Evaluation of crystal violet decolorization assay for minimal inhibitory concentration detection of primary antituberculosis drugs against Mycobacterium tuberculosis isolates

In this study we evaluated the crystal violet decolorization assay (CVDA) for detection of minimum inhibitory concentration (MIC) of antituberculosis drugs. 53 isolates were tested in this study and 13 of them were multidrug resistant (MDR) isolates. The antibiotics concentrations were 2-0.06 mg/L for isoniazid (INH) and rifampicin (RIF) and were 16-0.25 mg/L for streptomycin (STM) and ethambutol (EMB). Crystal violet (CV-25 mg/L) was added into the microwells on the seventh day of incubation and incubation was continued until decolorization. Decolorization of CV was the predictor of bacterial growth. Overall agreements for four drugs were detected as 98.1%, and the average time was detected as 9.5 ± 0.89 day after inoculation. One isolate for INH and two isolates for STM were determined resistant in the reference method, but susceptible by the CVDA. One isolate was susceptible to EMB by the reference method, but resistant by the CVDA. All results were concordant for RIF. This study shows that CVDA is a rapid, reliable and suitable for determination of MIC values of Mycobacterium tuberculosis. And it can be used easily especially in countries with limited-sources.

Ectasia ductal mamaria en niños, a propósito de un caso clínico: A case report / Mammary duct ectasia in children

Introducción: La telorragia es un síntoma poco frecuente en pacientes pediátricos, la causa más frecuente en esta población es la ectasia ductal mamaria (EDM), que es una afección benigna y autolimitada, caracterizada por la dilatación del conducto mamario, fibrosis e inflamación periductal. Objetivo: Presentar un caso de EDM, para facilitar el rápido reconocimiento por parte de los médicos, y evitar estudios y tratamientos agresivos. Caso clínico: Lactante de sexo masculino de 6 meses de edad, sano, alimentado por lactancia materna exclusiva; consultó por un nódulo retroareolar derecho y telorragia unilateral. Se realizó una ecografía Doppler que mostró una lesión multiquística, sugerente de una EDM. Se planteó tratamiento expectante y acudió a control a los 6 meses con excelente evolución. Conclusiones: La EDM es la principal causa de telorragia en niños, corresponde a una afección benigna, y la resolución generalmente es espontánea, antes de los 9 meses. Por lo que su conocimiento es de gran relevancia para el adecuado diagnóstico y manejo de estos pacientes.

Introduction: Bloody nipple discharge is an infrequent symptom during childhood. The most common cause in this population is mammary duct ectasia (MDE), which is a benign and self-limiting condition, that is characterized by dilatation of the mammary ducts, fibrosis and periductal inflammation. Objective: Report of a case of MDE in order to improve physicians’ diagnosis accuracy and avoid aggressive studies and treatments. Case report: Six-months old male healthy infant, exclusively breastfeeded, that visited our clinic with a lump beneath his right nipple and bloody discharge from the same nipple. An ultrasound was performed which showed a multicystic lesion suggestive of MDE. Watchful waiting was decided as treatment, with good evolution after six months of follow up. Conclusions: The MDE is the leading cause of bloody discharge in pediatric population, being a benign condition that resolves spontaneously before nine months. The knowledge of this condition is essential so as to accurately diagnose and treat it.

Methylparaben concentration in commercial Brazilian local anesthetics solutions

OBJECTIVE: To detect the presence and concentration of methylparaben in cartridges of commercial Brazilian local anesthetics. MATERIAL AND METHODS: Twelve commercial brands (4 in glass and 8 in plastic cartridges) of local anesthetic solutions for use in dentistry were purchased from the Brazilian market and analyzed. Different lots of the commercial brands were obtained in different Brazilian cities (Piracicaba, Campinas and São Paulo). Separation was performed using high performance liquid chromatography (HPLC) with UV-Vis detector. The mobile phase used was acetonitrile:water (75:25 - v/v), pH 4.5, adjusted with acetic acid at a flow rate of 1.0 ml.min-1. RESULTS: When detected in the solutions, the methylparaben concentration ranged from 0.01% (m/v) to 0.16% (m/v). One glass and all plastic cartridges presented methylparaben. CONCLUSION: 1. Methylparaben concentration varied among solutions from different manufacturers, and it was not indicated in the drug package inserts; 2. Since the presence of methylparaben in dental anesthetics is not regulated by the Brazilian National Health Surveillance Agency (ANVISA) and this substance could cause allergic reactions, it is important to alert dentists about its possible presence.

Improvement of XTT assay performance for studies involving Candida albicans biofilms

2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay has been used to study Candida biofilm formation. However, considering that the XTT reduction assay is dependent on cell activity, its use for evaluating mature biofilms may lead to inaccuracies since biofilm bottom cell layers tend to be relatively quiescent at later stages of biofilm formation. The aim of this study was to improve XTT reduction assay by adding glucose supplements to the standard XTT formulation. Candida albicans ATCC 90028 was used to form 24-, 48- and 72-h biofilms. The oxidative activity at 90, 180 and 270 min of incubation was evaluated. The control consisted of standard XTT formulation without glucose supplements, and was modified by the addition of 50, 100 and 200 mM of glucose. The XTT assay with 200 mM glucose showed more accurate and consistent readings correlating with biofilm development at 24, 48 and 72 h. Biofilm growth yield after 180 min incubation, when evaluated with the 200 mM glucose supplemented XTT, produced the most consistent readings on repetitive testing. It may be concluded that glucose supplementation of XTT could minimize variation and produce more accurate data for the XTT assay.

