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OBJECTIVES: Helicobacter pylori gastric infection strongly correlates with gastric diseases such as chronic gastritis, functional dyspepsia, and complications such as peptic ulcers and gastric cancer. In developing countries, systemic therapies are not usually successful due to elevated antibiotic resistance. Additionally, oral H. pylori infection and periodontal disease correlate with gastric treatment failures. This study aimed to explore the effect of an integral therapy, comprising oral hygiene and concomitant systemic treatment, to increase the eradication of gastric infection and recurrences. MATERIALS AND METHODS: A prospective, randomized, four-arm, parallel-group, open-label clinical trial was conducted to investigate the efficacy of integral therapy to eradicate gastric H. pylori infection and avoid recurrences in double-positive (real-time PCR oral and gastric infection) patients. Oral hygiene involved mouthwash with neutral electrolyzed water (NEW), with or without periodontal treatment. One hundred patients were equally distributed into four groups: NS, NS-PT, NEW, and NEW-PT. All patients had concomitant systemic therapy and additionally, the following oral treatments: mouthwash with normal saline (NS), periodontal treatment and mouthwash with normal saline (NS-PT), mouthwash with NEW (NEW), and periodontal treatment and mouthwash with NEW (NEW-PT). Gastric and oral infection and symptoms were evaluated one and four months after treatments. RESULTS: Integral therapy with NEW-PT increased gastric eradication rates compared with NS or NS-PT (84%-96% vs. 20%-56%; p < 0.001). Even more, a protective effect of 81.2% (RR = 0.1877; 95% CI: 0.0658-0.5355; p = 0.0018) against recurrences and 76.6% (RR = 0.2439; 95% CI: 0.1380-0.4310; p < 0.001) against treatment failure (eradication of infection and associated symptoms) was observed in patients from the NEW and NEW-PT groups. CONCLUSIONS: Implementation of oral hygiene and systemic treatment can increase the eradication of gastric infection, associated symptoms, and recurrences. NEW is recommended as an antiseptic mouthwash due to its efficacy and short- and long-term safety.
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Antibacterianos , Infecções por Helicobacter , Helicobacter pylori , Antissépticos Bucais , Higiene Bucal , Humanos , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/isolamento & purificação , Masculino , Feminino , Antissépticos Bucais/uso terapêutico , Antissépticos Bucais/administração & dosagem , Estudos Prospectivos , Adulto , Pessoa de Meia-Idade , Higiene Bucal/métodos , Antibacterianos/uso terapêutico , Antibacterianos/administração & dosagem , Resultado do Tratamento , Recidiva , Prevenção Secundária/métodos , Idoso , Terapia CombinadaRESUMO
Objective: Food insecurity (FI) is a priority for government and health organizations. Over 95% of the world's population has a carious lesion or will develop one before death. This study evaluated the association between FI and oral health in two rural communities in Chiapas, Mexico. Methods: The study was conducted with patients attending an oral health campaign for dental checkups. Data were collected between April and August 2017 using the Latin-American and Caribbean Scale of Food Security (ELCSA) and the International Caries Detection and Assessment System (ICDAS). We included 209 participants from Siltepec and Huehuetan, Mexico; 67% were women. Results: The results of the ELCSA were mild FI in 43% (n = 91), moderate FI in 22% (n = 45), and severe FI (n = 6) in 3%; 32% had food security. The ICDAS results were initial decay with a mean of 6.22, moderate decay with a mean of 1.81, and extensive decay with a mean of 1.77. Conclusions: FI is associated with dental caries, and food-insecure individuals have a higher probability of severe dental caries. In this study, the FI level was lower than in other rural populations in Mexico. Identifying these individuals and addressing the factors related to FI can be useful in the rural communities.
