Tuberculosis in pregnancy and adverse neonatal outcomes in two peruvian hospitals.

Background: According to the World Health Organization, tuberculosis (TB) ranks among the top 10 causes of death worldwide. The significance of TB during pregnancy lies in its symptoms, which can be mistaken for physiological changes associated with pregnancy. This confusion can lead to maternal-perinatal complications. Objective: To evaluate the association between pulmonary TB in pregnancy and adverse neonatal outcomes in two Peruvian hospitals. Methods: This is a retrospective cohort study. The target population consisted of pregnant women with and without pulmonary TB whose deliveries were attended at two public hospitals, located in Lima, Peru. The adverse neonatal outcomes were prematurity, low birth weight (LBW), and being small for gestational age (SGA). Crude and adjusted relative risks (RRa) were calculated with their respective 95% confidence intervals (95%CI). Results: Information from 212 patients was analyzed; 48.1% had TB during pregnancy, and 23.1% had adverse neonatal outcomes (8%, 11.3%, and 12.3% for LBW, prematurity, and SGA, respectively). In the adjusted model, pregnant women with pulmonary TB had a 3.52 times higher risk of having a newborn with at least one of the adverse outcomes than those who were not exposed (aRR, 3.52; 95%CI: 1.93-6.68). Conclusion: Pulmonary TB in pregnancy was jointly and independently associated with adverse neonatal outcomes, including LBW, prematurity, and being SGA.

Association between early sexual initiation and sexually transmitted infections among Peruvian reproductive-age women.

Background: Sexually transmitted infections (STIs) are a serious public health problem worldwide, especially among reproductive-age women. The early sexual onset of sexual intercourse (EOSI) has been suggested as a risk factor, although there is no data at the national level. Objective: To evaluate the association between EOSI and STIs in Peruvian women of childbearing age. Methods: Analytical cross-sectional study with secondary data analyzes of the Peruvian Demographic and Family Health Survey 2018. The outcome was the presence of STIs in the last 12 months and the exposure variable was EOSI (age < 15 years at the time of their first sexual experience). To evaluate the association of interest, crude and adjusted prevalence ratios (aPRs) were calculated using generalized linear models with Poisson family and logarithmic link function. Results: We analyzed data from 31,028 women of childbearing age. The 11.3% reported having STIs in the last 12 months and 20.2% of the participants had an EOSI. After adjusting for potential confounders, we found that EOSI was associated with STIs (aPR: 1.27; 95% CI: 1.08-1.50; p = 0.005). When conducting stratified analysis by area of residence and number of sexual partners, this association was maintained in women living in urban areas (aPR: 1.36; 95% CI: 1.11-1.66; p = 0.003) those who did not report having a history of multiple sexual partners (aPR: 1.27; 95% CI: 1.08-1.51; p = 0.005), and those in the middle (aPR: 1.42; 95% CI: 1.03-1.97; p = 0.034) and highest (aPR: 2.12; 95% CI: 1.33-3.39; p = 0.002) wealth quintiles. Conclusion: Among reproductive-age women from Peru, EOSI was associated with STIs, especially in women living in urban areas, with no history of multiple sexual partners, and belonging to the middle to higher wealth index. The implementation of measures to prevent EOSI and fostering appropriate sexual health counseling for women with EOSI is advised.

A Prospective Cohort of Alcohol Use and Alcohol-related Problems Before and After Metabolic and Bariatric Surgery in Adolescents.

OBJECTIVE: To describe alcohol use, alcohol-related harm, and alcohol-related problems preoperatively and up to 8 years following metabolic and bariatric surgery (MBS) in adolescents. BACKGROUND: Risk for alcohol use and alcohol use disorders (AUD) increases post-Roux-en-Y gastric bypass and vertical sleeve gastrectomy in adults. However, data are lacking in adolescents who undergo MBS. METHODS: This study includes 217 adolescents (aged 13-19 y) enrolled in a 5-center prospective cohort study who underwent Roux-en-Y gastric bypass or vertical sleeve gastrectomy (2007-2011) and reported alcohol use preoperatively and annually postoperatively for up to 8 years. Time to elevated Alcohol Use Disorder Identification Test-Consumption (AUDIT-C) score, alcohol-related harm, and alcohol-related problems were analyzed with Kaplan-Meier estimates of cumulative incidence. RESULTS: Preoperatively, the median age was 17 years; the median body mass index was 51 kg/m 2 . Alcohol use frequency and average quantity of drinks per drinking day increased postoperatively (2% consumed alcohol 2-4 times/month 6 months versus 24% 8 years postoperatively, P <0.001; 2% consumed≥3 drinks per drinking day 6 months versus 35% 8 years postoperatively, P <0.001). Cumulative incidence of postoperative onset elevated AUDIT-C score, alcohol-related harm, and alcohol-related problems at year 8 were 45% (95% CI:37-53), 43% (95% CI:36-51), and 47% (95% CI:40-55), respectively. CONCLUSIONS: Nearly half of those who underwent MBS as adolescents screened positively for AUD, symptoms of alcohol-related harm, or alcohol-related problems 8 years post-MBS, highlighting the risk for alcohol use and AUD after MBS in adolescents. AUD evaluation and treatment should be integrated into routine long-term care for adolescents undergoing MBS.

