Intrinsic Resistance Research Articles

Phenotypic changes and gene expression profiles of Vibrio parahaemolyticus in response to low concentrations of ampicillin.

Vibrio parahaemolyticus is a leading cause of seafood-associated gastroenteritis and possesses intrinsic resistance to ampicillin. While ampicillin can trigger transcriptional responses of global genes, the behavioral and molecular changes that occur in V. parahaemolyticus when exposed to ampicillin are not fully understood. In this work, we investigated the effects of low concentrations of ampicillin on the physiology and gene expression of V. parahaemolyticus by combining phenotypic assays and RNA sequencing (RNA-seq) analysis. Our results showed that the growth of V. parahaemolyticus were notably delayed, and both motility and c-di-GMP production were significantly inhibited in the response to low concentrations of ampicillin stress. In contrast, biofilm formation by V. parahaemolyticus was enhanced by exposure to low concentrations of ampicillin. However, low concentrations of ampicillin had no effect on the cytotoxicity or adherence activity of V. parahaemolyticus. The RNA-seq data revealed that a low concentration of ampicillin significantly affected the expression levels of 676 genes, including those involved in antibiotic resistance, virulence, biofilm formation, and regulation. This work contributes to our understanding of how V. parahaemolyticus alters its behavior and gene expression in response to ampicillin exposure.

Identification and preclinical evaluation of MMV676558 as a promising therapeutic candidate against Clostridioides difficile

Clostridioides difficile, a gram-positive, toxin-producing, spore-forming anaerobe, is a major cause of antibiotic-associated diarrhoea. The bacterium's intrinsic drug resistance limits current treatment options to fidaxomicin and vancomycin for initial episodes, with anti-toxin B monoclonal antibody or faecal microbiota transplantation recommended for complicated or recurrent cases. This underscores the urgent need for novel therapeutics. In this study, we screened the MMV Pathogen Box at a 10 µM concentration against C. difficile R20291. Primary hits were evaluated for minimum inhibitory concentrations (MIC), killing kinetics, and biofilm inhibition. Bacterial cytological profiling (BCP) and transmission electron microscopy (TEM) were employed to study the mode of action. MMV676558 was further tested in a mouse model to assess survival, histopathology, and gut microbiota effects. We identified nineteen hits that inhibited over 50 % of C. difficile growth. MIC assays revealed three hits with MICs below 16 µg/mL: MMV676558, MMV688755, and MMV690027. These hits were effective against various C. difficile ribotypes. Killing kinetics were comparable or superior to vancomycin and fidaxomicin, and biofilm assays showed inhibitory effects. BCP and TEM analyses suggested membrane function disruption as the mode of action. Furthermore, MMV676558 demonstrated a protective effect in mice, with favourable histopathology and gut microbiota profiles. Given the urgent threat posed by C. difficile antibiotic resistance, discovering new treatments is a top priority. Our study identified three promising hits from the MMV Pathogen Box, with MMV676558 showing significant in vivo potential for further evaluation.

Antibiotic Susceptibility Pattern of Pseudomonas aeruginosa and Detection of Virulence Genes in a Tertiary Care Hospital, Kathmandu, Nepal

Pseudomonas aeruginosa is one of the most common causes of hospital-acquired infections. It threatens worldwide public health because of its intrinsic antibiotic resistance and virulence. In this context, this research aimed to determine the occurrence of P. aeruginosa in a clinical setting, examine its susceptibility to antibiotics, and detect the presence of virulence genes, viz. exoY, oprL and toxA in all isolates. It was a cross-sectional study carried out from June 2022 to December 2022. A total of 1356 clinical specimens were collected and processed in the laboratory for Gram staining, culture techniques, and biochemical tests. The Kirby-Bauer disk diffusion method was applied to examine the antibiotic susceptibility pattern and conventional PCR was used to detect the virulence genes in all confirmed P. aeruginosa isolates. 40.5% of specimens showed bacterial growth, of which only 3.02% were identified as P. aeruginosa. Among them, 61.0% were multidrug resistant and 29.2% were β-lactamase producers. Aztreonam (70.7%) was the most effective antibiotic. Among all P. aeruginosa isolates, 68.3% isolates were exoY positive, 61.0% were oprL positive and 56.1% were toxA positive. Strategic interventions are required to stop the emergence and spread of antimicrobial resistance as a result of the isolation of multidrug resistant (MDR) isolates carrying these virulence genes. This study can benefit the medical staff and the entire community in building a surveillance system and enhancing infection control procedures.