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Purpose: Epidemiological studies have been conducted to improve the health and economic quality of life of indigenous communities in Mexico. These studies have found that infections cause frequent health problems. Helicobacter pylori are responsible for conditions ranging from gastritis to stomach cancer. This study determined the prevalence of H. pylori in families from Siltepec, Chiapas, Mexico. Patient and Methods: Ninety-nine dental plaque samples from 36 families were studied. Real-time PCR was performed to detect H. pylori using previously reported primers. The Mann-Whitney U-test was used for the statistical analysis. According to the family role of H. pylori-positive individuals, the VacA s1/m1 genotype and CagA gene correlated. Results: The mother had the highest expression of VacA s1/m1-/cagA- with 19% (8/42), followed by the first child with 14.3% (6/42). The major roles for the vacA s1/m1+/cagA- were the mother and first child with 9.5% (4/42), followed by the remaining children with 4.8% (2/42). The vacA s1/m1-/cagA+ genotype was 7.1% (3/42) for the mother and 4.8% (2/42) for the father. Finally, the vacA s1/m1+/cagA+ genotype only appeared in the mother, son I, and son III with 2.4% (1/42). Conclusion: The vacA s1/m1/cagA genotypes predominated in the mother, suggesting potential transmission between the mother and child during the first years of life.
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Culture conditions affect the production of secondary metabolites in endophytic fungi. Therefore, the aim of the present study was to evaluate the yield and anticancer and antioxidant activity of endophytic fungi extracts from the cactus Lophocereus marginatus, under different culture conditions. The strains Penicillium citrinum, Aspergillus versicolor, Metarhizium anisopliae, and Cladosporium sp. were fermented in different culture media (potato dextrose agar, Czapeck broth, and malt broth), types of inoculums (spore or mycelium), and shaking conditions (150 rpm or static) for one week. Methanol extracts were obtained from mycelia, which was followed by determining their yields and evaluating their effect on L5178Y-R murine lymphoma cells growth and human peripheral blood mononuclear cells (PBMCs) viability, using the 3-[4,5dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide reduction colorimetric assay. In addition, antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl test. We determined the half-maximal inhibitory concentration (IC50) values of tumor cell growth inhibition, the selectivity index (SI), and the antioxidant activity, as compared with the healthy cells control. The best yields were obtained with the Czapeck broth medium in all the evaluated strains, reaching values of 50.3%. Of the 48 extracts evaluated, only seven significantly (p < 0.01) inhibited tumor cell growth (IC50 < 250 µg/mL). A. versicolor extract showed the highest anticancer activity, after culturing spores (IC50 = 49.62 µg/mL; SI = 15.8) or mycelium (IC50 = 69.67 µg/mL; SI = 12.2) in malt broth, under static conditions. Extracts did not present significant antioxidant activity. In conclusion, we showed that culture conditions influenced the anticancer activity of L. marginatus endophytic fungi.
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Antioxidantes , Leucócitos Mononucleares , Humanos , Animais , Camundongos , Antioxidantes/farmacologia , Fungos , Meios de CulturaRESUMO
It is indisputable that every day it is demonstrated that natural products present diverse therapeutic benefits, which has boosted their incorporation within various products for clinical use. However, this must be accompanied by knowledge of their effect on cell lines to ensure their use is safe. The objective of this study was to evaluate the cytotoxic effect of two ethanolic extracts based on Peruvian natural products, on three human cell lines. Cervical cancer cell lines (HeLa), human gingival fibroblasts (HGF-1 - ATCC CRL-2014) (HGF-1) and peripheral blood mononuclear cells (PBMCs) were cultured and subsequently treated with preparations of ethanolic extracts of propolis (EEP) and Psidium guajava (EEG) from a concentration of 50 mg/mL to 0.024 mg/mL, by the 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazole bromide reduction assay. At a concentration of 0.24 mg/mL EEG, viability of 99.7±1.24%, 99.8±2.2% and 99.7±2.7% was observed in HeLa, HGF-1 and PBMCs, respectively; >90% cell viability values were observed with EPP at 0.024 mg/mL, with HGF-1 showing the highest viability (96.9±1.15%). A dose-dependent effect was observed for both extracts with a decrease in cell viability as concentrations increased (up to 50 mg/mL). EEP and EEG extracts at low concentrations do not show cytotoxicity in human cell lines, these findings are an advance in the preclinical evaluation on their safety and open a continuity to further studies for their potential applications in dentistry and medicine.
