceptibility profiles of 38 MAC clinical isolates from Thai patients and investigated the genetic basis that confers drug resistance using WGS analysis.Several studies have been conducted on drug resistance in MAC, including M. avium or M. intracellulare, were randomly selected and included in this study. The isolates were identified using the INNO-LiPA MYCOBACTERIA v2 line-probe assay  as recommended by the manufacturer [In total, 38 MAC isolates, identified as facturer . These ifacturer . All cliThe study protocol was approved by the Institutional Review Board (IRB) of Khon Kaen University Ethics Committee in Human Research (No. HE591454). Informed consent was waived for the use of medical data since patient information was anonymized and de-identified before analysis.The minimum inhibitory concentration (MIC) of each drug was determined using the Sensititre Slow Growing Myco SLOMYCOI assay  as recommended by the manufacturer and following the guidelines of the CLSI . The plaFor clarithromycin, amikacin, moxifloxacin, and linezolid, CLSI breakpoints  have beeGenomic DNA was extracted from multiple loopfuls of MAC colonies using the cetyl-trimethylammonium bromide sodium chloride (CTAB) method . The quaThe quality of sequence reads was assessed using FastQC version 0.11.9 . The avePaired-end filtered reads from each isolate were assembled using Unicycler version 0.4.8  with defMycobacterium avium 104 reference genome (GenBank accession number: CP000479.1) using BWA-MEM version 0.7.12 [Paired-end filtered reads of each isolate were mapped to the n 0.7.12 . The mapn 0.7.12 . GATK ven 0.7.12  was usedMycobacterium chelonae CCUG 47445 (CP007220.1) was used as the outgroup. Pairwise SNP distance matrices were analyzed using snp-dists version 0.8.2 [SAMtools mpileup, VCF and coverage files were used to generate the combined nucleotide frequencies among isolates of each positional SNP. The multiple sequence alignment CSV file was converted to FASTA format. The maximum-likelihood (ML) method with a general time-reversible (GTR) model of nucleotide substitution and a gamma-distribution model of rate heterogeneity was selected as the best model, implemented within MEGA version 10.0.5 . A phyloon 0.8.2 .Antimicrobial resistance genes were identified by conducting mass screening on the assembled MAC genomes using ABRicate software  with mulrrs, rrl, rplC, rplD, gyrA and gyrB genes were extracted from the assembled MAC genome sequences. Mutations in these genes were analyzed by performing multiple sequence alignments against corresponding sequences of the reference genome of M. avium 104 or M. intracellulare ATCC 13950 (CP003322.1) using the MUSCLE algorithm tool provided by MEGA.The sequences of Statistical analysis was performed using IBM SPSS Statistics for Windows, version 28.0 software . The number of resistant isolates was compared between the two species using the chi-squared or Fisher’s exact test. P-values less than 0.05 were considered statistically significant.M. avium (31.6%) and 26 isolates to M. intracellulare (68.4%). The majority of the isolates were collected from female patients (63.1%) with an average age of 57 years, ranging from 27 to 91 years. Patients were from many provinces in Northeast Thailand, including Buriram, Kalasin, Khon Kaen, Mahasarakham, Nakhon Phanom, Nong Khai and Yasothon. The characteristics of the MAC isolates are shown in Fig 1.Thirty-eight clinical isolates of MAC were recovered on LJ medium. Based on LPA analysis, 12 isolates were identified as S1 Table. The MIC distributions for all drugs were unimodal, except for clarithromycin, ethionamide and rifabutin, which displayed bimodal distributions . The MIC breakpoints for four drugs  were interpreted according to the approved guidelines established by the CLSI. The most common resistance phenotypes observed were those to linezolid (81.6%), followed by moxifloxacin (78.9%), clarithromycin (44.7%) and amikacin (36.8%). The two MAC species did not differ significantly in the proportion of isolates resistant to each drug.The drug-susceptibility profiles of all tested MAC isolates are presented in M. avium isolates, 91.7% (11/12) were phenotypically resistant to moxifloxacin, followed by linezolid , clarithromycin  and amikacin . The MIC50/MIC90 values of the isolates for amikacin, clarithromycin, linezolid and moxifloxacin were 16/32, 4/64, 32/64 and 8/>8 μg/mL, respectively.Among the M. intracellulare isolates, the most common resistance phenotypes were those to linezolid and moxifloxacin , followed by amikacin  and clarithromycin . The MIC50/MIC90 values of the isolates for amikacin, clarithromycin, linezolid and moxifloxacin were 32/64, 8/64, 32/64 and 4/8 μg/mL, respectively.Among the S2 Table).A total of 38 MAC isolates were sequenced and their genomes were annotated. The completeness and contamination of the genomes were on average >99% and <3%, respectively, which provides confidence in the results. The average genome size was 5,827,852.8 bp , and the average GC content was 67.8%. The N50 mean value was 457,039.5  (M. avium isolates (ranging from 115 to 415 SNPs) and 168 SNPs for M. intracellulare isolates, with a range of 89 to 290 SNPs (S2 Table). Based on the phylogenetic tree constructed using SNPs, the MAC isolates were not clonal strains .Among the 38 MAC clinical isolates there was an average pairwise difference of 191 SNPs, ranging from 89 to 415 SNPs. The pairwise differences averaged 243 SNPs for efpA, RbpA and rpoB2 as antimicrobial resistance genes (S3 Table).Based on ABRicate mass screening, five databases  were used to identify genes associated with antimicrobial resistance. No such genes were identified in the ARG-ANNOT, NCBI and ResFinder databases. However, three were identified in all isolates using the MEGARes database: drug and biocide MFS efflux pumps (EFPA), multi-drug RND efflux regulator (MTRAD) and RNA-polymerase binding protein (RBPA). The CARD database identified rrs mutation confers aminoglycoside resistance. A C464T mutation in the rrs gene was found in one M. avium isolate with intermediate resistance. Two mutations (T250A and G1453T) were found in the amikacin non-susceptible (intermediate and resistant) M. intracellulare isolates (S4 Table).For amikacin, the rrl confer macrolide resistance. Only clarithromycin-resistant M. avium isolates had rrl mutations at positions T321C, G1180A, A2058G, A2059C, A2059G and T2131G. For M. intracellulare, several mutations in the rrl gene were found in the non-susceptible isolates, including C304T, C1176G, G1240A, G1681A, A2058G, C2209G, G2215C, C2236T, C2402T, C2404T, C2825T and C2840T (S4 Table).For clarithromycin, mutations in rplC and rplD genes were investigated for resistance. This study did not include any M. avium isolates susceptible to linezolid, so a comparison between mutations in susceptible and non-susceptible isolates could not be made. However, several mutations were found in the rplC gene of non-susceptible M. avium isolates. Among M. intracellulare isolates, six mutations in the rplC gene were found, all of which were synonymous in linezolid non-susceptible isolates. In addition, we found some mutations in the rplD gene, two of which were non-synonymous (Arg148Lys and Thr153Ala) in linezolid non-susceptible M. intracellulare isolates (Tables ).For linezolid, the gyrA and gyrB were identified as being associated with fluoroquinolone resistance. Several mutations were found in gyrA and gyrB genes of M. avium isolates. However, due to the absence of moxifloxacin-susceptible M. avium isolates, mutations associated with moxifloxacin resistance could not be determined. In M. intracellulare, several mutations of the gyrA gene were found, all of which were synonymous, except for one causing the amino acid substitution Ala91Ser. Moreover, two amino acid substitutions (Thr521Ala and Thr521Ser) of the GyrB protein were found only in non-susceptible moxifloxacin isolates (Tables ).For moxifloxacin, mutations in the quinolone resistance-determining regions (QRDRs) of M. avium and M. intracellulare. The CLSI guidelines were used to investigate the MIC distribution for four drugs  and to define drug resistance. However, breakpoints for defining resistance against the other drugs have not yet been established [M. avium and M. intracellulare. Nevertheless, the rates of resistance to moxifloxacin, linezolid, clarithromycin and amikacin in this study appear to be higher than those reported in some previous studies [This study reports the MIC values (determined by broth microdilution) for 13 representative drugs in 38 MAC clinical isolates belonging to ablished . The maj studies , 13, 36.M. abscessus [M. avium and M. intracellulare isolates, respectively. A previous study found that most M. avium isolates were phenotypically susceptible to amikacin (85.9%), while no isolate was resistant [50/MIC90 values for amikacin were 16/32 and 32/64 μg/mL for M. avium and M. intracellulare isolates, respectively. To the best of our knowledge, the mechanism of high-level resistance in MAC isolates involves mutations in the rrs gene. A1408G, C1409T and G1419T are the most common mutations detected in this gene in kanamycin-resistant M. tuberculosis [M. intracellulare isolates, two (T250A and G1453T) were only in amikacin-intermediate and -resistant isolates. Therefore, we suggest that those mutations might be involved in amikacin resistance. Although these mutations were not present in all of our non-susceptible isolates, other mechanisms may contribute to aminoglycoside resistance. Previous literature suggests that aminoglycoside acetyltransferase and the drug efflux pump are associated with aminoglycoside resistance [Amikacin is an important parenteral drug for treating NTM, especially MAC and bscessus . We deteesistant . The MICrculosis . A recenrculosis . Among tsistance .rrl) [rrl mutation A2058G found in MAC isolates with clarithromycin resistance. Additional mutations A2059G and A2059C were also found only in clarithromycin-resistant M. avium isolates. Thus, rrl mutations  are potentially associated with clarithromycin resistance.Clarithromycin is a macrolide drug and the key therapeutic agent for NTM diseases  The CLSIrrl) –45. Our M. avium and 76.9% of M. intracellulare isolates. These results are consistent with a previous study in Korea [M. avium and M. intracellulare isolates, respectively. The rplC and rplD genes encode for the 50S ribosomal proteins L3 and L4, respectively, and mutations in these genes have been associated with resistance to linezolid in Gram-positive bacteria [M. avium and M. intracellulare isolates. In M. avium, some mutations, such as G237C and C459T in rplC and A489G in rplD, were concordant with previous studies [M. intracellulare, six mutations  were found in the rplC gene only in isolates non-susceptible to linezolid. In addition, several mutations were found in the rplD gene. One of these mutations (G443A) was found in non-susceptible M. intracellulare isolates, in agreement with a previous report [rplD gene were associated with resistance to linezolid [rplC and rplD genes have been associated with resistance to linezolid in MAC.Recent clinical guidelines recommend linezolid as a treatment option for drug-resistant TB and NTM infections , 47. Howin Korea , which fbacteria . In our  studies  on linezs report . Anotherinezolid , but we M. abscessus disease [gyrA and gyrB genes in the QRDRs, have been identified as associated with fluoroquinolone resistance [gyrA and gyrB in M. avium might be associated with this resistance [gyrA and gyrB were not found among the moxifloxacin-resistant M. avium and