O teste de redução do 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) tem sido utilizado para mensurar o desenvolvimento de biofilmes de Candida. Contudo, a reação de XTT é dependente da atividade celular e o seu uso para biofilmes maduros pode ser questionado, considerando que diferentes camadas celulares têm atividade metabólica diferenciadas. O objetivo deste estudo foi avaliar se a adição de glicose à formula de XTT diminuiria a variabilidade na mensuração da atividade metabólica. Biofilmes de Candida albicans ATCC 90028 com tempos de crescimento de 24, 48 e 72 h foram utilizados. Para avaliar o melhor tempo de incubação do XTT, este foi mantido a temperatura de 37 °C, em tempos de 90 180 e 270 min. A fórmula padrão do teste XTT (controle) foi modificada com a adição de 50, 100 e 200 mM de glicose para os grupos experimentais. Os melhores resultados para a incubação foi observado com tempo de 180 min e para a suplementação de glicose à concentração de 200 mM (p<0.001). Concluiu-se que a incubação de 180 min utilizando a suplementação de 200 mM de glicose apresenta resultados de atividade metabólica celular com a menor variação para o estudo de biofilmes de Candida albicans.

Comparative research of the influence factors of DNA extraction of bloodstain on the filter paper with Chelex-100 method / 法医学杂志

OBJECTIVE@#To improve DNA extraction from bloodstain on the filter paper and to establish a rapid, simple, and cost-effective method for DNA extraction suitable for database construction.@*METHODS@#Seven hundred and fifty two aged bloodstains on filter paper were randomly divided into four groups. The four different DNA extraction methods were compared with each other, and two DNA extraction methods used for 63 fresh bloodstains on filter paper were also compared with each other.@*RESULTS@#There were no statistically significant differences observed among the four DNA extraction methods (P > 0.05) for aged bloodstains on filter paper; But the difference between the two DNA extraction methods for fresh bloodstains on filter paper was obviously (P < 0.05).@*CONCLUSION@#Extraction of DNA samples from aged bloodstains on filter paper can be accomplished by using Chlex-100 methodology directly with no need to wash the bloodstains.

Detergent-mediated destaining of coomassie brilliant blue-stained SDS polyacrylamide gels.

A simple and one-step detergent-mediated destaining procedure for SDS Polyacrylamide gels for proteins is described. Suspension (5%, w/v) of a commercially available household detergent, Vim Ultra, has been found to be very efficient in destaining polyacrylamide gels without interfering with the resolution of proteins. As compared to the routinely used solvent (methanol-acetic acid-water)-mediated destaining procedure, the present method is economical and user-friendly.

Spectrophotometric analysis of resazurin reduction test and semen quality in men.

The resazurin reduction test (RRT) was performed on semen samples obtained from 225 untreated subfertile and 10 pregnancy confirmed fertile males. The results of RRT were determined visually using resazurin colour chart and again the extent of resazurin reduction in each sample was additionally read by spectrophotometer to assess the quality of samples. Absorption spectra was scanned for resazurin and resorufin and two most sensitive wavelengths (572 and 600 nm) were selected. The ratio of the two optical densities was used as a probe to discriminate the various grades of semen samples. In azoospermic samples, RRT ratio ranges from 0.7 to 1.16, in oligoasthenozoospermic samples from 1.10 to 1.35, in oligozoospermic samples from 1.5 to 2.0 (characterised visually as grades from 1-4) in normozoospermic and proven fertile samples from 2.25 to 5.9 (characterized visually as grades from 5 to 11). The highest correlation of RRT ratio was observed with sperm motility (r = 0.889, P < 0.001), followed by concentration (r = 0.848, P < 0.001) morphology (r = 0.660, P < 0.001) and viability (r = 0.544, P < 0.01). The test ratio had a positive predictive value of 95% for a sperm concentration of > 20 x 10(6)/ml and motility > 40% and a negative predictive value of 90% for a sperm concentration of < 20 x 10(6)/ml and motility < 40%. Therefore the evaluation of RRT results using spectrophotometric ratio method may provide a tool for obtaining a wider range of seminological diagnosis more accurately than the routine semen analysis. It is suggested that the method is simple and reliable, it can be performed in any andrology laboratories.

Comparative evaluation of determination of beta-lactam intermediates.

The Schiff's base formation between 6-aminopenicillanic acid, 7-aminodesacetoxycephalosporanic acid and 7-aminocephalosporanic acid and p-dimethylaminobenzaldehyde (PDAB) was investigated. The factors that affect the reaction such as concentration of PDAB, time and pH were studied and optimised for estimation of these intermediates.

Application of 3-methyl-1-phenyl-2-pyrazoline-4, s-dione as a novel reagent for development of latent fingerprints on paper

3-methyl- 1-phenyl- 2-pyrazoline- 4,5-dione is a new substance which has been prepared in the laboratory for detection of amino-acids. In this work, this substance of developing latent fingerprints on paper was used. It was compared with a well-known developer "ninhydrin" and was found to be the same as regard to sensitivity, clarity and contrast, whether the latent fingerprints were new or old, but the new substance is superior with regard to its preparation as it is easily prepared in the laboratory from available cheap substances i.e. it is economically better than ninhydrin