Development of Lipid Nanocarriers for Tuberculosis Treatment: Evaluation of Suitable Excipients and Nanocarriers.

BACKGROUND: Lipid nanocarriers have been widely tested as drug delivery systems to treat diseases due to their bioavailability, controlled release, and low toxicity. For the pulmonary route, the Food and Drug Administration favors the use of substances generally recognized as safe, as well as biodegradable and biocompatible to minimize the possibility of toxicity. Tuberculosis (TB) remains a public health threat worldwide, mainly due to the long treatment duration and adverse effects. Therefore, new drug delivery systems for treating TB are needed. OBJECTIVE: Physicochemical characterization of different lipid-based nanocarriers was used to optimize carrier properties. Optimized systems were incubated with Mycobacterium tuberculosis to assess whether lipid-based systems act as the energy source for the bacteria, which could be counterproductive to therapy. METHODS: Several excipients and surfactants were evaluated to prepare different types of nanocarriers using high-pressure homogenization. RESULTS: A mixture of trimyristin with castor oil was chosen as the lipid matrix after differential scanning calorimetry analysis. A mixture of egg lecithin and PEG-660 stearate was selected as an optimal surfactant system, as this mixture formed the most stable formulations. Three types of lipid nanocarriers, solid lipid nanoparticles, nanostructured lipid carriers (NLC), and nanoemulsions, were prepared, with the NLC systems showing the most suitable properties for further evaluation. It may provide the advantages of increasing the entrapment efficiency, drug release, and the ability to be lyophilized, producing powder for pulmonary administration as an alternative to entrap poor water-soluble molecules. CONCLUSION: Furthermore, the NLC system can be considered for use as a platform for the treatment of TB through the pulmonary route.

Cytotoxic and genotoxic effects of environmental relevant concentrations of bisphenol A and interactions with doxorubicin.

Bisphenol A (BPA) is one of the most widely utilized endocrine disruptors to which humans are exposed, particularity through ingestion. BPA is an aneugenic compound with a putative association to tumorigenesis. Although extensively studied in estrogen responsive cells, information regarding its effects on cells from the upper gastrointestinal tract exposed to free/active forms of BPA is still scarce. Similarly, BPA interactions with other drugs have been neglected, although it has been suggested to have a potential role in doxorubicin (DOX) chemoresistance. This study is intended to assess potential cytotoxic and genotoxic effects of BPA, as well as its interactions with DOX, in Human epithelial type 2 cells (Hep-2) originated from a human laryngeal carcinoma and in a DNA damage responsive cell line, the human lung fibroblasts (MRC-5). Cell viability was analyzed through the resazurin assay. The G protein-coupled estrogen receptor 1 (GPER) expression was visualized by immunodetection. Genotoxicity, namely DNA damage and oxidative DNA damage, were assessed by comet assay and micronuclei induction, and mitotic disruption was evaluated cytologically by fluorescent microscopy with DAPI staining. Cytotoxicity analysis showed that exposure to BPA per se does not affect cellular viability. Nevertheless, the genotoxic analysis showed that BPA induced an increase of DNA damage in the Hep-2 cell line and in oxidative damage in the MRC-5 cell line. An increase of micronuclei was also observed in both cell lines following BPA exposure. BPA and DOX co-exposures suggested that BPA acts as an antagonist of DOX effects in both cell lines. The interaction with DOX appears to be cell type dependent, exhibiting a non-monotonic response curve in MRC-5 cells, a GPER expressing cell line. Our study emphasizes the need for a deeper knowledge of BPA interactions, particularly with chemotherapeutic agents, in the context of risk assessment and public health.

Reliability and validity analyses of the North Star Ambulatory Assessment in Brazilian Portuguese.

