Molecular Biology Techniques Research Articles

Extracellular succinate derived from ectopic milieu drives adhesion and implantation growth of ectopic endometrial stromal cells via the SUCNR1 signal in endometriosis

BackgroundAs a dual-function metabolite, succinate has emerged in cell function and plays a key signaling role in linking mitochondrial function to other cellular functions. Succinate accumulation in the cytoplasm is commonly associated with hypoxia in the microenvironment and immune cell activation. Extracellular succinate released into the microenvironment is considered an inflammatory alarm that can be sensed by its membrane receptor SUCNR1, which boosts proinflammatory responses and acts akin to classical hormones and cytokines. Succinate plays an important role in the development of inflammatory diseases. Whether succinate facilitates the progression of endometriosis (EMs), characterized by chronic inflammation and peritoneal adhesion, is worth exploring.ObjectiveWe mimicked the ectopic milieu in vitro and in vivo to evaluate the main source and potential role of succinate in endometriosis. We assessed the molecular and functional effects of succinate on macrophages and peritoneal mesothelial cells in peritoneal cavity. The effect of succinate/SUCNR1 signaling on ectopic endometrial stromal cells (ESCs) was further explored in this study.MethodsIn this study, we used targeted organic acid metabolomics analysis and in vitro assays to assess the potential accumulation of succinate in the peritoneal fluid of EMs patients. We examined its correlation with disease severity, Visual Analogue Scale, and the Endometriosis Fertility Index. Flow cytometry, enzyme linked immunosorbent assay, western blot assay, quantitative real-time PCR, and other molecular biology techniques were used to explore the potential mechanisms.ResultsBy mimicking the ectopic milieu, we constructed an in vitro co-culture system and found that M1 polarized macrophages and that the peritoneal mesothelial cell line (HMrSV5) mainly released succinate into their microenvironment and activated the succinate receptor (SUCNR1) signal, which further polarized the macrophages and significantly enhanced the invasive survival of ESCs, and the adhesion to the peritoneum. We further investigated the pathological effects of extracellular succinate in vivo using a xenograft mouse models of endometriosis.ConclusionsSuccinate-SUCNR1 signaling facilitates the creation of inflammatory cells and plays a vital role in EMs progression and peritoneal adhesion. Our work on the molecular mechanisms underlying succinate accumulation and function will help elucidate the phenotypic mysteries of pain and infertility in EMs.BkiB34NQBDvkGHWZYD1GB2Video

CASE REPORT: POSSIBLE EMERGING OF SALMONELLA ENTERICA SUBSP. ENTERICA SEROVAR HAVANA INFECTION IN NEWBORN KID GOATS

Although salmonellosis has been considered an important problem in the ruminant farm from 10 weeks to 14 months old, there is no information about the disease in goats, especially in 2 to 3-day-old kids, in recent years. The goat farmers complained that the 2 to 3-day-old kids showed clinical signs of depression, weakness, lameness, and diarrhea. The kids might die within one week after the symptoms appeared. The mortality reached 60% (48/80). After many antimicrobial agents were dosed, no better outcome was expected. The farmer sent one kid to the official deputy for further investigation. By helping with bacterial and molecular biological techniques, autopsy, serotyping, and histopathological diagnosis, the kids were infected by Salmonella enterica subsp. enterica serovar Havana strain. After a farm visit, a moldy floor contaminated by 2 to 3[Formula: see text]cm thick feces was identified on the floor. After removing the mold and implementing disinfection, the outbreak was stopped. In conclusion, this report proves that salmonellosis has recently been identified in goat farms, especially in kids, within one week. Moreover, good hygiene and keeping feces out of barns are essential to preventing salmonellosis.

