Oral supplementation with postbiotics modulates the immune response produced by myxomatosis vaccination in wild rabbits.

Rabbits (Oryctolagus cuniculus) are vitally important species in the Iberian Peninsula ecosystem. However, since 1950, there has been a significant population decline, with major repercussions. This situation is mainly due to the presence of infectious diseases, such as myxomatosis, which is expanding and is characterized by severe and fatal clinical manifestations. Current control measures, mainly those based on vaccinations, are ineffective. Therefore, new strategies need to be developed and implemented. This study aimed to evaluate whether supplementation with postbiotic products modulates the immune response in wild rabbits vaccinated against myxomatosis. For this purpose, two groups of rabbits were established: a control group fed with standard feed ad libitum from weaning (28 days) until two months of age, and a treated group, which was fed under the same conditions but supplemented with postbiotics (3 kg/Tm). All the studied rabbits were vaccinated against this disease during weaning. In addition, a blood samples were obtained from all animals immediately before vaccination and 30 days later, which allowed us to evaluate the level of antibodies against myxomatosis virus (ELISA detection) and the relative expression of gene encoding to cytokines related to the immune response (IL6, TNFα and IFNγ), at both times of the experience. Weight and length measurements were also taken at both times to calculate body index and mean daily gain (MDG). No statistically significant differences in growth parameters were observed. There were also no differences in the serological response among groups. However, a relative underexpression of gene codifying to TNFα (p-value = 0.03683) and a higher expression on IFNγ (p-value = 0.045) were observed in the treated group. This modulation in cytokines could lead to less severe lesions in wild rabbit naturally infected with myxomatosis virus.

Infecciones de transmisión sexual (ITS) durante el estado de alarma por la pandemia de COVID-19 en España / Sexually Transmitted Infections During the First Wave of the COVID-19 Pandemic in Spain

Objetivo: Describir los cambios en el comportamiento de las ITS durante la situación de alerta sanitaria por la pandemia SARS-CoV-2. Material y métodos: Se recogieron datos demográficos, cronológicos y clínicos de todos los pacientes que solicitaron atención médica por ITS en los hospitales La Paz y Costa del Sol, y en los Centros Sandoval y de Diagnóstico Médico, entre el 14/03/2020 y el 30/06/2020. Resultados: Documentamos 674 casos de ITS. La mediana de edad fue de 33años. El mayor porcentaje de casos se dio en el rango de 30-40años y en hombres que tenían sexo con hombres. Los diagnósticos más frecuentes fueron: proctitis (36,5%), sífilis (16%), uretritis no gonocócica (13,3%) y gonocócica (11,3%), herpes genital (8,8%), vulvovaginitis/cervicitis (8,3%) y condilomas (4,2%). En el 77% de los casos hubo confirmación microbiológica, siendo los microorganismos más frecuentes Chlamydia trachomatis (35,7%), Neisseria gonorrhoeae (31,4%) y Treponema pallidum (17,2%). Se constató un incremento del número de casos de ITS tras el desconfinamiento, explicable por las mayores libertades y el aumento de consultas. Comparando los registros de ITS del Centro Sandoval y del Hospital La Paz en los cuatrimestres de marzo a junio de 2019 frente a 2020, se observó una disminución en 2020 de todos los diagnósticos, de hasta el 81% menos que en 2019.Conclusiones:Las medidas de distanciamiento y limitación de movilidad aparentemente generaron una disminución de la incidencia de ITS, pero sin llegar a una inhibición completa de las conductas sexuales de riesgo (AU)