M. intracellulare isolates [gyrA and gyrB genes in M. avium were observed and one amino acid substitution was identified, Thr521Ala (T521A) in GyrA. Some of these mutations were consistently present among moxifloxacin non-susceptible M. intracellulare isolates. Interestingly, we found that the GyrB substitutions Thr521Ala (T521A) and Thr521Ser (T521S) were present only in non-susceptible moxifloxacin isolates, similar to M. avium. Our finding suggests that the GyrB substitutions Thr521Ala (T521A) or Thr521Ser (T521S) were present only in M. avium and M. intracellulare isolates that were not susceptible to moxifloxacin, which might contribute to moxifloxacin resistance. However, such association should be further investigated with a larger sample size. Also, further studies are needed to better understand the molecular mechanisms of drug resistance in MAC isolates.As per the ATS/IDSA guidelines, fluoroquinolones such as moxifloxacin and levofloxacin are the recommended drugs for the treatment of macrolide-resistant MAC and pulmonary  disease , 46, 52. disease , 36. Mutsistance . Some musistance . In contsistance . AnotherThe genome of MAC was relatively large, consisting of approximately 5.8 Mbp. Studies of the MAC genome has identified numerous genes and pathways involved in virulence and drug resistance as well as variation in gene content and organization among different strains, which could contribute to differences in virulence and drug resistance , 56. UndLimitations of our study should be noted. First, the number of MAC isolates tested was limited. Future studies with larger sample sizes are required to draw more meaningful conclusions. Furthermore, several mutations observed in our tested isolates might have simply reflected the high genetic diversity of MAC. While some mutations have been reported previously, supporting the view that these are associated with drug resistance in MAC, further validation is required for certain novel mutations or amino acid substitutions that we found. Additional studies that evaluate the mutations found in non-susceptible isolates in-depth and investigate those associated with phenotypic resistance are warranted. The mutation databases associated with each of the antibiotics or MAC are still very limited. Thus, we could not properly compare the performance of genotypic drug-susceptibility determination with phenotypic drug-susceptibility testing. Instead, we analyzed mutations in genes known from other mycobacteria to be associated with phenotypic drug resistance to get an insight into the drug resistance mechanism of MAC.rrs mutation at C464T for amikacin intermediate resistance in M. avium, and two mutations (T250A and G1453T) in amikacin non-susceptible M. intracellulare. Mutations in rrl at A2058G, A2059C and A2059G were potentially linked to clarithromycin resistance. Non-susceptibility to linezolid was associated with mutations in rplC at G237C and C459T, as well as two rplD mutations at G443A and A489G. We also identified the GyrB substitution Thr521Ala (T521A) in moxifloxacin non-susceptible isolates, which may contribute to resistance against that drug. Our findings provide insights into the specific mutations associated with drug resistance in MAC clinical isolates. These results emphasize the importance of using both phenotypic and genotypic susceptibility testing to assist clinicians in selecting effective drugs for the treatment of MAC infections. Due the high resistance rate we found to the antibiotics used for treatment of MAC infection, especially moxifloxacin and linezolid, new treatment strategies and adjustment of antibiotic options are needed.In conclusion, our study evaluated the drug susceptibility of MAC isolates and revealed high rates of resistance to moxifloxacin, followed by linezolid, clarithromycin and amikacin. High resistance rates might be a consequence of the misuse of antibiotics in our region . SpecifiS1 Table(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S1 Fig(TIF)Click here for additional data file.


Protein-energy wasting (PEW) is a common complication in patients with peritoneal dialysis (PD). Few investigations involved risk factors identification and predictive model construction related to PEW. We aimed to develop a nomogram to predict PEW risk in patients with peritoneal dialysis.We collected data from end-stage renal disease (ESRD) patients who regularly underwent peritoneal dialysis between January 2011 and November 2022 at two hospitals retrospectively. The outcome of the nomogram was PEW. Multivariate logistic regression screened predictors and established a nomogram. We measured the predictive performance based on discrimination ability, calibration, and clinical utility. Evaluation indicators were receiver operating characteristic (ROC), calibrate curve, and decision curve analysis (DCA). The performance calculation of the internal validation cohort validated the nomogram.n = 210) and validation (n = 159) cohorts according to the proportion of 6:4. The incidence of PEW was 49.86%. Predictors were age, dialysis duration, glucose, C-reactive protein (CRP), creatinine clearance rate (Ccr), serum creatinine (Scr), serum calcium, and triglyceride (TG). These variables showed a good discriminate performance in development and validation cohorts . This nomogram was adequately calibrated. The predicted probability was consistent with the observed outcome.In this study, 369 enrolled patients were divided into development (This nomogram can predict the risk of PEW in patients with PD and provide valuable evidence for PEW prevention and decision-making. PEW is a prevalent complication in chronic kidney disease patients with peritoneal dialysis, manifesting as a disorder of multiple metabolism and nutrition and a decrease of protein and fat . PEW resPeritoneal dialysis is an exchange between blood and a glucose-containing peritoneal solution. In this process, proteins and amino acids transport into the dialysate effluent. Patients lose amino acids of 1–3.5 g/d and proteins of 5–10 g/d during peritoneal dialysis, mainly albumin . A previRecently nomogram has been a technique for clinicians to predict the probability of disease or outcome. This study aimed to develop a PEW nomogram in patients with PD. According to the nomogram, clinicians can calculate PEW risk and identify the influence factors. Practical intervention approaches help to reduce PEW occurrence.We retrospectively searched data on patients with PD from two hospitals in China. This study developed and validated a nomogram to predict PEW risk in patients with peritoneal dialysis.We enrolled patients who had received maintenance peritoneal dialysis between January 2011 and November 2022. The enrolled criterion was the following set: (1) >18 years old; (2) with dialysis duration at least more than 3 months; (3) with follow-up visits and medical data records. We excluded patients unable to oral dietary intake or with multiple organ failure.The Ethics Committee of Lishui Municipal Central Hospital has approved the study. The ethical approval number is 2021-111. We received written informed consent from participants in this study. Legally Authorized Representatives of illiterate participants provided informed consent for the study. The study was conducted following the Declaration of Helsinki.2 or 10% total body fat; (3) wasted muscle defined by the lean tissue index, calculated as a lean tissue mass normalized to the height-squared in the 10th percentile of the reference population; and (4) low dietary protein intake.We randomly divided the study cohort into training and validation sets with a proportion of 6:4. We collected twenty-six parameters from the peritoneal dialysis management system. These variables contained essential characteristics and clinical laboratory assessments. Basic features included gender, age, ≥65 years, dialysis duration, peritonitis occurrence number, Renin-Angiotensin-Aldosterone System inhibitors (RAASi) administration, and complications. Complications covered infection, peritonitis, hypertension, diabetes, and cardiovascular disease. Laboratory tests were ultrafiltration volume, KT/V, Ccr, BUN, Scr, serum potassium, sodium, calcium, phosphorus, CRP, glucose, TG, low-density lipoprotein (LDL), predialysis BUN, and PTH. PEW was the end-point. According to the PEW diagnosis proposed by ISRNM , we defiThis study examined some laboratory data from the last dialysis. PD patients infuse dialysate into the abdomen. After a dwell time of hours, patients discharge dialysis effluent to achieve a complete PD process. The ultrafiltration volume is the difference value of the dialysis effluent minusing the abdomen dialysate. PD patients have residual renal function and clearance performance from the dialysate. Therefore, Ccr in this research refers to a summation of the creatinine clearance rate from the kidney and dialysis. Before peritoneal dialysis, clinicians draw patients’ blood to estimate serum creatinine levels. After dialysis, they collected urine and dialyzate to measure the creatinine values in these two samples. Clinicians conducted an enzymatic method to test the level of serum creatinine. The peritoneal dialysis management system installed a calculation formula. We obtained total Ccr results by importing creatinine values, urine and dialysate volume, and serum creatinine.etc. Imaging examination and tumor markers evaluated if patients have cancer. Bone mineral density test estimated osteoporosis condition. The presence of at least two following points diagnoses patients as peritonitis (9/L (after a dwell time of at least 2 h), with polymorphonuclear leukocytes of more than 50%; (3) positive dialysis effluent culture. Normal blood pressure is a systolic pressure of 90–140 mmHg and a diastolic pressure of 60–90 mmHg. Patients over 140/90 mmHg under no medical administration are hypertensive. Patients consistent with at least one point can be diagnosed as diabetes: (1) fasting blood glucose more than 7.0 mmol/L; (2) 2-h postprandial blood glucose more than 11.1 mmol/L; (3) HbA1c ≥ 6.5%. We measured patients’ height and weight to calculate BMI. The calculation formula is the weight divided by the height square. Magnetic resonance imaging, bio-electrical impedance analysis, and dual-energy X-ray absorptiometry test the total body fat and lean tissue mass. The lean tissue index resulted from lean tissue mass divided by the height square. Low dietary protein intake is a daily protein intake of less than 0.8 g/kg/d. Clinicians can trace patients’ follow-up data in the peritoneal dialysis management system. This system recorded all essential information, biochemical parameters, and dialysis-related data in every examination.Researchers collected patients’ blood to assess biochemical parameters, including plasma glucose, CRP, inflammatory indicators, tumor markers, itonitis : (1) symp ≤ 0.05 was confirmed statistically significant. The statistical analysis tool is STATA 15.0 .All data consisted of continuous and categorical variables. Continuous data were described by mean n = 210) and validation (n = 159) sets. A total of 184 (49.86%) of 369 patients were PEW. Baseline characteristics are shown in A total of 369 patients were enrolled in this study. They were divided into development (p = 0.012), age , Ccr , dialysis duration , TG , serum calcium , glucose , CRP .Multiple logistic regression screened eight predictors associated with PEW . These pWe included eight predictors to develop a nomogram by multivariate logistic regression . Accordip = 0.404, and p = 0.220. This result presents the predicted outcomes were highly