RESUMO
Trichomonas vaginalis is the causative agent of the common sexually transmitted disease, trichomoniasis, which affects more than a hundred million people worldwide. Metronidazole and tinidazole, agents belonging to the 5-nitroheterocyclic class of antimicrobials, are most often used to treat infection, but increased resistance has been reported and adverse effects of these drugs can be significant. Consequently, an urgent need exists for the development of novel drug entities against trichomoniasis. Critical for antimicrobial drug development is the demonstration of in vivo efficacy. Murine models of vaginal T. vaginalis infection are unreliable for unknown reasons. Meanwhile, murine infections with the related bovine pathogen, Tritrichomonas foetus, tend to be more robust, although susceptibility to different antimicrobials might differ from T. vaginalis. Here, we explored the utility of T. foetus infection as a surrogate model for drug development against T. vaginalis. Four different T. foetus strains caused robust vaginal infection in young mice, while none of four diverse T. vaginalis strains did. Comparison of drug susceptibility profiles revealed that T. foetus and T. vaginalis were similarly susceptible to a range of 5-nitroheterocyclic and gold(I) compounds. By comparison, proteasome inhibitors were 10- to 15-fold less active against T. foetus than T. vaginalis, although one of the proteasome inhibitors, bortezomib, had low micromolar activity or better against multiple strains of both trichomonads. Different strains of T. foetus were used to demonstrate the utility of the murine vaginal infection models for in vivo efficacy testing, including for bortezomib and a gold(I) compound. The differences in susceptibility to proteasome inhibitors may be partially explained by differences in the proteasome subunit sequences between the two trichomonads, although the functional relevance of the proteasome was similar in both organisms. These findings indicate that T. foetus can serve as a reliable surrogate model for T. vaginalis in vitro and in murine infections in vivo, but caution must be exercised for specific drug classes with targets, such as the proteasome, that may display genetic divergence between the trichomonads.
Assuntos
Modelos Animais de Doenças , Trichomonas vaginalis , Tritrichomonas foetus , Animais , Tritrichomonas foetus/efeitos dos fármacos , Feminino , Trichomonas vaginalis/efeitos dos fármacos , Camundongos , Desenvolvimento de Medicamentos , Metronidazol/farmacologia , Tricomoníase/tratamento farmacológico , Tricomoníase/parasitologia , Vaginite por Trichomonas/tratamento farmacológico , Vaginite por Trichomonas/parasitologia , Vagina/parasitologia , Vagina/efeitos dos fármacosRESUMO
Introduction: The decreasing Helicobacter pylori eradication rate is primarily attributed to antibiotic resistance, and further exacerbated by uniform drug administration disregarding a host's metabolic capability. Consequently, applying personalized treatment based on antibiotic resistance-associated variants and the host's metabolic phenotype can potentially increase the eradication rate. Method: A custom next-generation sequencing panel for personalized H. pylori eradication treatment (NGS-PHET) was designed which targeted the regions for amoxicillin, clarithromycin, metronidazole, tetracycline, and levofloxacin-resistance in H. pylori and human proton-pump inhibitor (PPI) metabolism. The libraries were constructed following customized methods and sequenced simultaneously. The customized framework criteria, grounded in previously reported antibiotic resistance associated variants and the host's PPI metabolism, was applied to the NGS-PHET results and suggested a personalized treatment for each subject, which was validated through each subject's actual eradication outcome. Results: Both previously reported and novel variants were identified from H. pylori sequencing results. Concurrently, five CYP2C19 homozygous extensive metabolizers and three CYP3A4 intermediate metabolizers were identified. Among the total of 12 subjects, clarithromycin triple therapy was suggested for five subjects, bismuth quadruple therapy was suggested for six subjects, and rifabutin triple therapy was suggested for one subject by following the customized framework criteria. The treatment suggestion for nine of the 12 subjects was consistent with the treatment that each subject achieved eradication with. Discussion: Applying the methodology using the NGS-PHET and customized framework helps to perform eradication treatment quickly and effectively in most patients with antibiotic-resistant H. pylori strains, and is also useful in research to find novel antibiotic-resistance candidates.