Ghrelin/GHSR signaling in the lateral septum ameliorates chronic stress-induced depressive-like behaviors

Ghrelin is a gastrointestinal hormone on feeding and metabolism regulation, and acts through its receptor-growth hormone secretagogue receptor (GHSR), which is widely distributed throughout the central nervous system. Recent studies have suggested that ghrelin plays an important role in the regulation of depression, but the underlying mechanisms remain uncertain. Lateral septum (LS) is a critical brain region in modulating depression. Therefore, we investigated the role of ghrelin/GHSR signaling in the LS on the depressive-like behaviors of mice under conditions of chronic stress by using behavioral tests, neuropharmacology, and molecular biology techniques. We found that infusion of ghrelin into the LS produced antidepressant-like responses in mice. Activation of LS GABAergic neurons was involved in the antidepressant effect of ghrelin. Importantly, GHSR was highly expressed and distributed in the LS neurons. Blockade of GHSR in the LS reversed the ghrelin-induced antidepressant-like effects. Molecular knockdown of GHSR in the LS induced depressive-like symptoms in mice. Furthermore, administration of ghrelin into the LS alleviated depressive-like behaviors induced by chronic social defeat stress (CSDS). Consistent with the neuropharmacological results, overexpression of GHSR in the LS reversed CSDS-induced depressive-like behaviors. Our findings clarify a key role for ghrelin/GHSR signaling in the regulation of chronic stress-induced depressive-like behaviors, which could provide new strategies for the treatment of depression.

Rivaroxaban down-regulates pyroptosis and the TLR4/NF-κB/NLRP3 signaling pathway to promote flap survival

BackgroundFlap placement remains the primary method for wound repair, but postoperative ischemic flap necrosis is of major concern. This study explored whether rivaroxaban, a factor Xa inhibitor, enhanced flap survival. MethodsThirty-six rats were randomly divided into control, low-dose rivaroxaban (3 mg/kg/day), and high-dose rivaroxaban (7 mg/kg/day) groups. On postoperative day 7, the flap survival rate was analyzed and the average survival area calculated. After the rats were euthanized, immunological and molecular biological techniques were employed to assess vascular regeneration, pyroptosis, and inflammation. ResultsRivaroxaban upregulated VEGF expression, in turn enhancing angiogenesis, and it downregulated IL-1β, IL-6, and TNF-α expression, thereby mitigating inflammation. The drug also suppressed TLR4, NF-κB p65, NLRP3, caspase-1, and IL-18 syntheses, thus inhibiting pyroptosis. ConclusionsRivaroxaban enhanced random flap survival by down-regulating the TLR4/NF-κB/NLRP3 signaling pathway to suppress pyroptosis, promoting vascular regeneration and inhibiting inflammation.

The Dual Role of Sulforaphane-Induced Cellular Stress-A Systems Biological Study.

The endoplasmic reticulum (ER) plays a crucial role in cellular homeostasis. When ER stress is generated, an autophagic self-digestive process is activated to promote cell survival; however, cell death is induced in the case of excessive levels of ER stress. The aim of the present study was to investigate the effect of a natural compound called sulforaphane (SFN) upon ER stress. Our goal was to investigate how SFN-dependent autophagy activation affects different stages of ER stress induction. We approached our scientific analysis from a systems biological perspective using both theoretical and molecular biological techniques. We found that SFN induced the various cell-death mechanisms in a concentration- and time-dependent manner. The short SFN treatment at low concentrations promoted autophagy, whereas the longer treatment at higher concentrations activated cell death. We proved that SFN activated autophagy in a mTORC1-dependent manner and that the presence of ULK1 was required for its function. A low concentration of SFN pre- or co-treatment combined with short and long ER stress was able to promote cell survival via autophagy induction in each treatment, suggesting the potential medical importance of SFN in ER stress-related diseases.

Testosterone synthesis was inhibited in the testis metabolomics of a depression mouse model

IntroductionDepression is a common emotional disorder. Previous studies have suggested that depression is associated with the central nervous system. Recent studies have suggested that reduced testosterone level is the core inducement of depression. Testis is the vital organ for the synthesis of testosterone. How does testis mediate depression is still unknown. ObjectivesWe adopted a classical depression model of mouse caused through chronic mild stress (CMS). The metabolomics liquid chromatography-mass spectrometry was adopted to analyse the influence of CMS on testis metabolism. Then we confirmed the possible abnormal metabolism of the testis in depression mice by pathway analysis and molecular biological technique. ResultsCompared with control mice, 16 differential metabolites were found in CMS mice by multivariate statistical analysis. In comparison with control mice, CMS mice showed higher levels for campesterol, ribitol, citric acid, platelet activating factor, guanosine, cytosine and xanthine and lower levels for docosahexaenoic acid, hippuric acid, creatine, testosterone, dehydroepiandrosterone, progesterone, l-carnitine, acetyl carnitine and propionyl carnitine. The pathway analysis indicated that these differential metabolites are associated with steroid hormone synthesis, purine metabolism and phenylalanine metabolism. In addition, we also first discovered that testicular morphology in depression mice was damaged and steroid hormone synthetases (including steroidogenic acute regulatory protein and P450 cholesterol side chain cleavage) were inhibited. ConclusionThese findings may be helpful to parse molecular mechanisms of pathophysiology of depression. It also pointed out the direction to search for potential therapy schedules for male depression and provide novel insights into exploring the pathogenesis of male depression.