consistent with the actual effects. The ROCs in the development and validation sets performed a nomogram with an excellent discrimination ability    and 3. W−0.753])  and 5. T−0.753])  and 7.In this study, the PEW incidence in patients with peritoneal dialysis was 49.86%. The rate was within the range reported in the previous study . For theProtein and amino acids transport to the dialysate in the peritoneal dialysis process. Patients with PD tend to suffer from protein catabolism, malnutrition, and inflammation. ISRNM suggested the status as PEW . The PEWAlthough some variables’ odds ratios were very close to 1, such as SCr, age, Ccr, dialysis duration, and CRP, we still considered them PEW predictors according to previous research and clinical observation. The crucial impact of inflammation on the nutrition condition has been published . InflammUremia promotes protein catabolism, increases the generation of non-nutritional toxins, and decreases albumin levels . Toxins This study discovered that patients with short-term peritoneal dialysis had a higher PEW risk. No researcher reported the association between dialysis duration and PEW risk before. In the clinical, we found that patients with short-term peritoneal dialysis tend to present higher ultrafiltration. Higher ultrafiltration stimulated peritoneal protein loss, resulting in hypoalbuminemia . TherefoCreatine (Cr) is an energy precursor essential to cellular physiology. In patients with chronic kidney disease, skeletal and cardiac muscles are depleted of Cr with the dialysis duration. Cellular damage due to Cr depletion lead to a deterioration of musculoskeletal and neurological function and poor quality of life. Cr supplements improve patients’ musculoskeletal, brain, and peripheral nervous systems with chronic kidney disease. About 80–90% of total Cr exists in the human muscles. Approximately 2% of total Cr is converted to creatinine daily, which the kidney eliminates into the urine . Serum cThere is some limitation in this study. First, participants in this nomogram are patients with peritoneal dialysis. This nomogram is not suitable for patients with hemodialysis. Second, we will continue to recruit more participants to achieve external nomogram validation.We developed a predictive model for PEW in patients with peritoneal dialysis. Predictors were age, dialysis duration, glucose, CRP, Ccr, Scr, serum calcium, and TG. This nomogram provided valuable evidence for predicting PEW in patients with peritoneal dialysis. According to this nomogram, clinicians can identify factors and make specific intervention methods to prevent PEW occurrence in high-risk patients.10.7717/peerj.15507/supp-1Supplemental Information 1All information of patient characteristics.Click here for additional data file.


Gracilaria is cultivated worldwide, in part for its production of agar and other bioactive compounds with downstream pharmaceutical and industrial applications. This genus is also emerging as a model for algal evolutionary ecology. Here, we report new whole-genome assemblies for two species (Gracilaria chilensis and Gracilaria gracilis), a draft genome assembly of Gracilaria caudata, and genome annotation of the previously published Gracilaria vermiculophylla genome. To facilitate accessibility and comparative analysis, we integrated these data in a newly created web-based portal dedicated to red algal genomics (https://rhodoexplorer.sb-roscoff.fr). These genomes will provide a resource for understanding algal biology and, more broadly, eukaryotic evolution.Macroalgal (seaweed) genomic resources are generally lacking as compared with other eukaryotic taxa, and this is particularly true in the red algae (Rhodophyta). Understanding red algal genomes is critical to understanding eukaryotic evolution given that red algal genes are spread across eukaryotic lineages from secondary endosymbiosis and red algae diverged early in the Archaeplastids. The Gracilariales is a highly diverse and widely distributed order including species that can serve as ecosystem engineers in intertidal habitats and several notorious introduced species. The genus  Gracilaria species that will serve as a key resource for algal research on evolution, ecology, biotechnology, and aquaculture.The Gracilariales are an ecologically and economically important red algal order found throughout the coastal regions of the world. Understanding the biology, ecology, and evolution of species in this order, and that of red algae more broadly, has been hampered by the limited phylogenetic coverage of genomic resources. Here, we present whole-genome assemblies and gene annotations for four Red algae (Rhodophyta) represent a lineage of photosynthetic eukaryotes in the Archaeplastids that diverged from green algae around 1,700 Ma . Within The Florideophyceae have a life cycle in which haploid male and female gametophytes alternate with a diploid tetrasporophyte but see  online. Gracilaria  Gracilaria caudata, and 3) Gracilaria gracilis  Papenfuss is a successful invader in many of the bays and estuaries of the Northern Hemisphere  Steentoft, L.M. Irvine & Farnham is a long-lived species that inhabits tide pools along European coastlines. This species serves as model species to test hypotheses related to the evolution of sex . specieell wall ; they caell wall . The fougracilis . Gracilarvesting . The artrvesting  and lossrvesting . Gracilamisphere . These imisphere  and to bmisphere , as wellmisphere . Graciladal zone  and has dal zone . FinallyGracilaria species should aid in our understanding of the evolutionary ecology of red algae in their dynamic environment, during invasions of new habitats, under cultivation practices, and in response to climate change. Moreover, these new resources will add to the existing genomic data and illuminate key processes in eukaryotic evolution. The Rhodoexplorer Red Algal Genome Database currently includes the Gracilaria species discussed here but will include all the high-quality genomic resources available for the Rhodophyta , thereby providing a unique resource for comparative analyses.The availability of genomic and genetic resources for these four G. gracilis and G. chilensis, respectively. In addition, we created a draft genome assembly based on the Illumina sequencing only for G. caudata (30 Mb) and reassembled the genome of G. vermiculophylla  are the most contiguous red macroalgal genomes presently available in public databases, apart from G. vermiculophylla and Pyropia yezoensis where the addition of a HiC library enabled scaffolding nearly at the chromosome level . The G. caudata assembly was fragmented with a low N50 of 21 kb and 55,767/5,535 contigs/scaffolds. Despite the differences in assembly size, BUSCO scores were similar across the long-read–sequenced G. gracilis and G. chilensis (83.6% and 81.6% of conserved proteins present) and the more fragmented G. caudata genome  in red algal genomes at a median level of 69% . Among these genes, 70.6–76.6% did not contain any introns, as typical for the compact genomes of red algae (Gracilaria genes had homologous sequences in the Uniprot database (84.2–89.7%) and were annotated with at least one INTERPRO hit (91.7–93.6%). Between 47.9% and 54.4% of genes were associated with gene ontology (GO) annotations.Gene prediction yielded a total of 7,943, 8,737, and 9,460 protein-coding sequences for gracilis , which wed algae . Most GrG. chilensis to 18.2% for G. caudata. Both the annotated and the unknown species-specific genes constitute attractive targets to study their role in adaptation and speciation.OrthoFinder analyses identified 4,666 orthogroups present in all four genomes  online vhttps://rhodoexplorer.sb-roscoff.fr), which will include more red algal genomes in the future. The services provided include the following:In addition to depositing the raw reads and sequenced genome in a public repository, we integrated the data into the newly created Rhodoexplorer Red Algal Genome Database .Assembly and annotation metrics.Data downloads: genomic, genes and proteomic data sets, structural and functional annotations, orthology clusters, etc.A Blast interface with a selection of red algal genomes, predicted and de novo assembled transcriptomes and proteomes.Visualization tools: a genome browser to visualize the predicted genes and the RNA-sequence (RNAseq) data mapped on the genome and a web interface to visualize functional annotations and retrieve individual protein sequences.Gracilaria thalli, all bearing reproductive structures, were collected from natural populations: G. chilensis in Lenca , G. vermiculophylla in Charleston, SC , G. caudata in Paracuru, CE , and G. gracilis in Cape Gris-Nez . Gracilaria caudata and G. chilensis were maintained as clonal, unialgal cultures under laboratory conditions prior to nucleic acid extractions (see Culture conditions). Field-collected G. gracilis and G. vermiculophylla thalli were transported to the laboratory, examined under a microscope, and cleaned of contaminants. If visible, cystocarps were excised prior to preservation of the thalli at −80°C. Gracilaria species used in this study.Adult female and male Gracilaria caudata was grown in the modified von Stosch nutrient solution  or PacBio Sequel II with sheared gDNA large insert library (G. gracilis and G. chilensis) (Genomic DNA (gDNA) was extracted using DNeasy PowerPlant Pro Kit for ilensis)  online.G. chilensis, G. caudata, and G. gracilis using the RNeasy Mini Plant Kit (Qiagen) and Macherey Nagel NucleoSpin RNA Plant Kit for G. vermiculophylla, following the manufacturer's instructions. Paired-end 150-bp Illumina reads were generated with Illumina HiSeq 2500  using BlobTools v1.1.1 . Annotation of those de novo organellar genomes was done using the GeSeq web tool (https://chlorobox.mpimp-golm.mpg.de/geseq.html). Public sequences from G. caudata voucher SPF:57390 , G. chilensis voucher CNU050183 , G. gracilis voucher SPF:55734 , and G. vermiculophylla  were retrieved from NCBI and used as seeds and references for both assembly and annotation.Chloroplastic and mitochondrial genomes of each species were reconstructed from Illumina raw reads using NOVOPlasty  through Each reference genome was first masked using RepeatMasker v4.0.9  with Dfahttps://abims-sbr.gitlab.io/rhodoexplorer/doc/data_process/.All codes used for genomes assembly and annotation are available on the Gitpage dedicated to the genome database project https://rhodoexplorer.sb-roscoff.fr) has been implemented using the Python web framework Django, with data stored in a relational database (PostgreSQL).The main web portal (https://www.elastic.co/products/elasticsearch); and Galaxy, a browser-accessible workbench for scientific computing used as a data loading orchestrator for administrators (http://gitlab.sb-roscoff.fr/abims/e-infra/gga_load_data) allowing mass deployment of Docker containers and automated data loading through Galaxy with the Bioblend API  project . Each GGstrators . To facilend API .https://blast.sb-roscoff.fr/rhodoexplorer/) includes an implementation of the Blast algorithm using SequenceServer (The Blast interface (ceServer  graphicahttps://abims-sbr.gitlab.io/rhodoexplorer/doc/) is published from a GitLab repository, with Pages and MkDocs, a static site generator.The documentation website for navigating the platform web portal and resources (The entire informatic infrastructure is deployed and maintained on the ABiMS Bioinformatics platform of the Roscoff Biological Station, part of the national infrastructure French Bioinformatic Institute.evad124_Supplementary_DataClick here for additional data file.


