The concentration of glucose in the media influences the susceptibility of stallion spermatozoa to ferroptosis.

In brief: Although common in many commercial extenders, supraphysiological concentrations of glucose in the media may be detrimental to stallion spermatozoa. In this study, we present evidence that these elevated glucose levels may predispose spermatozoa to ferroptosis. Abstract: Stallion spermatozoa depend on oxidative phosphorylation as their major source of ATP; however, the metabolism of these cells is complex and a great degree of metabolic plasticity exists. The composition of the media in which the spermatozoa are extended, or exposed to in the mare's reproductive tract, exerts a profound effect on sperm function and may even accelerate cell demise. Recent research indicates that high concentrations of glucose in the media, although common in commercial extenders, may be detrimental. To determine if supraphysiological glucose concentration may induce or predispose to ferroptosis (a caspase-independent form of programmed cell death, triggered by oxidative stress), stallion spermatozoa were incubated under different concentrations of glucose, 67 mM (HG) or 1 mM plus 10 mM pyruvate (LG-HP), in the presence or absence of known inductors of ferroptosis. Furthermore, we developed a single-cell flow metabolic assay to identify different metabolic phenotypes in spermatozoa. Storage and incubation of spermatozoa under high glucose concentrations led to an increase in the percentage of necrotic spermatozoa (P < 0.0001). Moreover, ferroptosis was induced more intensely in sperm in media with high glucose concentrations (P < 0.0001). Finally, we observed that induction of ferroptosis modified two proteins (oxoglutarate dehydrogenase and superoxide dismutase 2) in spermatozoa incubated in media containing 67 mM glucose but not in media containing 1 mM glucose and 10 mM pyruvate. The composition of the media, especially the concentration of glucose, exerts a major impact on the functionality and life span of the spermatozoa. The results reported here may pave the way for improvements in existing semen extenders.

The SLC7A11: sperm mitochondrial function and non-canonical glutamate metabolism.

Spermatozoa are redox-regulated cells, and stallion spermatozoa, in particular, present an intense mitochondrial activity in which large amounts of reactive oxygen species (ROS) are produced. To maintain the redox potential under physiological conditions, sophisticated mechanisms ought to be present, particularly in the mitochondria. In the present study, we investigated the role of the SLC7A11 antiporter. This antiporter exchanges intracellular glutamate for extracellular cystine. In the spermatozoa, cystine is reduced to cysteine and used for GSH synthesis. The importance of the antiporter for mitochondrial functionality was studied using flow cytometry and UHPLC/MS/MS approaches. Intracellular GSH increased in the presence of cystine, but was reduced in the presence of Buthionine sulphoximine (BSO), a γ-glutamylcysteine synthetase inhibitor (P < 0.001). Inhibition of the SLC7A11 antiporter with sulfasalazine caused a dramatic drop in intracellular GSH (P < 0.001) and in the percentage of spermatozoa showing active mitochondria (P < 0.001). These findings suggest that proper functionality of this antiporter is required for the mitochondrial function of spermatozoa. We also describe that under some conditions, glutamate may be metabolized following non-conventional pathways, also contributing to sperm functionality. We provide evidences, that the stallion spermatozoa have important metabolic plasticity, and also of the relation between redox regulation and metabolic regulation. These findings may have important implications for the understanding of sperm biology and the development of new strategies for sperm conservation and treatment of male factor infertility.

Depletion of thiols leads to redox deregulation, production of 4-hydroxinonenal and sperm senescence: a possible role for GSH regulation in spermatozoa†.