Reduction in Long-term Mortality After Sleeve Gastrectomy and Gastric Bypass Compared to Nonsurgical Patients With Severe Obesity.

OBJECTIVE: To separately compare the long-term risk of mortality among bariatric surgical patients undergoing either Roux-en-Y gastric bypass (RYGB) or sleeve gastrectomy (SG) to large, matched, population-based cohorts of patients with severe obesity who did not undergo surgery. BACKGROUND: Bariatric surgery has been associated with reduced long-term mortality compared to usual care for severe obesity which is particularly relevant in the COVID-19 era. Most prior studies involved the RYGB operation and there is less long-term data on the SG. METHODS: In this retrospective, matched cohort study, patients with a body mass index ≥35 kg/m 2 who underwent bariatric surgery from January 2005 to September 2015 in three integrated health systems in the United States were matched to nonsurgical patients on site, age, sex, body mass index, diabetes status, insulin use, race/ethnicity, combined Charlson/Elixhauser comorbidity score, and prior health care utilization, with follow-up through September 2015. Each procedure (RYGB, SG) was compared to its own control group and the two surgical procedures were not directly compared to each other. Multivariable-adjusted Cox regression analysis investigated time to all-cause mortality (primary outcome) comparing each of the bariatric procedures to usual care. Secondary outcomes separately examined the incidence of cardiovascular-related death, cancer related-death, and diabetes related-death. RESULTS: Among 13,900 SG, 17,258 RYGB, and 87,965 nonsurgical patients, the 5-year follow-up rate was 70.9%, 72.0%, and 64.5%, respectively. RYGB and SG were each associated with a significantly lower risk of all-cause mortality compared to nonsurgical patients at 5-years of follow-up (RYGB: HR = 0.43; 95% CI: 0.35,0.54; SG: HR = 0.28; 95% CI: 0.13,0.57) Similarly, RYGB was associated with a significantly lower 5-year risk of cardiovascular-(HR = 0.27; 95% CI: 0.20, 0.37), cancer- (HR = 0.54; 95% CI: 0.39, 0.76), and diabetes-related mortality (HR = 0.23; 95% CI:0.15, 0.36). There was not enough follow-up time to assess 5-year cause-specific mortality in SG patients, but at 3-years follow-up, there was significantly lower risk of cardiovascular- (HR = 0.33; 95% CI:0.19, 0.58), cancer- (HR = 0.26; 95% CI:0.11, 0.59), and diabetes-related (HR = 0.15; 95% CI:0.04, 0.53) mortality for SG patients. CONCLUSION: This study confirms and extends prior findings of an association with better survival following bariatric surgery in RYGB patients compared to controls and separately demonstrates that the SG operation also appears to be associated with lower mortality compared to matched control patients with severe obesity that received usual care. These results help to inform the tradeoffs between long-term benefits and risks of bariatric surgery.

INCIDENCE OF INFLAMMATORY BOWEL DISEASE: A SINGLE CENTRE RETROSPECTIVE STUDY.