All data were analyzed using a custom pipeline called MyCodentifier. Sequences were matched against a custom hsp65 database (NGS-hsp65) and whole-genome database (NGS-WG) created based on the phylogeny presented by Tortoli et al. . Lastly, phylogenetic analysis was performed and correlated with clinical manifestation. In the explorative set, we observed 98.6% agreement between the line probe assay and the NGS-hsp65 database. In the validation set, 99.1% agreement between the NGS-WG and NGS-hsp65 databases was seen on the complex level. We identified a cluster of Mycobacterium marinum isolates not represented by the Tortoli et al. phylogeny. Phylogenetic analysis of M. avium complex isolates confirmed misclassification of M. timonense and M. bouchedurhonense and identified subclusters within M. avium although no correlation with clinical manifestation was observed. We performed routine NGS to identify NTM from MGIT enriched shotgun metagenomic data. Phylogenetic analyses identified subtypes of M. avium, but in our set of isolates no correlation with clinical manifestation was found. However, this NGS workflow paves a way for more personalized health care in the future.Currently, nontuberculous mycobacteria (NTM) are identified using small genomic regions, and species-level identification is often not possible. We introduce a next-generation sequencing (NGS) workflow that identifies mycobacteria to (sub)species level on the basis of the whole genome extracted from enriched shotgun metagenomic data. This technique is used to study the association between genotypes and clinical manifestations to pave the way to more personalized health care. Two sets of clinical isolates (explorative set [ Mycobacterium contains >200 different species, including the obligate pathogens Mycobacterium tuberculosis complex (MTBC) and M. leprae and the opportunistic pathogens classified as nontuberculous mycobacteria (NTM) (2The genus ia (NTM) , 2. NTM a (NTM) 24.M. avium complex (MAC), M. abscessus, M. malmoense, M. xenopi, and M. kansasii often indicates true NTM pulmonary disease , (multi)gene sequencing, or matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) –12. Beca–N-acetyl-l-cysteine (NaLC)-NaOH for 15 min to decrease contamination. After pretreatment, phosphate buffer was added to a final volume of 50 mL. For materials obtained in sterile settings, pretreatment was skipped and phosphate buffer was immediately added to a final volume of 50 mL. Tubes were then centrifuged at 3,000 × g for 20 min at 4°C, all supernatant was discarded, 2 mL phosphate buffer was added, and the remaining sediment was vortexed briefly. An 0.5-mL amount of the sediment was then added to a liquid culture mycobacterial growth indicator tube (MGIT) containing 0.8 mL BD Bactec MGIT oleic acid-albumin-dextrose-catalase (OADC) growth supplement and PANTA  antibiotic supplement , and 0.25 mL was inoculated onto Lowenstein-Jensen (LJ) solid medium. Liquid culture samples were then incubated in the Bactec MGIT until they flagged positive. All MGIT-positive samples underwent further culture for strain storage including inoculation on blood agar to ensure a monoculture. Stored Mycobacterium species were cultured in Middlebrook 7H9 broth with 10% OADC supplement before pellet collection. Finally, solid medium cultures were swabbed and resuspended in glycerol broth. Samples were heat inactivated for DNA isolation.New clinical samples were processed as previously described . In shorDNA isolation was performed using InstaGene matrix  in combination with bead beating. Two hundred microliters InstaGene matrix was added to bacterial pellets, vortexed, and incubated with shaking at 500 rpm for 30 min at 56°C and then another 30 min at 99°C. Samples were then transferred to a tube containing <106-μm glass beads  and subjected to two rounds of bead beating at maximum speed in a MagNA Lyser . DNA integrity was measured on a TapeStation 2200 , and concentration was determined using a Qubit fluorometer . Library preparation was performed using the Illumina Nextera DNA Flex kit .We generated a curated database containing assembled genome sequences of 148 NTM type strains, including the most recent characterization of mycobacterial taxonomic relationships, performed by Tortoli et al. .hsp65 gene from each strain. To this end, the ~1.6-kb fragment coding for the hsp65 gene was retrieved from each of the 148 genomes and an NGS-hsp65 typing database was constructed using KMA (version 1.3.2)  .All databases are made available via zenodo .https://jordycoolen.github.io/MyCodentifier/). In short, it is a Nextflow pipeline that cleans shotgun metagenomic NGS data and identifies the mycobacterial species up to sublineage level using the custom-built NGS-hsp65 and NGS-WG databases. Results are reported if hsp65 identity score and coverage are ≥70%; elsewise, they are reported as failed.For processing the NGS data, we used our custom-developed pipeline MyCodentifier (version 1.0 beta)  was obtained, the analysis was supplemented with partial hsp65 gene sequencing  to reacM. marinum and three M. avium subspecies retrieved from the NCBI genome database .The NGS-WG typing database was later expanded by adding additional reference genomes to improve NTM subspecies identification, one M. marinum-M. ulcerans group, M. avium complex, and M. abscessus using sequence reads obtained from MyCodentifier together with multiple reference genomes (Table S1)  parsimony trees of the able S1) –22.M. tuberculosis H37Rv andM. abscessus ATCC 19977 type strains were correctly identified; all M. avium ATCC 700898 type strains were identified as M. bouchedurhonense, a member of the Mycobacterium avium complex (MAC) almost indistinguishable from M. avium  and were typed using partial hsp65 Sanger sequencing. Of the remaining 212 isolates, 98.6% (n = 209) had the same outcome on the complex level , the NGS-hsp65 typing was able to identify (sub)species within the complex as reported by the LiPA . One NGS-hsp65 (M. intracellulare) species identification differed from the LiPA identification (M. chimaera). One M. simiae complex isolate identified by partial hsp65 Sanger sequencing as M. stomatepiae  was ideM. tuberculosis-M. intracellulare). Additional errors (M. fortuitum-M. intracellulare and M. timonense-M. chelonae) were untraceable but are likely due to similar human error.Discrepancy analysis, consisting of reviewing all steps from culture to analysis, of the three strains not explained by differences in databases was performed. One case was found to be due to a pipetting error  and NGS-hsp65 (M. mucogenicum) due to a mixed infection. The second discrepancy between NGS-WG (M. arupense) and NGS-hsp65 (M. gordonae) was due to the abundance calculation incorporated in the centrifuge typing tool, in which small genomes, like M. arupense, result in high abundance percentages despite lower numbers of reads. Upon human curation the isolate was identified as M. gordonae. On species level an accordance of 85.2% is found; the 14.8% observed discrepancies consist mainly of MAC species (11.4%) and M. marinum-M. pseudoshottsii (2.1%). One discrepancy (0.4%) is found between M. porcinum and M. boenickei, closely related strains within the M. fortuitum-smegmatis group. For M. abscessus, identification up to subspecies level was achieved for all isolates .As we aim to use NGS-WG in clinical practice, the NGS- strains . On the hsp65 and NGS-WG typing within the M. marinum group and MAC were studied further. We performed phylogenetic analysis of all five clinical strains belonging to the M. marinum group together with 25 reference genomes retrieved from NCBI  and one M. marinum strain from cluster II (M. marinum Europe), MyCodentifier identified all clinical M. marinum strains as M. pseudoshottsii; however, they should have been identified as a cluster I M. marinum  and 24 reference sequences from NCBI (Table S1). Phylogenetic analysis shows that MAC contains multiple well-defined clusters, largely related to existing (sub)species. Overall, the phylogenetic tree supports the findings from the NGS-WG and not the NGS-hsp65 typing , four clusters of clinical isolates are observed  and the presence of other relevant diseases  were recorded for all M. avium isolates, and no correlation was seen with specific clusters. Our validation set contained a number of strains sequenced sequentially from the same patient .We calculated the phylogenetic tree of all rpoB, or hsp65 sequencing allowed for identification of most species, it depends on the quality of sequences uploaded to noncurated online databases. The implementation of NGS-WG allows for highly specific and reproducible identifications, including identification of mixed infections, and it allows typing to subspecies level and beyond. In our current setting, the turnaround time (TAT) of WGS will not significantly delay the diagnosis in the context of a chronic infection with a slow-growing bacterium but will slightly delay species identification. With our current NGS-WG method, the TAT from positive MGIT to species determination is between 4 and 11 days, depending on the day the MGIT flags positive ; this is important because the incidence of macrolide susceptibility differs between M. abscessus subspecies and, as a result, so does the chance of successful antibiotic treatment , and the other includes M. marinum Europe (cluster II). Because our clinical M. marinum strains belonged to cluster I, they were initially identified as M. pseudoshottsii, the nearest ancestor present in the Tortoli et al. phylogeny . Interestingly, we see four distinct clusters within the sequenced M. avium strains. Previous studies have proposed that these clusters may represent strains of M. avium responsible for different disease manifestations or affecting different patient groups species is possible using typing on NGS-WG level and on NGS-s status , 23. We t groups , 26. We M. gordonae isolate as M. arupense, resulting from the black-and-white calculation methods underlying the bioinformatic pipeline. Human curation is also necessary to identify mixed infections based on the presence of reads mapping to multiple species. The current report not only highlights the most prevalent species but also includes a table with the 10 most dominant species, thereby allowing us to manually identify the presence of multiple species in one sample. We believe that this, together with laboratory sample exchange, is likely the reason for the small percentage of incorrectly typed strains relative to the LiPA. Furthermore, quality control of the WGS-NG workflow and pipeline is performed regularly by the inclusion of known positive and negative samples and monitored by the multidisciplinary team. In addition, although our current database is effective in identifying the majority of NTM isolates to subspecies level, 10 strains could not be identified. This is likely due to the limitations of our database. We included all strains in the database of Tortoli et al., because this is currently the most extensively described, curated database (Although automated analysis is convenient, it remains important that results are manually curated prior to reporting to the clinic. In our clinic manual curation is performed by a multidisciplinary team consisting of at least a specialized bioinformatician and medical microbiologist and sometimes a laboratory specialist when necessary. The need for manual curation is exemplified by the incorrect identification of an database . Howeverdatabase , 28, andM. tuberculosis complex organisms (M. abscessus (In contrast to NGS studies of rganisms , we haverganisms , predictbscessus , and in In conclusion, we present a novel methodology for identification of clinically isolated mycobacteria using NGS-WG that has multiple benefits over conventional typing methods. First, this method allows for more in-depth identifications to species or even subspecies level, often relevant for treatment regimens or understanding virulence. Second, fewer additional tests are needed to perform detailed identification and typing. The detection of antibiotic resistance in NTM, the presence of plasmids, and the associations between genotypes and clinical manifestations or virulence are important topics for future studies.