Objectivs: To describe changes in sexually transmitted infection (STI) diagnoses during the first wave of the COVID-19 pandemic in Spain. Material and methods: We collected demographic, chronological, and clinical data for all patients seen for a possible STI at Hospital La Paz, Centro Sanitario Sandoval, and Centro de Diagnóstico Médico in Madrid and Hospital Costa del Sol in Malaga between March 14, 2020 and June 30, 2020. Results: We identified 674 STI diagnoses. The median age of the patients was 33years. Most cases were observed among people aged 30 to 40years and among men who have sex with men. The most common diagnoses were proctitis (36.5%), syphilis (16%), nongonococcal (13.3%) and gonococcal (11.3%) urethritis, genital herpes (8.8%), vulvovaginitis/cervicitis (8.3%), and genital warts (4.2%). A microbiologically confirmed diagnosis was on record for 77% of cases. The main microorganisms identified were Chlamydia trachomatis (35.7%), Neisseria gonorrhoeae (31.4%) and Treponema pallidum (17.2%). The number of STI diagnoses increased after the easing of lockdown restrictions, which resulted in greater freedom of movement and more consultations. On comparing the 2019 and 2020 STI registries from Centro Sanitario Sandoval and Hospital La Paz for the period March to June, we observed reductions (of up to 81%) in all STI diagnoses. Conclusions: Physical distancing and movement restrictions appear to have resulted in a reduction in the incidence of STIs, although these measures did not completely eliminate sexual risk behaviors (AU)

[Translated article] Sexually Transmitted Infections During the First Wave of the COVID-19 Pandemic in Spain / Infecciones de transmisión sexual (ITS) durante el estado de alarma por la pandemia de COVID-19 en España

Objectivs: To describe changes in sexually transmitted infection (STI) diagnoses during the first wave of the COVID-19 pandemic in Spain. Material and methods: We collected demographic, chronological, and clinical data for all patients seen for a possible STI at Hospital La Paz, Centro Sanitario Sandoval, and Centro de Diagnóstico Médico in Madrid and Hospital Costa del Sol in Malaga between March 14, 2020 and June 30, 2020. Results: We identified 674 STI diagnoses. The median age of the patients was 33years. Most cases were observed among people aged 30 to 40years and among men who have sex with men. The most common diagnoses were proctitis (36.5%), syphilis (16%), nongonococcal (13.3%) and gonococcal (11.3%) urethritis, genital herpes (8.8%), vulvovaginitis/cervicitis (8.3%), and genital warts (4.2%). A microbiologically confirmed diagnosis was on record for 77% of cases. The main microorganisms identified were Chlamydia trachomatis (35.7%), Neisseria gonorrhoeae (31.4%) and Treponema pallidum (17.2%). The number of STI diagnoses increased after the easing of lockdown restrictions, which resulted in greater freedom of movement and more consultations. On comparing the 2019 and 2020 STI registries from Centro Sanitario Sandoval and Hospital La Paz for the period March to June, we observed reductions (of up to 81%) in all STI diagnoses. Conclusions: Physical distancing and movement restrictions appear to have resulted in a reduction in the incidence of STIs, although these measures did not completely eliminate sexual risk behaviors (AU)

Objetivo: Describir los cambios en el comportamiento de las ITS durante la situación de alerta sanitaria por la pandemia SARS-CoV-2. Material y métodos: Se recogieron datos demográficos, cronológicos y clínicos de todos los pacientes que solicitaron atención médica por ITS en los hospitales La Paz y Costa del Sol, y en los Centros Sandoval y de Diagnóstico Médico, entre el 14/03/2020 y el 30/06/2020. Resultados: Documentamos 674 casos de ITS. La mediana de edad fue de 33años. El mayor porcentaje de casos se dio en el rango de 30-40años y en hombres que tenían sexo con hombres. Los diagnósticos más frecuentes fueron: proctitis (36,5%), sífilis (16%), uretritis no gonocócica (13,3%) y gonocócica (11,3%), herpes genital (8,8%), vulvovaginitis/cervicitis (8,3%) y condilomas (4,2%). En el 77% de los casos hubo confirmación microbiológica, siendo los microorganismos más frecuentes Chlamydia trachomatis (35,7%), Neisseria gonorrhoeae (31,4%) y Treponema pallidum (17,2%). Se constató un incremento del número de casos de ITS tras el desconfinamiento, explicable por las mayores libertades y el aumento de consultas. Comparando los registros de ITS del Centro Sandoval y del Hospital La Paz en los cuatrimestres de marzo a junio de 2019 frente a 2020, se observó una disminución en 2020 de todos los diagnósticos, de hasta el 81% menos que en 2019.Conclusiones:Las medidas de distanciamiento y limitación de movilidad aparentemente generaron una disminución de la incidencia de ITS, pero sin llegar a una inhibición completa de las conductas sexuales de riesgo (AU)

[Sexually Transmitted Infections During the First Wave of the COVID-19 Pandemic in Spain]. / Infecciones de transmisión sexual (ITS) durante el estado de alarma por la pandemia de COVID-19 en España.