The North Star Ambulatory Assessment measures motor performance in ambulatory boys with Duchenne muscular dystrophy, a hereditary and degenerative muscle disorder. To use the North Star Ambulatory Assessment in Brazilian boys, we performed the cross-cultural adaptation to the Portuguese language spoken in Brazil and evaluated the reliability and validity of the instrument. Cross-cultural adaptation included: independent translations, synthesis, committee review, pre-testing in 12 boys, back-translation and comparison with the original instrument. Thirty-five boys with Duchenne muscular dystrophy and 38 healthy age-matched controls were recruited for further analyses. Reliability was assessed by internal consistency and reproducibility. Validity studies included face, content, construct and known-groups analyses. Cross-cultural adaptation resulted in an adequate instrument. Reliability studies demonstrated high internal consistency (Cronbach's alpha = 0.935) and adequate intra and inter-rater reproducibility (intraclass correlation coefficient = 0.988 and 0.962). Validation analyses indicated satisfactory content, face and convergent construct validities, with positive correlations with the Motor Function Measure total score (r = 0.863) and the 6-minute walk test (r = 0.433). The known group validity was demonstrated by higher scores in younger boys with Duchenne muscular dystrophy (p = 0.005). North Star Ambulatory Assessment in Brazilian Portuguese is a reliable and valid instrument to measure functional capacity in boys with Duchenne muscular dystrophy.

Identification of the Male-Produced Aggregation Pheromone of the Four-Spotted Coconut Weevil, Diocalandra frumenti.

The four-spotted coconut weevil, Diocalandra frumenti Fabricius (Coleoptera: Dryophthoridae), is a small weevil found attacking economically important palm species, such as coconut, date, oil, and Canary palms. Given the scarcity of detection and management tools for this pest, the availability of a pheromone to be included in trapping protocols would be a crucial advantage. Previous laboratory experiments showed evidence for aggregation behavior; thus, our main goal was to identify the aggregation pheromone in this species. The volatile profile of D. frumenti individuals was studied by aeration and collection of effluvia in Porapak-Q and also by solid phase microextraction (SPME) techniques. Moreover, solvent extraction of previously frozen crushed individuals was also performed. All resulting extracts and SPME fibers were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). The comparison of male and female samples provided the candidate compound, 5-ethyl-2,4-dimethyl-6,8-dioxabicyclo[3.2.1]octane (multistriatin), whose biological activity was evaluated in olfactometer and field assays.

Olive mill effluent depuration by ozonation and Fenton processes enhanced by iron wastes.

The aim of the present work was to compare the potential of iron industry wastes to enhance ozone and hydrogen peroxide action on the degradation of olive mill wastewaters (OMWs). The results attained show a higher efficiency for ozonation using a lower catalyst load. Nevertheless, Fenton's process led to a larger amount of chemical oxidation demand (COD) removed per mole of oxidant applied. It was concluded that hydroxyl radicals are responsible for the pollutant abatement. High eco-toxicity decay was observed after the treatments. Furthermore, a preliminary analysis of the iron shavings' stability was made by reusing it in two feed-batch trials. It was concluded that while activity was maintained for Fenton's, a decrease of about 20% was verified for catalytic ozonation. Comparing these results with the ones obtained for the same processes applied to an actual OMW, a lower percentage of COD abatement was achieved. However, when reporting the amount of COD removed per mole of oxidant used, the difference between effluents are not so high. This should be taken into account when deciding which process should be implemented at an industrial scale. With the outcomes of this research it was possible to conclude that integrating waste management with wastewater treatment was feasible.

[Prevalence of respiratory symptoms in areas of the Federal District, Brazil]. / Prevalência de sintomáticos respiratórios em regiões do Distrito Federal, Brasil.