Vibrio parahaemolyticus is a primary seafood-associated pathogen that could cause gastroenteritis. It can attach to various surfaces and form a biofilm, which poses serious threats to food safety. Hence, an effective strategy is urgently needed to control the biofilm formation of V. parahaemolyticus. Laurel essential oil (LEO) is used in food, pharmaceutical and other industries, and is commonly used as a flavoring agent and valuable spice in food industries. The potential antibiofilm effects of LEO against V. parahaemolyticus were examined in this study. LEO obviously reduced biofilm biomass at subinhibitory concentrations (SICs). It decreased the metabolic activity and viability of biofilm cells. Microscopic images and Raman spectrum indicted that LEO interfered with the structure and biochemical compositions of biofilms. Moreover, it also impaired swimming motility, decreased hydrophobicity, inhibited auto-aggregation and reduced attachment to different food-contact surfaces. RT-qPCR revealed that LEO significantly downregulated transcription levels of biofilm-associated genes of V. parahaemolyticus. These findings demonstrate that LEO could be potentially developed as an antibiofilm strategy to control V. parahaemolyticus biofilms in food industries. Vibrio parahaemolyticus (V. parahaemolyticus) is a rod-shaped, Gram-negative halophilic bacterium, which is highly abundant in estuarine and coastal waters. V. parahaemolyticus is recognized as an important foodborne pathogen transmitted mainly through contaminated seafood throughout the world  × 100.Bacterial adhesion to hydrocarbons (BATH) was performed using xylene . Xylene V. parahaemolyticus ATCC 17802 suspension was harvested, washed and resuspended with PBS to an OD600 nm (A0) of approximately 0.5. Then, the suspension was transferred to tubes. After 20 h, OD600 nm (A) was determined. The auto-aggregation (Aag) was calculated with the following formula: Aag% = (A0 – A)/A × 100%.According to Ling et al., the effect of LEO on auto-aggregation was measured . V. paraV. parahaemolyticus biofilms were determined using serial dilutions in PBS and were spread on 3% NaCl LB agar at 37 °C overnight.The biofilms on SS coupons, glass and food-grade silicone were as described previously, with minor modifications . Briefly5 cells per well) grown at 37 °C and 5% CO2 for 24 h. LEO at SICs were added for 12 h. The culture was eliminated. Meanwhile, 100 μL DMEM and 10 μL CKK-8 solution were added. After 1 h, OD450 nm was measured.The murine macrophage cell line, RAW 264.7 and human colon adenocarcinoma cell line, Caco-2 were used in the study. The cytotoxicity assay of LEO was detected using the CCK8 method . RAW 264.7 and Caco-2 cells were seeded in 96-well plate including DMEM with 10% fetal bovine serum. All cells were . The total RNA was reverse transcribed into cDNA with an Evo M-MLV RT Kit . −ΔΔCT) method was used to calculate relative quantity of gene expression. The 16 S rRNA gene was served as a reference gene for data normalization.The effect of LEO on the transcription of p < 0.05), ** (p < 0.01) and *** (p < 0.001).All treatments were conducted in triplicate. Data were presented as the mean ± SD. Differences were analyzed using one-way analysis of variance (ANOVA) using SPSS 19.0. Significance was indicated by *  was 1.17, 1.46 and 1.98 cm−1, respectively.The flagella-mediated motility plays a crucial role in bacterial initial attachment, which is an initial step in biofilm formation. The single polar flagellum is responsible for V. parahaemolyticus biofilms were presented in −1. Several other minor peaks were found at 1510, 1752 and 2647 cm−1. Furthermore, typical vibrational Raman bands of carbohydrates, proteins, carbohydrates and nucleic acid could be observed in the Raman spectra of V. parahaemolyticus biofilms. The distinct Raman bands were considered as carbohydrates: 563 cm−1 was attributed to C-O-C glycosidic ring deformation vibration, and 789 cm−1 was assigned to the O-P-O stretch of DNA [−1 and 1742 cm−1 belonged to carbohydrates and lipids, respectively [−1 was assigned to C=C stretching of carotenoids [−1 were due to a mix of C=C or C=O bond vibrations in fatty acids, DNA strand bonds and amide bonds in proteins [V. parahaemolyticus biofilm treated with LEO presented decreased Raman peak intensities. In particular, Raman peaks at 563 and 1088 cm−1 corresponding to carbohydrates showed the greatest reduction. The peak intensities of 2174 and 2412 cm−1 also exhibited a decreasing trend. The Raman peaks at 1510, 1742 and 2647 cm−1 disappeared. In addition, the peak intensity at 789 cm−1 remained unchanged.The Raman spectroscopy was carried out for biofilms formed on coverslips. Raman spectral features of h of DNA . The Ramectively . The peaotenoids . The Ramproteins . CompareV. parahaemolyticus to adhere to various surfaces. As displayed in V. parahaemolyticus untreated with LEO exhibited high CSH. CSH of V. parahaemolyticus, treatment with LEO  were reduced by 16.63%, 15.26% and 5.59%, respectively, compared to the control. Similarly, a significant inhibition in aggregation ability was also observed after LEO treatment  was shown in 2, and was 6.87 and 6.76 log CFU/cm2 on the SS and silicone surfaces, respectively. Compared to the control, LEO treatment  reduced the cell numbers on glass by 17.92%, 15.97% and 10.83%, respectively. The V. parahaemolyticus cell count on the SS surface was reduced by 22.13%, 17.90% and 7.60%, respectively. Similarly, V. parahaemolyticus count in the biofilm cells of the silicone surface was reduced by 21.45%, 17.60% and 8.14%, respectively, with LEO treatment .The effect of LEO on The effect of LEO on cell cytotoxicity is shown in V. parahaemolyticus flagellum , quorum sensing production regulators (aphA and opaR) and other biofilm-formation-related genes  were significantly downregulated after LEO treatment , which increase the difficulty of disinfection and eradication [Yersinia enterocolitica and Escherichia coli. It also has antifungal activity towards Penicillium digitatum, Monilinia laxa and Botrytis cinerea [V. parahaemolyticus biofilms has not been studied before.It is reported that al areas . V. paradication . In addidication . Therefodication . Moreove cinerea . NeverthV. parahaemolyticus biofilm formation. We used SICs to test the anti-biofilm potential of LEO to avoid selective pressure on growth, which may lead to the development of antibiotic resistance [Listeria monocytogenes biofilms, thereby destroying preformed biofilms [Our study suggests that LEO at SICs concentrations could effectively control sistance . Our resbiofilms . The probiofilms . FE-SEM biofilms ,41.flgM, flgL, flaA and flaE. Cao et al. reported that citral inhibited the swimming ability of V. parahaemolyticus and also downregulated the transcriptions of genes related to motility [flgM, flgL and flaA in V. parahaemolyticus [V. parahaemolyticus biofilms. Our study indicated that two maximum absorption peaks were at 563 and 1088 cm−1, which were assigned as polysaccharides, which confirmed that exopolysaccharides accounted for the majority of biofilm components. Moreover, LEO treatment led to a decreased amount of polysaccharides and other biochemical compositions , which could be assessed using the lower height of corresponding peaks.Bacteria could attach to various biotic and abiotic surfaces partly owing to flagellar-mediated motility, which successfully facilitates the initial adhesion and positively modulates biofilm formation. In this study, LEO at SICs significantly repressed swimming motility and further downregulated the expression of genes related to motility, including motility . In addiolyticus . EPS canolyticus . EPS areolyticus . We usedCronobacter sakazakii [grxB deletion mutant significantly inhibited biofilm formation by decreasing CSH and auto-aggregation. A significant positive correlation was found between auto-aggregation and biofilm formation in Myroides odoratus [V. parahaemolyticus partly due to reduced CSH and auto-aggregation.CSH is an important factor in the cell attachment stage, aggregation and colony assembly, thereby affecting biofilm formation. Furthermore, aggregation ability plays an essential role that interferes with the adhesion of pathogenic bacteria to various surfaces through competitive exclusion, leading to biofilm development and stability of the bacterial biofilm colonies . Ling etakazakii . In theiodoratus . These fV. parahaemolyticus. The reduced biofilms caused by LEO was associated with decreased bacterial swimming motility, CSH, auto-aggregation and decreased expression of biofilm-related genes of V. parahaemolyticus. These findings suggest that LEO could be potentially developed as an alternative strategy to control V. parahaemolyticus biofilms in food industries.In summary, LEO at SICs decreased biofilm biomass, disrupted biofilm architecture, reduced metabolic activity and changed biochemical composition of biofilms of