OBJECTIVE: To describe changes in sexually transmitted infection (STI) diagnoses during the first wave of the COVID-19 pandemic in Spain. MATERIAL AND METHODS: We collected demographic, chronological, and clinical data for all patients seen for a possible STI at Hospital La Paz, Centro Sanitario Sandoval, and Centro de Diagnóstico Médico in Madrid and Hospital Costa del Sol in Malaga between March 14, 2020 and June 30, 2020. RESULTS: We identified 674 STI diagnoses. The median age of the patients was 33 years. Most cases were observed among people aged 30 to 40 years and among men who have sex with men. The most common diagnoses were proctitis (36.5%), syphilis (16%), nongonococcal (13.3%) and gonococcal (11.3%) urethritis, genital herpes (8.8%), vulvovaginitis/cervicitis (8.3%), and genital warts (4.2%). A microbiologically confirmed diagnosis was on record for 77% of cases. The main microorganisms identified were Chlamydia trachomatis (35.7%), Neisseria gonorrhoeae (31.4%) and Treponema pallidum (17.2%). The number of STI diagnoses increased after the easing of lockdown restrictions, which resulted in greater freedom of movement and more consultations. On comparing the 2019 and 2020 STI registries from Centro Sanitario Sandoval and Hospital La Paz for the period March to June, we observed reductions (of up to 81%) in all STI diagnoses. CONCLUSIONS: Physical distancing and movement restrictions appear to have resulted in a reduction in the incidence of STIs, although these measures did not completely eliminate sexual risk behaviors.

Sexually Transmitted Infections During the First Wave of the COVID-19 Pandemic in Spain.

OBJECTIVE: To describe changes in sexually transmitted infection (STI) diagnoses during the first wave of the COVID-19 pandemic in Spain. MATERIAL AND METHODS: We collected demographic, chronological, and clinical data for all patients seen for a possible STI at Hospital La Paz, Centro Sanitario Sandaval, and Centro de Diagnóstico Médico in Madrid and Hospital Costa del Sol in Malaga between March 14, 2020 and June 30, 2020. RESULTS: We identified 674 STI diagnoses. The median age of the patients was 33 years. Most cases were observed among people aged 30 to 40 years and among men who have sex with men. The most common diagnoses were proctitis (36.5%), syphilis (16%), nongonococcal (13.3%) and gonococcal (11.3%) urethritis, genital herpes (8.8%), vulvovaginitis/cervicitis (8.3%), and genital warts (4.2%). A microbiologically confirmed diagnosis was on record for 77% of cases. The main microorganisms identified wereChlamydia trachomatis (35.7%), Neisseria gonorrhoeae (31.4%), and Treponema pallidum (17.2%). The number of STI diagnoses increased after the easing of lockdown restrictions, which resulted in greater freedom of movement and more consultations. On comparing the 2019 and 2020 STI registries from Centro Sanitario Sandoval and Hospital La Paz for the period March to June, we observed reductions (of up to 81%) in all STI diagnoses. CONCLUSIONS: Physical distancing and movement restrictions appear to have resulted in a reduction in the incidence of STIs, although these measures did not completely eliminate sexual risk behaviors.