The identification of individuals with respiratory symptoms (RS) is important for the early detection of tuberculosis. The aim of this study was to estimate the prevalence of RS in three administrative regions of the Federal District, Brazil. For this, we used the 30 by 7 cluster sampling technique proposed by the World Health Organization. Individuals with RS were defined as those aged 15 years or older living in the administrative regions of Estrutural, Itapoã, or Varjão and reporting a cough lasting at least 3 weeks at the date of the interview. The prevalence of RS was 5.7% in Estrutural and Varjão (95% CI: 2.4-9.0) and 4.8% in Itapoã (95%CI: 1.6-7.9), with a design effect close to 1.0. In Estrutural and Itapoã, fewer years of schooling, and in Itapoã and Varjão, lower income, were associated with RS. Cigarette smoking was associated with the presence of RS in all regions. The prevalence of RS in the three administrative regions investigated is consistent with that of other areas with a similar socioeconomic profile.

Prevalência de sintomáticos respiratórios em regiões do Distrito Federal, Brasil / Prevalence of respiratory symptoms in areas of the Federal District, Brazil

A captação de sintomáticos respiratórios (SR) é importante para a detecção precoce de tuberculose. O objetivo deste estudo foi estimar a prevalência de SR em três regiões administrativas (RA) do Distrito Federal, Brasil. Para tanto, foi utilizada a técnica de amostragem por conglomerados (30 por 7) proposta pela Organização Mundial da Saúde. Foram definidos como SR os indivíduos com 15 anos ou mais residentes nas RAs Estrutural, Itapoã ou Varjão e que apresentassem tosse há pelo menos 3 semanas na data da entrevista. A prevalência de SR em Estrutural e Varjão foi de 5,7 por cento (IC95 por cento: 2,4 a 9,0), e em Itapoã, de 4,8 por cento (IC95 por cento: 1,6 a 7,9), com efeito de desenho próximo de 1,0. Em Estrutural e Itapoã, menos anos de estudo, e em Itapoã e Varjão, menor renda, estiveram associados com ser SR. O tabagismo esteve associado com a presença de sintomas respiratórios em todas as RAs. A prevalência de SR está de acordo com aquela encontrada em outras áreas com perfil socioeconômico semelhante ao das áreas estudadas.

The identification of individuals with respiratory symptoms (RS) is important for the early detection of tuberculosis. The aim of this study was to estimate the prevalence of RS in three administrative regions of the Federal District, Brazil. For this, we used the 30 by 7 cluster sampling technique proposed by the World Health Organization. Individuals with RS were defined as those aged 15 years or older living in the administrative regions of Estrutural, Itapoã, or Varjão and reporting a cough lasting at least 3 weeks at the date of the interview. The prevalence of RS was 5.7 percent in Estrutural and Varjão (95 percent CI: 2.4-9.0) and 4.8 percent in Itapoã (95 percentCI: 1.6-7.9), with a design effect close to 1.0. In Estrutural and Itapoã, fewer years of schooling, and in Itapoã and Varjão, lower income, were associated with RS. Cigarette smoking was associated with the presence of RS in all regions. The prevalence of RS in the three administrative regions investigated is consistent with that of other areas with a similar socioeconomic profile.

Biopsy of peripheral lymph nodes: a useful tool to diagnose opportunistic diseases in HIV-infected patients.

Peripheral lymphadenopathy is commonly present in HIV-infected patients and has a wide spectrum of differential diagnoses. We carried out a cross-sectional study of peripheral lymph node biopsies performed from January 2004 to December 2008 in HIV-infected patients who attended a tertiary-care hospital in southern Brazil. Only 60 of 210 peripheral lymph node biopsies performed (28%) were non-diagnostic. The most common diagnoses included: mycobacteriosis (105 cases; 50.2%); lymphoma (19 cases; 9.0%); systemic mycosis (12 cases; 5.7%) including histoplasmosis, cryptococcosis and histoplasmosis; and metastatic cancer (2.9%). Peripheral lymph node biopsy is a simple and useful tool to diagnose opportunistic diseases in HIV-infected patients.

Continuous infusion of amphotericin B deoxycholate in the treatment of cryptococcal meningoencephalitis: analysis of safety and fungicidal activity.