Assuntos
Antibacterianos , Infecções por Helicobacter , Helicobacter pylori , Sequenciamento de Nucleotídeos em Larga Escala , Medicina de Precisão , Humanos , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/genética , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Medicina de Precisão/métodos , Inibidores da Bomba de Prótons/uso terapêutico , Claritromicina/farmacologia , Claritromicina/uso terapêutico , Masculino , Farmacorresistência Bacteriana/genética , Pessoa de Meia-Idade , Feminino , Adulto , Quimioterapia Combinada , Metronidazol/farmacologia , Metronidazol/uso terapêutico , Amoxicilina/uso terapêutico , Amoxicilina/farmacologia , Citocromo P-450 CYP2C19/genética , Testes de Sensibilidade Microbiana , Levofloxacino/farmacologia , Levofloxacino/uso terapêutico , Tetraciclina/farmacologia , Tetraciclina/uso terapêutico , Resultado do TratamentoRESUMO
OBJECTIVES: Aim of this study was to analyse causal microbiological agents and their bacterial resistance in orofacial infections requiring hospital admission. MATERIALS AND METHODS: Presented is a 10-year retrospective study of patients hospitalised at a single department in 2014-2023. 744 patients were involved. In the statistical analysis, following data was evaluated: causal microbes and their resistance to Penicillin, Amoxicillin-Clavulanate, Clindamycin and Metronidazole. RESULTS: Most frequent aetiology was odontogenic with causal tooth in socket (n = 468; 62,9%), followed by odontogenic - post extraction (n = 152; 20.4%), jaw fracture (n = 41; 5.5%), sialadenitis n = 31 (4.2%), osteonecrosis n = 22 (3.0%), oncological diagnosis in head and neck (n = 17; 2.3%), unknown (n = 10; 1.3%) and multiple factors (n = 3; 0.4%). 408 patients (54.8%) underwent extraoral abscess revision, 336 patients (45.2%) patients were treated locally without extraoral revision. In odontogenic group with tooth still present, superior CRP (m = 145.8 mg/l; SD = 117.7) and leukocyte values (m = 13.6*109l; SD = 6.6) were observed in comparison to other groups. There were 698 cultivated bacteria in 362 patients. Most frequent bacteria were Streptococci (n = 162; 23.2%), Prevotella (n = 83; 11.2%) and Parvimonas (n = 65; 9.3%). Clindamycin resistance was highest (n = 180 resistant bacteria; 25.8%), followed by Metronidazole (n = 178; 25.5%), Penicillin (n = 107; 15.3%) and Amoxicillin-Clavulanate (n = 34; 4.9%). CONCLUSIONS: Orofacial infections in head and neck region are mostly of odontogenic origin with causal tooth still in socket. Causal bacteria show a high antibiotic resistance rate, especially to Clindamycin and Metronidazole. CLINICAL RELEVANCE: Acquired data will be used to determine guidelines for empirical antibiotic prescription in cases of orofacial infections.
Assuntos
Antibacterianos , Humanos , Estudos Retrospectivos , Masculino , Feminino , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Pessoa de Meia-Idade , Adulto , Testes de Sensibilidade Microbiana , Farmacorresistência Bacteriana , Idoso , Metronidazol/uso terapêutico , Metronidazol/farmacologia , Adolescente , Clindamicina/uso terapêutico , Clindamicina/farmacologia , Criança , Penicilinas , Combinação Amoxicilina e Clavulanato de Potássio/uso terapêutico , Doenças da Boca/microbiologia , HospitalizaçãoRESUMO
BACKGROUND: Metronidazole is a first-line antibiotic to treat Helicobacter pylori infections. However, the Clinical Laboratory Standards Institute guidelines recommend against using antimicrobial susceptibility test (AST) to test metronidazole resistance, due to the unreliable predictive power which can result in treatment failure. OBJECTIVES: The aim of this study was to establish an 8-h, metabolic-phenotype based AST for H. pylori metronidazole susceptibility using D2O-probed Raman microspectroscopy. METHODS: Minimal inhibitory concentration (MIC) measured by conventional AST (E-test) were compared with expedited MIC via metabolic activity (eMIC-MA) for 10 H. pylori isolates. Raman barcodes of cellular-response to stress (RBCS) incorporating protein and carbohydrate Raman bands, were utilized to identify a biomarker to distinguish metronidazole susceptibility. RESULTS: Specifically, eMIC-MA produces metronidazole susceptibility results showing 100% agreement with E-test, and determines the bactericidal dosage for both high- and low-level resistant H. pylori strains. In addition, RBCS not just reliably distinguish between metronidazole-susceptible and -resistant strains, but reveal their distinct mechanisms in bacterial responses to metronidazole. CONCLUSION: The speed, accuracy, low cost, and rich information content that reveals the mode-of-action of drugs suggest the method's value in guiding metronidazole prescriptions for H. pylori eradication and in rapid screening based on drug-resistance mechanism.
Assuntos
Antibacterianos , Infecções por Helicobacter , Helicobacter pylori , Metronidazol , Testes de Sensibilidade Microbiana , Análise Espectral Raman , Helicobacter pylori/efeitos dos fármacos , Metronidazol/farmacologia , Análise Espectral Raman/métodos , Testes de Sensibilidade Microbiana/métodos , Humanos , Antibacterianos/farmacologia , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/diagnóstico , Análise de Célula Única/métodos , Farmacorresistência BacterianaRESUMO
OBJECTIVE: The primary goal is to evaluate the effects of two different intracanal medicaments, calcium hydroxide [Ca(OH)2] and double antibiotic paste (DAP), on radiographic outcomes during regenerative endodontic procedures (REP) of immature permanent mandibular first molars with symptomatic irreversible pulpitis and symptomatic apical periodontitis (SIP/SAP). Additionally, the secondary goal was to evaluate MMP-8 levels during REP using two different intracanal medicaments. METHODOLOGY: The study included 20 patients with immature mandibular first molars exhibiting SIP/SAP. Participants were randomly assigned into two groups based on the applied intracanal medicament. Ca(OH)2 (n=10) was prepared by mixing it with sterile distilled water, while the same amount of powdered metronidazole and ciprofloxacin were mixed and combined with sterile distilled water for DAP (n=10). MMP-8 in periapical samples were measured at baseline and on the 14th day using immunofluorometric assay. Image-J software with TurboReg plug-in was utilized to determine changes in root length, root width, radiographic root area (RRA) during the 12-month follow-up period. Data were analyzed by SPSS 25.0 (p<.05). RESULTS: Significant increase in MMP-8 on the 14th day compared to baseline in both groups (p<0.001). There was no significant difference between the two groups in terms of the increase in MMP-8 (p>0.05). Root length significantly increased in both groups (p=0.001), with Ca(OH)2 showing a greater increase (p=0.046). Root width and RRA increased similarly in both groups at 12th month. CONCLUSION: Both Ca(OH)2 and DAP applications resulted in a significant increase in periapical MMP-8 levels. Increase in radiographic root width and root area was similar between two groups, but Ca(OH)2 led to a significantly greater increase in root length. Further studies with larger sample sizes are necessary to validate our findings during REP of vital immature permanent mandibular molars. Clinical Trials database: NCT05581706.