We hypothesized that thiols and particularly glutathione (GSH) are essential for the regulation of stallion sperm functionality. To test this hypothesis, we initially investigated the relationship between sperm function and GSH content, revealing highly significant correlations between GSH, sperm viability, motility, and velocity parameters (P < 0.001). Furthermore, the deleterious effects of GSH depletion using menadione and 1,3 dimethoxy 1,4, naphtoquinone (DMNQ) were able to be prevented by the addition of cysteine, but no other antioxidant. Pre-incubation with cysteine prevented menadione and DMNQ induced damage to sperm membranes after 1 h (P < 0.001; P < 0.05) and after 3 h of incubation (P < 0.001, P < 0.05). Pre-incubation with cysteine ameliorated both the menadione- and DMNQ-induced increase in 4-hydroxynonenal (P < 0.001). As cysteine is a precursor of GSH, we hypothesized that stallion spermatozoa are able to synthesize this tripeptide using exogenous cysteine. To test this hypothesis, we investigated the presence of two enzymes required to synthesize GSH (GSH and GCLC) and using western blotting and immunocytochemistry we detected both enzymes in stallion spermatozoa. The inhibition of GCLC reduced the recovery of GSH by addition of cysteine after depletion, suggesting that stallion spermatozoa may use exogenous cysteine to regulate GSH. Other findings supporting this hypothesis were changes in sperm functionality after BSO treatment and changes in GSH and GSSG validated using HPLC-MS, showing that BSO prevented the increase in GSH in the presence of cysteine, although important stallion to stallion variability occurred and suggested differences in expression of glutamate cysteine ligase. Mean concentration of GSH in stallion spermatozoa was 8.2 ± 2.1 µM/109 spermatozoa, well above the nanomolar ranges per billion spermatozoa reported for other mammals.

[Diagnosis of Mycoplasma genitalium by MgPa and rRNA 16S gene amplification]. / Diagnóstico de Mycoplasma genitalium por amplificación de los genes MgPa y ARN ribosomal 16S.

OBJECTIVE: Mycoplasma genitalium has been associated with nongonococcal urethritis (NGU). Diagnosis by PCR has become the primary detection method for this organism. Thus, diagnosis by DNA amplification using the PCR technique should be utilized. MATERIAL AND METHODS: GMF/GMR and MgpF/MgpR primer pairs, complementary to the M. genitalium 16S rRNA and MgPa genes, respectively, were selected. Specificity and sensibility assays were conducted and clinical samples were studied. RESULTS: The PCR with each primer pair was specific only for M. genitalium, and the sensibility was higher with the GMF/GMR primers. In the study of 34 clinical samples, 18,5% were positive for M. genitalium, with more positive samples when the MgpF/MgpR primers were used. CONCLUSIONS: DNA amplification by PCR should be applied in clinical practice to the diagnosis of M. genitalium in patients with NGU should using.

Identification of sperm morphometric subpopulations in two different portions of the boar ejaculate and its relation to postthaw quality.

A statistical approach using sequentially principal component analysis (PCA), clustering, and discriminant analyses was developed to identify sperm morphometric subpopulations in well-defined portions of the fresh boar ejaculate. Semen was obtained as 2 portions (the first 10 mL of the sperm-rich fraction and the rest of the ejaculate, respectively) and frozen using a conventional protocol. Before freezing, an aliquot was used for computer-assisted sperm morphometry analysis (ASMA). Postthaw quality was evaluated using computer-assisted sperm analysis (CASA), and an annexin-V/PI assay evaluated sperm membranes. The PCA revealed that 3 variables represented more than 78% of the cumulative variance in sperm subpopulations. The clustering and discriminant analyses, based on 5780 individual spermatozoa, revealed the existence of 4 sperm subpopulations. The relative percentage of these subpopulations varied between boar and ejaculate portions. Linear regression models based on measured morphometric characteristics could account for up to 36% of the percentage of intact sperm membranes postthaw. The ASMA protocol used in our study was useful to detect subtle morphometric differences between spermatozoa, and the combination of this analysis with a multivariate statistical procedure gave new information on the biological characteristics of boar ejaculates that is not given by conventional sperm analysis.

Animal model of Mycoplasma fermentans respiratory infection.