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Própole , Psidium , Fibroblastos , Células HeLa , Humanos , Leucócitos Mononucleares , Peru , Extratos Vegetais/farmacologia , Própole/farmacologiaRESUMO
BACKGROUND: Stage-specific embryonic antigen-4 (SSEA-4) is a marker for the identification of multipotent embryonic cells. It is also positive in neuroepithelial cells, precursor neural cells (NPC), and human dental pulp cells. The aim of this study was to evaluate the potential morphodifferentiation and histodifferentiation to NPC of SSEA-4 positive stem cells from human exfoliated deciduous teeth (SHED). METHODS: A SHED population in culture, positive to SSEA-4, was obtained by magnetic cell separation. The cells were characterized by immunohistochemistry and flow cytometry. Subsequently, a neurosphere assay was performed in a medium supplemented with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF); afterward, cells were neurodifferenciated with a neurobasal medium. Finally, indirect immunohistochemistry was performed to identify neuronal markers. RESULTS: The morphological and histological changes in the SSEA-4 positive SHEDs were observed after induction with epidermal and fibroblast growth factors in neurobasal culture medium. At the end of induction, the markers Nestin, TuJ-1, and GFAP were identified. CONCLUSIONS: The findings show that SSEA-4 positive SHEDs have a behavior similar to neuronal precursor cells. Our findings indicate that the dental pulp of deciduous teeth is a promising source for regeneration therapies associated with neurodegenerative diseases or peripheral nerve alterations.
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Polpa Dentária , Células-Tronco Neurais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Antígenos Embrionários Estágio-Específicos , Dente DecíduoRESUMO
Objective: Mouthwash is effective in maintaining oral hygiene in patients; however, there is concern that it may adversely affect human oral mucosa. We evaluated a pH-neutral electrolyzed super-oxidized solution (ESS, tradename OxOral®) combined with dental scaling in periodontitis patients. This longitudinal study was conducted with 34 patients divided into three groups. The control group treated with scaling plus saline, the second with scaling plus ESS mouthwash, and another with scaling plus ESS mouthwash and gel. The plaque index (PI), gingival index (GI), and probing depth (PD) were determined before and after periodontal treatment. Results: The final PI and GI decreased compared with the initial measurements in the three treatment groups (p < 0.05). Scaling plus ESS mouthwash and gel significantly reduced the final PI, GI, and DP compared to the control group (p < 0.05). Conclusion: Our study shows the antiseptic properties of ESS with mouthwash and gel. Further studies are needed to verify the results.
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Introducción: El SARS-CoV-2 afecta el sistema respiratorio en diferentes grados. La cavidad oral es el lugar más colonizado por bacterias, por lo tanto, al no tener una adecuada higiene pueden presentarse diferentes enfermedades secundarias, lo que ha causado alerta en el gremio odontológico, ya que puede contribuir a complicaciones posteriores en los pacientes. Material y métodos: El estudio fue conformado por 47 pacientes voluntarios recuperados de SARS-CoV-2, residentes de Montemorelos, Nuevo León, México, donde fueron atendidos en Bucalia Dent, consultorio dental. Después del consentimiento informado de cada paciente, se realizó una historia clínica para conocer los síntomas, enfermedades sistémicas, ausencia de dientes y nivel de inflamación gingival de acuerdo al índice de Loe y Silness. A continuación, se tomó una muestra de biofilm microbiano (placa dentobacteriana), la cual se suspendió en una solución buffer de fosfato, posteriormente fue llevada al Centro de Investigación y Desarrollo en Ciencias de la Salud (CIDICS), Monterrey, N.L, México. Se extrajo DNA y se purificó, después se realizó PCR para detectar los patógenos orales; la PCR se visualizó en gel de agarosa (1.5%) por tinción de bromuro de etidio. Resultados: Se detectó 80.85% Porphyromona gingivalis y 68.09% Fusobacterium nucleatum en pacientes recuperados de SARS-CoV-2; 23.4% presentaron inflamación leve