OBJETIVO: Describir los cambios en el comportamiento de las ITS durante la situación de alerta sanitaria por la pandemia SARS-CoV-2. MATERIAL Y MÉTODOS: Se recogieron datos demográficos, cronológicos y clínicos de todos los pacientes que solicitaron atención médica por ITS en los hospitales La Paz y Costa del Sol, y los Centros Sandoval y de Diagnóstico Médico entre el 14/03/2020 y el 30/06/2020. RESULTADOS: Documentamos 674 casos de ITS. La mediana de edad fue de 33 años. El mayor porcentaje de casos se dio en el rango de 30-40 años y en hombres que tenían sexo con hombres. Los diagnósticos más frecuentes fueron: proctitis (36,5%), sífilis (16%), uretritis no gonocócica (13,3%) y gonocócica (11,3%), herpes genital (8,8%), vulvovaginitis/cervicitis (8,3%) y condilomas (4,2%).En 77% de los casos hubo confirmación microbiológica, siendo los microorganismos más frecuentes Chlamydia trachomatis (35,7%), Neisseria gonorrhoeae (31,4%) y Treponema pallidum (17,2%).Se constató un incremento del número de casos de ITS tras el desconfinamiento, explicable por las mayores libertades y el aumento de consultas. Comparando los registros de ITS del Centro Sandoval y el Hospital La Paz en los cuatrimestres de marzo a junio de 2019 frente a 2020, se observó una disminución en 2020 de todos los diagnósticos, de hasta un 81% menos que en 2019. CONCLUSIONES: Las medidas de distanciamiento y limitación de movilidad aparentemente generaron una disminución de la incidencia de ITS, pero sin llegar a una inhibición completa de las conductas sexuales de riesgo.

Ultrastructural study of the denervated diaphragm in rats / Estudio ultraestructural del diafragma desnervado en ratas

The structural alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into fragments, which were used for analysis transmission electron microscopy. During the initial phase of denervation (4 weeks), ultrastructural muscle modifications appeared in scattered fibers and in foci along these fibers. Muscle fibers with foci of less dense and loosely arranged myofibrils, disorganized Z line, displaced T tubules, and central nucleus exhibiting reentrances and fragmented aspect were observed. After 8 weeks, formation of large aggregates of small elongated mitochondria showing altered cristae, matrix inclusions and increased electron density was noted. At 12 weeks of denervation the alterations were found to be more drastic. Nuclei with internal deposits of myofibrillar or amorphous material were observed. In these fibers, vacuoles harbored enormous myeloid structures in the subsarcolemmal or intermyofibrillar region.

Fueron estudiadas las alteraciones ultra estructurales de las fibras musculares del diafragma denervado de ratas. Fueron utilizadas 15 ratas albinas (Rattus norvegicus), machos, adultas, con peso promedio de 200 g de aproximadamente 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, que fueron utilizados para el estudio en microscopia electrónica de transmisión. Durante los períodos iniciales de denervación (4 semanas), las modificaciones en la ultraestructura del músculo se disponen en fibras dispersas y en focos a lo largo de las mismas. Se observan fibras musculares con focos de miofibrillas rarefactas y laxamente dispuestas; línea Z desorganizada; túbulos T dislocados; núcleo central con aspecto fragmentado. Después de 8 semanas de denervación, se observa la formación de numeroso agregados de pequeñas mitocondrias alargadas, con alteraciones en las crestas, inclusiones en la matriz y aumento de la electrodensidad. Con 12 semanas de denervación, las alteraciones se muestran más drásticas; se observan núcleos con depósitos internos de material miofibrillar o amorfo. En estas fibras, las vacuolas presentan grandes estructuras mieloides en la región subsarcolemal o intermiofibrillar.

Morphologic alterations resulting from denervation of the diaphragm in rats / Alteraciones morfológicas resultantes de la denervación del diafragma en ratas

The morphological alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into two fragments, which were used for histological (H.E.) and histoenzymological (NADH-TR and myofibrillar ATPase). After 4 weeks, denervated muscle fibers showed important light microscopic alterations: atrophy with angular profiles in cross-sections, cytoplasm containing vacuoles, enlarged interstitial space with increased connective tissue, cellular infiltration, and muscle fibers without defined contours. The most marked alterations being observed for type IIb and IIa fibers. Eight and 12 weeks after denervation, the NADH-TR reaction showed that it was impossible to characterize the muscle fibers based on their metabolic profile...