BACKGROUND: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder affecting the gastrointestinal tract. The etiology of this alarming condition is multifactorial. A Recently increasing trend in IBD is noted in our country. OBJECTIVE: The present study was designed with the main objective to assess the incidence and to identify the associated risk factors including demographic, geographical areas, and dietary patterns of IBD population of Northern of Karnataka viz. Hubli-Dharwad city. METHODS: A retrospective investigation was conducted on a cohort of 226 patients with a working diagnosis of IBD and those who were admitted between 2015 to 2019 the department of gastroenterology, SDMCMS&H. The diagnosis of IBD was made based on clinical, radiological, endoscopic, and histopathologic findings. The patients were categorized into IBD and those who have symptoms suggestive of IBD but did not fit into the diagnostic criteria into, non-IBD groups. The data about of on demography, diet patterns, and laboratory parameters were recorded. RESULTS: Among 226 patients enrolled in this study 2015-2019, IBD was confirmed in 54 Ulcerative colitis - 44 (19.46%), Crohn's disease - 10 (4.42%) patients with varying distribution of disease among different age groups and both genders, Ulcerative colitis (UC) [M: F: 28 (63.6%): 16 (36.4%)] and Crohn's disease (CD) [M: F: 07 (70.0%):03 (30.0%)]. Dietary pattern and other habitats had no significant contribution to illness and its symptoms. Urban (U) and Rural (R) divide was UC [U: R: 32 (72.7%): 12 (27.3%)], CD [U:R:07(70.0%):03(30.0%)] maintained. CONCLUSION: Incidence of IBD was high with UC as compared to CD. The incidence of IBD among patients presenting with symptoms suggestive of IBD is 19.46% with UC being major as compared to CD (4.42%). Male predominant patterns of IBD incidences were noted. Year by year increasing trend in disease burden was observed. The Dietary pattern has no direct correlation with IBD disease prevalence and incidences.

Incidência de doença inflamatória intestinal: estudo retrospectivo de um único centro / Incidence of inflammatory bowel disease: a single centre retrospective study

ABSTRACT Background: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder affecting the gastrointestinal tract. The etiology of this alarming condition is multifactorial. A Recently increasing trend in IBD is noted in our country. Objective: The present study was designed with the main objective to assess the incidence and to identify the associated risk factors including demographic, geographical areas, and dietary patterns of IBD population of Northern of Karnataka viz. Hubli-Dharwad city. Methods: A retrospective investigation was conducted on a cohort of 226 patients with a working diagnosis of IBD and those who were admitted between 2015 to 2019 the department of gastroenterology, SDMCMS&H. The diagnosis of IBD was made based on clinical, radiological, endoscopic, and histopathologic findings. The patients were categorized into IBD and those who have symptoms suggestive of IBD but did not fit into the diagnostic criteria into, non-IBD groups. The data about of on demography, diet patterns, and laboratory parameters were recorded. Results: Among 226 patients enrolled in this study 2015-2019, IBD was confirmed in 54 Ulcerative colitis - 44 (19.46%), Crohn's disease - 10 (4.42%) patients with varying distribution of disease among different age groups and both genders, Ulcerative colitis (UC) [M: F: 28 (63.6%): 16 (36.4%)] and Crohn's disease (CD) [M: F: 07 (70.0%):03 (30.0%)]. Dietary pattern and other habitats had no significant contribution to illness and its symptoms. Urban (U) and Rural (R) divide was UC [U: R: 32 (72.7%): 12 (27.3%)], CD [U:R:07(70.0%):03(30.0%)] maintained. Conclusion Incidence of IBD was high with UC as compared to CD. The incidence of IBD among patients presenting with symptoms suggestive of IBD is 19.46% with UC being major as compared to CD (4.42%). Male predominant patterns of IBD incidences were noted. Year by year increasing trend in disease burden was observed. The Dietary pattern has no direct correlation with IBD disease prevalence and incidences.

