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1.
World J Microbiol Biotechnol ; 40(9): 273, 2024 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-39030443

RESUMO

Helicobacter pylori is a common resident in the stomach of at least half of the world's population and recent evidence suggest its emergence in other organs such as the pancreas. In this organ, the presence of H. pylori DNA has been reported in cats, although the functional implications remain unknown. In this work, we determined distinct features related to the H. pylori manifestation in pancreas in a rodent model, in order to analyse its functional and structural effect. Gerbils inoculated with H. pylori exhibited the presence of this bacterium, as revealed by the expression of some virulence factors, as CagA and OMPs in stomach and pancreas, and confirmed by urease activity, bacterial culture, PCR and immunofluorescence assays. Non-apparent morphological changes were observed in pancreatic tissue of infected animals; however, delocalization of intercellular junction proteins (claudin-1, claudin-4, occludin, ZO-1, E-cadherin, ß-catenin, desmoglein-2 and desmoplakin I/II) and rearrangement of the actin-cytoskeleton were exhibited. This structural damage was consistent with alterations in the distribution of insulin and glucagon, and a systemic inflammation, event demonstrated by elevated IL-8 levels. Overall, these findings indicate that H. pylori can reach the pancreas, possibly affecting its function and contributing to the development of pancreatic diseases.


Assuntos
Gerbillinae , Infecções por Helicobacter , Helicobacter pylori , Junções Intercelulares , Pâncreas , Animais , Helicobacter pylori/patogenicidade , Helicobacter pylori/genética , Infecções por Helicobacter/microbiologia , Pâncreas/microbiologia , Pâncreas/patologia , Junções Intercelulares/microbiologia , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Antígenos de Bactérias/metabolismo , Antígenos de Bactérias/genética , Fatores de Virulência/metabolismo , Fatores de Virulência/genética , Estômago/microbiologia , Estômago/patologia , Modelos Animais de Doenças , Masculino , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas da Membrana Bacteriana Externa/genética
2.
Microb Pathog ; 165: 105494, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35314281

RESUMO

In this study, the genomes of two lytic bacteriophages, vB_EcoS-phiEc3 and vB_EcoS-phiEc4, were sequenced and characterized using bioinformatics approaches. Whole-genome analysis showed that both phages belonged to the Kagunavirus genus, Guernseyvirinae subfamily and Siphoviridae family. Moreover, their genomes had 45, 288 bp and 44,540 bp, and G + C content of 48.42% and 50.04%, respectively. The genome of vB_EcoS-phiEc3 harbored 80 protein coding sequences (CDSs), whereas vB_EcoS-phiEc4 harbored 75 CDSs. Among them, 50 CDSs in vB_EcoS-phiEc3 and 44 CDSs in vB_EcoS-phiEc4 were considered as functional genes. Their lytic activity against multidrug-resistant uropathogenic Escherichia coli (UPEC) strains, as well as the absence of antibiotic resistance genes, lysogenic and virulence genes, enable vB_EcoS-phiEc3 and vB_EcoS-phiEc4 as a safe therapy option against UPEC infections.


Assuntos
Bacteriófagos , Infecções por Escherichia coli , Siphoviridae , Escherichia coli Uropatogênica , Bacteriófagos/genética , Genoma Viral , Humanos , Siphoviridae/genética , Escherichia coli Uropatogênica/genética
3.
BMC Microbiol ; 19(1): 184, 2019 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-31395006

RESUMO

BACKGROUND: Helicobacter pylori recurrence after successful eradication is an important problem. Children are particularly vulnerable to reinfection, by intrafamilial transmission which facilitates the acquisition or recombination of new genetic information by this bacterium. We investigated the evolutionary dynamics of 80 H. pylori strains isolated from two paediatric patients with recurrent infection (recrudescence and reinfection). RESULTS: We characterized the virulence genes vacA (s1, m1, s2, and m2), cagA, cagE, and babA2 and performed multilocus sequence typing (MLST) on 7 housekeeping genes (atpA, efp, ureI, ppa, mutY, trpC, and yphC) to infer the evolutionary dynamics of the H. pylori strains through phylogenetic and genealogic inference analyses, genetic diversity analysis and the exploration of recombination events during recurrent infections. The virulence genotype vacAs1m1/cagA+/cagE+/babA2 was present at a high frequency, as were the EPIYA motifs EPIYA-A, -B and -C. Furthermore, the housekeeping genes of the H. pylori strains exhibited high genetic variation, comprising 26 new alleles and 17 new Sequence Type (ST). In addition, the hpEurope (76.5%) and hspWAfrica (23.5%) populations predominated among the paediatric strains. All strains, regardless of their ancestral affiliation, harboured western EPIYA motifs. CONCLUSIONS: This study provides evidence of the evolutionary dynamics of the H. pylori strains in two paediatric patients during recrudescence and reinfection events. In particular, our study shows that the strains changed during these events, as evidenced by the presence of different STs that emerged before and after treatment; these changes may be due to the accumulation of mutations and recombination events during the diversification process and recolonization of the patients by different genotypes.