Fueron estudiadas las alteraciones morfológicas de las fibras musculares del diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus) machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Se denervó el diafragma y después de 4, 8 y 12 semanas los animales fueron sacrificados. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, utilizados para el estudio histológico (H-E), histoenzimológico (NADH-TR y ATPasa miofibrilar). Después de 4 semanas las fibras musculares denervadas presentaron alteraciones importantes en lamicroscopía de luz: atrofia con perfiles angulados en secciones transversales; citoplasma con vacuolas; aumento del espacio intersticial con aumento de tejido conjuntivo; infiltración celular y fibras musculares sin contornos definidos, siendo las alteraciones más marcadas en las fibras tipo IIb e IIa. Después de 8 y 12 semanas de denervación la reacción para NADH-TR demuestra que es imposible caracterizar a las fibras musculares a través de su perfil metabólico...

Morphometric and morphological analysis of neuromuscular junction alterations in the denervated rat diaphragm / Evaluación de las alteraciones morfológicas y morfométricas de las uniones neuromusculares del diafragma denervado en ratas

The morphological and structural alterations that occur in the neuromuscular junctions of the denervated rat diaphragm were studied. Fifteen adult male albino rats (Rattus norvegicus) aged about 60 days and with a mean weight of 200 g were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by section of the phrenic nerve and the right antimere was used as control. Each antimere was divided into three fragments: one was used for histochemical (nonspecific esterase) and morphometric study of neuromuscular junctions, and the other two were used for transmission and scanning electron microscopy (SEM) analysis. Histochemical analysis of the diaphragm neuromuscular junctions after denervation showed only small changes in junction morphology. However, these junctions became smaller and elongated and presented less visible contours with increasing time of denervation. Ultrastructural analysis of neuromuscular junctions after 12 weeks showed more or less organized junctional folds on the muscle fiber surface. The junctional cytoplasm exhibited important alterations such as mitochondrial degeneration and the presence of numerous filaments. SEM revealed the presence of deep primary synaptic grooves with peripheral excavations which housed the nerve terminal boutons and exhibited internally the secondary synaptic clefts present among the junctional folds of the sarcolemma. This study showed that some of the morphological changes demonstrated in other denervated striated skeletal muscles are not repeated at the same intensity or in the same temporal pattern in the rat diaphragm.

En este trabajo se estudiaron las alteraciones morfológicas y estructurales de las uniones neuromusculares en el diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus), machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos y los animales se sacrificaron después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en 3 fragmentos: uno fue utilizado para el estudio histoquímico (esterasa inespecífica) y morfométrico. Los otros dos se destinaron al estudio de microscopía electrónica de transmisión (MET) y microscopia electrónica de barrido (MEB) de las uniones neuromusculares. El estudio histoquímico de las uniones neuromusculares posterior a la denervación, muestra que la morfología de esas uniones sufre pequeñas alteraciones. Con la evolución del tiempo de denervación esas uniones muestran tamaños menores, son alargadas y con contornos menos nítidos. La ultra-estructura de las uniones neuromusculares después de 12 semanas, demostró que la superficie de la fibra muscular exhibe pliegues de unión más o menos organizados. La región del citoplasma de unión exhibe alteraciones importantes, con degeneración mitocondrial y presencia de muchos filamentos. En MEB se observa que los botones sinápticos primarios son profundos, presentan escavaciones periféricas donde estaban alojados los botones de las terminaciones nerviosas y exhiben internamente, los espacios sinápticos secundarios presentes entre los pliegues de unión del sarcolema. Este estudio mostró que algunos patrones morfológicos demostrados en otros músculos estriados esqueléticos denervados no se repiten con la misma intensidad y curso temporal en el diafragma de ratas.