Experimental Investigation on Shear Strength at the Permeable Concrete–Fine-Grained Soil Interface for Slope Stabilization Using Deep Socket Counterfort Drains

In slopes where high pore water pressure exists, deep counterfort drains (also called drainage trenches or trench drains) represent one of the most effective methods for improving stability or mitigating landslide risks. In the cases of deep or very deep slip surfaces, this method represents the only possible intervention. Trench drains can be realized by using panels or secant piles filled with coarse granular material or permeable concrete. If the trenches are adequately “socket” into the stable ground (for example sufficiently below the sliding surface of a landslide or below the critical slip surface of marginally stable slopes) and the filling material has sufficient shear strength and stiffness, like porous concrete, there is a further increase in shear strength due to the “shear keys” effect. The increase in shear strength is due both to the intrinsic resistance of the concrete on the sliding surface and the resistance at the concrete–soil interface (on the lateral surface of the trench). The latter can be very significant in relation to the thickness of the sliding mass, the “socket depth”, and the spacing between the trenches. The increase in shear strength linked to the “shear keys effect” depends on the state of the porous concrete–soil interface. For silty–clayey base soils, it is very significant and is of the same order of magnitude as the increase in shear resistance linked to the permanent reduction on the slip surface in pore water pressure (draining effect). This paper presents the results of an experimental investigation on the shear strength at the porous interface of concrete and fine-grained soils and demonstrates the high significance and effectiveness of the “shear keys” effect.

Enhancing electrical conductivity of RuO2 nanosheet-coated films by enlarging the nanosheet area

This study focused on enhancing the electrical conductivity of metal oxide nanosheet films by increasing the size of the nanosheets to promote the electron flow in the desired in-plane direction. We developed a method for increasing the size of RuO2 metal oxide nanosheets, a 2D material, by controlling the heat-treatment process. The size of the RuO2 nanosheets increased from 0.7 to 3.5 μm by modifying the heat-treatment conditions of KxRuO2, which was used as the precursor material. As a consequence, the electrical conductivity of the large-sized-RuO2 nanosheet-coated films was eight times higher than that of their small-sized counterparts. This lower resistance was attributed to the minimal number of contacts required for electron flow; the contact resistance between the nanosheets being higher than the intrinsic resistance. The proposed approach for enhancing the conductivity of metal oxide 2D materials is very promising and is expected to significantly contribute to the development of advanced flexible electronic devices.

Abstract B089: Identification of novel drivers of gemcitabine resistance in pancreatic cancer

Abstract Pancreatic cancer is an aggressive disease with limited therapeutic options. Gemcitabine is a clinically used chemotherapy for pancreatic cancer treatment but has limited efficacy. The molecular mechanisms responsible for gemcitabine resistance in pancreatic cancer are still unclear. Here, we analyzed the genomics of drug sensitivity in cancer dataset and identified various pancreatic cancer cells with distinct sensitivity to gemcitabine. Validation with proliferation and clonogenic assays confirmed that while CFPAC1 and PANC0327 are highly gemcitabine-sensitive, ASPC1 and PANC1 display intrinsic gemcitabine resistance. Analysis of the gene expression profile of these cells identified several genes that are differentially upregulated in resistant versus sensitive cells. Pathway enrichment analysis using the differential genes revealed the upregulation of Hippo, Wnt, MAPK signaling pathways as well as autophagy and glutathione metabolism in the resistant cells. Using quantitative PCR, immunoblotting and biochemical assays, and extracellular flux analysis methods, we have characterized the molecular distinctions between gemcitabine sensitive and resistant cells. Our results provide insight on the molecular mechanisms driving gemcitabine resistance in pancreatic cancer and could lead to novel therapeutic strategies to overcome resistance and improve patients’ treatment. Citation Format: Amina D Bilalbegovic, Bowen Fu, Zeribe Nwosu. Identification of novel drivers of gemcitabine resistance in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B089.