Assuntos
Infecções por Helicobacter/microbiologia , Helicobacter pylori/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Genótipo , Helicobacter pylori/classificação , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/isolamento & purificação , Humanos , Lactente , Masculino , Pediatria , Filogenia , Recidiva , Fatores de Virulência/genética
4.
Indian J Microbiol ; 59(3): 295-303, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31388206

RESUMO

The three-component apsXRS system senses and responds to cationic antimicrobial peptides (CAMPs), which induces the expression of the dlt operon and the genes mprF and vrafG, modifying the surface net charge in Staphylococcus epidermidis, resulting in the repulsion of CAMPs. The apsXRS system has been only studied in the S. epidermidis 1457 strain, and there are no studies of prevalence and level of expression of apsXRS in commensal and clinical isolates. From 60 isolates, those selected from commensal healthy skin (n = 20), commensal healthy conjunctive (n = 10), and clinical ocular infection (n = 30) presented the apsX, apsR, and apsS genes in their genomes. Constitutive expression of apsX, apsR, and apsS genes was determined by RT-qPCR in all isolates. It was found that expression of apsX, apsR, and apsS was 3.3-5.9-fold higher in commensal isolates stimulated with LL-37 (15 µg/mL) than in clinical isolates. Similarly, expression of the dlt operon and the genes mprF, and vraFG was 8-10-fold higher in commensal isolates than in clinical. However, LL-37 did not increase the addition of lysine in the phospholipids of the cytoplasmic membrane in any of the isolates. Mutations in the apsS loop region, apsR, and their promoter sequence were not found. These results demonstrated that apsXRS system is essential in all isolates for its constitutive expression; however, LL-37 caused an increase of apsXRS expression in commensal isolates, suggesting that S. epidermidis isolates do not respond in the same way to the presence of LL-37.

5.
BMC Infect Dis ; 18(1): 463, 2018 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-30219037

RESUMO

BACKGROUND: Helicobacter pylori is a major aetiologic agent associated with gastritis. H. pylori infections increase the expression of the Toll-like receptor (TLR), which in turn modulates the expression of microRNA (miRNA)-146a and miRNA-155. The objective of this study was to compare the expression of miRNA-146a and miRNA-155 in gastric lesions of paediatric and adult patients with different pathologies and in Mongolian gerbils (Meriones unguiculatus) infected with H. pylori 26,695. METHODS: Quantification of miRNA expression was performed by quantitative real-time polymerase chain reaction (qRT-PCR) of paraffin-embedded gastric lesions of children with or without an infection (n = 25), adults with follicular gastritis and metaplasia (n = 32) and eight-week-old M. unguiculatus males (Hsd:MON) infected with H. pylori 26,695 for 0, 3, 6, 12 and 18 months (n = 25). The genes RNU48 and RNU6 were used as endogenous controls for data normalization. Statistical analyses were performed using Kruskal-Wallis, Mann-Whitney, ANOVA and Student's t-test. RESULTS: The expression of miRNA-146a and miRNA-155 in infected children increased by 247.6- and 79.4-fold (on average), respectively, compared to that observed in the control group. However, these results were not significant (p = 0.12 and p = 0.07 respectively). In some children a gradual increase in expression was observed, while in others, expression was very high. Additionally, the expression levels of miRNA-146a and miRNA-155 increased by an average of 21.7- and 62-fold, respectively, in adult patients with follicular gastritis when compared to those of the controls. In M. unguiculatus infected with H. pylori 26,695, the expression of both miRNAs increased as the infection progressed. CONCLUSION: This is the first report to show differences in the expression of miRNA-146a and miRNA-155 in paediatric and adult patients with gastritis who were infected with H. pylori. In addition, in M. unguiculatus infected with H. pylori, miRNA expression was associated with the progression of infection and the ability of the bacteria to adapt to the host.


