Depending on the presence of mutations that determine isoniazid (INH) susceptibility (katG and inhA), Mycobacterium tuberculosis may be susceptible to high doses of INH or ethionamide (ETH).
To ...describe the INH resistance profile and association of katG mutation with previous INH treatment and level of drug resistance based on rapid molecular drug susceptibility testing (DST) in southern Brazil and central Mozambique.
Descriptive study of 311 isolates from Ribeirão Preto, São Paulo, Brazil (2011-2014) and 155 isolates from Beira, Mozambique (2014-2015). Drug resistance patterns and specific gene mutations were determined using GenoType(®) MTBDRplus.
katG gene mutations were detected in 12/22 (54.5%) Brazilian and 32/38 (84.2%) Mozambican isolates. inhA mutations were observed in 9/22 (40.9%) isolates in Brazil and in 4/38 (10.5%) in Mozambique. Both katG and inhA mutations were detected in respectively 1/22 (5%) and 2/38 (5.2%). The difference in the frequency of katG mutations in Brazil and Mozambique was statistically significant (P = 0.04). katG mutations were present in 68.8% (33/48) of patients previously treated with INH and 31.2% (15/48) of patients without previous INH. This difference was not statistically significant (P = 0.223).
INH mutations varied geographically; molecular DST can be used to guide and accelerate decision making in the use of ETH or high doses of INH.
Mortality and adverse neurologic sequelae from HIV-associated cryptococcal meningitis (HIV-CM) remains high due to raised intracranial pressure (ICP) complications. Cerebrospinal fluid (CSF) high ...opening pressure occurs in more than 50% of HIV-CM patients. Repeated lumbar puncture with CSF drainage and external lumbar drainage might be required in the management of these patients. Usually, there is a high grade of uncertainty and the basis for clinical decisions regarding ICP hypertension tends to be from clinical findings (headache, nausea and vomiting), a low Glasgow coma scale score, and/or fundoscopic papilledema. Significant neurological decline can occur if elevated CSF pressures are inadequately managed. Various treatment strategies to address intracranial hypertension in this setting have been described, including: medical management, serial lumbar punctures, external lumbar and ventricular drain placement, and either ventricular or lumbar shunting. This study aims to evaluate the role of a non-invasive intracranial pressure (ICP-NI) monitoring in a critically ill HIV-CM patient.
Resistance to Mycobacterium tuberculosis is a reality worldwide, and its diagnosis continues to be difficult and time consuming. To face this challenge, the World Health Organization has recommended ...the use of rapid molecular tests. We evaluated the routine use (once a week) of a line probe assay (Genotype MTBDRplus) for early diagnosis of resistance and for assessment of the main related risk factors over 2 years. A total of 170 samples were tested: 15 (8.8%) were resistant, and multidrug resistance was detected in 10 (5.9%). The sensitivity profile took 3 weeks (2 weeks for culture and 1 week for rapid testing). Previous treatment for tuberculosis and the persistence of positive acid-fast smears after 4 months of supervised treatment were the major risk factors observed. The use of molecular tests enabled early diagnosis of drug-resistant bacilli and led to appropriate treatment of the disease. This information has the potential to interrupt the transmission chain of resistant M. tuberculosis.
Objective
To evaluate whether colchicine treatment was associated with the inhibition of NLRP3 inflammasome activation in patients with COVID-19.
Methods
We present a post hoc analysis from a ...double-blinded placebo-controlled randomized clinical trial (RCT) on the effect of colchicine for the treatment of COVID-19. Serum levels of NOD-like receptor protein 3 (NLRP3) inflammasome products—active caspase-1 (Casp1p20), IL-1β, and IL-18—were assessed at enrollment and after 48–72 h of treatment in patients receiving standard-of-care (SOC) plus placebo vs. those receiving SOC plus colchicine. The colchicine regimen was 0.5 mg tid for 5 days, followed by 0.5 mg bid for another 5 days.
Results
Thirty-six patients received SOC plus colchicine, and thirty-six received SOC plus placebo. Colchicine reduced the need for supplemental oxygen and the length of hospitalization. On Days 2–3, colchicine lowered the serum levels of Casp1p20 and IL-18, but not IL-1β.
Conclusion
Treatment with colchicine inhibited the activation of the NLRP3 inflammasome, an event triggering the ‘cytokine storm’ in COVID-19.
Trial registration numbers
RBR-8jyhxh
Polymerase chain reaction (PCR) has been widely investigated for the diagnosis of tuberculosis. However, before this technique is applied on clinical samples, it needs to be well standardized. We ...describe the use of McFarland nephelometer, a very simple approach to determine microorganism concentration in solution, for PCR standardization and DNA quantitation, using Mycobacterium tuberculosis as a model. Tuberculosis is an extremely important disease for the public health system in developing countries and, with the advent of AIDS, it has also become an important public health problem in developed countries. Using Mycobacterium tuberculosis as a research model, we were able to detect 3 M. tuberculosis genomes using the McFarland nephelometer to assess mycobacterial concentration. We have shown here that McFarland nephelometer is an easy and reliable procedure to determine PCR sensitivity at lower costs.