Abstract C031: Keratin 17 promotes pancreatic cancer chemoresistance through mitochondrial translocation and stabilization of dihydroorotate dehydrogenase (DHODH)

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) has emerged as the third leading cause of cancer-related deaths in the U.S., in part, due to intrinsic resistance to main-line chemotherapeutic protocols. Our prior data showed that the upregulation of pyrimidine but not purine de novo biosynthesis is associated with shortened survival of patients whose tumors expresses high keratin 17 (K17). Furthermore, we reported that K17 expression confers chemoresistance by impacting the inner mitochondrial membrane protein, dihydroorotate dehydrogenase (DHODH), the rate-limiting enzyme in the de novo pyrimidine synthesis pathway1. Here, we explore the mechanisms through which K17 enters mitochondria, stabilizes DHODH, and thereby, enhances downstream de novo pyrimidine biosynthesis and chemoresistance. Methods: We generated a KPC K17 stably expressing cell line and a L3.6 K17 knockout cell line to assess the effects of K17 on cell viability and performed western blot, immunoprecipitation, mass spectrometry, immunofluorescence imaging, and cellular sub- fractionation. We also generated wild type K17, MLS mutant K17 and coil 2 deletion K17 overexpression in L3.6 K17 KO and KPC cell lines to study the importance of MLS and coil 2 domains. Results: Our previous studies suggested that K17 drives chemoresistance by increasing mitochondrial DHODH, elevating intracellular pyrimidines. In our current report, we identified by Mitoprot a mitochondria localization signal (MLS) of K17 with a probability of 86% and validated that it is required for K17 to enter the mitochondria and impact DHODH. Mechanistically, we found that K17 enters the mitochondria by specifically interacting with the translocase of the outer mitochondrial membrane complex (TOM20/TOM40). Additionally, we uncovered a coil 2 domain of K17 that mediates DHODH stability at the inner mitochondrial membrane. Our data showed that K17 with deletion of the coil 2 domain fails to interact with DHODH suggesting that this domain is critical for mediating interaction and DHODH stabilization. By contrast, MLS mutant K17 did not enter the mitochondria and promote DHODH stabilization, thus having no impact in altering the downstream de novo pyrimidine synthesis. Furthermore, we found that MLS mutant or coil 2 deleted K17 expressing cells have increased sensitivity towards gemcitabine, compared to wild type K17 expressing cells. Conclusions: We report for the first time that K17 encodes an MLS, enabling K17 to enter mitochondria, and that the coil 2 domain of K17 stabilizes DHODH at the inner mitochondrial membrane, thereby upregulating de novo pyrimidine biosynthesis and chemoresistance to gemcitabine. Further studies are warranted to explore whether a therapeutic strategy targeting K17 mitochondrial transport or interaction with DHODH could enhance the therapeutic efficacy of gemcitabine or other pyrimidine analog-based pharmacologic agents. 1. Pan et. al. Targeting keratin 17- mediated reprogramming of de novo pyrimidine biosynthesis to overcome chemoresistance in pancreatic cancer. bioRxiv, 2022 Citation Format: Yinghuan Lyu, Monisankar Ghosh, Chun-Hao Pan, Shayan Sarkar, Girish H Rajacharya, Bo Chen, Natalia Marchenko, Pankaj K Singh, Kenneth R. Shroyer, Luisa F. Escobar-Hoyos. Keratin 17 promotes pancreatic cancer chemoresistance through mitochondrial translocation and stabilization of dihydroorotate dehydrogenase (DHODH) [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C031.

Simultaneous Targeting of NQO1 and SOD1 Eradicates Breast Cancer Stem Cells via Mitochondrial Futile Redox Cycling.