Assuntos
Antibacterianos , Hidróxido de Cálcio , Ciprofloxacina , Metaloproteinase 8 da Matriz , Metronidazol , Periodontite Periapical , Endodontia Regenerativa , Irrigantes do Canal Radicular , Humanos , Hidróxido de Cálcio/uso terapêutico , Hidróxido de Cálcio/farmacologia , Metaloproteinase 8 da Matriz/análise , Metronidazol/uso terapêutico , Metronidazol/farmacologia , Ciprofloxacina/farmacologia , Masculino , Feminino , Resultado do Tratamento , Irrigantes do Canal Radicular/uso terapêutico , Irrigantes do Canal Radicular/farmacologia , Periodontite Periapical/terapia , Periodontite Periapical/diagnóstico por imagem , Endodontia Regenerativa/métodos , Fatores de Tempo , Adolescente , Pulpite/terapia , Pulpite/diagnóstico por imagem , Dente Molar/efeitos dos fármacos , Estatísticas não Paramétricas , Valores de Referência , Reprodutibilidade dos Testes , CriançaRESUMO
Helicobacter pylori infection constitutes a silent pandemic of global concern. In the last decades, the alarming increase in multidrug resistance evolved by this pathogen has led to a marked drop in the eradication rates of traditional therapies worldwide. By using a high-throughput screening strategy, in combination with in vitro DNA binding assays and antibacterial activity testing, we identified a battery of novel drug-like HsrA inhibitors with MIC values ranging from 0.031 to 4 mg/L against several antibiotic-resistant strains of H. pylori, and minor effects against both Gram-negative and Gram-positive species of human microbiota. The most potent anti-H. pylori candidate demonstrated a high therapeutic index, an additive effect in combination with metronidazole and clarithromycin as well as a strong antimicrobial action against Campylobacter jejuni, another clinically relevant pathogen of phylum Campylobacterota. Transcriptomic analysis suggests that the in vivo inhibition of HsrA triggers lethal global disturbances in H. pylori physiology including the arrest of protein biosynthesis, malfunction of respiratory chain, detriment in ATP generation, and oxidative stress. The novel drug-like HsrA inhibitors described here constitute valuable candidates to a new family of narrow-spectrum antibiotics that allow overcoming the current resistome, protecting from dysbiosis, and increasing therapeutic options for novel personalized treatments against H. pylori.
Assuntos
Antibacterianos , Proteínas de Bactérias , Helicobacter pylori , Testes de Sensibilidade Microbiana , Helicobacter pylori/efeitos dos fármacos , Antibacterianos/farmacologia , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/metabolismo , Humanos , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Claritromicina/farmacologia , Metronidazol/farmacologiaRESUMO
The core intent of the existing effort was to explore a triple therapy to eradicate Helicobacter pylori. A hard gelatin capsule filled with metronidazole (MNZ) floating microspheres aided with Plantago ovata seed mucilage (POSM) and Clarithromycin (CMN) floating microspheres aided with Abelmoschus esculentus fruit mucilage (AEFM). These mucilages were adopted as they have gastro-protective actions. These microspheres were designed by a central composite design. The influence of polymers used was checked towards the drug entrapment efficacy and floating time was tallied as a response. The capsule also contains Pantoprazole sodium (PZS) enteric-coated mini-tablets. These mini-tablets were checked for the coating thickness as a response (Design Expert). The microspheres and the mini-tablets were gauged for tests and a positive response was reported. The study summarizes that microspheres of MNZ & CMN and PZS enteric-coated mini-tablets can be used to eradicate H. pylori effectively. POSM and AEFM can aid MNZ and CMN microspheres formulations and have ulcer-curing and gastric-protective abilities.
Assuntos
Abelmoschus , Claritromicina , Infecções por Helicobacter , Helicobacter pylori , Metronidazol , Microesferas , Plantago , Helicobacter pylori/efeitos dos fármacos , Claritromicina/administração & dosagem , Metronidazol/administração & dosagem , Metronidazol/farmacologia , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Plantago/química , Abelmoschus/química , Antibacterianos/administração & dosagem , Pantoprazol/administração & dosagem , Pantoprazol/farmacologia , Cápsulas , Polímeros/química , Sementes/química , Comprimidos com Revestimento Entérico , Quimioterapia Combinada , Frutas , Gelatina/química , Humanos , Antiulcerosos/administração & dosagem , Antiulcerosos/farmacologiaRESUMO
INTRODUCTION: Bacteria and their byproducts are key contributors to the onset and perpetuation of pulpoperiapical pathosis. Intracanal medication is vital in achieving successful endodontic outcomes as it targets and eradicates remaining microorganisms following biomechanical preparation. AIM AND OBJECTIVE: The aim of the study was to compare and evaluate the antimicrobial efficacy of calcium hydroxide (CH) paste, triple antibiotic paste (TAP), and probiotics (PBs) as intracanal medicament in 12-17-year-old children undergoing root canal treatment for the management of infected pulpal tissues in young permanent teeth. MATERIALS AND METHODS: A total of 30 patients aged 12-17 years indicated for endodontic therapy in maxillary incisors and with no systemic complications were selected. They were randomly divided into three groups, i.e., Group I - CH group, Group II - TAP, and Group III - PB allocating 10 teeth in each group. After access opening, the first sample (S1) was collected by inserting a paper point into the root canal, the second sample (S2) was collected immediately after biomechanical preparation, and the third sample (S3) was collected after 7 days, i.e., postintracanal medication. Samples were sent for microbiological analysis to assess the microbial count, and statistical analysis was done for the obtained data. RESULTS: The three intracanal medicaments were successful in reducing the microbial counts of Enterococcus faecalis in the infected root canals. However, according to the results of the study, the PB group demonstrated greater effectiveness against E. faecalis compared to the CH group and displayed similar antimicrobial efficacy as the TAP group. CONCLUSION: PB exhibited antimicrobial efficacy comparable to TAP but greater than Ca (OH) 2 paste. Hence, PB can be utilized as an intracanal medicament in young permanent teeth.