Assuntos
Gastrite/genética , Infecções por Helicobacter/genética , Helicobacter pylori/fisiologia , MicroRNAs/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Pré-Escolar , Modelos Animais de Doenças , Progressão da Doença , Feminino , Mucosa Gástrica/metabolismo , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Gastrite/microbiologia , Perfilação da Expressão Gênica , Gerbillinae , Infecções por Helicobacter/complicações , Infecções por Helicobacter/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
6.
Rev Argent Microbiol ; 49(1): 15-23, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28017482

RESUMO

Coagulase-positive staphylococci (CoPS) are opportunistic pathogens carrying various mechanisms of resistance that have a large number of virulence factors, and whose ability to induce illness is associated with the host. This study aimed to investigate the presence of environmental coagulase-positive staphylococci, their susceptibility profile, clonal relationship and ability to form biofilm. The 16S rRNA genes from CoPS isolates were analyzed, and their antibiotic susceptibility was evaluated using the agar dilution method in accordance with Clinical and Laboratory Standards Institute guidelines. The clonal profile was obtained by pulsed-field gel electrophoresis (PFGE) and biofilm formation was measured by a crystal violet retention assay. A total of 72 Staphylococcus spp. strains were isolated from air, metal surfaces, and nostrils from humans, dogs, cats, and birds. Three species were identified: Staphylococcus aureus (17%), Staphylococcus intermedius (63%), and Staphylococcus pseudintermedius (21%). Ninety three percent (93%) of the strains were resistant to at least one of 13 tested antibiotics. S. pseudintermedius strains were the only resistant ones to methicillin while most of these isolates were multidrug-resistant, had significantly higher ability to form biofilm and PFGE grouped into seven different patterns, without showing clonal dispersion among animals and environmental isolates. This study suggests that dogs, cat, and air are environmental sources potentially carrying multidrug-resistant S. pseudintermedius, which survives in different environments through biofilm formation and multidrug resistance, characteristics that can be transmitted horizontally to other bacteria and exacerbate the problem of antibiotic resistance in humans.


Assuntos
Biofilmes , Coagulase , Fatores R , Staphylococcus , Animais , Antibacterianos , Gatos , Humanos , Testes de Sensibilidade Microbiana , RNA Ribossômico 16S , Infecções Estafilocócicas , Staphylococcus/isolamento & purificação
7.
Helicobacter ; 20(3): 231-7, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25735460

RESUMO

BACKGROUND: The genes jhp0940, jhp0945, jhp0947, and jhp0949 belong to the plasticity region of the Helicobacter pylori genome. Due to their prevalence in isolates from patients with gastritis, duodenal ulcer, and gastric cancer, they have been proposed as markers of gastroduodenal diseases. These genes are associated with pro-inflammatory cytokine induction through the NF-κB activation pathway. Nevertheless, the status of these genes is unknown in H. pylori isolates from children. The aim of the present work was to determine the frequency of the jhp0940-jhp0945-jhp0947-jhp0949 genes in H. pylori isolates from children. MATERIALS AND METHODS: We identified the jhp0940, jhp0945, jhp0947, and jhp0949 genes and the relationship of each with the virulence factors cagA, cagPAI, and dupA by PCR in 49 isolates of H. pylori from children. The results were corroborated using dot blots. In addition, we compared the prevalence of these genes with the prevalence in adults. RESULTS: The prevalence of jhp0940 (53.1%), jhp0945 (44.9%), jhp0947 (77.6%), and jhp0949 (83.7%) was determined in the isolates from children, as was the prevalence of the virulence genes cagA (63.3%), cagPAI (71.4%), and dupA (37.5%). No association was found between the four genes of the plasticity region and the virulence genes. The presence of the intact locus integrated by jhp0940-jhp0945-jhp0947-jhp0949 was very common among the isolates from children. CONCLUSION: The genes jhp0940, jhp0947, and jhp0949 were present in more than 50% of the H. pylori isolates, and the joint presence of jhp0940-jhp0945-jhp0947-jhp0949 was very frequent. The frequency of these genes in isolates from children could contribute to the virulence of H. pylori and the evolution of the infection.


