Analysis of the structure of mycolic acids of Mycobacterium simiae reveals a particular composition of alpha-mycolates in strain 'habana' TMC 5135, considered as immunogenic in tuberculosis and leprosy.

Structural analysis of mycolic acids from Mycobacterium simiae (including some 'habana' strains) was carried out using (1)H-NMR and MS. Results indicated that this species presents a general pattern of alpha-, alpha'- and keto-mycolates. alpha-Mycolates were composed of a complex mixture of 82 to 89 carbon atoms (C82-C89), with the predominant molecular species containing two di-substituted cyclopropane rings. Among keto-mycolates (C84-C89), those containing one trans di-substituted cyclopropane ring were the most abundant. The alpha'-mycolates were monounsaturated (C64, C66). According to MS and (1)H-NMR data, the strains studied differed in fine structural details of alpha-mycolates and keto-mycolates. Notably, strain 'habana' TMC 5135 (belonging to the 'habana' group, and considered as highly immunogenic in tuberculosis and leprosy) presented a particular composition of alpha-mycolates, with a major component (C87) containing one cis plus one trans di-substituted cyclopropane ring, unlike the type strain of M. simiae and other strains of the 'habana' group (IPK-220 and IPK-337R), in which the major component (C84) contained two cis di-substituted cyclopropane rings. In spite of this finding, the 'habana' strains were closely related to each other and mainly differed from the type strain of M. simiae in some details of the fine structure of keto-mycolates. The present work indicated that within an identical general pattern of mycolic acids, there is a complex composition in M. simiae and structural variation among different strains, as reported for pathogenic species of the genus. Noteworthy was the particular composition of alpha-mycolates in strain 'habana' TMC 5135.

Drug-resistant tuberculosis in Cuba. Results of the three global projects.

SETTING: The spread of multidrug-resistant tuberculosis (MDR TB) in the world remains a major public health problem. Surveillance of anti-TB drug resistance is therefore an essential tool for monitoring the effectiveness of TB control program and, through policy development, for improving national and global TB control. OBJECTIVE: To determine the prevalence of anti-TB drug resistance in Cuba during the execution of the three global projects. DESIGN: Drug-resistance was determined using the proportion method in 1528 Mycobacterium tuberculosis strains to first line anti-TB drugs. RESULTS: Resistance in new cases was 8.3%, 4.6% and 5.0%; MDR was 0.7%, 0% and 0.3% in the first (1997), second (2000) and third (2004) global projects, respectively. In new cases, none showed resistance to the four drugs. There were among previously treated cases statistically significant decreases in the prevalence of resistance to at least one drug when comparing the results obtained in the three global projects. CONCLUSIONS: The contributed data through Cuba demonstrated that our country is relatively free of MDR strains, reflecting the good National Control Program and the possibility of TB elimination in Cuba.

[Surveillance of resistance of Mycobacterium tuberculosis to anti-TB drugs in Cuba, 1995-1998]. / Vigilancia de la resistencia de Mycobacterium tuberculosis a las drogas antituberculosas en Cuba, 1995-1998.

Drug-resistance surveillance by means of culture and drug susceptibility test (DST) allows the estimation of the regional and global magnitude of multidrug-resistance (MDR). Two reports on anti-tuberculosis drug resistance by the WHO/IUATLD showed that drug-resistance TB varied widely across regions. This work determined the prevalence of drug resistance among new cases and among previously treated cases of tuberculosis in Cuba during the period 1995-1998. Drug resistance was determined using the proportion method in 1379 Mycobacterium tuberculosis strains to first line antituberculosis drugs (isoniazid, streptomycin, ethambutol and rifampicin). The resistance in new cases was 8.3% and 6.5%; MDR was 0.7% and 0% in the first and second study respectively. The contributed data through Cuba demonstrated that our country is relatively free of MDR strains, recognizing it to world level the good tuberculosis-control programs and the success of the application in our country of the directly-observed-treatment strategy from the year 1971.