Estudo das relações anatômicas e suas variações entre o nervo ciático e o músculo piriforme / Study on anatomical relationships and variations between the sciatic nerve and piriform muscle

CONTEXTO: A síndrome do músculo piriforme pode ter como causa a passagem anormal do nervo ciático ou de uma de suas partes pelo ventre do músculo piriforme. OBJETIVO: Analisar as relações anatômicas e métricas entre o músculo piriforme e o nervo ciático, contribuindo com o conhecimento anátomo-clínico da região glútea. MÉTODO: Foram utilizados 20 cadáveres adultos de ambos os sexos. O nervo ciático e o músculo piriforme foram dissecados, medidos e fotodocumentados. RESULTADOS: Observou-se que 85 por cento das 40 regiões glúteas apresentaram o nervo como tronco único, passando pela borda inferior do músculo piriforme, e 15 por cento mostraram uma variação bilateral, caracterizada pela passagem do nervo fibular comum através do músculo piriforme. Os dados obtidos não revelaram diferenças estatisticamente significantes.

CONTEXT: Piriform muscle syndrome can be caused by abnormal passage of the sciatic nerve or one of its parts through the belly of the piriform muscle. OBJECTIVE: To analyze the anatomical and measurement relationships between the piriform muscle and the sciatic nerve in order to contribute towards better anatomoclinical understanding of the gluteal region. METHOD: Twenty adult cadavers of both sexes were used. The sciatic nerve and piriform muscle were dissected, measured and photodocumented. RESULTS: The sciatic nerve was seen to be a single trunk passing through the lower margin of the piriform muscle in 85 percent of the 40 gluteal regions, and 15 percent showed bilateral variation characterized by the passage of the common fibular nerve through the piriform muscle. The data obtained did not show any statistically significant differences.

In vitro effects of gamma radiation from 60Co and 137Cs on plasmid DNA.

The effects of gamma radiation from (60)Co and (137)Cs on DNA in aqueous solution are studied experimentally. Using an improved plasmid purification protocol and improved electrophoretic gel analysis techniques provided results with relatively small uncertainties. The results are compared with both theoretical and experimental results. In particular, the results obtained here are discussed in the light of recent discussion on supposed differences of the effects induced by gamma radiation from (60)Co and (137)Cs. We find that the effects of both types of gamma radiation are similar.

Pectinolytic activity secreted by yeasts isolated from fermented citrus molasses.

AIMS: The aim of this study was to obtain improved strains of pectinolytic yeasts adapted to the conditions of an industrial fermentation process, which was continuously operated to convert citrus molasses into ethanol. METHODS AND RESULTS: The starter yeast of the industrial fermentation process was a commercial baker's yeast, which was capable of growing without forming any secretion halo of pectinase activity on solid medium. Nevertheless, isolates showing secretion of pectinolytic activity on plates were obtained from the fermentation process. The secretion of pectin-degrading activity by isolates on plates was repressed by galactose and improved as the result of colony aging on polygalacturonic acid plates at 30 degrees C. Liquefaction of polygalacturonate gels as well as the splitting of the pectin-degrading activity into a wall-linked and a supernatant fraction were also observed when the starter yeast was propagated under agitation in liquid medium containing pectin. CONCLUSIONS: Isolates capable of secreting pectinolytic activity on plates were predominant at the end of the citrus molasses fermentation. Nevertheless, the sizes of the secretion haloes on plates were not necessarily an indication of the levels of pectinolytic activity secreted in the liquid medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Improved pectinolytic strains of Saccharomyces can be used as a source of pectinases for a variety of applications. This organism also participates in plant deterioration processes.

Weight evolution in girls treated for idiopathic central precocious puberty with GnRH analogues.