The neutrophil-to-lymphocyte ratio (NLR) is recognized as a biomarker for systemic inflammation and immune activation. However, its connection with the mortality risk in individuals with rheumatoid arthritis (RA) is not well understood. This study aimed to investigate the association between NLR and all-cause and cardiovascular mortality risk in U.S. adults with RA.Data were gathered from the National Health and Nutrition Examination Survey (NHANES) cycles spanning 1999 to March 2020. We included adults aged ≥20 years. The NLR was computed by dividing the neutrophil count by the lymphocyte count from complete blood counts. The maximally selected rank statistics method helped identify the optimal NLR cutoff value associated with significant survival outcomes. Multivariable logistic regression models were performed to investigate the relationship between the NLR and the all-cause and cardiovascular mortality of RA. Restricted cubic spline (RCS) analyses were utilized to detect whether there were linear or non-linear relationships between NLR and mortality.In this study, 2002 adults with RA were included, with 339 having a higher NLR (≥3.28) and 1663 having a lower NLR (<3.28). During a median follow-up of 84 months, 79 RA individuals died. Participants with higher NLR had a 2-fold increased risk of all-cause  and cardiovascular mortality  versus lower NLR, after adjusting for demographics, socioeconomic status, and lifestyle factors. Kaplan-Meier analysis revealed that the survival rate for the higher NLR group was significantly lower than the lower NLR group, in terms of both all-cause and cardiovascular mortality (both P<0.0001). The RCS curve demonstrated a positive linear association between the NLR and all-cause and cardiovascular mortality.A higher NLR was independently predictive of elevated long-term mortality risk in U.S. adults with RA. The NLR may serve as an inexpensive, widely available prognostic marker in RA. RA is aA perfect prognostic scoring system should offer prognostic parameters that are easily discernible during diagnosis and be cost-effective in clinical practice, ensuring accessibility and affordability for widespread use. The neutrophil-to-lymphocyte ratio (NLR) stands as a burgeoning biomarker, offering insights into systemic inflammation and immune system activation . It provIn RA, previous studies have touched upon the relationship between NLR and the presence of the disease , 11. RecIn light of the necessity for affordable and accessible prognostic markers in RA, our objective was to investigate the association between NLR and the risk of all-cause and cardiovascular mortality. We conducted these analyses using a vast, nationally representative sample of U.S. adults with RA obtained from the National Health and Nutrition Examination Survey (NHANES) dataset.22.1http://www.cdc.gov/nchs/nhanes.htm, accessed on 1 October 2023).NHANES is an ongoing program conducted by the Centers for Disease Control and Prevention (CDC) of America to assess the health and nutritional status of US adults and children. The survey combines interviews, physical examinations, and laboratory tests. Detailed methods can be extracted from the official website  documenting death from any cause. These mortality data were sourced from the National Death Index (NDI) database, accessible at 2.32), normal (18.50-24.99 kg/m2), overweight (25.00-29.99 kg/m2), or obese (≥30.00 kg/m2); smoking status (current smoker vs nonsmoker); and health insurance status (insured vs uninsured). All multivariable models sequentially adjusted for demographic factors, socioeconomics, lifestyle factors, comorbidities, and medications where appropriate. Missing covariate data were addressed through listwise deletion.Demographic covariates included age (continuous), sex , and race/ethnicity . Socioeconomic status was assessed using educational attainment  and poverty-income ratio . Lifestyle and health-related factors comprised body mass index (BMI) categorized as underweight (<18.49 kg/m2.4Baseline characteristics were summarized using the mean (standard deviation) for continuous variables and the number (percentage) for categorical variables. Differences by the presence of RA or NLR groups were compared using chi-squared tests for categorical variables and t tests for continuous variables. Cox proportional hazards models were used to investigate the association between NLR and all-cause and cardiovascular mortality in individuals with RA. Three models were constructed with incremental adjustment for potential confounders. Model 1 was unadjusted. In model 2, age, sex, race/ethnicity, and educational attainment were additionally adjusted; Model 3 was additionally adjusted by age, sex, race/ethnicity, educational attainment, BMI, poverty income ratio, smoking status, and health insurance. Kaplan-Meier curves were created to illustrate the survival probabilities based on different NLR categories. Subgroup analyses were performed to assess effect modification by pertinent clinical confounders. Restricted cubic spline (RCS) model was constructed to visualize the relationship between continuous NLR and outcomes. Analyses were performed using R version 4.3.1 . Two-sided P<0.05 was considered statistically significant.33.1Of 2002 participants with RA, 339 had higher NLR (≥3.28) while 1663 had lower NLR (<3.28)  than expected (67.3). In the unadjusted model (Model 1), higher NLR as a continuous variable was significantly associated with increased risk of all-cause mortality  and cardiovascular mortality . When NLR was evaluated as a categorical variable, participants in the higher NLR group had a 2-fold higher risk of all-cause mortality  and a 3-fold higher risk of cardiovascular mortality  compared to the lower NLR group. Model 2 was adjusted for demographic factors including age, sex, race/ethnicity, and education. The associations remained statistically significant. Model 3 was further adjusted for BMI, poverty income ratio, smoking, and insurance in addition to demographics. The positive associations between higher NLR and elevated mortality risks persisted. As a continuous variable, each 1-unit NLR increase was linked to 20% and 22% higher all-cause and cardiovascular mortality. Categorically, the higher NLR group had 2.02 and 2.48 times the risk of all-cause and cardiovascular mortality versus lower NLR participants (3.3In subgroup analyses stratified by sex, age, race/ethnicity, education, BMI, and smoking status, higher NLR was consistently associated with increased all-cause and cardiovascular mortality risk across all subgroups (3.44In this large nationally representative cohort of US adults, we found that a higher NLR was independently predictive of increased all-cause and cardiovascular mortality risk among adults with RA. Kaplan-Meier curves illustrated significantly lower survival probabilities for those with higher NLR. RCS analyses demonstrated linear positive associations between NLR and all-cause and cardiovascular mortality. These data highlight the potential utility of the NLR as an inexpensive and widely available marker for RA risk stratification and prognostication in clinical practice.A number of prior studies have examined the association between NLR and RA with inconsistent findings. Several studies found significantly higher NLR values in RA patients than in healthy controls, suggesting that the NLR may be a useful biomarker for RA diagnosis –18. OthePrior studies have reported associations between higher NLR and increased mortality risk across the general population and various conditions marked by significant inflammatory components , 26. In Moreover, determining specific NLR cutoffs tailored to individual patient populations could improve the predictive accuracy of this marker in clinical settings. While a consensus on normal NLR values is lacking, there is a suggested range of 1–2 for normal values, 2–3 as a gray area indicating subclinical inflammation, and values above 3 indicating inflammation . Our stuThe intricate pathological processes underlying autoimmune diseases have been meticulously explored, elucidating the interplay between innate and adaptive immunity. The NLR, comprising neutrophil and lymphocyte counts, provides valuable insights into the body’s inflammatory status and immune response. A higher neutrophil count suggests an ongoing nonspecific inflammatory process, while a lower lymphocyte count indicates a relatively compromised immune system . SeveralSeveral limitations should be noted. First, the observational design of this study limit causal inferential ability. We identified associations between NLR and outcomes but cannot confirm a definitive causal relationship. Second, residual confounding is possible despite adjustments for an extensive set of potential confounders. Unmeasured factors such as comorbidities, smoking intensity, medications, and RA characteristics could influence the observed relationships. Third, the NLR was measured at a single timepoint and may not reflect variability over time or with interventions. Serial NLR measurements could provide a more accurate representation of inflammatory status. Finally, the study population comprised U.S. adults, which may restrict generalizability to other populations. Further studies in diverse cohorts are warranted to validate our findings. However, the strengths of a large nationally representative sample, long follow-up, and robust adjustment for confounders enhance the reliability of the results.5In summary, the NLR provides a simple readout of the balance between pathological innate inflammation and protective adaptive immunity. Our study of a large nationally representative cohort demonstrates that a higher NLR is independently predictive of increased risks of both all-cause and cardiovascular mortality among adults with RA. These findings highlight the potential clinical utility of the NLR as an inexpensive and widely available biomarker that can be integrated into routine care to improve risk stratification and prognostication in RA patients.The original contributions presented in the study are included in the article/supplementary materials, further inquiries can be directed to the corresponding author/s.The original survey protocol underwent rigorous ethical scrutiny and received approval from the Institutional Review Board of CDC. The studies were conducted in accordance with the local legislation and institutional requirements. Written informed consent for participation in this study was provided by the participants’ legal guardians/next of kin.JW: Conceptualization, Writing – review & editing. EZ: Data curation, Writing – original draft. XZ: Methodology, Writing – review & editing. YY: Supervision, Writing – original draft, Writing – review & editing.