Triple negative breast cancer (TNBC) contains the highest proportion of cancer stem-like cells (CSCs), which display intrinsic resistance to currently available cancer therapies. This therapeutic resistance is partially mediated by an antioxidant defense coordinated by the transcription factor NRF2 and its downstream targets including NQO1. Here, we identified the antioxidant enzymes NQO1 and SOD1 as therapeutic vulnerabilities of ALDH+ epithelial-like CSCs and CD24-/loCD44+/hi mesenchymal-like CSCs in TNBC. Effective targeting of these CSC states was achieved by utilizing IB-DNQ, a potent and specific NQO1-bioactivatable futile redox cycling molecule, which generated large amounts of reactive oxygen species (ROS) including superoxide and hydrogen peroxide. Furthermore, the CSC killing effect was specifically enhanced by genetic or pharmacological inhibition of SOD1, a copper-containing superoxide dismutase highly expressed in TNBC. Mechanistically, a significant portion of NQO1 resided in the mitochondrial intermembrane space, catalyzing futile redox cycling from IB-DNQ to generate high levels of mitochondrial superoxide, and SOD1 inhibition markedly potentiated this effect resulting in mitochondrial oxidative injury, cytochrome c release, and activation of the caspase 3-mediated apoptotic pathway. Treatment with IB-DNQ alone or together with SOD1 inhibition effectively suppressed tumor growth, metastasis, and tumor-initiating potential in xenograft models of TNBC expressing different levels of NQO1. This futile oxidant-generating strategy, which targets CSCs across the epithelial-mesenchymal continuum, could be a promising therapeutic approach for treating TNBC patients.

Genomics of Staphylococcus aureus and Enterococcus faecalis isolated from the ocular surface of dry eye disease sufferers

Ocular surface inflammatory disorders, such as dry eye, are becoming increasingly prevalent. Developing new treatment strategies targeting harmful bacteria could provide significant therapeutic benefits. The purpose of this study was to characterize the common ocular pathogen Staphylococcus aureus and the rarer endophthalmitis-associated species Enterococcus faecalis isolated from the ocular surface of dry eye disease patients in Norway. Together the 7 isolates (5 S. aureus and 2 E. faecalis) comprise the complete set of members of each species isolated in our previous study of the ocular microbiome of 61 dry eye sufferers. We aimed to investigate the pathogenic potential of these isolates in relation to ocular surface health. To this end, we used whole genome sequencing, multiplex PCR directed at virulence genes and antibiotic susceptibility tests encompassing clinically relevant agents. The E. faecalis isolates showed resistance to only gentamicin. S. aureus isolates displayed susceptibility to most of the tested antibiotics, except for two isolates which showed resistance to trimethoprim/sulfamethoxazole and three isolates which were resistant to ampicillin. Susceptibilities included sensitivity to several first-line antibiotics for treatment of ocular infections by these species. Thus, treatment options would be available if required. However, spontaneous resistance development to gentamicin and rifampicin occurred in some S. aureus which could be a cause for concern. Whole genome sequencing of the isolates showed genome sizes ranging from 2.74 to 2.83 Mbp for S. aureus and 2.86 Mbp for E. faecalis, which is typical for these species. Multilocus sequence typing and phylogenetic comparisons with previously published genomes, did not suggest the presence of eye-specific clusters for either species. Genomic analysis indicated a high probability of pathogenicity among all isolates included in the study. Resistome analysis revealed the presence of the beta-lactamase blaZ gene in all S. aureus isolates and the dfrG gene in two of them; while E. faecalis isolates carried the lsa(A) gene which confers intrinsic resistance to lincosamides and streptogramin A in this species. Screening for virulence factors revealed the presence of various pathogenicity associated genes in both S. aureus and E. faecalis isolates. These included genes coding for toxin production and factors associated with evading the host immune system. Some of the identified genes (tst, hylA &hylB) are suggested to be linked to the pathophysiology of dry eye disease. Lastly, the presence of specific S. aureus virulence genes was confirmed through multiplex PCR analysis.