UNLABELLED: Data concerning the effects of GnRHa on weight gain are scarce. OBJECTIVE: To assess the variation of the body mass index (BMI) in girls during GnRHa treatment for idiopathic central precocious puberty (CPP). PATIENTS AND METHODS: Semestral anthropometric data from 176 girls treated with goserelin or leuprorelin were analyzed. RESULTS: BMI z-score increased from 1.5 +/- 0.1 SD before treatment (n = 176) to 1.7 +/- 0.2 SD after 24 months (n = 61, p = 0.008). In girls with normal weight before treatment, this variation was greater (n = 112, 0.2 +/- 0.1 SD, p = 0.01) than in those who were overweight (n = 63, -0.9 +/- 0.2 SD, p = 0.7). In the goserelin group the weight change adjusted for bone age was greater (n = 28, 0.4 +/- 0.1 SD) than in the leuprorelin group (n = 5, 0.04 +/- 0.1 SD, p = 0.05). CONCLUSIONS: A slight increase in BMI was noted, mainly in girls with normal weight before treatment. The influence of different GnRHa on weight must be further investigated.

Evaluation of alternative reporter genes for the yeast two-hybrid system

The yeast two-hybrid system is a powerful tool for screening protein-protein interactions and has also been used for large-scale studies. We evaluated two protein-coding sequences as reporter genes for the yeast two-hybrid system, to determine if it was suitable as an alternative screening strategy. Aspergillus awamori glucoamylase activity results in clear haloes around colonies producing this enzyme after growth on starch plates and staining with iodine vapors. However, transcription activation by Gal4 on Gal-regulated promoters was insufficient for this type of phenotypic visualization. A modified gene of Aequoria victoria enhanced green fluorescent protein (EGFP) was tested to determine its suitability for interaction screenings with flow cytometry. When the EGFP reporter gene system was incorporated into the cells, Gal4 transcriptional activation produced sufficient fluorescence for detection with the flow cytometer, especially when there were strong interactions

Phenotypic characterization of Trichophyton rubrum isolates from two geographic locations in Brazil.

To characterize possible Trichophyton rubrum phenotypes, which circulate in two Brazilian localities, we tested 53 isolates of this dermatophyte for their ability to assimilate several carbon sources, for keratinase, proteinase, phospholipase, lipase and desoxiribonuclease (DNase) secretions, and for their susceptibility to the antifungals fluconazole, ketoconazole and itraconazole. For each method, the isolates were submitted to similarity analysis and the methods were evaluated for their discriminatory indexes. None of the isolates were capable of assimilating arabinose, dulcitol, lactose, melibiose, ribose and xylose, while all of the isolates assimilated maltose, sucrose and sorbitol. However, adonitol, cellobiose, dextrin, erythritol, fructose, galactose, inulin, mannitol, mannose, raffinose, rhamnose and trehalose were assimilated by some isolates but not by others. All isolates secreted keratinase and DNase, while none secreted phospholipase. Proteinase and lipase were secreted only by some isolates. All but four isolates were resistant to fluconazole, most of them were sensitive to ketoconazole and all were sensitive to itraconazole. Carbohydrate assimilation was the method that presented the highest discriminatory index, and also the method that displayed the largest number of biotypes. Taken together, these data suggest that significant phenotypic variations exist among T. rubrum isolates. They seem to occur independently from their geographic origins.

Studies on polyhydroxyalkanoate (PHA) accumulation in a PHA synthase I-negative mutant of Burkholderia cepacia generated by homogenotization.

In the genome of Burkholderia cepacia strain IPT64, which accumulates a blend of the two homopolyesters poly(3-hydroxybutyrate), poly(3HB), and poly(3-hydroxy-4-pentenoic acid), poly(3H4PE), from sucrose or gluconate as single carbon source, the polyhydroxyalkanoate (PHA) synthase structural gene was disrupted by the insertion of a chloramphenicol-resistant gene cassette (phaC1::Cm). The suicide vector pSUP202 harboring phaC1::Cm was transferred to B. cepacia by conjugation. The inactivated gene was integrated into the chromosome of B. cepacia by homologous recombination. This mutant and also 15 N-methyl-N'-nitrosoguanidine (NMG)-induced mutants still accumulated low amounts of PHAs and expressed low PHA synthase activity. The analysis of the mutant phaC1::Cm showed that it accumulated about 1% of PHA consisting of 68.2 mol% 3HB and 31.8 mol% 3H4PE from gluconate. The wild-type, in contrast, accumulated 49.3% of PHA consisting of 96.5 mol% 3HB and 3. 5 mol% 3H4PE. Our results indicated that the genome of B. cepacia possesses at least two PHA synthase genes, which probably have different substrate specificities.