Aspergillus felis.I read with sadness the report of an immunocompetent 18-year-old man with fatal fungal rhinosinusitis (FRS) and cerebral abscess caused by A. felis was described a decade ago after its isolation from pet cats in Australia with invasive fungal rhinosinusitis (FRS) characterized by sinonasal and sino-orbital involvement [The parallels between this case and infections reported in immunocompetent domestic cats are striking. olvement . As in tolvement .A. felis is an environmental saprophyte that has been isolated from diverse environments on most continents [et al. state that there are six pathogenic species within the Aspergillus viridinutans complex: A. udagawae, A. felis, A. pseudofelis, A. parafelis, A. pseudoviridinutans and A. wyomingensis. However, the complex has undergone extensive revision after sequencing of clinical and environmental strains from six continents [A. felis, A. parafelis and A. pseudofelis are, in fact, one species (A. felis). Ten species make up the A. viridinutans complex, two of which have only been isolated from Australia – A. viridinutans [A. frankstonensis [ntinents . Parkes-ntinents . It is ndinutans  and A. ftonensis . The forAspergillus species in cats [I have wondered why it is that apparently immunocompetent cats develop invasive FRS. Over-representation of Persian-related cat breeds raises the possibility of an uncharacterized genetic defect in fungal immunity or of impaired sinus drainage due to foreshortening of the skull . In favour of the former, these same breeds of cats are also susceptible to other invasive fungal infections (IFIs), such as dermatophytic pseudomycetoma. There has been very limited veterinary research to investigate putative immunological defects. The findings from one study did not support a role for single-nucleotide polymorphisms (SNPs) in the coding regions of pattern recognition receptor genes, specifically Toll-like receptor (TLR) genes 1, 2 and 4, in the pathogenesis of fungal rhinosinusitis caused by  in cats  .Aspergillus flavus, has been reported for decades in immunocompetent or mildly immunosuppressed people from Sudan, India, Pakistan and Saudia Arabia [A. flavus conidia are high in these regions [Regardless, invasive FRS also occurs in immunocompetent crossbred cats with no known underlying risk factors. An analogous condition, that of chronic granulomatous invasive fungal sinusitis, caused by a Arabia . The env regions . ExposurA. felis type strain  showed significantly higher virulence in Galleria mellonella larvae than Aspergillus fumigatus. In immunosuppressed BALB/c mice, A. felis showed slightly higher virulence than A. fumigatus. However, in a chronic granulomatous disease murine model, inoculation with A. fumigatus, unlike A. felis, was uniformly fatal, demonstrating decreased virulence in hosts deficient in production of reactive oxygen species [Aspergillus species, where the infecting isolate had been identified by PCR and sequencing of the internal transcribed spacer (ITS) regions (ITS1–5.8S-ITS2), partial β-tubulin and/or partial calmodulin genes [A. fumigatus confined to the sino-nasal cavity, while cryptic species infections (mostly caused by A. felis and A. udagawae) were associated with invasive sino-orbital disease, including orbital and paranasal soft tissue involvement and lysis of the orbital bone.A role for species-specific fungal virulence factors in the development of invasive FRS in immunocompetent individuals must also be considered. In experimental models the  species . Anotherin genes . There wA. viridinutans complex members are heterothallic but, unlike A. fumigatus, readily produce teleomorphs [A further consideration is that eomorphs . WhetherA. felis isolate reported by Parkes-Smith et al. is typical of most A. viridinutans complex members, i.e. low MICs/MECs of posaconazole and echinocandins, respectively, elevated MICs of voriconazole and itraconazole and variable MICs of amphotericin B. The combination of posaconazole and liposomal amphotericin B used in the report was also used to cure a cat with FRS caused by A. felis [A. udagawae and A. felis were cured with combined therapy including posaconazole, caspofungin and terbinafine and remained asymptomatic more than 2 years after presentation [A. udagawae was treated successfully using posaconazole and caspofungin [The antifungal susceptibility profile of the A. felis . More reentation . A humanpofungin .Finally, a shout-out to the One Health approach of Australian microbiologists and to the International Society of Human and Animal Mycoses (ISHAM), where sharing of comparative human and animal data has long been actively encouraged. Together, we can learn more.


Nanocomposites, formed by combining a matrix (commonly polymer or ceramic) with nanofillers (nano-sized inclusions like nanoparticles or nanofibers), possess distinct attributes attributed to their composition. Their unique physicochemical properties and interaction capabilities with microbial cells position them as a promising avenue for infectious disease treatment. The escalating prevalence of multi-drug resistant bacteria intensifies the need for alternative solutions. Traditional approaches involve antimicrobial agents like antibiotics, antivirals, and antifungals, targeting specific microbial aspects. This review presents a comprehensive overview of diverse nanocomposite types and highlights the potential of tailored matrix and antibacterial agent selection within nanocomposites to enhance treatment efficacy and decrease antibiotic resistance risks. Challenges such as toxicity, safety, and scalability in clinical applications are also acknowledged. Ultimately, the convergence of nanotechnology and infectious disease research offers the prospect of enhanced therapeutic strategies, envisioning a future wherein advanced materials revolutionize the landscape of medical treatment. In the global scenario, the escalating emergence of antibiotic-resistant organisms presents a formidable global menace, jeopardizing lives worldwide . InfectiThe relentless proliferation of multi-drug resistant bacteria (MDR) exacerbates this crisis, compelling the exploration of alternative therapeutic avenues. Consequently, scientists have embarked on the development of exceptionally potent drugs capable of combating a diverse array of bacterial systems, thereby making it exceedingly arduous for bacteria to develop resistance mechanisms. One promising strategy to address the shortcomings of traditional antibiotics is the integration of antimicrobial nanoscale materials .A nanocomposite is a solid material characterized by multiple phases, with at least one phase exhibiting dimensions in one, two, or three directions that are smaller than 100 nm. Alternatively, it can have structures with nano-scale repeating distances between the various phases composing the material . They inWithin this context, nanocomposites have emerged as a propitious frontier in the battle against infectious diseases, capitalizing on their distinctive physicochemical attributes and their ability to effectively engage with microbial cells. The subsequent sections of this review delve into an extensive spectrum of nanocomposites, encompassing metallic elements and phyto-molecules, with the aim of combatting infections and combatting pathogens that have demonstrated resistance to conventional drug therapies .Metal nanocomposites offer a multifaceted approach to tackling pathogens. The nanocomposites of metals and metal oxides can be integrated into various matrices, such as polymers, ceramics, hydrogel matrices, etc., creating a synergistic effect that enhances their antimicrobial properties . The intSilver nanocomposites (Ag-NC) offer an innovative strategy in the battle against microbial pathogens and present numerous advantages compared to traditional antimicrobial agents. The efficacy of silver (Ag) as an antimicrobial agent stems from its capacity to disrupt essential cellular mechanisms across a wide array of microorganisms. Through the controlled release of silver ions from these nanocomposites, a consistent and localized antimicrobial impact is achieved. The approach minimizes potential harm to host cells and diminishes the overall necessary dosage. Categorically, silver-based nanocomposites fall into distinct groups: i) Silver-polymer nanocomposites, ii) Silver-biopolymer nanocomposites, iii) Silver-inorganic compound nanocomposites, and iv) Silver (I) complexes.E. coli, S. aureus, and C. albicans , was thoroughly explored, with the bacterial cellulose/Ag (BC-Ag0.05) nanocomposite demonstrating the most pronounced antibacterial properties. The outcome implies a direct correlation between the concentration of silver and the antimicrobial effectiveness of the nanocomposites. The noteworthy ability of bacterial cellulose nanocomposites to impede the proliferation of diverse bacteria and fungi, encompassing E. coli, S. aureus, B. subtilis, and C. albicans, underscores their potential as versatile materials with promising prospects for medical applications , synthesized utilizing the E. coli free-cell supernatant, was harnessed as an antifungal agent to counteract B. cinerea infection in faba bean (Vicia faba L.). The Ag/SiO2NC, with positively charged silver nanoparticles, demonstrated potent in-vitro antifungal potency, displaying a minimal inhibitory concentration (MIC) of 40 ppm. The in-vitro efficacy of the fabricated Ag/SiO2NC against B. cinerea underscores its disease-control potential. Corroborating these findings, in vivo tests demonstrated that plants subjected to Ag/SiO2NC treatment displayed increased resistance to fungal infection. The enhancement was accompanied by biochemical and ultrastructural modifications that are closely associated with the activation of plant defense mechanisms  of 60 g/mL and 550 g/mL, respectively (Silicon (Si) and its nanomaterial derivatives have been acknowledged for their capacity to aid plants in alleviating the adverse effects of both abiotic and biotic stressors. A silver/silicon dioxide nanocomposite  complexes demonstrate a distinct antimicrobial range compared to the ligand and the hydrated silver (I) ion. A well-known example is the use of the silver sulfadiazine complex for managing burns and wounds. Moreover, these silver complexes are frequently employed as additives in diverse polymer and biopolymer composites to address infectious ailments .E. coli and S. aureus. When compared to utilizing chitosan or metal oxides separately, these CS-MO nanocomposites demonstrated increased antibacterial efficacy. The superior antibacterial effectiveness of the chitosan/nickel oxide (CS-NiO) nanocomposite, which reduced S. aureus and E. coli viabilities to 2–8 percent after a 12-h incubation period, was particularly remarkable. Based on these outcomes, it was concluded that CS/NiO nanocomposites hold the potential to act as antibacterial agents against harmful bacterial pathogens. The polymeric materials based on metal oxides can effectively differentiate between mammalian and bacterial cells, offering prolonged antibacterial growth suppression. The particle sizes observed were 18.3 nm for CS-NiO and 25.5 nm for CS-MgO. All samples exhibited antibacterial activity against both E. coli and S. aureus strains, with a stronger effect on S. aureus compared to E. coli. When treated with 15 mg/mL of CS-NiO, inhibition rates reached 98% for S. aureus and 92.3% for E. coli cells. Additionally, the photocatalytic degradation of E. coli and S. aureus was investigated using ZnO, AgCl, and AgCl/ZnO nanocomposite-incorporated chitosan hydrogel beads. The process involved the breakdown of these gram-negative and positive bacteria using a chitosan/AgCl/ZnO (CS/AgCl/ZnO) nanocomposite hydrogel beads system as a photocatalyst, specifically under visible light irradiation  nanoparticles, also display photocatalytic properties. However, ZnO nanoparticles offer improved biocompatibility and stronger UV light absorption compared to TiO2. Regarding the generation of reactive oxygen species (ROS), ZnO nanoparticles share antimicrobial capabilities with TiO2 nanoparticles. Notably, ZnO photocatalytic processes under UV irradiation have been found to potentially produce hydrogen peroxide (S. aureus diminished due to the photoactivation of TiO2 released from TiO2 encapsulated within polylactic (PLA) microspheres (2-PTFE) exhibited inhibitory effects against E. coli and S. aureus. The antibacterial and anti-adhesion properties against these bacterial strains were attributed to the synergistic interplay between TiO2 and PTFE. Furthermore, both TiO2 and PTFE coatings demonstrated exceptional corrosion resistance and enhanced biocompatibility , a single-atom-thick lattice of carbon atoms, exhibits exceptional properties. It is being explored for diverse applications such as nanoelectronics, conductive films, supercapacitors, biosensors, and nanomedicine and has recently gained attention for its antibacterial properties. While challenges remain, GO’s unique attributes hold promise for innovative antibacterial applications. Derivatives of graphene found in nanocomposites include graphene oxide, reduced graphene oxide (rGO), and graphene quantum dots (GQDs)  Table 2Table 2. 