Assuntos
Antibacterianos , Hidróxido de Cálcio , Irrigantes do Canal Radicular , Humanos , Adolescente , Criança , Hidróxido de Cálcio/uso terapêutico , Hidróxido de Cálcio/farmacologia , Antibacterianos/uso terapêutico , Antibacterianos/farmacologia , Irrigantes do Canal Radicular/farmacologia , Irrigantes do Canal Radicular/uso terapêutico , Probióticos/uso terapêutico , Dentição Permanente , Ciprofloxacina/farmacologia , Ciprofloxacina/uso terapêutico , Incisivo , Masculino , Metronidazol/farmacologia , Metronidazol/uso terapêutico , Feminino , Tratamento do Canal Radicular/métodos , Combinação de MedicamentosRESUMO
Objective: This study aimed to evaluate antibiotic susceptibility and antimicrobial resistance trends among clinically significant anaerobes in Kuwait hospitals from 2013 to 2022, comparing these findings with data from 2002 to 2012. Methods: The study prospectively collected 2,317 anaerobic isolates from various body sites across four Kuwaiti hospitals between January 2013 and December 2022. The minimum inhibitory concentrations for 11 antianaerobic antibiotics were determined using E-test methodology. The study analyzed trends and resistance rates across two periods: 2013-2017 and 2018-2022, using statistical analysis for resistance comparison. Results: Of the 2,317 isolates, most were from wounds (42.2%), fluids (28.0%), and tissues (20.5%). Bacteroides fragilis was the most common pathogen (34.0%), followed by Prevotella bivia (13.4%). Over 90% of isolates were susceptible to imipenem, meropenem, tigecycline, and metronidazole, whereas lower susceptibility was observed for penicillin, amoxicillin-clavulanic acid, and clindamycin. Notable differences in resistance profiles since 2002 were observed, especially in amoxicillin-clavulanic acid, piperacillin, piperacillin-tazobactam, and clindamycin. Conclusion: Owing to detected resistance to all antibiotics, susceptibility testing for anaerobic isolates is recommended in severe infections to ensure effective antimicrobial therapy. Continuous surveillance is crucial for developing antibiotic policies to manage invasive anaerobic infections.
Assuntos
Antibacterianos , Bactérias Anaeróbias , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Bactérias Anaeróbias/efeitos dos fármacos , Kuweit/epidemiologia , Humanos , Estudos Prospectivos , Tigeciclina/farmacologia , Farmacorresistência Bacteriana , Bacteroides fragilis/efeitos dos fármacos , Bacteroides fragilis/isolamento & purificação , Metronidazol/farmacologia , Metronidazol/uso terapêuticoRESUMO
BACKGROUND: Helicobacter pylori (H. pylori)is a Gram-negative, microaerophilic, and spiral shape bacterium that resides inside the human stomach. The human stomach serves as its primary reservoir. Complaints about stomach complication due to H. pylori infections are reported in the majority of populations around the globe. Chronic gastritis and intestinal metaplasia of the gastric mucosa are major complications of a long-term H. pylori infections that can lead to gastric cancer in severe cases. This study aims to characterize H. pylori isolates from gastroenteritis patients and to determine the resistance of H. pylori to various antibiotics. METHODS: In the current study, a total of (n = 80) gastric biopsy samples were randomly collected from gastroenteritis patients in brain-heart infusion broth. These were inoculated on Columbia blood agar supplemented with Helicobacter pylori selective supplement (DENT). After culturing, Microscopy and biochemical tests were performed. The susceptibility profile of H. pylori isolates was evaluated using the Kirby Bauer disk diffusion method. On the basis of the drug resistance profile, a total of (n = 20) isolates including (n = 10) from females and (n = 10) from males were selected for the detection and characterization of resistant genes. After confirmation of H. pylori using 16s rRNA, polymerase chain reaction (PCR) was done for the detection of resistance genes including Metronidazole resistance (rdxA gene), Clarithromycin resistance (23s rRNA gene) and Amoxicillin resistance (Penicillin-binding protein A1 (pbpA1) gene). RESULTS: In a total of (n = 80) samples, H. pylori was isolated from 72.5% (n = 58) samples. Among the positive patients, there were 62% (n = 36) of female positive patients while in males, its ratio was 38% (n = 22). It was more common in the age between 30-50 years 55.17% (n = 32). It has shown the highest resistance towards Metronidazole 90% (n = 52), and the lowest toward Levofloxacin 65% (n = 38). Metronidazole resistance gene (rdxA gene) was detected in (n = 13) isolates including (n = 9) isolates from females and (n = 4) from males. In the case of, the Clarithromycin resistance gene (23s rRNA) (n = 10) was positive for H. pylori including (n = 6) isolates from females and (n = 7) were positive for Amoxicillin (pbpA1 gene) including (n = 2) in female and (n = 5) from male patients. CONCLUSION: This study highlights the increasing incidence of H. pylori infections in both male and female patients. It also revealed the current status of antibiotic resistance and its resistance genes in patients facing gastrointestinal issues. Continuous surveillance of resistant clones will help in formulating strategies that can help in combating of resistant clone. It will also help clinician in proper prescription and management of H. pylori infections.