Molecular biology techniques for assessing the loss of HLA heterozygosity after allogeneic hematopoietic stem cell transplantation in children with acute leukemia

According to several observations, up to a third of post-transplant relapses in childhood acute leukemia are associated with the loss of heterozygosity of the major histocompatibility complex (HLA). Furthermore, the inefficacy of the graft-versus-leukemia reaction, as evidenced by the lack of therapeutic effect from the infusion of donor lymphocytes, indicates the need for timely detection of this marker to change the treatment strategy in the post-transplant period. To detect the loss of HLA heterozygosity, the method using the commercial KMR-HLA system and analysis using next-generation sequencing (NGS), as well as the method based on the analysis of highly polymorphic STR and VNTR markers located in the HLA loci region on the short arm of chromosome 6, are widely used. The primary objective of our study was to compare the informativeness of these approaches in diagnosing HLA heterozygosity loss in children during the post-transplant period. The obtained data on the frequency of detecting HLA heterozygosity loss were comparable to the literature data and constituted 23 % of cases of post-transplant relapse of B-cell acute lymphoblastic leukemia, 33 % of cases of T-cell acute lymphoblastic leukemia, and 23% of cases of acute myeloid leukemia. We also demonstrated that the method based on STR marker analysis has sensitivity comparable to allele-specific PCR and NGS sequencing methods. Meanwhile, preliminary sorting of the blast population increases the sensitivity of STR analysis and can be recommended in routine practice.

Transcriptomic analysis to elucidate the molecular mechanisms underlying the growth advantage of Kluyveromyces marxianus XZ1 and the development of resistance in apple induced by XZ1

The preceding study demonstrated that Kluyveromyces marxianus XZ1 (XZ1) exhibits potential as a biological agent for combating disease by P. expansum in apple fruit. This potential is primarily attributed to its capacity for competing for resources and space, as well as activating certain genes involved in the transduction of plant hormone signals. In this study, transcriptomic analysis was performed on the interaction system of XZ1 and apple to investigate the detail molecular mechanisms of the yeast to develop competitive advantage and the molecular mechanisms of the apple to develop defense response action at the gene expression level. The transcriptome analysis of XZ1 in apple indicated that the meiosis pathway was activated to promote rapid proliferation of the yeast in apple. Autophagy, ubiquitin-mediated proteolysis, and the starch and sucrose metabolism pathway are induced to generate vital nutrients and energy for yeast proliferation. The transcriptome analysis of apple revealed upregulation in the expression levels of genes associated with transcription factors (TFS) involved in pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) following treatment with XZ1. This finding suggests that XZ1 activates PTI and ETI in apple, thereby promoting disease resistance. Moreover, the yeast treatment induced the activation of ethylene and jasmonate signal transduction pathways, which play a crucial role in regulating disease resistance development in apple. Additionally, the yeast treatment resulted in the enhancement of lignin biosynthesis, serving as a protective mechanism for the apple against potential threats, as observed in the response to XZ1 treatment. The findings of this study establish a theoretical foundation for elucidating the molecular mechanism underlying the growth advantage of antagonistic yeast in the host and the induced resistance in apple. Additionally, this study offers a substantial repertoire of gene resources that can be utilized to enhance yeast biocontrol efficacy and bolster the disease resistance of apple through the application of molecular biology techniques.

Bacterial genome engineering using CRISPR-associated transposases.