Resumo Contexto: A Doença inflamatória intestinal (DII) é uma doença inflamatória crônica que afeta o trato gastrointestinal. A etiologia desta condição alarmante é multifatorial. Uma tendência recentemente crescente na DII é notada em nosso país. Objetivo: O presente estudo foi desenhado com o objetivo principal de avaliar a incidência e identificar os fatores de risco associados, incluindo demográficos, áreas geográficas e padrões alimentares da população com DII do Norte de Karnataka viz. Cidade de Hubli-Dharwad. Metodos: Uma investigação retrospectiva foi realizada em uma coorte de 226 pacientes com diagnóstico de DII e que foram admitidos entre 2015 e 2019 no departamento de gastroenterologia, SDMCMS&H. O diagnóstico de DII foi feito com base em achados clínicos, radiológicos, endoscópicos e histopatológicos. Os pacientes foram categorizados em DII e aqueles que apresentam sintomas sugestivos de DII, mas não se enquadraram nos critérios diagnósticos em grupos sem DII. Os dados sobre a demografia, padrões de dieta e parâmetros laboratoriais foram registrados. Resultados: Entre os 226 pacientes inscritos neste estudo entre 2015-2019, DII foi confirmada em 54 [RCUI - 44 (19,46%), DC - 10 (4,42%)] com distribuição variada da doença entre diferentes faixas etárias e ambos os sexos, colite ulcerativa (RCUI) [M: F: 28 (63,6%):16 (36,4%)] e doença de Crohn (DC) [M: F: 07 (70,0%): 03 (30,0%)]. O padrão alimentar e outros hábitos não tiveram contribuição significativa para a doença e seus sintomas. Urbanos (U) e rurais (R) dividiram-se em RCUI [U: R: 32 (72,7%):12 (27,3%)], DC [07 (70,0%): 03 (30,0%)]. Conclusão A incidência de DII foi elevada para RCUI em relação a DC. A incidência de DII entre os pacientes com sintomas sugestivos de DIB é de 19,46% com a RCUI sendo maior em relação a DC (4,42%). Foram observados padrões predominantes masculinos de incidência de DII. Ano a ano foi observada tendência crescente de carga da doença. O padrão dietético não tem correlação direta com a prevalência e incidências da DII.

Long-term Satisfaction with Roux-en-Y Gastric Bypass Surgery: Results From a Multicenter Prospective Cohort Study.

OBJECTIVE: The aim of this study was to examine change in overall satisfaction with Roux-en-Y gastric bypass (RYGB) surgery over 3 to 7 years post-surgery and identify pre-surgery predictors and post-surgery factors associated with not being satisfied. BACKGROUND: It is unclear how satisfaction with RYGB surgery changes over time following surgery and factors associated with not being satisfied are not well understood. METHODS: Participants of a multicenter prospective cohort study of bariatric surgery were followed annually < 7 years. A total of 1423 participants of the 1770 who underwent RYGB had data on satisfaction with surgery (81% female; median age 47 years; median body mass index 46 kg/m 2 ). RESULTS: The percentage of participants who were not satisfied with RYGB surgery significantly increased from 15.4% 3 years post-surgery to 23.0% 7 years post-surgery ( P = 0.01). Pre-surgery younger age, lower BMI, higher percent weight loss needed to reach dream weight, poorer physical and mental health, and less social support independently predicted higher risk of not being satisfied with surgery. When examining pre- to post-surgery changes, less post-surgery weight loss, worsening physical and mental health status, less social support, and greater depressive symptomology were associated with higher risk of not being satisfied with surgery. CONCLUSIONS: Level of satisfaction with RYGB surgery significantly decreased 3 to 7 years following surgery. Several pre- and post-surgery characteristics were associated with not being satisfied with surgery and provide potentially useful insight into individual patient experiences following RYGB. Knowledge of these characteristics may be useful in communication between surgeons and patients regarding post-surgical expectations and the impact of surgery on patients' lives.

Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors.

Calcium/Calmodulin-dependent Protein Kinase Kinase 2 (CAMKK2) acts as a signaling hub, receiving signals from various regulatory pathways and decoding them via phosphorylation of downstream protein kinases - such as AMPK (AMP-activated protein kinase) and CAMK types I and IV. CAMKK2 relevance is highlighted by its constitutive activity being implicated in several human pathologies. However, at present, there are no selective small-molecule inhibitors available for this protein kinase. Moreover, CAMKK2 and its closest human homolog, CAMKK1, are thought to have overlapping biological roles. Here we present six new co-structures of potent ligands bound to CAMKK2 identified from a library of commercially-available kinase inhibitors. Enzyme assays confirmed that most of these compounds are equipotent inhibitors of both human CAMKKs and isothermal titration calorimetry (ITC) revealed that binding to some of these molecules to CAMKK2 is enthalpy driven. We expect our results to advance current efforts to discover small molecule kinase inhibitors selective to each human CAMKK.

Proximal Roux-en-Y gastric bypass: Addressing the myth of limb length.