Vigilancia de la resistencia de Mycobacterium tuberculosis a las drogas antituberculosas en Cuba, 1995-1998

L a vigilancia de la resistencia a fármacos a través del cultivo y de las pruebas de susceptibilidad in vitro permite conocer la magnitud regional y mundial de la resistencia en tuberculosis. En el presente trabajo se determinó la prevalencia de la resistencia a fármacos antituberculosos en Cuba, durante el periodo 1995-1998 en casos nuevos y en aquéllos que han recibido tratamiento previo. Los resultados incluidos en este estudio forman parte de los dos proyectos mundiales organizados por la OMS/UICTER. La resistencia a los medicamentos se evaluó usando el método de las proporciones en 1.379 cepas de Mycobacterium tuberculosis a los fármacos de primera línea (isoniacida, rifampicina, estreptomicina y etambutol). La resistencia en casos nuevos fue del 8,3% y 6,5% y la resistencia múltiple a fármacos ( multidrug-resistance, MDR) fue del 0,7% y 0% en el primer y segundo estudio, respectivamente. Estos resultados permitieron demostrar la escasa circulación de cepas MDR en Cuba; se reconoció a nivel mundial el buen funcionamiento del Programa Nacional de Control y el éxito de la aplicación en nuestro país de la estrategia del tratamiento estrictamente supervisado desde 1971.

Surveillance of resistance of Mycobacterium tuberculosis to anti-TB drugs in Cuba, 1995-1998 Drug-resistance surveillance by means of culture and drug susceptibility test (DST) allows the estimation of the regional and global magnitude of multidrug-resistance (MDR). Two reports on anti-tuberculosis drug resistance by the WHO/IUATLD showed that drug-resistance TB varied widely across regions. Thecurrent study determined the prevalence of drug resistance among new cases and among previously treated cases of tuberculosis in Cuba during the period 1995-1998. Using the proportion method, drug resistance was determined in 1,379 Mycobacterium tuberculosis strains to four front-line anti- tuberculosis drugs (isoniazid, streptomycin, ethambutol and rifampicin). In two separate studies, the overall resistance in new cases was 8.3% and 6.5%, and the MDR was 0.7% and 0%, respectively. The Cuba data demonstrated that Colombia, in comparison, is relatively free of MDR strains, indicating effective tuberculosis control programs in Colombia and the success of the direct observation-treatment strategy that has been in continuous use since 1971.

The number and organization of the rRNA genes of several strains of Mycobacterium simiae.

The type strain of Mycobacterium simiae and four Cuban strains, each representing a group of variants sharing a characteristic pattern of glycopeptidolipids, were investigated. Each of the five strains was found to have a single rRNA (rrn) operon per genome. Each rrn operon was found to be located downstream from murA. Unusually for slow-growing mycobacteria, three transcription start points were identified for each operon. Gene sequences were established extending from near to the 3'-ends of murA, the intergenic regions and the 5'-ends of the 16S rDNAs. Characteristic strain differences were identified.

[Hepatitis C virus infection in female sexual workers from northern Lima]. / Hepatitis C en trabajadoras sexuales del cono norte de Lima.

UNLABELLED: Hepatitis C is the second major endemic viral infection in the world after HIV. In the USA it is the first liver transplant indication. Currently, the main risk factors to develop this disease are history of blood transfusions and IV drug usage. Sexual transmission remains controversial and accounts probably for less than 5% of case transmissions. PURPOSE: Determine the prevalence of hepatitis C virus infection in female sexual workers from Northern Lima and identify other risk factors. MATERIAL AND METHODS: A prospective study was performed in female sexual workers from Northern Lima. Risk factors were assessed in a survey; at the time of the assessment they were HIV negative. The COBAS CORE Anti-HCV EIA II Test was used. RESULTS: HCV serology was negative in all 98 sexual workers. 90% used protection with condoms, none used IV drugs and only one had a previous history of blood transfusion. CONCLUSIONS: HCV prevalence in female sexual workers evaluated was zero. They have few risk factors, sexual transmission of this virus is very low and even lower in a population where over 90% of the people uses condoms.

[Chylous ascites as evidence of a retroperitoneal lymphoma]. / Ascitis quilosa como manifestación de una linfoma retroperatoneal.

A case of Chylous Ascites was reported in a 64-year-old male patient who underwent a diagnostic laparoscopy, through which its etiology was determined. It is a retroperitoneal lymphoma with liver infiltration, an unusual presentation of this disease, where diagnosis could be determined.