Assuntos
Proteínas de Bactérias/genética , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/genética , Adolescente , Criança , Pré-Escolar , Úlcera Duodenal/epidemiologia , Úlcera Duodenal/microbiologia , Feminino , Gastrite/epidemiologia , Infecções por Helicobacter/epidemiologia , Helicobacter pylori/patogenicidade , Humanos , Masculino , México/epidemiologia , NF-kappa B/metabolismo , Úlcera Péptica/epidemiologia , Úlcera Péptica/microbiologia , Prevalência , Proteínas Serina-Treonina Quinases/genética , Neoplasias Gástricas/microbiologia , Virulência , Fatores de Virulência
8.
Clin Lab ; 60(8): 1277-85, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25185412

RESUMO

BACKGROUND: Adenovirus (AdV) causes respiratory infection; recent observations suggest that some subtypes have more ability to develop fatal disease. AdV infection has been associated with co-infection with human bocavirus (HBoV). We analysed the frequency of AdV infection, its subtypes and the presence of co-infection with HBoV, as well the clinical characteristics of such co-infection in Mexican paediatric immunosuppressed (IP) and non-immunosuppressed patients (non-IP) diagnosed with pneumonia. METHODS: A total of 5185 nasopharyngeal swabs from two groups of children with pneumonia, one IP and the other non-IP, were analysed for the detection of AdV by immunofluorescence and confirmed by PCR and culture. HBoV was identified by PCR. Positive samples for AdV and AdV/HBoV were typed using PCR sequencing, the clinical characteristics of the AdV/HBoV co-infection were analysed. RESULTS: Thirty-seven of the 5185 (0.71%) samples were positive for AdV, of those 27/37 (73%) were detected in non-IP and 10/37 (27%) in the IP group. Twelve were typed as follows: 9/12 (75%) as Species B1 subtype 3, of those 8/9 (88.9%) in non-IP and 1/9 in the IP group. One of twelve AdV2 subtype B11a was identified in one non-IP and the remaining two out of 12 successfully typed, were identified as Species C subtypes 2 and 6 in the group of non-IP. The presence of both AdV and HBoV1 in co-infection was observed in 2/37 (5.4%) non-IP with a syndrome like influenza. CONCLUSIONS: In this 5 year analysis of samples from non-IP and IP hospitalized paediatric patients with a diagnosis of pneumonia, a low incidence of AdV was found. B1 was the most frequent subtype and frequently found in non-IP, and two cases of co-infection AdV/HBoV1 were detected in two non-IP with a influenza-like syndromes. This is the first report of HBoV and AdV co-infection in Mexico. The frequency of AdV and HBoV co-infection was lower than that reported in other populations.


Assuntos
Infecções por Adenoviridae/complicações , Adenoviridae/genética , Bocavirus/genética , Coinfecção/virologia , Infecções por Parvoviridae/complicações , Pneumonia/complicações , Adolescente , Sequência de Bases , Criança , Pré-Escolar , Estudos Transversais , Genótipo , Humanos , Hospedeiro Imunocomprometido , Lactente , México , Dados de Sequência Molecular , Pneumonia/virologia , Prevalência , Análise de Sequência de DNA
9.
Microorganisms ; 11(11)2023 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-38004759

RESUMO

Dysbiosis plays an important role in the development of bacterial infections in the gastric mucosa, particularly Helicobacter pylori. The international guidelines for the treatment of H. pylori infections suggest standard triple therapy (STT). Nevertheless, because of the increasing resistance rates to clarithromycin, metronidazole has been widely considered in several countries. Unfortunately, the non-justified administration of antibiotics induces dysbiosis in the target organ. We characterized the gastric microbiota of patients diagnosed with follicular gastropathy and pangastropathy attributed to H. pylori infection, before and after the administration of STT with metronidazole. Dominant relative abundances of Cutibacterium were observed in pre-treatment patients, whereas H. pylori was observed at <11%, suggesting the multifactor property of the disease. The correlation of Cutibacterium acnes and H. pylori with gastric infectious diseases was also evaluated using quantitative real-time polymerase chain reaction. The dominance of C. acnes over H. pylori was observed in gastritis, gastropathies, and non-significant histological alterations. None of the microorganisms were detected in the intestinal metaplasia. Post-treatment alterations revealed an increase in the relative abundances of Staphylococcus, Pseudomonas, and Klebsiella. Non-H. pylori gastrointestinal bacteria can be associated with the initiation and development of gastric diseases, such as pathobiont C. acnes.