Long-term persistence of diverse clones shapes the transmission landscape of invasive Listeria monocytogenes

BackgroundThe foodborne bacterium Listeria monocytogenes (Lm) causes a range of diseases, from mild gastroenteritis to invasive infections that have high fatality rate in vulnerable individuals. Understanding the population genomic structure of invasive Lm is critical to informing public health interventions and infection control policies that will be most effective especially in local and regional communities.MethodsWe sequenced the whole draft genomes of 936 Lm isolates from human clinical samples obtained in a two-decade active surveillance program across 58 counties in New York State, USA. Samples came mostly from blood and cerebrospinal fluid. We characterized the phylogenetic relationships, population structure, antimicrobial resistance genes, virulence genes, and mobile genetic elements.ResultsThe population is genetically heterogenous, consisting of lineages I–IV, 89 clonal complexes, 200 sequence types, and six known serogroups. In addition to intrinsic antimicrobial resistance genes (fosX, lin, norB, and sul), other resistance genes tetM, tetS, ermG, msrD, and mefA were sparsely distributed in the population. Within each lineage, we identified clusters of isolates with ≤ 20 single nucleotide polymorphisms in the core genome alignment. These clusters may represent isolates that share a most recent common ancestor, e.g., they are derived from the same contamination source or demonstrate evidence of transmission or outbreak. We identified 38 epidemiologically linked clusters of isolates, confirming eight previously reported disease outbreaks and the discovery of cryptic outbreaks and undetected chains of transmission, even in the rarely reported Lm lineage III (ST3171). The presence of animal-associated lineages III and IV may suggest a possible spillover of animal-restricted strains to humans. Many transmissible clones persisted over several years and traversed distant sites across the state.ConclusionsOur findings revealed the bacterial determinants of invasive listeriosis, driven mainly by the diversity of locally circulating lineages, intrinsic and mobile antimicrobial resistance and virulence genes, and persistence across geographical and temporal scales. Our findings will inform public health efforts to reduce the burden of invasive listeriosis, including the design of food safety measures, source traceback, and outbreak detection.

Antibiotic heteroresistance and persistence: an additional aid in hospital acquired infections by Enterococcus spp.?

Enterococcus, particularly E. faecium and E. faecalis, are responsible for many hospital-acquired infections. With their intrinsic antibiotic resistance and ability to form biofilms, enterococcal infections are already challenging to manage. However, when heterogenous populations are present, such as those exhibiting heteroresistance and persistence, the complexity of these infections increases exponentially not only due to their treatment but also due to their difficult diagnosis. In this study, we provide a summary of the current understanding of both heteroresistance and persistence in terms of mechanisms, diagnosis and treatment and subsequently review recent literature pertaining to these susceptibility types specifically in enterococci.

Pbp4 provides transpeptidase activity to the FtsW-PbpB peptidoglycan synthase to drive cephalosporin resistance in Enterococcus faecalis.

Enterococci exhibit intrinsic resistance to cephalosporins, mediated in part by the class B penicillin-binding protein (bPBP) Pbp4 that exhibits low reactivity toward cephalosporins and thus can continue crosslinking peptidoglycan despite exposure to cephalosporins. bPBPs partner with cognate SEDS (shape, elongation, division, and sporulation) glycosyltransferases to form the core catalytic complex of peptidoglycan synthases that synthesize peptidoglycan at discrete cellular locations, although the SEDS partner for Pbp4 is unknown. SEDS-bPBP peptidoglycan synthases of enterococci have not been studied, but some SEDS-bPBP pairs can be predicted based on sequence similarity. For example, FtsW (SEDS)-PbpB (bPBP) is predicted to form the catalytic core of the peptidoglycan synthase that functions at the division septum (the divisome). However, PbpB is readily inactivated by cephalosporins, raising the question-how could the FtsW-PbpB synthase continue functioning to enable growth in the presence of cephalosporins? In this work, we report that the FtsW-PbpB peptidoglycan synthase is required for cephalosporin resistance of Enterococcus faecalis, despite the fact that PbpB is inactivated by cephalosporins. Moreover, Pbp4 associates with the FtsW-PbpB synthase and the TPase activity of Pbp4 is required to enable growth in the presence of cephalosporins in an FtsW-PbpB-synthase-dependent manner. Overall, our results implicate a model in which Pbp4 directly interacts with the FtsW-PbpB peptidoglycan synthase to provide TPase activity during cephalosporin treatment, thereby maintaining the divisome SEDS-bPBP peptidoglycan synthase in a functional state competent to synthesize crosslinked peptidoglycan. These results suggest that two bPBPs coordinate within the FtsW-PbpB peptidoglycan synthase to drive cephalosporin resistance in E. faecalis.