Study objectives: The detection of Mycobacterium sp on a culture remains the âgold standardâ technique for the diagnosis of mycobacterial infections.
A small percentage of these cultures, ...however, may be contaminated by other nonfastidious microorganisms, making accurate
diagnosis difficult. We evaluated the use of a polymerase chain reaction (PCR) protocol that was specific for the genus Mycobacterium,
and specifically for Mycobacterium tuberculosis, Mycobacterium avium , and Mycobacterium intracellulare , for the identification of Mycobacterium sp growing on contaminated cultures.
Design: This prospective study was designed to identify Mycobacterium sp growing on mycobacterial cultures contaminated with other
microorganisms.
Samples and patients: Twenty-six samples, taken from 23 patients with probable mycobacterial disease, that resulted in Mycobacterium growth but
were contaminated during their processing were evaluated in this study. Clinical data and the clinical status of each patient
were used to ascertain the final diagnosis.
Results: All samples studied here exhibited Mycobacterium growth on solid media but were contaminated by nonfastidious bacteria, compromising
the biochemical identification of the Mycobacterium sp. PCR correctly identified the genus Mycobacterium in all samples. M tuberculosis was identified in 14 samples, and M avium in 10 samples. No amplification of M intracellulare was obtained, and in two samples there was amplification only for the genus Mycobacterium. In the cultures of those patients
in whom a mycobacterial infection was evident, PCR identified M avium and M tuberculosis in samples from 6 and 12 patients, respectively. However, PCR identified M avium (two patients) and M tuberculosis (two patients) in the cultures of four patients for whom a mycobacterial disease could not be confirmed by our case definition.
Finally, in two samples from one patient only the genus Mycobacterium was amplified by PCR.
Conclusion: PCR, with its advantages of greater speed and effectiveness than conventional detection methods, was successfully used to
identify the Mycobacterium sp growing on contaminated cultures.
The recent increase in the number of tuberculosis cases has called the world's attention once again to a perennial health problem, especially prevalent in developing countries. The time elapsed ...between the diagnosis and the institution of therapy is an obstacle to tuberculosis control and there is an urgent need for the development of techniques for the disease's rapid diagnosis. To achieve this goal, molecular biology techniques have been exhaustively investigated. This work describes the use of a polymerase chain reaction for rapid diagnosis of tuberculosis in a developing country. The sensitivity and specificity of this technique is compared to standard techniques used in the microbiology laboratory.
This study was undertaken in Ribeirão Preto, S. Paulo State, Brazil. Forty-two sputum samples from suspected cases of tuberculosis attending the municipal health care centers were sent to the microbiology laboratory. The samples were processed for the detection of Mycobacterium tuberculosis by acid-fast bacilli determination, culture in Lowenstein-Jensen medium, and by a polymerase chain reaction that amplified a fragment of 123 base pairs of the Mycobacterium tuberculosis genome.
Of the forty-two samples studied, one was contaminated and excluded from the study, ten were culture positive, ten were positive for the presence of acid-fast bacilli, and sixteen were polymerase chain reaction positive. The sensitivity and specificity of this technique were 90% and 81%, respectively.
The polymerase chain reaction presented a sensitivity comparable to the culture and the whole procedure took only one day to complete. The results presented here make it a strong candidate for rapid diagnosis of tuberculosis in clinical settings making it possible to begin the specific therapy early in the course of the disease. However, standardization of the technique is necessary, and the correlation with clinical findings is of paramount importance due to the high sensitivity of this technique.
To describe the process of integration and revision of a pediatric program curriculum which resulted in the creation of a competency-based framework recommended in the Brazilian National Curricular ...Guidelines.
Quali-quantitative analysis of an intervention evaluating the students and professors' perception of the pediatric program curriculum (focus groups and semi-structured interviews). Results were discussed during teaching development workshops. A competency-based framework was suggested for the pediatric program from the 3rd to the 6th year. The new curriculum was approved, implemented, and reevaluated six months later.
Twelve students (12%) from the 3rd to the 6th year participated in the focus groups, and 11 professors (78.5%) answered the questionnaire. Most participants reported lack of integration among the courses, lack of knowledge about the learning goals of the internships, few opportunities of practice, and predominance of theoretical evaluation. In the training workshops, a competency-based curriculum was created after pediatrics and collective health professors reached an agreement. The new curriculum was focused on general competency, learning goals, opportunities available to learn these goals, and evaluation system. After six months, 93% (104/112) of students and 79% (11/14) of professors reported greater integration of the program and highlighted the inclusion of the clinical performance evaluation.
The collective creation of a competency-based curriculum promoted higher satisfaction of students and professors. After being implemented, the new curriculum was considered to integrate the teaching practices and contents, improving the quality of the clinical performance evaluation.
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