Antimicrobial resistance patterns of Haemophilus influenzae isolated from patients with meningitis in São Paulo, Brazil.

From 1989 to 1995, a total of 391 Haemophilus influenzae isolates were recovered from the cerebrospinal fluid (CSF) of hospitalized patients in São Paulo, Brazil. The majority of strains were isolated from infants aged less than 5 years. Strains belonging to biotype I (64.7%), biotype II (34.5%) and biotype IV (0.76%) were detected. Ninety-nine percent of these strains were serotype b. Minimal inhibitory concentration (MIC) was determined for ampicillin, chloramphenicol and ceftriaxone. The ss-lactamase assay was performed for all strains. The rate of ss-lactamase producer strains ranged from 10 to 21.4% during a period of 7 years, with an overall rate of 13.8%. Of the 391 strains analyzed, none was ss-lactamase negative ampicillin resistant (BLNAR). A total of 9.7% of strains showed resistance to both ampicillin and chloramphenicol; however, 4% of them were resistant to ampicillin only and 2% to chloramphenicol. All strains were susceptible to ceftriaxone and the MIC90 was 0.007 microg/ml, suggesting that ceftriaxone could be an option for the treatment of bacterial meningitis in pediatric patients who have not been screened for drug sensitivity.

Antimicrobial resistance patterns of Haemophilus influenzae isolated from patients with meningitis in São Paulo, Brazil

From 1989 to 1995, a total of 391 Haemophilus influenzae isolates were recovered from the cerebrospinal fluid (CSF) of hospitalized patients in São Paulo, Brazil. The majority of strains were isolated from infants aged less than 5 years. Strains belonging to biotype I (64.7 per cent), biotype II (34.5 per cent) and biotype IV (0.76 per cent) were detected. Ninety-nine percent of these strains were serotype b. Minimal inhibitory concentration (MIC) was determined for ampicillin, chloramphenicol and ceftriaxone. The ß-lactamase assay was performed for all strains. The rate of ß-lactamase producer strains ranged from 10 to 21.4 per cent during a period of 7 years, with an overall rate of 13.8 per cent. Of the 391 strains analyzed, none was ß-lactamase negative ampicillin resistant (BLNAR). A total of 9.7 per cent of strains showed resistance to both ampicillin and chloramphenicol; however, 4 per cent of them were resistant to ampicillin only and 2 per cent to chloramphenicol. All strains were susceptible to ceftriaxone and the MIC90 was 0.007 µg/ml, suggesting that ceftriaxone could be an option for the treatment of bacterial meningitis in pediatric patients who have not been screened for drug sensitivity.

Further characterization of the yeast pso4-1 mutant: interaction with rad51 and rad52 mutants after photoinduced psoralen lesions.

The pso4-1 mutant was characterized as deficient in some types of recombination, including gene conversion, crossing over, and intrachromosomal recombination. The mode of interaction between pso4-1 and rad51 and between pso4-1 and rad52 mutants indicated that the PSO4 gene belongs to the RAD52 epistasis group for strand-break repair. Moreover, the presence of the pso4-1 mutation decreased 8-MOP-photoinduced mutagenesis of the rad51 and rad52 mutants. Complementation tests using heterozygous diploid strains showed that the pso4 protein might interact with the rad52 protein during repair of 8-mop photolesions. The pso4-1 mutant, even though defective in inter- and intra-chromosomal recombination, conserves the ability for plasmid integration of circular and linear plasmid DNA. On the other hand, similar to the rad51 mutant, pso4-1 was able to incise but did not restore high-molecular-weight DNA during the repair of cross links induced by 8-MOP plus UVA. These results, together with those of previous reports, indicate that the PSO4 gene belongs to the RAD52 DNA repair group and its product participates in the DNA rejoining step of the repair of cross-link lesions, which are crucial for induced mutagenesis and recombinogenesis.