BACKGROUND: Some studies suggest that changes in weight or metabolic outcomes are affected by the lengths of the gastrointestinal limbs in the Roux-en-Y gastric bypass. METHODS: Participants (N = 1,770) underwent primary Roux-en-Y gastric bypass and were followed ≤7 years in the Longitudinal Assessment of Bariatric Surgery-2, a multicenter US cohort study. Alimentary limb and biliopancreatic limb lengths were measured according to research protocol; common channel was measured in a subsample (N = 547). Aimentary limb, biliopancreatic limb, and common channel ratio to total small bowel length were calculated. RESULTS: Median presurgery body mass index was 46 (25th-75th percentile: 43-51) kg/m2. Medians (25th-75th percentiles) for alimentary limb length were 125 cm (100-150), for biliopancreatic limb length were 50 cm (50-60), and common channel length were 410 cm (322-520). Statistics for ratios to the small bowel length were 0.23 (0.18-0.27) for alimentary limb, 0.09 (0.07-0.10) for biliopancreatic limb, and 0.69 (0.63-0.73) for common length. There were no significant associations between alimentary limb, biliopancreatic limb, common channel, alimentary limb ratio, biliopancreatic limb ratio or common channel ratio, and either weight loss or improvement in cardiometabolic outcomes. CONCLUSION: The common channel length in Roux-en-Y gastric bypass is highly variable between individuals. None of the limb lengths in this study, nor alimentary limb, biliopancreatic limb, or common channel ratios, seem to be related to weight loss or metabolic improvements >7 years.

Alcohol Use Thresholds for Identifying Alcohol-related Problems Before and Following Roux-en-Y Gastric Bypass.

OBJECTIVE: The aim of this study was to determine the sensitivity and specificity of potential thresholds of alcohol use for identifying alcohol-related problems in women post-Roux-en-Y gastric bypass (RYGB). BACKGROUND: Despite evidence that RYGB alters alcohol pharmacokinetics and is associated with an increased risk for alcohol-related problems, the level of alcohol use that should prompt further screening for alcohol-related problems following RYGB is unclear. METHODS: The Longitudinal Assessment of Bariatric Surgery-2 is a prospective cohort study. Before surgery and annually for ≤7 years following surgery, participants completed the 10-item Alcohol Use Disorder Identification Test (AUDIT), which assesses past-year frequency and quantity of alcohol, frequency of consuming ≥6 drinks, and alcohol-related problems (ie, symptoms of alcohol dependence and/or alcohol-related harm). The AUDIT-Consumption (AUDIT-C) score was determined from the first 3 AUDIT items. RESULTS: Post-RYGB, 835 women reported current drinking at 1 or more annual assessment(s). Compared with higher frequency thresholds, drinking ≥2 times/month had the highest combined sensitivity (85.3%) and specificity (61.4%) for identifying alcohol-related problems. Compared with higher quantity thresholds, drinking ≥3 drinks/drinking day had the highest combined sensitivity (64.2%) and specificity (87.2%). An AUDIT-C score ≥3, versus other thresholds, had the highest combined sensitivity (76.4%) and specificity (81.6%). CONCLUSION: The sensitivity and specificity of these thresholds indicate assessment of alcohol consumption alone may be inadequate for identifying women at risk for alcohol-related problems post-RYGB. Additional screening tools for alcohol-related problems, which assess symptoms of alcohol-related problems, should be conducted in this population.

High Helicobacter pylori resistance to metronidazole and clarithromycin in Brazilian children and adolescents.

OBJECTIVE: The aim of the present study was to assess the primary and secondary resistance of Helicobacter pylori strains to clarithromycin, amoxicillin, furazolidone, tetracycline, and metronidazole, the conventional antibiotics presently used in Brazilian children and adolescents. METHODS: Seventy-seven consecutive H pylori strains, 71 of 77 strains obtained from patients without previous eradication treatment for H pylori infection, and 6 strains from patients in whom previous eradication treatment had failed. RESULTS: Global rate of resistance was 49.3% (38/77): 40% of strains were resistant to metronidazole, 19.5% to clarithromycin, and 10.4% to amoxicillin. All of the tested H pylori strains were susceptible to furazolidone and tetracycline. Multiple resistance were detected in 18.2% (14/77 patients) of the strains: 6 of 14 (43%) simultaneously resistant to clarithromycin and metronidazole; 5 of 14 (36%) to amoxicillin and metronidazole; 2 of 14 (14%) to amoxicillin, clarithromycin, and metronidazole; and 1 of 14 (7%) to clarithromycin and amoxicillin. CONCLUSIONS: The high resistance rate to metronidazole and clarithromycin observed in clinical H pylori isolates can exclude these antimicrobials in empirical eradication treatment in Brazil. Otherwise, furazolidone and tetracycline presented no resistance. Properly assessing the risks and benefits, these 2 antimicrobials and their derivatives could be used in empirical eradication schedules, both associated with amoxicillin, which showed a low resistance rate despite its wide use in pediatric patients.