de acuerdo al índice de Loe y Silness, 54.5% fueron masculinos y 45.5% femeninos. Por otro lado, 36.4% de los pacientes con inflamación leve tenían de cuatro a seis dientes ausentes. En estos pacientes se detectó 18.18% únicamente con Fusobacterium nucleatum y 27.27% sólo con Porphyromona gingivalis; el sexo masculino tuvo predisposición en 66.6% y el femenino en 33.33%. Se observó infección con los dos patógenos presentes en 45.45%; y 60% de estos pacientes fueron masculinos. Conclusiones: Los pacientes recuperados de SARSCoV- 2 analizados en esta investigación mostraron mala higiene oral y alta prevalencia de los patógenos mencionados altamente relacionados a inflamación gingival o enfermedad periodontal, lo que nos indica que es indispensable la intervención del odontólogo al finalizar el periodo de infección de cada paciente (AU)
Introduction: SARS-CoV-2 affects the respiratory system to different degrees. The oral cavity is a colonized place by bacterias, therefore, by not having good hygiene, different secondary diseases can occur; this has caused an alert in the dental industry, since it can contribute to later complications in patients. Material and methods: The study was conducted in 47 SARS-CoV-2 recovered volunteers from the Montemorelos city of the Nuevo León state, Mexico, who were attended at the Bucalia Dent dental clinic. An informed consent was obtained from each of the patients, then their clinical history was documented in order to know the symptoms, previous systemic diseases, absence of teeth and degree of gingival inflammation, as suggested by Loe and Silness. Subsequently, a dental plaque sample was taken from all patients, which was suspended in a phosphate buffered solution and shipped to The Center for Research and Development in Health Sciences (CIDICS), Monterrey, NL, Mexico for storage. DNA extraction and purification was performed and PCR was carried out for the oral pathogens detection. All PCR products were visualized on 1.5% agarose gel by ethidium bromide staining. Results: Porphyromona gingivalis and Fusobacterium nucleatum were detected in 80.85% and 68.09% of SARS-CoV-2 recovered patients, respectively. 23.4% showed mild inflammation based on the Loe and Silness criteria, 54.5% were male and 45.5% female. On the other hand, 36.4% of patients with mild inflammation had between 4 to 6 missing teeth. A single infection by Fusobacterium nucleatum was detected in 18.18% and by Porphyromona gingivalis in 27.27%; the male sex had a predisposition with 66.66% and 33.33% female; coinfection of both pathogens was observed in 45.45% where 60% were male. Conclusions: SARS-CoV-2 recovered patients show poor oral hygiene and a high prevalence of oral pathogens related to the development of inflammatory gingival or periodontal disease, this suggests the need for an odontological clinical intervention at the end of the course of infection or disease caused by SARS-CoV-2 (AU)
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Humanos , Masculino , Feminino , Adulto , Higiene Bucal , Fusobacterium nucleatum , Porphyromonas gingivalis , SARS-CoV-2 , DNA , Índice de Higiene Oral , Índice Periodontal , Reação em Cadeia da Polimerase , Placa Dentária/microbiologia , Eletroforese em Gel de Ágar , Distribuição por Idade e Sexo , Gengivite/epidemiologia , MéxicoRESUMO
OBJECTIVE: To evaluate the effect of the combination of calcium hydroxide (Ca(OH)2) and a novel electrolyzed superoxidized solution at neutral pH, known as OxOral® on Enterococcus faecalis growth in root canals. METHODS: Sixty human teeth were used, from which root canals were infected and randomly divided into the following treatment groups: saline solution, saline solution plus Ca(OH)2, OxOral®, and OxOral® plus Ca(OH)2. RESULTS: A permanent reduction in bacterial growth was observed at days 1, 6, 12, and 18 after OxOral® plus Ca(OH)2 treatment from 4.4 ± 0.074 log10 CFU/mL to 0.0 ± 0.001 log10 CFU/mL. In addition, alkaline conditions maintenance was observed from application time (pH = 12.2 ± 0.033) to 18 d posttreatment (pH = 12.6 ± 0.083). CONCLUSION: The combination of OxOral® and Ca(OH)2 provides an alkaline pH and inhibits E. faecalis growth into the root canals. Our study opens the possibility for further research on the use of OxOral® in endodontic therapy.