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated transposases have the potential to transform the technology landscape for kilobase-scale genome engineering, by virtue of their ability to integrate large genetic payloads with high accuracy, easy programmability and no requirement for homologous recombination machinery. These transposons encode efficient, CRISPR RNA-guided transposases that execute genomic insertions in Escherichia coli at efficiencies approaching ~100%. Moreover, they generate multiplexed edits when programmed with multiple guides, and function robustly in diverse Gram-negative bacterial species. Here we present a detailed protocol for engineering bacterial genomes using CRISPR-associated transposase (CAST) systems, including guidelines on the available vectors, customization of guide RNAs and DNA payloads, selection of common delivery methods, and genotypic analysis of integration events. We further describe a computational CRISPR RNA design algorithm to avoid potential off-targets, and a CRISPR array cloning pipeline for performing multiplexed DNA insertions. The method presented here allows the isolation of clonal strains containing a novel genomic integration event of interest within 1-2 weeks using available plasmid constructs and standard molecular biology techniques.

Non-negligible N2O emission hotspots: Rivers impacted by ion-adsorption rare earth mining

Rare earth mining causes severe riverine nitrogen pollution, but its effect on nitrous oxide (N2O) emissions and the associated nitrogen transformation processes remain unclear. Here, we characterized N2O fluxes from China's largest ion-adsorption rare earth mining watershed and elucidated the mechanisms that drove N2O production and consumption using advanced isotope mapping and molecular biology techniques. Compared to the undisturbed river, the mining-affected river exhibited higher N2O fluxes (7.96 ± 10.18 mmol m−2d−1 vs. 2.88 ± 8.27 mmol m−2d−1, P = 0.002), confirming that mining-affected rivers are N2O emission hotspots. Flux variations scaled with high nitrogen supply (resulting from mining activities), and were mainly attributed to changes in water chemistry (i.e., pH, and metal concentrations), sediment property (i.e., particle size), and hydrogeomorphic factors (e.g., river order and slope). Coupled nitrification-denitrification and N2O reduction were the dominant processes controlling the N2O dynamics. Of these, the contribution of incomplete denitrification to N2O production was greater than that of nitrification, especially in the heavily mining-affected reaches. Co-occurrence network analysis identified Thiomonas and Rhodanobacter as the key genus closely associated with N2O production, suggesting their potential roles for denitrification. This is the first study to elucidate N2O emission and influential mechanisms in mining-affected rivers using combined isotopic and molecular techniques. The discovery of this study enhances our understanding of the distinctive processes driving N2O production and consumption in highly anthropogenically disturbed aquatic systems, and also provides the foundation for accurate assessment of N2O emissions from mining-affected rivers on regional and global scales.

Molecular characterization of whitefly on different host plants

The objective of this work was to determine the whitefly (Bemisia tabaci Genn.) biotypes that occur in the state of Paraná. For the determination of the biotypes, whiteflies were collected in different host plants during 2016 in 21 cities of the state of Paraná, and molecular biology techniques were used for the analysis of the samples. Total insect DNA extraction was performed with Chelex® 100 resin, followed by PCR amplification of the mitochondrial cytochrome oxidase 1 (mtCOI) gene region. The PCR products were sequenced and analyzed with Mega 7.0.21 software along with the ClustalW2 site. The dendrogram of the samples generated along with the patterns of the other species showed a clustering of all the samples with those of the MEAM1 species (biotype B), being the only species found in the studied regions. The identity of the nucleotides among the whiteflies ranged from 96.94% to 100%. Biotype A was not found in regions where it was previously occurring, probably due to the MEAM 1 species. The whiteflies of the MEAM 1 species (biotype B) were found in 14 different plant species. These results are important in the current scenario of Paraná and could contribute to better efficiency in the management of this pest.

Evaluation of Various Diagnostic Strategies for Bacterial Vaginosis, Including a New Approach Based on MALDI-TOF Mass Spectrometry.