10.
Antibiotics (Basel) ; 12(11)2023 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-37998810

RESUMO

Citral is a monoterpene constituted by two isomers known as neral and geranial. It is present in different plant sources and recognized as safe (GRAS) by the Food and Drug Administration (FDA). In recent years, investigations have demonstrated that this compound exhibited several biological activities, such as antibacterial, antifungal, antibiofilm, antiparasitic, antiproliferative, anti-inflammatory, and antioxidant properties, by in vitro and in vivo assays. Additionally, when incorporated into different food matrices, citral can reduce the microbial load of pathogenic microorganisms and extend the shelf life. This compound has acceptable drug-likeness properties and does not present any violations of Lipinski's rules, which could be used for drug development. The above shows that citral could be a compound of interest for developing food additives to extend the shelf life of animal and vegetable origin foods and develop pharmaceutical products.

11.
Intervirology ; 55(4): 318-20, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-21734356

RESUMO

We used nested multiplex polymerase chain reaction to determine the distribution of cytomegalovirus gB genotype in 30 Mexican children undergoing bone marrow transplantation. The genotypes were: gB1, 9/30 (30%); gB2, 8/30 (27%); gB3, 4/30 (13%), and gB4, 1/30 (3%); mixed genotypes were identified in 8/30 patients (27%). We conclude that the predominant genotypes identified were gB2 and gB1. The gB5 genotype was not detected, and a high proportion of mixed genotype was found.


Assuntos
Transplante de Medula Óssea/efeitos adversos , Infecções por Citomegalovirus/virologia , Citomegalovirus/classificação , Citomegalovirus/genética , Transplante Homólogo/efeitos adversos , Proteínas do Envelope Viral/genética , Adolescente , Criança , Pré-Escolar , Coinfecção/virologia , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/epidemiologia , Genótipo , Humanos , Lactente , México/epidemiologia , Epidemiologia Molecular , Reação em Cadeia da Polimerase Multiplex , Reação em Cadeia da Polimerase , Prevalência
12.
Pediatr Blood Cancer ; 59(5): 950-2, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22434694

RESUMO

Haploidentical hematopoietic cell transplantation using CD34(+) cells depleted of T lymphocytes by the CliniMACS is a treatment for hematological malignancy. We report on four Mexican children, three with acute lymphocytic leukemia and one with chronic myelocytic leukemia, who was transplanted with 12 × 10(6) CD34(+) stem cells/kg body weight (98% of purity) with a follow-up of 9½ years. The engraftment was successful in three of the four children. All showed cytomegalovirus reactivation, and one died because of graft rejection and infectious complication. The risk of infections was a major problem.


Assuntos
Transplante de Medula Óssea , Transplante de Células-Tronco Hematopoéticas , Leucemia Mielogênica Crônica BCR-ABL Positiva/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Adolescente , Pré-Escolar , Feminino , Seguimentos , Rejeição de Enxerto , Sobrevivência de Enxerto , Teste de Histocompatibilidade , Humanos , Masculino , México , Transplante Homólogo
13.
PLoS One ; 17(5): e0266742, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35536836