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Anti-Infecciosos/administração & dosagem , Hidróxido de Cálcio/administração & dosagem , Cavidade Pulpar/efeitos dos fármacos , Cavidade Pulpar/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Peróxido de Hidrogênio/administração & dosagem , Enterococcus faecalis/crescimento & desenvolvimento , Humanos , Peróxido de Hidrogênio/química , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Irrigantes do Canal Radicular/administração & dosagem , Irrigantes do Canal Radicular/química , Tratamento do Canal Radicular/métodos , SoluçõesRESUMO
OBJECTIVE: To evaluate the antimicrobial activity of a silver nanoparticles/carboxymethyl-cellulose (AgNPs/CMC) composite on in vitro and dentine disc heterogeneous biofilms. DESIGN: AgNPs/CMC composite effect on normal human gingival fibroblast cells (HGF) viability was determined by the MTT reduction assay. In addition, we evaluated the antimicrobial effect of AgNPs/CMC composite on Candida albicans, Enterococcus faecalis, and Fusobacterium nucleatum growth in vitro and heterogeneous biofilms, as well as dentine disc biofilms. RESULTS: Quasi-spherical AgNPs/CMC composites, with a mean 22.3â¯nm particle-size were synthesized. They were not toxic to HGF cells at concentrations tested that were antimicrobial, however they caused significant cytotoxicity (89 %, pâ¯<⯠0.05) at concentrations > 15 µg/mL. In vitro, they inhibited up to 67 %, 66 %, and 96 % C. albicans, E. faecalis, and F. nucleatum growth at concentrations ranging from 1.2 µg/mL to 9.6 µg/mL, as compared with untreated control. We also demonstrated significant (pâ¯<⯠0.05) 58 % biofilm reduction by 4.8 µg/mL AgNPs/CMC composite on human dentine discs. CONCLUSION: AgNPs/CMC composite showed anti biofilm activity on monocultures, heterogenous cultures, and dentine discs, resulting a potentially effective alternative to prevent and eliminate infections after endodontic treatment.
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Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Dentina/microbiologia , Nanopartículas Metálicas , Prata/farmacologia , Carboximetilcelulose Sódica/química , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Gengiva/citologia , Humanos , Testes de Sensibilidade MicrobianaRESUMO
The titanium alloy, Ti6Al4V, is used in dentistry for dental implants because of its excellent resistance to corrosion and its high biocompatibility. However, periimplantitis is considered the main reason for treatment failure. The Ti6Al4V alloy was used to study the corrosion behavior for dental implant applications, using an experimental arrangement of three electrodes with the bacteria Streptococcus gordonii and Fusobacterium nucleatum, in addition to Ringer's lactate as electrolytes, at 37 °C and a pH of 5.6. Their electrochemical behavior was studied by open circuit potential (OCP) and cyclic potentiodynamic polarization (CPP) according to ASTM G3-14 and ASTM G61-11, respectively. Scanning electron microscopy (SEM) was employed to determine the morphology of the alloy studied. An experimental model, in situ, was established with the bacteria present in an oral environment to understand the electrochemical behavior of the alloy used in dental implants. The greatest corrosion in Ti6Al4V alloy was produced by the medium that contained the bacterium Streptococcus gordonii, which is considered a primary colonizer. In addition, the Ti6Al4V alloy presented uniform corrosion in the three solutions at the different exposure times showing a negative hysteresis in CPP.
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Human papilloma virus (HPV) is a DNA virus associated with the development of cervical, penile, anal, vulvar, and oral cancers. In recent years, there has been an increase in oral cancer, which could be due to changes in sexual behavior in the general population. In México, there is scarce information on this regard, which prompted us to study HPV infection prevalence in the oral cavity of an indigenous community from the municipality of Siltepec, Chiapas, Mexico. Oral samples from 198 individuals were obtained with cytobrush for virus detection by nested PCR, using MY09/MY11 and GP5+/GP6+ primers, and positive samples were sequenced for HPV genotyping. We observed 12.1% HPV infection prevalence, which depended on gender, number of sexual partners, lack of using condoms, and oral sex practices. In contrast, no significant association between HPV infection and tobacco or alcohol consumption was detected. Furthermore, sequencing analyzes were performed where HPV-13 (21/24), -16 (2/24), -32 (1/24), -81 (1/24), and -83 (1/24) were evidenced and HPV-16 European/Asian and Asian/American E6 variants identified. These results demonstrated an important prevalence of HPV infection in the oral cavity of a Mexican indigenous community, where the predominant genotypes were associated with benign pathologies, and showed that high-risk genotype variants derived from different lineages.