We measured fungicidal activity of continuous infusion of amphotericin B deoxycholate plus 5'flucytosine using quantitative cultures of cerebrospinal fluid (CSF) obtained from lumbar punctures of human immunodeficiency virus (HIV)-infected patients with neurocryptococcosis during 14 days of treatment. Glomerular renal function was preserved in all patients. Mycological efficacy with progressive reduction in CSF cryptococcal colony-forming units was comparable to standard 4-h infusion of amphotericin B.

Chronic slowly progressive monoarthritis tuberculosis of the hip without systemic symptoms mimicking osteoarthritis: a case report.

The authors report and discuss the clinical and radiological features of a immunocompetent patient with chronic progressive monoarthritis tuberculosis of the hip without systemic symptoms such as fever, and weight loss presenting as caseating abscess and severe destruction of the hip joint, treated with resection arthroplasty.

Determination of vitamin E and carotenoid pigments by high performance liquid chromatography in shell of Chionoecetes opilio.

This study reports the optimization of a method for the determination of vitamin E and carotenoids in shells of Chionoecetes opilio samples by online HPLC coupled with UV-vis and fluorescence detectors. The carotenoids were determined with diode-array detector (lambda 450 nm) and vitamin E with fluorescence detection (lambda(ex) 288, lambda(em) 331 nm). Two extractions methods were compared, saponification followed by an extraction step and a simple extraction with acetone. The last one was selected because allows to determine all compounds. Linearity, precisions and recoveries achieved for all compounds were satisfactory. Mean concentrations (mg per 100 g dry weight) were; 23.3 for vitamin E, 9.49 for astaxanthin and 0.2 mg for beta-carotene.

Laboratory tests in the detection of extended spectrum beta-lactamase production: National Committee for Clinical Laboratory Standards (NCCLS) screening test, the E-test, the double disk confirmatory test, and cefoxitin susceptibility testing

Extended spectrum beta-lactamase (ESBL) production by Klebsiella sp. and E. coli is an emerging problem. In this study, 107 clinical isolates (53 E. coli, 47 K. pneumoniae and 7 K. oxytoca) screened as ESBL producers by the NCCLS disk diffusion procedure were submitted to a double disk confirmatory test (DDT) and to the E-test double strip for confirmation of ESBL production by demonstration of clavulanic acid inhibition effect (CAIE). Only 72/107 (67 percent) of the isolates were confirmed as ESBL producers by DDT, with diverse results among species. By the E-test, 58/107 (54 percent) isolates were confirmed as ESBL producers, and 18/107 (17 percent) were not determinable. Susceptibility to cefoxitin was found in 57/68 (83 percent) of strains that did not show CAIE. ESBL detection remains a controversial issue and clinical laboratories are in need of a simple and effective way to recognize strains with this kind of resistance.

Laboratory tests in the detection of extended spectrum beta-lactamase production: National Committee for Clinical Laboratory Standards (NCCLS) screening test, the E-test, the double disk confirmatory test, and cefoxitin susceptibility testing.

Extended spectrum beta-lactamase (ESBL) production by Klebsiella sp. and E. coli is an emerging problem. In this study, 107 clinical isolates (53 E. coli, 47 K. pneumoniae and 7 K. oxytoca) screened as ESBL producers by the NCCLS disk diffusion procedure were submitted to a double disk confirmatory test (DDT) and to the E-test double strip for confirmation of ESBL production by demonstration of clavulanic acid inhibition effect (CAIE). Only 72/107 (67%) of the isolates were confirmed as ESBL producers by DDT, with diverse results among species. By the E-test, 58/107 (54%) isolates were confirmed as ESBL producers, and 18/107 (17%) were not determinable. Susceptibility to cefoxitin was found in 57/68 (83%) of strains that did not show CAIE. ESBL detection remains a controversial issue and clinical laboratories are in need of a simple and effective way to recognize strains with this kind of resistance.