Assuntos
Amoxicilina , Antibacterianos , Claritromicina , Farmacorresistência Bacteriana , Gastroenterite , Infecções por Helicobacter , Helicobacter pylori , Metronidazol , Humanos , Helicobacter pylori/genética , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/isolamento & purificação , Claritromicina/farmacologia , Feminino , Amoxicilina/farmacologia , Masculino , Metronidazol/farmacologia , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/complicações , Gastroenterite/microbiologia , Gastroenterite/tratamento farmacológico , Farmacorresistência Bacteriana/genética , Adulto , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Proteínas de Bactérias/genéticaRESUMO
Within the penile microbiome, bacteria associated with seroconversion, immunology, and cells (BASIC species) enhance HIV susceptibility in heterosexual uncircumcised men by inducing foreskin inflammation and HIV target cell recruitment. This phase 1/2 clinical trial randomizes HIV-uninfected Ugandan men (n = 125) to either oral tinidazole, topical metronidazole, topical clindamycin, or topical hydrogen peroxide to define impact on ex vivo foreskin HIV susceptibility, penile immunology, and BASIC species density. Antimicrobials are well tolerated, and 116 (93%) participants complete the protocol. Topical metronidazole and oral tinidazole reduce the inner foreskin tissue density of HIV-susceptible CD4+ T cells (predefined primary endpoint). Antimicrobials also have varying but substantial effects on reducing prepuce inflammation and BASIC species density, reducing density of foreskin T cell subsets, and increasing foreskin epithelial integrity. Immune alterations correlate strongly with changes in the abundance of BASIC species. Clinical interventions targeting the penile microbiota, particularly topical metronidazole, may reduce HIV susceptibility in uncircumcised men.
Assuntos
Infecções por HIV , Pênis , Humanos , Masculino , Infecções por HIV/imunologia , Infecções por HIV/tratamento farmacológico , Adulto , Pênis/imunologia , Pênis/microbiologia , Pênis/efeitos dos fármacos , Pênis/patologia , Anti-Infecciosos/farmacologia , Anti-Infecciosos/uso terapêutico , Prepúcio do Pênis/imunologia , Suscetibilidade a Doenças , Circuncisão Masculina , Adulto Jovem , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Metronidazol/farmacologia , Metronidazol/administração & dosagem , UgandaRESUMO
The hypoxic microenvironment of solid tumors can lead to reduced therapeutic DNA damage to the tumor cells, thus diminishing tumor sensitivity to radiotherapy. Although hypoxic radiosensitizers can improve radiotherapy efficacy by enhancing the role of oxygen, their effects are limited by the uneven distribution of oxygen within solid tumor tissues. In this study, a novel radiosensitizer via leveraging gold complexes and metronidazole (MN) was synthesized to improve radiotherapeutic efficacy. The gold atoms incorporated in the radiosensitizer enabled efficient deposition of high-energy radiation; the hydrophobic metronidazole was reduced to hydrophilic aminoimidazole under hypoxia conditions and further promoted radiotherapy sensitization. The results of CCK-8 assays, Live/Dead assays, γ-H2AX immunofluorescence indicated that metronidazole-modified Au@BSA nanocomposites (NCs) exhibited excellent antitumor effects. The in vivo antitumor tests further showed an inhibition rate of 100%. These results demonstrated that the NCs successfully enhanced radiotherapy efficacy by the dual sensitization strategy. Overall, we believe this multimodal radiosensitizing nanocomplex can significantly inhibit tumor growth and metastasis, with their hypoxia-oriented characteristics ensuring a higher efficacy and safety.
Assuntos
Antineoplásicos , Ouro , Metronidazol , Nanocompostos , Radiossensibilizantes , Ouro/química , Ouro/farmacologia , Metronidazol/química , Metronidazol/farmacologia , Radiossensibilizantes/química , Radiossensibilizantes/farmacologia , Nanocompostos/química , Animais , Humanos , Camundongos , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/síntese química , Proliferação de Células/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Ensaios de Seleção de Medicamentos Antitumorais , Tamanho da Partícula , Neoplasias/tratamento farmacológico , Camundongos Nus , FemininoRESUMO
Trichomonas vaginalis is the etiologic agent of trichomoniasis, a worldwide distributed sexually transmitted infection (STI) that affects the genitourinary tract. Even though this disease already has a treatment in the prescription of drugs of the 5-nitroimidazole class, described low treatments adhesion, adverse side effects and cases of resistant isolates demonstrate the need for new formulations. With this in mind, chalcones emerge as a potential alternative to be tested, being compounds widely distributed in nature, easy to chemically synthesize and presenting several biological activities already reported. In this experiment, we evaluated the antiparasitic activity of 10 chalcone at a concentration of 100 µM against ATCC 30236 T. vaginalis isolates, considering negative (live trophozoites), positive (Metronidazole 100 µM) and vehicle (DMSO 0.6%) controls. Compounds 3a, 3c, 3 g and 3i showed promising results, with MICs set at 70 µM, 80 µM, 90 µM and 90 µM, respectively (p < 0,05). Cytotoxicity assays were performed on VERO and HMVII cell lines and revealed low inhibition rates at concentrations bellow 20 µM. To elucidate a possible mechanism of action for these molecules, the DPPH, ABTS and FRAP assays were performed, in which none of the four compounds presented antioxidant activity. Assays to verify ROS and lipid peroxidation in the parasite membrane were performed. None of the tested compounds identified ROS accumulation after incubation with trophozoites. 3 g molecule promoted an increase in MDA production after incubation. Results presented in this paper demonstrate the promising trichomonicidal profile, although further tests are still needed to optimize their performance and better elucidate the mechanisms of action involved.