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Boca/virologia , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Adulto , Idoso , Estudos Transversais , Feminino , Genótipo , Neoplasias de Cabeça e Pescoço/virologia , Humanos , Povos Indígenas/estatística & dados numéricos , Masculino , México/epidemiologia , México/etnologia , Pessoa de Meia-Idade , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Prevalência , Comportamento Sexual , Parceiros Sexuais , Carcinoma de Células Escamosas de Cabeça e Pescoço/virologiaRESUMO
BACKGROUND: The oral microenvironment provides the conditions for the establishment of microorganisms not usually considered residents of the normal oral microbiota. Sexually transmitted microorganisms such as Chlamydia trachomatis can adhere to any mucosal surface and ascend to reach appropriate locations to survive and develop symptomatic infections. MATERIALS AND METHODS: To determine the presence of C. trachomatis, direct immunofluorescence of this microorganism was carried out in 76 randomly selected patients attending a periodontal clinic during a period of 1 year. Samples from the gingival sulcus and the pharynx were collected for detection of C. trachomatis. Patients who attended the periodontal clinic were divided into two groups: those without periodontitis and those with periodontitis. For the purpose of performing other statistical analyses, all patients were also divided by gender and age. RESULTS: From the total of 76 patients, in the group without periodontitis, 61% were positive for C. trachomatis in the gingival sulcus and 63.4% in the pharynx; in the periodontitis group, 45.7% were positive in the sulcus and 40% in the pharynx. When we compared patients by gender or age, no statistical difference was found. CONCLUSIONS: The prevalence of C. trachomatis in this group was 53.9% in the gingival sulcus and pharynx of the studied patients.
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Infecções por Chlamydia/epidemiologia , Chlamydia trachomatis/isolamento & purificação , Gengiva/microbiologia , Periodontite/microbiologia , Faringe/microbiologia , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Infecções por Chlamydia/diagnóstico , Infecções por Chlamydia/microbiologia , Estudos Transversais , Feminino , Técnica Direta de Fluorescência para Anticorpo , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Periodontite/diagnóstico , Periodontite/patologia , Prevalência , Adulto JovemRESUMO
BACKGROUND/PURPOSE: Helicobacter pylori (H. pylori) infection is the most common in the world and is associated with various gastrointestinal pathologies, including chronic gastritis, peptic ulcers, and gastric cancer. The prevalence is associated with socioeconomic conditions, with this infection being more common in developing countries than in developed countries. The presence and permanence of H. pylori in the oral cavity has been reported, but its role is controversial. The aim of this study was to determine the prevalence of H. pylori in dental plaque of patients with periodontitis. MATERIALS AND METHODS: A cross-sectional study was carried out and Periodontal Screening and Recording (PSR) index was determined. 38 dental plaque samples were taken and total DNA was extracted and qPCR was performed. RESULTS: 60.5% of the samples (nâ¯=â¯23) were positive for the presence of H. pylori by the amplification of the 16S rRNA and vacA genes. In addition, cagA gene was detected in 21.7% (nâ¯=â¯5) of H. pylori-positive. A significant relationship between periodontal status and H. pylori oral infection was found (Pâ¯≤â¯0.05); patients with initial and moderate periodontitis were the most affected with 39.1% and 30.4%, respectively. CONCLUSION: Our results suggest that the prevalence of H. pylori in the oral cavity could be related to the progression of periodontal disease. Therefore, oral hygiene and treatment for the elimination of oral H. pylori could stop the progression of periodontal disease.