2O) nanocomposite, utilizing the electrostatic interaction and unique electronic transition between rGO and Cu2O. The design showcased remarkable antibacterial capabilities and persistent effectiveness. The results of antibacterial testing demonstrated that the rGO-CuO nanocomposites exhibited potent and enduring antibacterial potential against E. coli and S. aureus. Through stability testing, copper ion release analysis, and ROS detection, the enhanced antibacterial mechanisms of rGO-Cu2O nanocomposites were systematically unveiled. These mechanisms were attributed to a synergistic effect involving the sustained release of copper ions, increased production of reactive oxygen species, and excellent dispersion of the rGO-Cu2O nanocomposites. Additional proposed mechanisms of action encompassed the prolonged release of copper ions due to cellular encapsulation, damage to bacterial cell membranes caused by the presence of sharp edges on rGO nanosheets, and the generation of ROS  in terms of their antibacterial and antibiofilm effectiveness, rGO-AgO displayed significant antibiofilm activity against S. aureus (91.72%), C. albicans (91.17%), and E. coli (90.36%). After subjecting S. aureus to treatment with rGO-AgO, scanning electron microscopy revealed noticeable changes in morphology, including complete lysis of the outer surface and deformation of bacterial cells. The release of cellular protein from S. aureus increased proportionally with the concentration of rGO-AgO, rGO-NiO, and rGO-ZnO nanocomposites, shedding light on the antibacterial mechanism of action (S. aureus and E. coli. Synergistic enhancement of carboxymethyl-cellulose (CMC) and sodium alginate nanocomposite films was observed through the incorporation of graphene oxide and nano silver, leading to the inhibition of E. coli and S. aureus growth. The CMC/SA/Ag-GO2% nanocomposite displayed the highest zone of inhibition, measuring 0.70 mm against E. coli and 1.00 mm against S. aureus. The finding implies that the CMC/SA/Ag-GO2% nanocomposite exhibited superior antibacterial activity against E. coli compared to S. aureus . The nanocomposite system of rGO–IONP was synthesized to capitalize on the synergistic effects that result in the simultaneous physical and chemical harm to MRSA. The rGO efficiently absorbs and converts near-infrared energy into localized heat, leading to physical damage to the bacteria. Concurrently, the IONP catalyzes the breakdown of H2O2, which is significantly elevated due to local inflammation, producing highly reactive •OH radicals through Fenton reactions. This chemical process further contributes to the damage inflicted on MRSA  and silver nanoparticles has been extensively explored for their antibacterial properties. Numerous studies have demonstrated the capacity of MWCNTs to induce severe membrane damage and cell death in various pathogens, underscoring their potential as antimicrobial agents. The incorporation of silver nanoparticles onto MWCNTs has shown as a valuable substance with antibacterial efficacy across diverse medical applications and water disinfection strategies aimed at curtailing the spread of infectious diseases. Ag-MWCNTs have exhibited broad-spectrum activity against a range of Gram-positive and Gram-negative bacteria, including nas spp. . The fabproperty .S. aureus, B. subtilis, P. aeruginosa, E. coli, and C. albicans. It was noteworthy to observe the substantial escalation in antimicrobial activity at low concentrations of MWCNTs in combination with ZnO and Ag  (PVA), is a biodegradable, water-soluble polymer and biocompatible with low cytotoxicity, used for various biological applications. Methylated melamine grafted polyvinyl benzyl chloride was used as an additive in PVA and graphene nanosheets were used to reinforce the mechanical strength (S. aureus (99.7%) and E. coli (97.1%) with an increased activity recorded in the highest graphene-loaded region. PLA (Polylactic acid) is one of the most used synthetic biodegradable polymers for medical applications. It has been combined with different G-based nanomaterials and their mixtures, resulting in nanocomposites with higher antibacterial activity against S.aureus and E.coli. Compared with PLA, the addition of GO inhibited the proliferation of bacteria and cell adhesion of S. aureus and E. coli (Poly (N-vinyl carbazole) based materials are used majorly in biological applications due to their antimicrobial properties. In this regard, researchers  developestrength . The res E. coli .co-acrylic acid) hybrid hydrogels with poly aspartic acid penetrating polymer networks with graphene, and metal nanoparticles   . The antE. faecalis  and Gram-negative (E. coli and Vibrio parahaemolyticus) bacteria were used to test the Ag/PLA-NC films’ antibacterial properties. The findings showed that when the amount of Ag-NPs in the PLA increased, the antibacterial activity of Ag/PLA-NC films increased (E.coli and S.aureus. Yet another cationic polymer polyethyleneimine in combination with poly (glycidyl methacrylate) and AgNPs-based multifunctional nanocomposite was synthesized to detect and kill Gram-negative bacteria, paving the way to creating smart biomaterials with high selectivity  sulfate polyacrylonitrile (PAN) composite nanofibers were incorporated with copper oxide and copper hydroxide nanowires, copper nanoparticles and were tested for their antimicrobial activity. The result showed that all copper composite nanofibers exhibited good antimicrobial activity against  unclear .S. aureus and P. aeruginosa. The formulation prevented the attachment of bacteria to the substrate due to the biofilm disruption. Cationic acrylate copolyvidone–iodine NPs showed no bacterial growth and demonstrated dose dependent effect. Nanocomposite maintained antibacterial effects for 11 days and it has been reported that the growth inhibition of S. aureus was lower than E. coli (Polyethylene glycol (PEG) and poly (lactide-co-glycolide) (PLGA) are biocompatible polymers approved by the U.S. Food and Drug Administration . PEG–PLG E. coli .L. monocytogenes, S. aureus, E. coli and P. aeruginosa. Due to the interaction with the bacterial membrane lectins, the mannose functionalization greatly reduced bacterial growth, and this growth suppression was more pronounced in Gram-negative than in Gram-positive bacteria. Compared to unmodified CS NPs, mannose functionalized nanocomposites showed the most antibiofilm ability, especially against E.coli and P.aeruginosa  nanocomposites were synthesized through the utilization of leaf extract from 73 mg/mL .Camellia sinensis and Prunus africana bark against carbapenem-resistant bacteria. Dynamic Light Scattering analysis revealed a net zeta potential of +12.5 mV. The CuONPs synthesized through this phyto-synthesis exhibited notably extensive growth inhibiting zones, showcasing robust antibacterial potency against MRSA, carbapenem-resistant E. coli, and K. pneumoniae at low MIC  from Punica granatum, in conjunction with Chitosan (CS), to function as a reducing agent for generating gold nanoparticles . During Despite the promising potential of various nanocomposites in the treatment of infectious diseases, their clinical-scale implementation faces persistent challenges. Nanotoxicology, a specialized field, addresses the toxicity of all nanomaterials, encompassing nanoparticles and nanocomposites, on living organisms. Drawbacks associated with the use of nanocomposites emerge from their synthesis through to application. Factors influencing the toxicity of nanocomposites include both exogenous  and endogenous (degradation) factors .2, carbon nanotubes, and carbon nanoparticles show potential as antibacterial agents with low cytotoxicity in vitro, in vivo experiments have demonstrated lung inflammation, hyperplasia, and genotoxicity upon inhalation (During manufacturing, nanocomposites release nano and micro-scale fragments due to thermal and mechanical stress, photodegradation, and incineration. These fragments pose occupational hazards as they can be inhaled. Their pervasive nature allows them to bypass the body’s natural defenses, disrupting crucial cellular processes and contributing to a range of diseases. For instance, while TiOhalation . Preventhalation . Althoughalation . The AgNhalation .in vivo studies are essential to examine targeted drug delivery, nanocomposite stability within the body, leakage mechanisms, and their fate. Furthermore, a systematic investigation of the mechanism of action on drug-resistant pathogens, impact on gene expression, and metabolic pathways is imperative before considering clinical applications of nanocomposites as antibacterial agents. Finally, selecting an appropriate synthesis method, solvent, and polymer is crucial for upscaling production and eventual commercialization for therapeutic purposes (To mitigate the toxicity associated with nanocomposites, common strategies include altering the type of polymer used or employing a capping agent like PEG or chitosan . Howeverpurposes .The review encompasses the synthesis of various nanocomposites, combining metals and phytomolecules, aimed at combating infections and tackling multidrug-resistant pathogens. Going beyond nanoparticles, nanocomposites represent a significant breakthrough across biotechnology, microbiology, and biomedical research. Their nanoscale dimensions confer distinctive benefits like augmented surface area and reactivity, thus holding potential to transform the medical field. With the escalating threat of infectious diseases and the rise of hospital and community-acquired infections, innovative solutions are imperative. Nanocomposites emerge as promising candidates due to their remarkable properties. Exploiting their antimicrobial potential could potentially lead to breakthroughs in addressing antimicrobial resistance. Consequently, nanocomposites can evolve into biosensors, smart biomaterials for bacterial eradication, and customized drug delivery systems, enhancing treatment outcomes and mitigating antibiotic resistance risks . However