Assuntos
Chalconas , Trichomonas vaginalis , Trichomonas vaginalis/efeitos dos fármacos , Animais , Chalconas/farmacologia , Chalconas/química , Chlorocebus aethiops , Células Vero , Humanos , Linhagem Celular , Espécies Reativas de Oxigênio/metabolismo , Metronidazol/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Testes de Sensibilidade MicrobianaRESUMO
The understanding of how central metabolism and fermentation pathways regulate antimicrobial susceptibility in the anaerobic pathogen Bacteroides fragilis is still incomplete. Our study reveals that B. fragilis encodes two iron-dependent, redox-sensitive regulatory pirin protein genes, pir1 and pir2. The mRNA expression of these genes increases when exposed to oxygen and during growth in iron-limiting conditions. These proteins, Pir1 and Pir2, influence the production of short-chain fatty acids and modify the susceptibility to metronidazole and amixicile, a new inhibitor of pyruvate: ferredoxin oxidoreductase in anaerobes. We have demonstrated that Pir1 and Pir2 interact directly with this oxidoreductase, as confirmed by two-hybrid system assays. Furthermore, structural analysis using AlphaFold2 predicts that Pir1 and Pir2 interact stably with several central metabolism enzymes, including the 2-ketoglutarate:ferredoxin oxidoreductases Kor1AB and Kor2CDAEBG. We used a series of metabolic mutants and electron transport chain inhibitors to demonstrate the extensive impact of bacterial metabolism on metronidazole and amixicile susceptibility. We also show that amixicile is an effective antimicrobial against B. fragilis in an experimental model of intra-abdominal infection. Our investigation led to the discovery that the kor2AEBG genes are essential for growth and have dual functions, including the formation of 2-ketoglutarate via the reverse TCA cycle. However, the metabolic activity that bypasses the function of Kor2AEBG following the addition of phospholipids or fatty acids remains undefined. Overall, our study provides new insights into the central metabolism of B. fragilis and its regulation by pirin proteins, which could be exploited for the development of new narrow-spectrum antimicrobials in the future.
Assuntos
Antibacterianos , Bacteroides fragilis , Metronidazol , Bacteroides fragilis/genética , Bacteroides fragilis/efeitos dos fármacos , Bacteroides fragilis/enzimologia , Bacteroides fragilis/metabolismo , Metronidazol/farmacologia , Metronidazol/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Testes de Sensibilidade Microbiana , Regulação Bacteriana da Expressão GênicaRESUMO
BACKGROUND: Giardiasis, caused by the protozoan parasite Giardia intestinalis, often presents a treatment challenge, particularly in terms of resistance to metronidazole. Despite extensive research, markers for metronidazole resistance have not yet been identified. METHODS: This study analysed 28 clinical samples of G. intestinalis from sub-assemblage AII, characterised by varying responses to metronidazole treatment. We focussed on copy number variation (CNV) of the multi-copy flavohemoprotein gene, analysed using digital polymerase chain reaction (dPCR) and next generation sequencing (NGS). Additionally, chromosomal ploidy was tested in 18 of these samples. Flavohemoprotein CNV was also assessed in 17 samples from other sub-assemblages. RESULTS: Analyses revealed variable CNVs of the flavohemoprotein gene among the isolates, with no correlation to clinical metronidazole resistance. Discrepancies in CNVs detected from NGS data were attributed to biases linked to the whole genome amplification. However, dPCR helped to clarify these discrepancies by providing more consistent CNV data. Significant differences in flavohemoprotein CNVs were observed across different G. intestinalis sub-assemblages. Notably, Giardia exhibits a propensity for aneuploidy, contributing to genomic variability within and between sub-assemblages. CONCLUSIONS: The complexity of the clinical metronidazole resistance in Giardia is influenced by multiple genetic factors, including CNVs and aneuploidy. No significant differences in the CNV of the flavohemoprotein gene between isolates from metronidazole-resistant and metronidazole-sensitive cases of giardiasis were found, underscoring the need for further research to identify reliable genetic markers for resistance. We demonstrate that dPCR and NGS are robust methods for analysing CNVs and provide cross-validating results, highlighting their utility in the genetic analyses of this parasite.