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Helicobacter pylori is an infectious agent that colonizes the gastric mucosa of half of the population worldwide. This bacterium has been recognized as belonging to group 1 carcinogen by the World Health Organization for the role in development of gastritis, peptic ulcers, and cancer. Due to the increase in resistance to antibiotics used in the anti-H. pylori therapy, the development of an effective vaccine is an alternative of great interest, which remains a challenge. Therefore, a rational, strategic, and efficient vaccine design against H. pylori is necessary where the use of the most current bioinformatics tools could help achieve it. In this study, immunoinformatics approach was used to design a novel multiepitope oral vaccine against H. pylori. Our multiepitope vaccine is composed of cholera toxin subunit B (CTB) that is used as a mucosal adjuvant to enhance vaccine immunogenicity for oral immunization. CTB fused to 11 epitopes predicted of pathogenic (UreB170-189, VacA459-478, CagA1103-1122, GGT106-126, NapA30-44, and OipA211-230) and colonization (HpaA33-52, FlaA487-506, FecA437-456, BabA129-149, and SabA540-559) proteins from H. pylori. CKS9 peptide (CKSTHPLSC) targets epithelial microfold cells to enhance vaccine uptake from the gut barrier. All sequences were joined to each other by proper linkers. The vaccine was modeled and validated to achieve a high-quality three-dimensional structure. The vaccine design was evaluated as nonallergenic, antigenic, soluble, and with an appropriate molecular weight and isoelectric point. Our results suggest that our newly designed vaccine could serve as a promising anti-H. pylori vaccine candidate.
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Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Epitopos/imunologia , Infecções por Helicobacter/prevenção & controle , Helicobacter pylori/imunologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/farmacologia , Administração Oral , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/química , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/química , Toxina da Cólera/administração & dosagem , Toxina da Cólera/imunologia , Biologia Computacional , Epitopos/administração & dosagem , Infecções por Helicobacter/imunologia , Helicobacter pylori/química , Humanos , Camundongos , Modelos Moleculares , Estrutura Secundária de ProteínaRESUMO
Helicobacter pylori is a spiral Gram-negative bacterium associated with inflammation of the gastric mucosa, peptic ulcer, and gastric adenocarcinoma, whose treatment has failed due to antibiotic resistance and side effects. Furthermore, because there are no vaccines effective against H. pylori, an appropriate vaccine design targeting conserved/essential genes must be identified. In the present study, a H. pylori 50-52 kDa immunogen-derived peptide antigen with the sequence Met-Val-Thr-Leu-Ile-Asn-Asn-Glu (MVTLINNE) was used to immunize against H. pylori infection. For this, mice received an intraperitoneal injection of 100 µg of H. pylori peptide on the first week, followed by two weekly subcutaneous reinforcements and further 109 bacteria administration in the drinking water for 3 weeks. Thymic cells proliferative responses to concanavalin A, serum levels of IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ, and TNF-α cytokines, and IgG1, IgG2a, IgG2b, IgG3 IgM, and IgA immunoglobulins were evaluated. Significant (p < 0.05) increases on lymphoproliferation and spleen weights after immunization were observed. In contrast, infection significantly (p < 0.05) decreased lymphoproliferation, which was recovered in immunized mice. In addition, levels of serum TH1 and TH2 cytokines were not altered after immunization, except for the significant increase in IL-6 production in immunized and/or infected animals. Moreover, immunization correlated with plasma secretory IgA and IgG, whereas infection alone only elicited IgM antibodies. Peptide immunization protected 100% of mice against virulent H. pylori. MVTLINNE peptide deserves further research as an approach to the prophylaxis of H. pylori infection.