Análisis de ácidos grasos de cepas de Mycobacterium habana y Mycobacterium simiae / Analysis of fatty acids from strains of Mycobacterium habana and Mycobacterium simiae

A comparative analysis of mycobacterial fatty acid fractions of Mycobacteriun habana and Mycobacterium sineae strains was made. This study used the gas-liquid chromatography coupled with mass spectrometry. Chromatographic profiles obtained from this technique were exposed and compared. This technique proves to be valuable as an alternative element in mycobacterial characterization and makes it possible to analyze the possible differences that may exist among mycobacterial species and to identify the present fatty acid fractions. The outcome proved that the studied strains had quantifiable quantities of over 20 C atom chain fatty acids. There are small differences among the strains in terms of these organic components. It was confirmed that each of then describes a characteristic chromatographic pattern although the composition of present fatty acids is very similar in both studied species.

Análisis cromatográfico de cepas de Mycobacterium tuberculosis aisladas de un brote en pacientes VIH en Cuba

Se realiza el estudio mediante técnicas cromatográficas, de un grupo de cepas de Mycobacterium tuberculosis aisladas de un brote en pacientes infectados con el virus VIH. Se utilizaron como referencia un grupo de cepas de M. tuberculosis de pacientes sintomáticos respiratorios (SR+14) y cepas patrones de la colección del laboratorio, con el objetivo de comparar cualitativamente los patrones cromatográficos descritos por las cepas aisladas de pacientes. Se obtuvieron y compararon los perfiles cromatográficos de las cepas aisladas de pacientes (ST+) y de VIH+ por la técnica de cromatografía en capa delgada. Se identificó cada uno de los ácidos grasos presentes por la técnica de cromatografía de gases acoplada a espectrometría de masas. Todas las cepas estudiadas fueron clasificadas como Mycobacterium tuberculosis. Por resultados obtenidos se demuestra la utilidad de las técnicas cromatográficas como técnicas alternativas para el diagnóstico micobacteriano.

A group of strains of Mycobacterium tuberculosis isolated from an outbreak in HIV-infected patients was studied by chromatographic techniques.A group of strains of M. Tuberculosis from symptomatic respiratory patients (SR + 14) and patterns strains from the laboratory collection were used as a reference aimed at making a qualitative comparison of the chromatographic patterns described by the strains isolated from patients. The chromatographic profiles of the strains isolated from patients (SR +) and fro HIV + were obtained and comparede by thin layer chromatography (TLC). Each of the present fatty acids was identified by using the gas chromatography technique (GC) coupled to mass spectrum analysis. All the studied strains were classified as Mycobacterium tuberculosis. According to the results attained, the usefulness of the chromatographic techniques as alternative techniques for the mycobacterial diagnosis is demonstrated.

Analysis of lipids reveals differences between 'Mycobacterium habana' and Mycobacterium simiae.

Fatty and mycolic acids and the pattern of glycolipids were studied in a collection of 34 strains of 'Mycobacterium habana' and in two strains of Mycobacterium simiae. Major glycolipids of these micro-organisms were assigned to the glycopeptidolipid (GPL) structural type, but both mycobacteria differed in the patterns obtained by TLC. The strains of 'M. habana' were separated into four groups (A-D), taking into account the presence or absence of several polar GPLs: group A contained GPL-I, GPL-II and GPL-III; group B contained GPL-I, GPL-II', GPL-II and GPL-III; group C contained GPL-II', GPL-II and GPL-III; group D did not contain any of these compounds. Fatty acids of both bacteria were similar, and ranged from 14 to 26 carbon atoms, hexadecanoic, octadecenoic and tuberculostearic acids being predominant. Mycolic acids were also similar by TLC and HPLC, and consisted of alpha-, alpha'- and ketomycolates. Partial structural analysis by MS carried out in strains 'M. habana' TMC 5135 and M. simiae ATCC 25275T revealed that alpha- and ketomycolates ranged, in general, from 79 to 87 carbon atoms, and alpha'-mycolates from 58 to 67 carbon atoms. The alpha- and ketomycolates belonged to several structural series, and minor variations were found between the two strain examined. The data obtained justified the synonymy between 'M. habana' and M. simiae but indicated, in turn, that the former can be distinguished on the basis of GPL analysis. Most strains of 'M. habana' can be defined by the presence of GPL-II and GPL-III, a finding that could be useful in the quality control of potential vaccine strains.

Utilización del medio de cultivo UIT-A en la obtención de biomasa para la identificación bioquímica micobacteriana

Se compara el medio de cultivo sólido UIT-A con el Lowenstein Jensen, para la obtención de biomasa en el montaje del test bioquímico de identificación de micobacterias "no tuberculosas" (MNT), con el objetivo de valorar la posibilidad de utilización como medio de cultivo para el montaje y análisis de dichas pruebas bioquímicas. Los resultados obtenidos en los 2 medios de cultivo utilizados fueron iguales, por lo que se recomienda el uso del medio de cultivo UIT-A fundamentalmente para aquellas cepas con crecimiento pobre o escaso.