Coccidioidomycosis in elderly persons.

BACKGROUND: Coccidioidomycosis is a fungal infection acquired via inhalation of airborne fungal arthrospores of Coccidioides species in regions of endemicity in the deserts of the southwestern United States and northern Mexico. In recent years, the incidence of coccidioidomycosis has increased in areas of endemicity, and previous studies have found the highest incidence of coccidioidal infection in Arizona among persons in older age groups. METHODS: We conducted a retrospective review of data for all patients with coccidioidomycosis who were treated at our institution that compared clinical manifestations of coccidioidomycosis in patients aged >or=60 years with those in patients aged <60 years. RESULTS: We compared 210 patients aged >or=60 years with 186 patients aged <60 years. No significant differences were observed with regard to manifestations of coccidioidomycosis, even after adjustment for comorbid conditions, excluding immunosuppression. Regardless of age, when coccidioidal illnesses in immunosuppressed patients were compared with those in nonimmunosuppressed patients, immunosuppressed patients were significantly more likely to have extrapulmonary dissemination of infection, to require hospitalization, and to have progressive infection or to die of coccidioidomycosis. Univariate logistic regression identified immunosuppression as the only marker that increased risk of extrapulmonary dissemination of infection (odds ratio, 2.13;P=.05), hospitalization (odds ratio, 2.68; P<.001), and death (odds ratio, 8.39; P<.001). Multivariate analysis revealed that neither age nor an interaction of age and immunosuppression had a significant impact on coccidioidal manifestations. CONCLUSIONS: Coccidioidomycosis is a serious illness in all patients, but its different manifestations in older-aged persons, compared with those in younger-aged persons, may be related to immunosuppression rather than age alone.

[Analysis of molecular fingerprint and virulence factors of Helicobacter pylori strains]. / Análise das impressões digitais de DNA e de fatores de virulência de linhagens de Helicobacter pylori.

BACKGROUND: Helicobacter pylori is now accepted as the most important agent of gastritis in humans, as well as a risk factor for peptic ulcer disease and gastric carcinoma. The outcome of the infection is related to several factors, among them bacterial ones such as cagA and vacA s1m1 genotype. Random amplified polymorphic DNA (RAPD)-PCR, has been used to generate DNA fingerprints to evaluate similarity among strains within a bacterial species. AIM: To assess the association between RAPD fingerprinting, virulence factors and the disease. METHODS: H. pylori was isolated from 112 patients (41 with gastritis; 19 with gastric ulcers; 38 with duodenal ulcer disease; and 14 with gastroesophageal reflux disease). Allelic variants of cagA and vacA were identified using the polymerase chain reaction (PCR) and the fingerprints were generated by RAPD-PCR. RESULTS: There was a strong association between the genotype vacA s1m1 and duodenal ulcers. Although RADP-PCR is a very useful tool in genotyping H. pylori, no significant correlation between the diseases studied and DNA fingerprint was detected neither with fingerprint and different vacA and, cagA genotypes. CONCLUSIONS: The extension of our analysis to patients with well-characterized gastric diseases may provide significant information on the relationship between vacA genotypes and clinical outcomes of H. pylori infection.

Differentially expressed genes in response to amoxicillin in Helicobacter pylori analyzed by RNA arbitrarily primed PCR.

Because the molecular mechanism of amoxicillin resistance in Helicobacter pylori seems to be partially explained by several mutational changes in the pbp1A gene, the aim of the present study was to evaluate the gene expression pattern in response to amoxicillin in the Amx(R) Hardenberg strain using RNA arbitrarily primed PCR (RAP-PCR). In the experiments, c. 100 differentially expressed RAP-PCR products were identified using five arbitrary primers. The cDNAs that presented the highest levels of induction or repression were cloned and sequenced, and the sequences were compared with those present in databases using the blast search algorithm. The differential expression of the isolated cDNAs was confirmed by real-time PCR. The preliminary results showed that amoxicillin alters the expression of five cDNAs involved in biosynthesis, two involved with pathogenesis, four related to cell envelope formation, two involved in cellular processes, three related with transport and binding proteins, one involved with protein degradation, one involved with energy metabolism and seven hypothetical proteins. Further analysis of these cDNAs will allow a better comprehension of both the molecular mechanism(s) of amoxicillin resistance and the adaptative mechanism(s) used by H. pylori in the presence of this antibiotic.