Bacterial vaginosis (BV) is a common dysbiosis of unclear etiology but with potential consequences representing a public health problem. The diagnostic strategies vary widely. The Amsel criteria and Nugent score have obvious limitations, while molecular biology techniques are expensive and not yet widespread. We set out to evaluate different diagnostic strategies from vaginal samples using (1) a combination of abnormal vaginal discharge and vaginal pH > 4.5; (2) the Amsel-like criteria (replacing the "whiff test" with "malodorous discharge"); (3) the Nugent score; (4) the molecular quantification of Fannyhessea vaginae and Gardnerella vaginalis (qPCR); (5) and MALDI-TOF mass spectrometry (we also refer to it as "VAGI-TOF"). Overall, 54/129 patients (42%) were diagnosed with BV using the combination of vaginal discharge and pH, 46/118 (39%) using the Amsel-like criteria, 31/130 (24%) using qPCR, 32/130 (25%) using "VAGI-TOF", and 23/84 (27%) using the Nugent score (not including the 26 (31%) with intermediate flora). Of the 84 women for whom the five diagnostic strategies were performed, the diagnosis of BV was considered for 38% using the combination of vaginal discharge and pH, 34.5% using the Amsel-like criteria, 27% using the Nugent score, 25% using qPCR, and 25% using "VAGI-TOF". When qPCR was considered as the reference, the sensitivity rate for BV was 76.2% for the combination of vaginal discharge and pH, 90.5% for the Amsel-like criteria, 95.2% for the Nugent score, and 90.5% for "VAGI-TOF", while the specificity rates were 74.6%, 84.1%, 95.3%, and 95.3%, respectively. When the Nugent score was considered as the reference, the sensitivity for BV was 69.6% for the combination of vaginal discharge and pH, 82.6% for the Amsel-like criteria, 87% for qPCR, and 78.7% for "VAGI-TOF", while the specificity rates were 80%, 94.3%, 100%, and 97.1%, respectively. Overall, the use of qPCR and "VAGI-TOF" provided a consistent diagnosis of BV, followed by the Nugent score. If qPCR seems tedious and for some costly, "VAGI-TOF" could be an inexpensive, practical, and less time-consuming alternative.

Whole-Mount RNA In Situ Hybridization of Zebrafish Embryos.

A commonly employed technique in molecular biology to evaluate the temporal and spatial expression of a certain gene is in situ hybridization. This method is an effective strategy to construct synexpression groups, co-expressed genes acting in shared biological processes, and to find new members of genes engaged in the same signaling pathways to discover similar spatial and temporal expression patterns in zebrafish embryos. The major disadvantage of this method is that RNA probes can penetrate within 2days of post-fertilization embryos, and therefore, in later developmental stages, the probe can only reach the surface tissues. Further application of the method in histological sections will be required for a complete and accurate gene expression investigation. However, this method is highly effective at late embryogenesis and early larval stages for observing gene expression in endodermal derivatives and sensory organs. RNA probes for in situ hybridization can be prepared through in vitro transcription from plasmids carrying specific promoter elements and mRNA-specific cDNA, or an alternative polymerase chain reaction (PCR) method can be used through PCR amplification. This chapter describes the procedures for detecting gene expression in zebrafish embryos using whole-mount RNA in situ hybridization.

Evaluating the neuroprotective potential of MSC secretome for retinal neurodegenerative diseases

Retinal neurodegenerative diseases (RND), including age‐related macular degeneration, glaucoma, diabetic retinopathy, retinitis pigmentosa, and others, are the most common cause of irreversible low vision and blindness. Although the aetiology and clinical characteristics of retinal RND can differ phenotypically, the neurodegenerative mechanisms of these diseases share common cellular and molecular signalization pathways. No cures are available for most RND, and cell‐based therapy may be a potential treatment through the paracrine properties of mesenchymal stromal cells (MSC). Furthermore, developing cell‐free compositions based on the production of “MSC secretome cocktails” is a promising therapeutic strategy and has significant advantages over using living cells in clinical practice.Ex vivo neuroretina models are adequate tools for evaluating retinal physiology and pathobiology. Neuroretina explant cultures closely resemble in vivo conditions, thus, preserving complex in vivo neuronal connections and retaining the functionality of non‐neuronal cells of the retina. Furthermore, cellular and molecular biology techniques can be applied to retinal explant cultures.In the ex vivo spontaneous neuroretinal degeneration model, the MSC secretome preserves retinal morphology and limits pro‐apoptotic‐ and pro‐necroptotic‐related genes and protein expression. Furthermore, the MSC secretome modulates autophagy‐related genes and proteins and stimulates the activation of antioxidant‐associated genes. Therefore, the neuroprotective ability of the MSC secretome is associated with the inhibition of apoptosis and necroptosis, modulation of autophagy, and activation of the antioxidant machinery during retinal degeneration.MSC secretome shows neuroprotective properties and the potential to modulate retinal responses to neurodegeneration, thus reinforcing the idea that MSC secretome may be a therapeutic strategy for treating RND.