Resistencia a drogas antimicrobianas, en cepas de Haemophilus influenzae tipo b, aisladas de pacientes con síndrome neurológico infeccioso / Antimicrobial drug resistance in Haemophilus influenzae type b strains isolated from patients with infectious neurologic syndrome

Se estudió por primera vez la sensibilidad de 92 cepas de Haemophilus influenzae tipo b, a diferentes antimicrobianos : ampicillina, cloranfenicol, rifampicina, ceftriaxone (rocephin) y cefotaxime (claforán), por el método de dilución en placas de agar. Se encontró una mayor resistencia de las cepas estudiadas frente al ampicillina (42,39 por ciento ) y al cloranfenicol (58,69 por ciento ) y una alta sesibilidad frente a la rifampicina (100 por ciento ), Ceftriaxone (100 por ciento ) y Cefotaxime (100 por ciento )

Utilización de una técnica de aglutinación al latex (Finlatex-Staph) para la identificación de cultivos de Staphylococcus aureus / Use of a latex agglutination technique (Finlatex-Staph) for the identification of staphylococus aureus cultures

Se compara una forma rápida de identificar cultivos de Staphylococcus aureus con la utilización de la técnica de aglutinación al látex y algunas de las técnicas utilizadas tradicionalmente para la clasificación de dichos cultivos. Se clasificaron mediante las técnicas tradicionales 90 cepas como staphylococcus aureus y 10 como no Staphylococcus aureus. La técnica de aglutinación al látex identificó correctamente 86 de las cepas identificadas inicialmente como Staphylococcus aureus por lo que se considera una prueba rápida y sensible

Crecimiento micobacteriano en el medio UIT modificado (UIT-L), modificación líquida: ensayo I / Mycobacterial growth in a modificated medium UIT (UIL-L), liquid modification: assay I

Diferentes especies micobacterianas no tuberculosas fueron sembradas en la modificación líquida del medio UIT, la cual denominamos UIT-L. Se midió el crecimiento micobacteriano al realizar lecturas de las densidades ópticas cada 18 horas. Se obtuvo crecimiento en 14 de las 16 especies micobacterianas estudiadas. En las especies Mycobacterium terrae y Mycobacterium xenopi el crecimiento fue pobre, en el resto, sus curvas favorecen un aumento de la concentración celular

Ensayo de un esquema de clasificación de cepas de mycobacterium tuberculosis por marcadores fagotípicos / Assay of a scheme for the classification of strain of mycobacterium tuberculosis by phagotypin

Se ensaya un sistema de clasificación para el marcaje fagotípico de 18 cepas de Mycobacterium tuberculosis, utilizando un juego de 10 micobacteriofágos, que incluye 4 fagos mayores y 6 fagos auxiliares o de subdivisión. De las 18 cepas se obtuvieron 5 del tipo A0, 3 del tipo A2, 3 de los tipos A4, A5 y B, que no habían sido informadas anteriormente en el país y I del tipo A3. Se logró una apertura en la diferenciación de las cepas de M. tuberculosis en el número de 16 subtipos diferentes, de los 6 tipos incialmente encontrados por el esquema, con los fagos mayores

Estudio comparativo de diferentes medios para micobacterias, que incluye la nueva modificación UIT-A / Comparative study of different media for mycobacteria, including new UIT-A modification

Se estudia la eficiencia de la producción de biomasa micobacteriana entre diferentes medios de cultivos, éstos son: Sauton agar, Middlebrook, UIT y la modificación UIT-A. Los resultados obtenidos después del análisis estadístico son satisfactorios con respecto a la modificación UIT-A

Aplicación de la técnica de cromatografía en capa delgada a 32 cepas del M- habana / Application of thin-layer chromatography to 32 strains of M. habana

Se informa que se estudiaron 32 cepas del M. habana aisladas de pacientes sintomáticos respiratorios y se comparan por la técnica de cromatografía en capa delgada. Se plantea que la extracción de lípidos se hizo aplicando el método de diacetilación. Se trabaja con los lípidos intactos y los lípidos diacetilados. Se concluye que esta especie micobacteriana pertencente al Grupo I (según Brennan), pues sus lípidos son álcalis resistentes