RESUMO

Pseudomonas aeruginosa has different resistant mechanisms including the constitutive MexAB-OprM efflux pump. Single nucleotide polymorphisms (SNPs) in the mexR, nalC, and nalD repressors of this efflux pump can contribute to antimicrobial resistance; however, it is unknown whether these changes are mainly related to genetic lineages or environmental pressure. This study identifies SNPs in the mexR, nalC, and nalD genes in clinical and environmental isolates of P. aeruginosa (including high-risk clones). Ninety-one P. aeruginosa strains were classified according to their resistance to antibiotics, typified by multilocus sequencing, and mexR, nalC, and nalD genes sequenced for SNPs identification. The mexAB-oprM transcript expression was determined. The 96.7% of the strains were classified as multidrug resistant. Eight strains produced serine carbapenemases, and 11 strains metallo-ß-lactamases. Twenty-three new STs and high-risk clones ST111 and ST233 were identified. SNPs in the mexR, nalC, and nalD genes revealed 27 different haplotypes (patterns). Sixty-two mutational changes were identified, 13 non-synonymous. Haplotype 1 was the most frequent (n = 40), and mainly identified in strains ST1725 (33/40), with 57.5% pan drug resistant strains, 36.5% extensive drug resistant and two strains exhibiting serin-carbapenemases. Haplotype 12 (n = 9) was identified in ST233 and phylogenetically related STs, with 100% of the strains exhibiting XDR and 90% producing metallo-ß-lactamases. Haplotype 5 was highly associated with XDR and related to dead when compared to ST1725 and ST233 (RRR 23.34; p = 0.009 and RRR 32.01; p = 0.025). A significant relationship between the mexR-nalC-nalD haplotypes and phylogenetically related STs was observed, suggesting mutational changes in these repressors are highly maintained within genetic lineages. In addition, phylogenetically related STs showed similar resistant profiles; however, the resistance was (likely or partly) attributed to the MexAB-OprM efflux pump in 56% of the strains (only 45.05% showed mexA overtranscription), in the remaining strains the resistance could be attributed to carbapenemases or mechanisms including other pumps, since same SNPs in the repressor genes gave rise to different resistance profiles.


Assuntos
Nucleotídeos , Pseudomonas aeruginosa , Antibacterianos/metabolismo , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Genes Reguladores , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Testes de Sensibilidade Microbiana , Nucleotídeos/metabolismo , Pseudomonas aeruginosa/metabolismo , beta-Lactamases/genética
14.
World J Gastroenterol ; 28(29): 3886-3902, 2022 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-36157534

RESUMO

BACKGROUND: The high prevalence and persistence of Helicobacter pylori (H. pylori) infection, as well as the diversity of pathologies related to it, suggest that the virulence factors used by this microorganism are varied. Moreover, as its proteome contains 340 hypothetical proteins, it is important to investigate them to completely understand the mechanisms of its virulence and survival. We have previously reported that the hypothetical protein HP0953 is overexpressed during the first hours of adhesion to inert surfaces, under stress conditions, suggesting its role in the environmental survival of this bacterium and perhaps as a virulence factor. AIM: To investigate the expression and localization of HP0953 during adhesion to an inert surface and against gastric (AGS) cells. METHODS: Expression analysis was performed for HP0953 during H. pylori adhesion. HP0953 expression at 0, 3, 12, 24, and 48 h was evaluated and compared using the Kruskal-Wallis equality-of-populations rank test. Recombinant protein was produced and used to obtain polyclonal antibodies for immunolocalization. Immunogold technique was performed on bacterial sections during adherence to inert surfaces and AGS cells, which was analyzed by transmission electron microscopy. HP0953 protein sequence was analyzed to predict the presence of a signal peptide and transmembrane helices, both provided by the ExPASy platform, and using the GLYCOPP platform for glycosylation sites. Different programs, via, I-TASSER, RaptorX, and HHalign-Kbest, were used to perform three-dimensional modeling. RESULTS: HP0953 exhibited its maximum expression at 12 h of infection in gastric epithelium cells. Immunogold technique revealed HP0953 localization in the cytoplasm and accumulation in some peripheral areas of the bacterial body, with greater expression when it is close to AGS cells. Bioinformatics analysis revealed the presence of a signal peptide that interacts with the transmembrane region and then allows the release of the protein to the external environment. The programs also showed a similarity with the Tip-alpha protein of H. pylori. Tip-alpha is an exotoxin that penetrates cells and induces tumor necrosis factor alpha production, and HP0953 could have a similar function as posttranslational modification sites were found; modifications in turn require enzymes located in eukaryotic cells. Thus, to be functional, HP0953 may necessarily need to be translocated inside the cell where it can trigger different mechanisms producing cellular damage. CONCLUSION: The location of HP0953 around infected cells, the probable posttranslational modifications, and its similarity to an exotoxin suggest that this protein is a virulence factor.