Emergence of multidrug-resistant Providencia rettgeri clone in food-producing animals: A public health threat

The occurrence of carbapenemases encoding genes in Providencia rettgeri is a critical public health concern since this species has intrinsic resistance to several antimicrobials, including polymyxins. The identification of this multidrug-resistant (MDR) pathogen outside the hospital setting has become increasingly frequent, and raises an alert for the global health agencies, as they indicate a possible spread of such pathogens. Herein, we described three MDR P. rettgeri isolates carrying a diversity of antimicrobial resistance genes (ARGs) isolated from stool samples of swine and bovine in Brazil. Molecular analysis revealed that all isolates belonged to the same clone. The whole genome sequencing (WGS) of a representative isolate (PVR-188) was performed by MiSeq Illumina® platform, while the assembling and annotation was achieved using SPAdes and Prooka, respectively. The WGS analyses indicated the presence of ARGs that confer resistance to β-lactams (blaNDM-1, blaCTX-M-2), quinolones (qnrD1), aminoglycosides (aadA2, aadA1, aph(3′)-Via), phenicol (catB2), sulfonamides (sul1, sul2), and trimethoprim (dfrA12, dfrA1). The presence of three plasmid replicons (Col3M, IncQ1, and IncT) was detected, but no phage sequences were found. The phylogenetic analyses confirmed the genomic relationship of the PVR-188 with P. rettgeri isolates recovered from animals and humans in the USA and Malaysia. In conclusion, we report the occurrence of MDR P. rettgeri clone colonizing the gut microbiota of food-producing animals in Brazil, revealing the spread of this pathogen beyond hospital boundaries.

Intrinsic self-sensing piezoresistive behaviors of ultra-high strength alkali-activated concrete

In this work, the self-sensing piezoresistive behaviors of carbon fiber (CF)-reinforced ultra-high strength concrete (UHSC) are investigated using alternating current impedance spectroscopy (ACIS) technique. The effects of different fiber contents, fiber lengths, and curing ages are considered. The electrical behaviors of alkali-activated slag (AAS)-based UHSC (AAS-UHSC) and ordinary Portland cement (OPC)-based UHSC (OPC-UHSC) at similar strength from 80 to 110 MPa are compared. The findings reveal that the intrinsic electrical resistivity of AAS-UHSC is significantly lower than that of OPC-UHSC, approximately 1/30 of the latter. The addition of CF results in a substantial reduction in electrical resistivity for both UHSC types, with reductions of up to 96 %, and shows better compatibility with the AAS-UHSC matrix. The piezoresistive properties of CF-reinforced UHSC are investigated using ACIS at a fixed frequency and further evaluated by parameters including sensitivity, linearity, repeatability, and hysteresis. The results indicate that plain OPC-UHSC does not exhibit piezoresistivity, whereas the addition of CF enhances both the sensitivity and linearity of piezoresistivity. In contrast, AAS-UHSC demonstrates inherent piezoresistive behaviors even without CF. The introduction of 0.5 wt% CF improves the sensitivity (by up to 50 %) and linearity (R2 increased from 0.76 to above 0.90) of piezoresistivity in AAS-UHSC. However, increasing the CF content to 1.0 wt% diminishes the sensitivity (by up to 62.5 %) due to decreased fiber dispersion uniformity. Moreover, the addition of 1.0 wt% 8-mm CF reduces the hysteresis of UHSC regardless of the binder type, with the maximum reduction reaching 62.2 %.