Assuntos
Antiprotozoários , Variações do Número de Cópias de DNA , Resistência a Medicamentos , Giardia lamblia , Giardíase , Metronidazol , Giardia lamblia/genética , Giardia lamblia/efeitos dos fármacos , Metronidazol/farmacologia , Resistência a Medicamentos/genética , Humanos , Giardíase/parasitologia , Giardíase/tratamento farmacológico , Antiprotozoários/farmacologia , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Protozoários/genéticaRESUMO
Bacterial vaginosis (BV) is an imbalance of the vaginal microbiome in which there are limited lactobacilli and an overgrowth of anaerobic and fastidious bacteria such as Gardnerella. The propensity for BV recurrence is high, and therapies involving multiple treatment modalities are emerging to meet this need. However, current treatments requiring frequent therapeutic administration are challenging for patients and impact user compliance. Three-dimensional (3D)-printing offers a novel alternative to customize platforms to facilitate sustained therapeutic delivery to the vaginal tract. This study designed a novel vehicle intended for dual sustained delivery of both antibiotic and probiotic. 3D-printed compartmental scaffolds consisting of an antibiotic-containing silicone shell and a core containing probiotic Lactobacillus were developed with multiple formulations including biomaterials sodium alginate (SA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyethylene oxide (PEO), and kappa-carrageenan (KC). The vehicles were loaded with 50 µg of metronidazole/mg polymer and 5 × 107 CFU of L. crispatus/mg scaffold. Metronidazole-containing shells exhibited cumulative drug release of 324.2 ± 31.2 µg/mL after 14 days. Multiple polymeric formulations for the probiotic core demonstrated cumulative L. crispatus recovery of >5 × 107 CFU/mg scaffold during this timeframe. L. crispatus-loaded polymeric formulations exhibited ≥2 log CFU/mL reduction in free Gardnerella in the presence of VK2/E6E7 vaginal epithelial cells. As a first step towards the goal of facilitating patient compliance, this study demonstrates in vitro effect of a novel 3D-printed dual antibiotic and probiotic delivery platform to target BV.
Assuntos
Lactobacillus crispatus , Metronidazol , Impressão Tridimensional , Probióticos , Silicones , Humanos , Silicones/química , Metronidazol/farmacologia , Metronidazol/administração & dosagem , Metronidazol/química , Feminino , Probióticos/administração & dosagem , Vaginose Bacteriana/tratamento farmacológico , Vaginose Bacteriana/microbiologia , Vaginose Bacteriana/terapia , Antibacterianos/farmacologia , Antibacterianos/administração & dosagem , Antibacterianos/química , Preparações de Ação Retardada , Liberação Controlada de Fármacos , Sistemas de Liberação de Medicamentos/métodosRESUMO
OBJECTIVES: Clostridioides difficile ranks among the primary sources of healthcare-related infections and diarrhoea in numerous nations. We evaluated the drug susceptibility and resistance mechanisms of C. difficile isolates from a hospital in Chongqing, China, and identified resistance rates and resistance mechanisms that differed from previous findings. METHODS: The toxin genes and drug resistance genes of clinical strains were detected using Polymerase Chain Reaction (PCR), and these strains were subjected to Multilocus Sequence Typing (MLST). The agar dilution technique was employed for assessing susceptibility of antibiotics. Clinical data collection was completed through a review of electronic medical records. RESULTS: A total of 67 strains of toxin-producing C. difficile were detected. All C. difficile isolates demonstrated susceptibility to both metronidazole and vancomycin. However, resistance was observed in 8.95%, 16.42%, 56.72%, 56.72%, 31.34% and 5.97% of the isolates for tigecycline, tetracycline, clindamycin, erythromycin, moxifloxacin and rifampin, respectively. Among the strains with toxin genotypes A + B + CDT - and belonging to the ST3, six strains exhibited reduced susceptibility to tigecycline (MIC=0.5mg/L) and tetracycline (MIC=8mg/L). The tetA(P) and tetB(P) genes were present in these six strains, but were absent in tetracycline-resistant strains. Resistance genes (ermB, tetM, tetA(P) and tetB(P)) and mutations (in gyrA, gyrB, and rpoB) were identified in resistant strains. CONCLUSIONS: In contrast to prior studies, we found higher proportions of ST3 isolates with decreased tigecycline sensitivity, sharing similar resistance patterns and resistance genes. In the resistance process of tigecycline and tetracycline, the tetA(P) and tetB(P) genes may play a weak role.
Assuntos
Antibacterianos , Clostridioides difficile , Infecções por Clostridium , Hospitais de Ensino , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Clostridioides difficile/genética , Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/isolamento & purificação , Clostridioides difficile/classificação , China , Humanos , Antibacterianos/farmacologia , Infecções por Clostridium/microbiologia , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Toxinas Bacterianas/genética , Tigeciclina/farmacologia , Adulto , Farmacorresistência Bacteriana/genética , Genótipo , Metronidazol/farmacologia , Vancomicina/farmacologia , Reação em Cadeia da Polimerase , Farmacorresistência Bacteriana Múltipla/genéticaRESUMO
The development of amoebic liver abscess (ALA) leads to liver necrosis, accompanied by an exacerbated inflammatory response and the formation of multiple granulomas. Adequate management of the infection through the administration of treatment and the timely response of the organ to the damage allows the injury to heal with optimal regeneration without leaving scar tissue, which does not occur in other types of damage such as viral hepatitis that may conducts to fibrosis or cirrhosis. The Hedgehog signaling pathway (Hh) is crucial in the embryonic stage, while in adults it is usually reactivated in response to acute or chronic injuries, regeneration, and wound healing. In this work, we characterized Hh in experimental hepatic amoebiasis model, with the administration of treatment with metronidazole, as well as a pathway inhibitor (cyclopamine), through histological and immunohistochemical analyses including an ultrastructure analysis through transmission electron microscopy. The results showed an increase in the percentage of lesions obtained, a decrease in the presence of newly formed hepatocytes, a generalized inflammatory response, irregular distribution of type I collagen accompanied by the presence of fibroblast-type cells and a decrease in effector cells of this pathway. These results constitute the first evidence of the association of the activation of Hh with the liver regeneration process in experimental amebiasis.