Diplopia as the presenting symptom of a post‐transplant lymphoproliferative disorder in a bilateral inferior lim transplanted young adult

Aims/Purpose: To present a unique case of post‐transplant lymphoproliferative disorder (PTLD) after bilateral inferior limb transplantation, which is still controversial due to the absence of precedents in the literature and the unclear risk–benefit ratio.Methods: Case report.Results: A 22‐year‐old male presented to the emergency room complaining of double vision. He had a history of bilateral inferior limb amputation 8 years ago and received a bilateral lower limb transplantation 2 years later. He had normal graft function and was receiving cytomegalovirus prophylaxis. During neurological examinations left abducens nerve (CN VI) palsy was noted. Other cranial nerves affected produced facial hypoesthesia (CN V) and mild paresis (CN VII), hearing loss and tinnitus in left side (CN VIII) and soft palate asymmetry (CN X). He was otherwise neurologically intact, alert and orientated. MRI revealed a lesion located at the facial colliculus at the floor of 4th ventricle. In the context of immunosuppression, the differential diagnosis included abscess versus PTLD. Biopsy of the lesion was performed. Morphologically atypical lymphocytic cells suggestive of B lymphoma were identified. Monoclonal rearrangement for heavy chain Igen and immunoglobulins were identified with molecular biology techniques. The histology features were consistent with a diagnosis of CNS B lineage primary lymphoma. Two days later, bilateral supracondylar amputation was performed. Following the explant, immunosuppression was withdrawn. His underlying condition was finally diagnosed as PTLD, for which he started a chemotherapeutic regimen with radical intention (methotrexate and cytosine arabinoside). The patient recovered fully and up to date, 10 years later, is doing well.Conclusions: Due to its diverse aetiologias, a proper workup must be done of the patients presenting with diplopia to the emergency room. PLTD must be a condition to always be considered in patients with history of solid organ transplant; since the early diagnosis is crucial for a successful management of this often‐fatal condition.

Flying to the moon: Impactful accounts of triatomines invasion from the 2nd to the 13th floor of an urban residential building in the municipality of Rio Branco, Acre, Brazil.

Vectorial transmission through hematophagous triatomine insects remains the primary mode of Chagas Disease contagion. These insects have become increasingly common in urban environments. Therefore, this study aimed to report an encounter of triatomines with trypanosomatid infection in a vertical residential condominium in Rio Branco, the capital of the state of Acre, in the western Brazilian Amazon. Triatomines were collected from residents and sent to the municipality's Entomological Surveillance sector. Trypanosomatid positivity was evaluated using optical microscopy, followed by species and genotype identification using molecular biology techniques. Twenty-five adult triatomine specimens were collected from two of three condominium buildings invading apartments from the 2nd to 13th floors. Six specimens were identified as Rhodnius sp. and 19 as R. montenegrensis. Among these, molecular tests were conducted on seven specimens, with five testing positive for Trypanosoma cruzi, all belonging to genotype TcI. These findings underscore the need for further studies to better understand the invasive capacity of these insects in these environments and the mechanisms involved in this process.

Assessing the Public Health Implications of Virulent and Antibiotic-Resistant Bacteria in Côte d'Ivoire's Ready-to-Eat Salads.

In Côte d'Ivoire, the popularity of ready-to-eat salads has grown substantially. Despite their convenience, these products often face criticism for their microbiological safety. This research was conducted to assess the virulence and antibiotic resistance profiles of Escherichia coli (E. coli), Salmonella spp., and Staphylococcus aureus (S. aureus) isolated from salads available in hypermarkets across Abidjan. The study utilized a combination of microbiological and molecular biology techniques. Results indicated that E. coli isolates harbored virulence genes such as stx2 (50%) and ST (62.50%), though genes stx1 and LT were absent in the samples tested. In S. aureus, virulence genes detected included sea (55.55%), sec (11.110%), and sed (44.44%). The antibiotic resistance assessment revealed high resistance in E. coli to β-lactam antibiotics, with all isolates resistant to cefuroxime (100%) and the majority to ampicillin and cefoxitin (87.5%). Most Salmonella spp. isolates were sensitive to the antibiotics tested, except for cefoxitin and ampicillin, showing resistance rates of 42.85% and 57.15%, respectively. Staphylococcus aureus demonstrated considerable resistance, particularly to cefoxitin (44.44%), benzylpenicillin (100%), and ampicillin (55.55%). In addition, resistance to aminoglycosides (55.55% to both kanamycin and gentamicin) and macrolides (66.66% to erythromycin and 55.55% to clindamycin) was noted. Resistance to various fluoroquinolones ranged between 33.33% and 55.55%. The presence of resistance genes such as blaTEM (10.52%), qnrA (2.26%), qnrB (5.26%), qnrS (5.26%), and mecA (13.15%) in E. coli and S. aureus underscores the challenge of multidrug resistance, exhibiting phenotypes such as ESBL (50%), Meti-R (55.55%), KTG (44.44%), MLSB (44.44%), and FQ-R (25%). These results carry significant epidemiological and public health implications, highlighting the urgent need for improved safety regulations and practices regarding ready-to-eat salads in urban food markets.