Genetic traits and transmission of antimicrobial resistance characteristics of cephalosporin resistant Escherichia coli in tropical aquatic environments

This study investigates the genetic traits and transmission mechanisms of cephalosporin-resistant Escherichia coli in tropical aquatic environments in Singapore. From 2016 to 2020, monthly samples were collected from wastewater treatment plants, marine niches, community sewage, beaches, reservoirs, aquaculture farms, and hospitals, yielding 557 isolates that were analyzed for antimicrobial resistance genes (ARGs) and mobile genetic elements (MGEs) using genomic methods. Findings reveal significant genotypic similarities between environmental and hospital-derived strains, particularly the pandemic E. coli ST131. Environmental strains exhibited high levels of intrinsic resistance mechanisms, including mutations in porins and efflux pumps, with key ARGs such as CMY-2 and NDM-9 predominantly carried by MGEs, which facilitate horizontal gene transfer. Notably, pathogenic EPEC and EHEC strains were detected in community sewage and aquaculture farms, posing substantial public health risks. This underscores the critical role of these environments as reservoirs for multidrug-resistant pathogens and emphasizes the interconnectedness of human activities and environmental health.

EmbB and EmbC regulate the sensitivity of Mycobacterium abscessus to echinomycin.

Treatment of Mycobacterium abscessus (Mab) infections is very challenging due to its intrinsic resistance to most available drugs. Therefore, it is crucial to discover novel anti-Mab drugs. In this study, we explored an intrinsic resistance mechanism through which Mab resists echinomycin (ECH). ECH showed activity against Mab at a minimum inhibitory concentration (MIC) of 2 µg/ml. A ΔembC strain in which the embC gene was knocked out showed hypersensitivity to ECH (MIC: 0.0078-0.0156 µg/ml). The MICs of ECH-resistant strains screened with reference to ΔembC ranged from 0.25 to 1 µg/ml. Mutations in EmbB, including D306A, D306N, R350G, V555I, and G581S, increased the Mab's resistance to ECH when overexpressed in ΔembC individually (MIC: 0.25-0.5 µg/ml). These EmbB mutants, edited using the CRISPR/Cpf1 system, showed heightened resistance to ECH (MIC: 0.25-0.5 µg/ml). The permeability of these Mab strains with edited genes and overexpression was reduced, as evidenced by an ethidium bromide accumulation assay, but it remained significantly higher than that of the parent Mab. In summary, our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab's sensitivity to ECH. Mutation in EmbB may partially compensate for a loss of EmbC function.

ScarfingTool: An Advanced tool to repair Carbon Fiber Reinforced plastic

Carbon Fiber Reinforced Plastic (CFRP) is going to assume more and more importance in mechanical and aerospace engineering in the near future. This is due to its intrinsic high lightness and resistance, if compared to traditional metallic materials. CFRP is characterized by a proper production and repair technology. The tool showed in this paper is conceived to repair and verify the restoration of such kind of material. Its application is limited to the first part of the repair process: in jargon the scarfing operation, otherwise called “defect removal”. This operation is similar to a high-speed grinding.For this reason, the designed tool can be easily applied to other contexts, such as sanding, polishing or deburring.Basically, the ScarfingTool is composed by a high-speed electro-spindle and a 3D smart snapshot sensor. This tool is conceived to be as light as possible to be installed on a light collaborative robot. Moreover, its specific design assures great compliance that, as needed, can be regulated. A high efficiency suction inlet completes the tool and guarantees a clean environment during operation. The scarfing process has already been dealt with in [1] to which the interested reader is addressed.This work is licensed under CC BY-SA 4.0. (https://creativecommons.org/licenses/by-sa/4.0)

Identification and confirmation of powdery mildew (Erysiphe polygoni) resistance in pea germplasm