Assuntos
Modelos Animais de Doenças , Proteínas Hedgehog , Regeneração Hepática , Transdução de Sinais , Regeneração Hepática/fisiologia , Proteínas Hedgehog/metabolismo , Animais , Abscesso Hepático Amebiano/patologia , Masculino , Fígado/patologia , Fígado/metabolismo , Metronidazol/farmacologia , Metronidazol/uso terapêutico , Alcaloides de Veratrum/farmacologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Helicobacter pylori (H. pylori) infections are on the rise, presenting a significant global health challenge. Mass Galla chinesis et camelliae Fermentata (Chinese gall leaven, CGL), a traditional Chinese medicinal product made from the fermentation of Rhus chinensis Mill., is frequently employed to address digestive system ailments. Contemporary pharmacological research reveals that CGL exhibits anti-inflammatory, anti-diarrheal, and enzyme-inhibitory activities and holds potential as a treatment for H. pylori infections. However, the precise mechanisms underlying CGL's efficacy against H. pylori remain to be fully elucidated. AIM: The objective of the study is to evaluate CGL's ability to disrupt the H. pylori biofilm and to explore its synergistic potential with antibiotics in targeting the biofilm-efflux pump system, a mechanism implicated in bacterial resistance. METHORDS: The study determined the Minimum Inhibitory Concentration (MIC) of CGL and metronidazole against H. pylori and evaluated their effects on H. pylori biofilms using an in vitro model. Structural changes induced by drug interventions were compared to those in untreated and antibiotic-treated models through scanning electron microscopy and laser confocal microscopy. The accumulation of H33342 dye in planktonic and biofilm H. pylori before and after drug treatment was assessed to evaluate cell viability and biofilm disruption. The study also involved adding experimental drugs to a biofilm H. pylori medium containing D-glucose, measuring glucose concentrations post-intervention using the glucose oxidase method, and calculating changes in glucose uptake. Finally, the relative expression levels of several genes in planktonic and biofilm H. pylori treated with CGL alone or in combination with antibiotics were measured to understand the impact on the biofilm-efflux pump system. RESULTS: Both CGL alone and in combination with metronidazole demonstrated effective disruption of H. pylori biofilms. The combination therapy was particularly effective in reducing the biofilm transfer-enhancing effect of metronidazole and decreasing SpoT expression in the 'SpoT-(p)ppGpp' pathway, especially in biofilms. It showed a greater inhibition of the 'σ54-gluP-sugar uptake' pathway, with significant reductions in rpoN and gluP expression under biofilm conditions compared to CGL or metronidazole alone. The treatment also suppressed H. pylori proliferation and may have altered glucose uptake mechanisms. Moreover, it significantly inhibited the 'hp0939/hp0497/hp0471-RND efflux pump' pathway, with a notable reduction in gene expression compared to the 1/2 MIC metronidazole treatment. CONCLUSION: This study demonstrates that CGL effectively hinders the development of drug resistance in H. pylori by targeting biofilm formation and critical molecular pathways associated with antibiotic resistance. The synergistic effect of combining CGL with metronidazole notably enhances biofilm disruption and inhibits the bacterium's metabolic and reparative mechanisms. Further in vivo studies are needed to confirm these results and to investigate additional mechanisms of CGL's action.
Assuntos
Antibacterianos , Biofilmes , Helicobacter pylori , Testes de Sensibilidade Microbiana , Biofilmes/efeitos dos fármacos , Helicobacter pylori/efeitos dos fármacos , Antibacterianos/farmacologia , Metronidazol/farmacologia , Rhus/química , Farmacorresistência Bacteriana/efeitos dos fármacos , Sinergismo Farmacológico , Extratos Vegetais/farmacologia , Medicamentos de Ervas Chinesas , TaninosRESUMO
Objective: Trichomonas vaginalis is a sexually transmitted protozoan parasite that usually causes infections in women. Metronidazole is used as the first choice in the treatment of this parasitic disease, but there is a need for new drugs since 1980's with increasing numbers of reported resistance. In this study, it was aimed to determine the antitrichomonal activity of the major components of Cinnamomum zeylanicum (cinnamon) and Thymus vulgaris (thyme) essential oils, cinnamaldehyde, carvacrol and thymol against metronidazole resistant and susceptible T. vaginalis strains, and to determine their interaction with metronidazole by checkerboard method. Methods: Cinnamaldehyde, carvacrol, thymol and metronidazole were obtained commercially. Two clinical isolates and one metronidazole resistant T. vaginalis reference strain were used in the study. MIC50 and MLC values of essential oil components and metronidazole were determined by broth microdilution method. The combinations of essential oil components with metronidazole were determined by the checkerboard method. Results: According to in vitro activity tests, cinnamaldehyde was determined to be most effective essential oil component. Clinical isolates were susceptible to metronidazole. In combination study, metronidazole showed synergy with cinnamaldehyde and carvacrol, and partial synergy with thymol. Conclusion: It was determined that cinnamaldehyde, carvacrol and thymol, which are known to have high antimicrobial activity, also have strong activity against T. vaginalis isolates and show a synergistic interaction with metronidazole. The use of metronidazole at lower doses in the synergistic interaction may contribute to the literature in terms of reducing drug side effects, creating a versatile antimicrobial target, and reducing the rate of resistance development.