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The presence of Helicobacter pylori in the oral cavity has been associated to the failure of antimicrobial therapy in patients with gastrointestinal infection and the development of oral diseases. However, it has been reported that the maintenance of good oral hygiene can improve the therapeutic success rates, where the use of mouthwashes with anti-Helicobacter activity would help to achieve it. The aim was to evaluate the antimicrobial activity of OxOral® mouthwash against H. pylori and its effect on biofilm formation. The minimum inhibitory concentration (MIC) of OxOral® (pH = 6.4-7.5, ORP = 650-900 mV) against H. pylori was calculated testing serial dilutions 0.117-15 ppm against 1 × 108 CFU/mL of H. pylori (ATCC® 700824™) by broth microdilution method using 96-well plates. The H. pylori biofilm formation was determined by the optical density measurement at 600 nm from coverslips stained with 0.1% crystal violet. The gene expression of ureA, luxS, flaA, omp18, and lpxD were analyzed by RT-qPCR. OxOral® cytotoxicity was evaluated in a human gingival fibroblast cell line by MTT assay. MIC was of 3.75 ppm, with 99.7 ± 7.7% bacterial growth inhibition. In the negative control, the biofilm formation was observed, whereas when bacteria were treated with OxOral® at 0.234, 0.469, and 0.938 ppm, an inhibition of 35.5 ± 0.9%, 89.1 ± 1.2%, and 99.9 ± 5.5% were obtained, respectively. The gene expression analysis showed that flaA, omp18, and lpxD genes were down-regulated with OxOral® compared with control (p < 0.05). Low cytotoxicity of 16.5 ± 7.6% was observed at the highest dose (15 ppm); no significant differences were observed from 15 to 0.469 ppm compared to the control of untreated cells (p > 0.05). Our results reveal an important anti-Helicobacter activity of OxOral® and open the possibility of its therapeutic use new studies, which would increase the success rate of conventional therapies against H. pylori.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Helicobacter pylori/efeitos dos fármacos , Antissépticos Bucais/farmacologia , Linhagem Celular , Linhagem Celular Tumoral , Fibroblastos/microbiologia , Gengiva/microbiologia , Infecções por Helicobacter , Humanos , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana/métodosRESUMO
[This corrects the article DOI: 10.1155/2017/2052938.].
RESUMO
The variability in Helicobacter pylori vacA and cagA genes has been related to the progression of the gastrointestinal disease; also the presence of H. pylori in the oral cavity has been associated with periodontal disease in adults, but, in children without dyspeptic symptoms, little is known about this. We evaluated the prevalence of H. pylori and the presence of vacA/cagA genotypes in the oral cavity of Mexican children without dyspeptic symptoms. The gingival status was measured, and dental plaque samples (n = 100) were taken. 38% of children were positive for H. pylori 16S rRNA gene by qPCR. A significant association between H. pylori oral infection and gingival status was observed (P < 0.001). In 34.6% (9/26) of mild gingivitis cases, s1m2 genotype was found, while s1m1 was typed in 50% (3/6) of moderate gingivitis. The cagA prevalence among H. pylori-positive children was 80.8% (21/26), 83.3% (5/6), and 16.7% (1/6) of cases of mild gingivitis, moderate gingivitis, and nongingivitis, respectively (P < 0.001). The s1m1/cagA+ combinational genotype was the most detected in children with gingivitis. Our results suggest that the prevalence of H. pylori and detection of vacA/cagA genotypes-associated gastrointestinal disease in the oral cavity could be related to the progression of gingivitis in asymptomatic children.
Assuntos
Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Placa Dentária/microbiologia , Infecções por Helicobacter/epidemiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/genética , Adolescente , Criança , Pré-Escolar , Feminino , Genótipo , Humanos , Masculino , México , Prevalência , RNA Ribossômico 16S/genéticaRESUMO
Αlpha-solanine (α-solanine) is a glycoalkaloid present in potato (Solanum tuberosum). It has been of particular interest because of its toxicity and potential teratogenic effects that include abnormalities of the central nervous system, such as exencephaly, encephalocele, and anophthalmia. Various types of cell culture have been used as experimental models to determine the effect of α-solanine on cell physiology. The morphological changes in the mesenchymal stem cell upon exposure to α-solanine have not been established. This study aimed to describe a reliable and reproducible model for assessing the structural changes induced by exposure of mouse bone marrow mesenchymal stem cells (MSCs) to different concentrations of α-solanine for 24 h. The results demonstrate that nonlethal concentrations of α-solanine (2-6 µM) changed the morphology of the cells, including an increase in the number of nucleoli, suggesting elevated protein synthesis, and the formation of spicules. In addition, treatment with α-solanine reduced the number of adherent cells and the formation of colonies in culture. Immunophenotypic characterization and staining of MSCs are proposed as a reproducible method that allows description of cells exposed to the glycoalkaloid, α-solanine.