GinmappeR: an unified approach for integrating gene and protein identifiers across biological sequence databases.

The proliferation of biological sequence data, due to developments in molecular biology techniques, has led to the creation of numerous open access databases on gene and protein sequencing. However, the lack of direct equivalence between identifiers across these databases difficults data integration. To address this challenge, we introduce ginmappeR, an integrated R package facilitating the translation of gene and protein identifiers between databases. By providing a unified interface, ginmappeR streamlines the integration of diverse data sources into biological workflows, so it enhances efficiency and user experience. from Bioconductor: https://bioconductor.org/packages/ginmappeR.

Viabilidade da substituição da técnica de isolamento viral em camundongos pela RT-qPCR para o diagnóstico da raiva

ABSTRACT: Rabies is a viral encephalitis that affects mammals, including humans. Rapid and effective laboratory diagnosis of the rabies virus is critical for public health. This study evaluated the operational, technical, and financial viability of the RT-qPCR in replacement of inoculation in mice for diagnosing rabies in the laboratory routine. A total of 316 samples of mammalian brains were analyzed, 121 positives and 195 negatives, previously diagnosed by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The samples were submitted to the duplex TaqMan RT-qPCR technique. The accuracy, sensitivity, and specificity of RT-qPCR were analyzed. We analyzed the costs for performing the RT-qPCR technique and compared it with the cost of MIT. The results showed 99.37% accuracy, 99.17% sensitivity, and 99.49% specificity by RT-qPCR when related to DIF and MIT results, which proved to be a robust and repeatable technique. The minimum time for a positive diagnosis was reduced in RT-qPCR (1 day) if compared to MIT (9.64 days), with a 17.7% reduction in the cost of the molecular technique. The present study demonstrated that the molecular biology technique is an efficient tool to diagnose rabies in the laboratory routine, being able to replace MIT.

Oleate alters the immune response in non-small cell lung adenocarcinoma through regulation of HMGB1 release.

Background: Cancer cell evasion of the immune response is critical to cancer development and metastases. Clinicians' ability to kickstart the immune system to target these rogue cells is an ever-growing area of research and medicine. This study delved into the relationship between lipid metabolism, High Mobility Group Box 1 protein (HMGB1)-a pro-inflammatory damage-associated molecular pattern protein-and immune regulation within non-small cell lung adenocarcinoma (NSCLC). Method: To address this question, we used a combination of proteomics, molecular biology, and bioinformatic techniques to investigate the relationship between fatty acids and immune signals within NSCLC. Results: We found that the expression of stearoyl CoA desaturase 1 (SCD1) was decreased in NSCLC tumors compared to normal tissues. This emphasized the critical role of lipid metabolism in tumor progression. Interestingly, monounsaturated fatty acid (MUFA) availability affected the expression of programmed death ligand-1 (PD-L1), a pivotal immune checkpoint target in lung cancer cells and immune cells, as well as HMGB1, suggesting a novel approach to modulating the immune response. This study uncovered a complex interplay between SCD1, PD-L1, and HMGB1, influencing the immunological sensitivity of tumors. Conclusion: Our work underscores the critical importance of understanding the intricate relationships between lipid metabolism and immune modulation to develop more effective NSCLC treatments and personalized therapies. As we continue to explore these connections, we hope to contribute significantly to the ever-evolving field of cancer research, improving patient outcomes and advancing precision medicine in NSCLC.