Análise das impressões digitais de DNA e de fatores de virulência de linhagens de Helicobacter pylori / Analysis of molecular fingerprint and virulence factors of Helicobacter pylori strains

RACIONAL: Helicobacter pylori é hoje aceito como o principal agente etiológico de gastrite em seres humanos e fator de risco para úlcera péptica e câncer gástrico. A evolução da infecção está relacionada a diversos fatores, inclusive bacterianos, como presença do gene cagA e o genótipo vacA s1m1, associados ao desenvolvimento de úlcera e adenocarcinoma gástrico. A técnica de RAPD ("random amplified polimorphic") tem sido amplamente utilizada para obtenção de impressões digitais de DNA para examinar a similaridade entre linhagens. OBJETIVOS: Avaliar a presença de cagA e alelos do vacA em amostras de H. pylori e associar os achados com a doença apresentada e também investigar possível clonicidade entre os fatores de virulência e as doenças com a impressão digital de DNA gerada pelo RAPD-PCR. MÉTODOS: Foram incluídas 112 amostras provenientes de pacientes com diferentes laudos endoscópicos: gastrite (n = 41), esofagite de refluxo (n = 14), úlcera gástrica (n = 19) e úlcera duodenal (n = 38). A análise dos fatores de virulência da bactéria foi feita por PCR e as impressões digitais de DNA foram estabelecidas pelo método de RAPD-PCR. RESULTADOS: Os resultados obtidos indicam que houve uma associação significativa entre úlcera duodenal e o mosaico vacA s1m1. Analisando-se os padrões de bandas geradas pelo RAPD-PCR, sete diferentes dendogramas foram construídos e não foi possível detectar associação significativa entre os agrupamentos, sugerindo que as amostras não possuem perfil clonal. CONCLUSÃO: Os resultados reforçam a importância do gene vacA como um marcador de virulência do H. pylori. O RAPD da impressão digital de DNA realizado foi incapaz de associar o padrão de bandas com as enfermidades e os genótipos de vacA e cagA.

BACKGROUND: Helicobacter pylori is now accepted as the most important agent of gastritis in humans, as well as a risk factor for peptic ulcer disease and gastric carcinoma. The outcome of the infection is related to several factors, among them bacterial ones such as cagA and vacA s1m1 genotype. Random amplified polymorphic DNA (RAPD)-PCR, has been used to generate DNA fingerprints to evaluate similarity among strains within a bacterial species. AIM: To assess the association between RAPD fingerprinting, virulence factors and the disease. METHODS: H. pylori was isolated from 112 patients (41 with gastritis; 19 with gastric ulcers; 38 with duodenal ulcer disease; and 14 with gastroesophageal reflux disease). Allelic variants of cagA and vacA were identified using the polymerase chain reaction (PCR) and the fingerprints were generated by RAPD-PCR. RESULTS: There was a strong association between the genotype vacA s1m1 and duodenal ulcers. Although RADP-PCR is a very useful tool in genotyping H. pylori, no significant correlation between the diseases studied and DNA fingerprint was detected neither with fingerprint and different vacA and, cagA genotypes. CONCLUSIONS: The extension of our analysis to patients with well-characterized gastric diseases may provide significant information on the relationship between vacA genotypes and clinical outcomes of H. pylori infection.

The influence of endoscopic procedures upon the contamination of Helicobacter pylori cultures.

BACKGROUND: Among the various diagnostic methods for the detection of Helicobacter pylori infection, histological examination and microbiological processing of gastric biopsy samples are assumed to be the gold standard techniques. AIMS: Since H. pylori culture can be affected by the presence of non-H. pylori bacteria, we evaluated the efficacy of endoscope disinfection and the influence of endoscopic procedures on culture contamination. PATIENTS AND METHODS: The procedures used during the first two routine endoscopies were evaluated during 28 consecutive days. Endoscopy room, forceps and endoscopic channel were analyzed before and after the beginning of normal procedures. After disinfection, a biopsy simulation was performed to verify the gastric bacteria. RESULTS: Endoscope disinfection removed all organisms from forceps and endoscopic channel with 100% efficacy. The most frequent non-H. pylori bacteria detected were Streptococcus bovis, Enterobacter hormaechei, and Staphylococcus aureus. The sensibility of the H. pylori culture was affected by the presence of non-H. pylori bacteria. CONCLUSION: The risk of transmission of microorganisms was not detectable when sterilized biopsy forceps and stringent disinfection standards were employed. Whilst S. bovis and E. hormaechei may be common in gastric microbial flora, the presence of P. aeruginosa and S. aureus indicated that the manipulation of biopsies could be responsible for culture contamination with these bacteria.