Assuntos
Infecções por Helicobacter , Helicobacter pylori , Proteínas de Bactérias/metabolismo , Células Epiteliais/metabolismo , Epitélio/metabolismo , Exotoxinas/metabolismo , Mucosa Gástrica/patologia , Infecções por Helicobacter/microbiologia , Humanos , Sinais Direcionadores de Proteínas , Proteoma/metabolismo , Proteínas Recombinantes/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fatores de Virulência/metabolismo
15.
Microorganisms ; 10(8)2022 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-36013951

RESUMO

Recent multidrug resistance in Pseudomonas aeruginosa has favoured the adaptation and dissemination of worldwide high-risk strains. In June 2018, 15 P. aeruginosa strains isolated from patients and a contaminated multi-dose meropenem vial were characterized to assess their association to an outbreak in a Mexican paediatric hospital. The strains were characterized by antibiotic susceptibility profiling, virulence factors' production, and biofilm formation. The clonal relationship among isolates was determined with pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) sequencing. Repressor genes for the MexAB-OprM efflux pump were sequenced for haplotype identification. Of the strains, 60% were profiled as extensively drug-resistant (XDR), 33% as multidrug-resistant (MDR), and 6.6% were classified as sensitive (S). All strains presented intermediate resistance to colistin, and 80% were sensitive to aztreonam. Pyoverdine was the most produced virulence factor. The PFGE technique was performed for the identification of the outbreak, revealing eight strains with the same electrophoretic pattern. ST235 and ten new sequence types (STs) were identified, all closely related to ST233. ST3241 predominated in 26.66% of the strains. Twenty-five synonymous and seventeen nonsynonymous substitutions were identified in the regulatory genes of the MexAB-OprM efflux pump, and nalC was the most variable gene. Six different haplotypes were identified. Strains from the outbreak were metallo-ß-lactamases and phylogenetically related to the high-risk clone ST233.

16.
Pediatr Transplant ; 15(1): 103-11, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21199205

RESUMO

We aimed to identify those pediatric patients undergoing ABMT with CMV EOD who developed GCV resistance. Forty-seven patients were analyzed following ABMT. Prospective post-transplant CMV monitoring was performed weekly for the detection of viral leukocyte DNAaemia, viral plasma DNAaemia, and viral DNAuria by PCR. Plasma DNAaemia was confirmed from whole blood by the detection of CMV pp67 late mRNA using NASBA technology. In the cases of persistence of viral DNA in plasma, and positive viral RNA detection in blood, CMV drug resistance screening by comprehensive PCR-based RFLP and sequencing of the viral UL97 gene were performed retrospectively. Thirty of the 47 (63.82%) patients showed active CMV infection with 27/30 (74.4%) patients belonging to the D+R+ group and 25/30 with proven viral replication. In total, 2/30 (6.6%) children developed CMV pneumonia proven by immunohistochemistry. Screening of the viral UL97 gene revealed in one of these two cases (1/30, 3.3%) the simultaneous presence of two point mutations in codon 460 (M460V, M460I) conferring GCV resistance. The CMV seroprevalence (81%) and the incidence of active infection (63.8%) in Mexican children undergoing ABMT are very high.


Assuntos
Transplante de Medula Óssea/métodos , Infecções por Citomegalovirus/complicações , Farmacorresistência Viral , Viroses/complicações , Adolescente , Transplante de Medula Óssea/efeitos adversos , Criança , Pré-Escolar , Códon , DNA Viral/sangue , Ganciclovir/farmacologia , Humanos , Imuno-Histoquímica/métodos , Lactente , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Mutação Puntual , Condicionamento Pré-Transplante
17.
Front Microbiol ; 12: 557902, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33746909

RESUMO

Outer membrane vesicles (OMVs) from Gram-negative bacteria were first described more than 50 years ago. However, the molecular mechanisms involved in biogenesis began to be studied only in the last few decades. Presently, the biogenesis and molecular mechanisms for their release are not completely known. This review covers the most recent information on cellular components involved in OMV biogenesis, such as lipoproteins and outer membrane proteins, lipopolysaccharide, phospholipids, quorum-sensing molecules, and flagella.