Erysiphe polygoni DC, the causative agent of powdery mildew, poses a significant threat to pea crop, resulting in substantial yield and economic losses. To address this menace, an exploration of intrinsic resistance within pea germplasm becomes imperative for effective disease management. In our study, phenotyping of 1095 pea accessions, sourced from both indigenous and exotic origins, under natural epiphytotic conditions during Rabi 2021-22, revealed a prevailing susceptibility, with over 60 per cent disease severity in majority of accessions. Remarkably, only 22 accessions, including IC208327, IC220109, IC220378, IC258401, IC262762, IC262849, IC267140, IC267174, IC267727, IC274039, IC274040, IC310074, IC311066, IC345548, IC424898, IC552779, IC613133, IC629544, IC629687, NC57793, IC220193 and EC598669, exhibited resistance with a disease score of <1. Subsequent, pot screening of these 22 resistant accessions under artificial epiphytotic conditions during Rabi 2022-23 confirmed their resistance showing disease severity ranging from 5.83 - 17.50 per cent. Recognizing the pivotal role of resistant varieties in disease management, these identified accessions hold immense potential as resistant donors in crop improvement programs.. KEYWORDS :Germplasm, pea, powdery mildew, resistance

Enhanced electrochemical performances of ternary polypyrrole/MXene/Gum arabic composites as supercapacitors electrode materials

The development of supercapacitor electrodes for high energy storage applications requires the use of ternary composites made of PPy, Mxene, Gum Arabic (PPy/Mxene/GA) these materials are special because they have new characteristics like cyclic stability, and high specific capacitance. The pure PPy, PPy/Mxene and PPy/Mxene/Gum Arabic show the shifting and appearance of new peaks in UV-Visible spectra, which are further supported by the Fourier transform infrared spectroscopy (FT-IR). The appearance of new peaks in X-ray diffraction (XRD) of PPy confirms presence of Mxene and Gum Arabic, which is in agreement with the energy dispersive X-ray (EDX) analysis. The electrochemical techniques such as Cyclic Voltammetry (CV), Galvanostatic Charge Discharge (GCD) and Electrochemical Impedance Spectroscopy (EIS) show that the (PPy/Mxene/GA) ternary composite maintains 94 % of the specific capacitance retention after 2000 cycles and; reaches a maximum specific capacitance of 657.64 F/g at 1 A/g. As compared to Pure PPy and PPy/Mxene composite the PPy/Mxene/GA composite show small intrinsic resistance Rs and charge transfer Rct values 0.11Ω and 6.15Ω respectively. The outstanding electrochemical properties of PPy/Mxene/GA composite are ascribed to the blending of three different type compounds PPy, Mxene and GA and theirs mutual beneficial effects. These findings show that ternary composite material holds great promise for use in these condition involving portable electronic storage devices.

Insight into Virulence and Mechanisms of Amphotericin B Resistance in the Candida haemulonii Complex.

The Candida haemulonii complex includes emerging opportunistic human fungal pathogens with documented multidrug-resistance profiles. It comprises Candida haemulonii sensu stricto, Candida haemulonii var. vulnera, Candida duobushaemulonii, Candida pseudohaemulonii, and Candida vulturna. In recent years, rates of clinical isolation of strains from this complex have increased in multiple countries, including China, Malaysia, and Brazil. Biofilm formation, hydrolytic enzymes, surface interaction properties, phenotype switching and cell aggregation abilities, extracellular vesicles production, stress response, and immune evasion help these fungi to infect the host and exert pathological effects. Multidrug resistance profiles also enhance the threat they pose; they exhibit low susceptibility to echinocandins and azoles and an intrinsic resistance to amphotericin B (AMB), the first fungal-specific antibiotic. AMB is commonly employed in antifungal treatments, and it acts via several known mechanisms. Given the propensity of clinical Candida species to initiate bloodstream infections, clarifying how C. haemulonii resists AMB is of critical clinical importance. This review outlines our present understanding of the C. haemulonii complex's virulence factors, the mechanisms of action of AMB, and the mechanisms underlying AMB resistance.