The influence of endoscopic procedures upon the contamination of Helicobacter pylori cultures

RACIONAL E OBJETIVOS: Dentre os vários métodos diagnósticos empregados na detecção da infecção por Helicobacter pylori, o diagnóstico histológico e a análise microbiológica de biopsia gástrica são consideradas as técnicas mais sensíveis. Entretanto, a sensibilidade da cultura de H. pylori pode ser reduzida pela presença de outras bactérias. Desse modo, avaliou-se a eficácia da desinfecção do endoscópio e a influência dos procedimentos endoscópicos na contaminação da cultura bacteriana. Para tal, as duas primeiras endoscopias durante 28 dias consecutivos foram estudadas. A sala de endoscopia, o fórceps e o canal do endoscópio foram analisados antes e depois do início dos procedimentos endoscópicos rotineiros. Depois da desinfecção, uma simulação de coleta de biopsia foi realizada para verificar a presença das bactérias gástricas. RESULTADOS: A desinfecção do endoscópio foi capaz de remover todos os organismos do fórceps e do canal do endoscópio. As bactérias não-H. pylori mais freqüentemente detectadas foram Streptococcus bovis, Enterobacter hormaechei e Staphylococcus aureus. Em alguns casos a sensibilidade da cultura do H. pylori foi reduzida pela presença de bactérias contaminantes. CONCLUSAO: Não houve risco de transmissão de microorganismos quando fórceps esterilizados e desinfecção adequada foram empregadas. A presença de S. bovis e E. hormaechei parece ser comum na microflora gástrica; por outro lado, a detecção de P. aeruginosa e S. aureus indica que a manipulação de biopsias pode ser responsável pela contaminação da cultura por essas bactérias.

Detection of high-level tetracycline resistance in clinical isolates of Helicobacter pylori using PCR-RFLP.

Tetracycline is one of four antibiotics commonly used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing as the incidence of tetracycline resistance is increasing. In five Brazilian tetracycline-resistant (Tet(R)) H. pylori isolates, high-level tetracycline resistance is mediated by the triple-base-pair substitution AGA(926-928)-->TTC in both 16S rRNA genes, as was previously observed in two independent high-level Tet(R) H. pylori strains. A polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) assay was developed for the detection of the AGA(926-928)-->TTC substitution, and confirmed the presence of the aforementioned triple-base-pair substitution in all five Brazilian Tet(R) isolates. This PCR-RFLP-based approach distinguishes the high-level Tet(R) isolates from low-level Tet(R) and Tet(S) H. pylori strains and thus allows the direct detection of Tet(R) H. pylori isolates.

The transcription factor Snf1p is involved in a Tup1p-independent manner in the glucose regulation of the major methanol metabolism genes of Hansenula polymorpha

Hansenula polymorpha is a methylotrophic yeast widely employed in biotechnology as a ''protein factory''. Most promoters used for heterologous protein expression, like MOX (methanol oxidase) and DAS (di-hydroxy acetone synthase), are involved in the peroxisomal methanol metabolism (C1 metabolism) and are under strong glucose repression. Interestingly, the MOX promoter is subjected to glucose regulation also in Saccharomyces cerevisiae, a non-methylotrophic yeast in which this phenomenon is well studied. In this species, the transcription factor Tup1p plays an essential role in glucose repression of several genes. This effect is counteracted by the activator Snf1p when glucose is exhausted from medium. Therefore, to test whether this regulatory circuit has been conserved in H. polymorpha, HpTUP1 and HpSNF1 were partially cloned and disrupted. Deletion of HpTUP1 did not affect glucose repression of the major C1 metabolism genes (MOX, DAS). Thus, though conserved, HpTUP1 does not seem to take part in a general glucose repression in H. polymorpha. In contrast, the deletion of HpSNF1 led to significant decreases in the activation of these genes in the absence of glucose. Therefore, the effect of HpSNF1 in transcriptional activation may be through an HpTUP1- independent circuit.