18.
Pathogens ; 10(9)2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34578235

RESUMO

Microbiomes are defined as complex microbial communities, which are mainly composed of bacteria, fungi, and viruses residing in diverse regions of the human body. The human stomach consists of a unique and heterogeneous habitat of microbial communities owing to its anatomical and functional characteristics, that allow the optimal growth of characteristic bacteria in this environment. Gastric dysbiosis, which is defined as compositional and functional alterations of the gastric microbiota, can be induced by multiple environmental factors, such as age, diet, multiple antibiotic therapies, proton pump inhibitor abuse, H. pylori status, among others. Although H. pylori colonization has been reported across the world, chronic H. pylori infection may lead to serious consequences; therefore, the infection must be treated. Multiple antibiotic therapy improvements are not always successful because of the lack of adherence to the prescribed antibiotic treatment. However, the abuse of eradication treatments can generate gastric dysbiotic states. Dysbiosis of the gastric microenvironment induces microbial resilience, due to the loss of relevant commensal bacteria and simultaneous colonization by other pathobiont bacteria, which can generate metabolic and physiological changes or even initiate and develop other gastric disorders by non-H. pylori bacteria. This systematic review opens a discussion on the effects of multiple environmental factors on gastric microbial communities.

19.
Clin Invest Med ; 32(1): E57-63, 2009 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-19178880

RESUMO

PURPOSE: To determine MRSA carriage rates and genetic relationships of S. aureus strains in children attending day care centres in 14 cities from three geographic regions in Mexico. MATERIALS AND METHODS: Cross-Sectional Study performed in apparently healthy children aged from 6 mo to 6 yr attending day care centres (DCCs). From September 2002 To January 2003, 2345 nasopharyngeal specimens from a similar number of children were collected. Nasopharyngeal samples for bacterial isolation were obtained by standard methods. Antimicrobial susceptibility was determined and genetic relatedness of all MRSA isolates was determined by pulsed field gel electrophoresis (PFGE). RESULTS: S. aureus was identified in 237 children (10.1%), twenty-two children had MSRA for an overall prevalence of MRSA carriage of 0.93%. Children attending DCCs from cities located in the north and south of Mexico showed higher prevalence than children from DCCs in the central region; 1.75%, and 1.71 vs. 0.08%, respectively (P < 0.05). PFGE demonstrated six different restriction profiles of MRSA with a predominant pattern. CONCLUSIONS: We documented the presence of MRSA strain colonizing children attending DCCs in Mexico, mainly in the south and north regions of the country. Clone A and B which are closely related represented 45 % of the total of MRSA isolates. We found both, SCCmec type II and type IV strains in the three regions.


Assuntos
Creches , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Portador Sadio , Criança , Pré-Escolar , Estudos Transversais , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Lactente , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , México , Testes de Sensibilidade Microbiana , Nasofaringe/microbiologia
20.
Metallomics ; 11(7): 1265-1276, 2019 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-31173034

RESUMO

Streptococcus pneumoniae is a causal agent of otitis media, pneumonia, meningitis and severe cases of septicemia. This human pathogen infects elderly people and children with a high mortality rate of approximately one million deaths per year worldwide. Antibiotic-resistance of S. pneumoniae strains is an increasingly serious health problem; therefore, new therapies capable of combating pneumococcal infections are indispensable. The application of gold nanoparticles has emerged as an option in the control of bacterial infections; however, the mechanism responsible for bacterial cell lysis remains unclear. Specifically, it has been observed that gold nanoparticles are capable of crossing different structures of the S. pneumoniae cells, reaching the cytosol where inclusion bodies of gold nanoparticles are noticed. In this work, a novel process for the separation of such inclusion bodies that allowed the analysis of the biomolecules such as carbohydrates, lipids and proteins associated with the gold nanoparticles was developed. Then, it was possible to separate and identify proteins associated with the gold nanoparticles, which were suggested as possible candidates that facilitate the interaction and entry of gold nanoparticles into S. pneumoniae cells.


Assuntos
Antibacterianos/farmacologia , Ouro/farmacologia , Nanopartículas Metálicas , Infecções Pneumocócicas/tratamento farmacológico , Streptococcus pneumoniae/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Metabolismo dos Carboidratos/efeitos dos fármacos , Resistência Microbiana a Medicamentos , Ouro/química , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Viabilidade Microbiana/efeitos dos fármacos , Streptococcus pneumoniae/fisiologia
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