BACKGROUND: Mycoplasma fermentans has been associated with respiratory, genitourinary tract infections and rheumatoid diseases but its role as pathogen is controversial. The purpose of this study was to probe that Mycoplasma fermentans is able to produce respiratory tract infection and migrate to several organs on an experimental infection model in hamsters. One hundred and twenty six hamsters were divided in six groups (A-F) of 21 hamsters each. Animals of groups A, B, C were intratracheally injected with one of the mycoplasma strains: Mycoplasma fermentans P 140 (wild strain), Mycoplasma fermentans PG 18 (type strain) or Mycoplasma pneumoniae Eaton strain. Groups D, E, F were the negative, media, and sham controls. Fragments of trachea, lungs, kidney, heart, brain and spleen were cultured and used for the histopathological study. U frequency test was used to compare recovery of mycoplasmas from organs. RESULTS: Mycoplasmas were detected by culture and PCR. The three mycoplasma strains induced an interstitial pneumonia; they also migrated to several organs and persisted there for at least 50 days. Mycoplasma fermentans P 140 induced a more severe damage in lungs than Mycoplasma fermentans PG 18. Mycoplasma pneumoniae produced severe damage in lungs and renal damage. CONCLUSIONS: Mycoplasma fermentans induced a respiratory tract infection and persisted in different organs for several weeks in hamsters. This finding may help to explain the ability of Mycoplasma fermentans to induce pneumonia and chronic infectious diseases in humans.

Diversidad de Vanilla spp. (Orchidaceae) y sus perfiles bioclimáticos en México / Diversity and bioclimatic profiles of Vanilla spp. (Orchidaceae) in Mexico

Resumen: El género Vanilla comprende alrededor de 110 especies, distribuidas en las partes tropicales del mundo, y es el continente americano donde se encuentra la mayor cantidad de especies reportadas. En México, el cultivo de la vainilla está relacionado con diversas culturas como la totonaca, maya, chinanteca, mazateca, entre otras. En la actualidad, este cultivo presenta factores condicionantes tales como: técnicos, económicos, sociales, ecológicos y climáticos, que limitan su producción y la conservación de especies silvestres y cultivadas, por lo que es necesario, conocer su estado actual, en relación con su diversidad, así como algunos de los principales indicadores del perfil bioclimático de cada una de las especies, que ayuden en la toma de decisiones para su conservación y mejoramiento genético. Durante 2008, se realizaron consultas a herbarios de IPN, MEXU, XAL, base de datos de la Red Mundial de Información sobre la Biodiversidad de la CONABIO (REMIB), Global Biodiversity Information Facility (GBIF) y datos de accesiones vivas del banco de germoplasma de vainilla de la BUAP, formado del 2008 al 2014. Se realizaron mapas de distribución mediante un sistema de información geográfica. Se obtuvo el perfil bioclimático de cada especie considerando 19 variables de World Clim, y altitud a una resolución espacial de aproximadamente 1 Km2. Se calcularon las medias, desviaciones estándar y varianzas de las 19 variables en cada uno de los puntos registrados. Se obtuvieron los intervalos de las condiciones ambientales extremas (mínimo, promedio y máximo) para cada una de las especies de vainilla. Para determinar las variables de mayor importancia en la distribución de las especies se realizó un análisis de componentes principales, y a las variables que resultaron significativas se les realizó pruebas de Kruskal-Wallis y Dunn. Los resultados indicaron que en México se tienen registros de V. planifolia, V. pompona, V. insignis, V. inodora V. odorata, V. cribbiana y V. sprucei distribuidas en nueve estados. V. planifolia presentó intervalos amplios de temperatura y precipitación; V. pompona, V. odorata, V. insignis y V. inodora tuvieron intervalos intermedios. La amplitud de los datos extremos de cada especie puede considerarse para ubicar los sitios donde se puedan llevar a cabo estrategias regionales de conservación ex situ y el establecimiento de cultivos. El perfil bioclimático encontrado permite inferir de manera indirecta la condición genética de cada especie que podría ser utilizada en programas de mejoramiento genético como: la alta altitud y tolerancia a bajas temperaturas (V. odorata), la tolerancia a altas temperaturas (V. inodora) y tolerancia a baja precipitación (V. odorata, V. pompona y V. planifolia).

Abstract: The genus Vanilla comprises around 110 species distributed throughout Earth's tropical regions, with the largest number of reported species growing in the American continent. Vanilla farming is associated with many Mexican cultures such as the Totonac, Mayan, Chinantec, and Mazatec, among others. Currently, this crop is threatened by technical, social, ecological, and climatic conditioning factors, limiting its production and the preservation of wild and cultivated species. It is therefore necessary to ascertain the current diversity status of each of these species, as well as some of their main bioclimatic profile indicators, in order to help decision-making, aimed at preserving and genetically improve these species. During 2008, we gathered data from IPN, MEXU, and XAL herbaria, as well as from CONABIO's World Information Network on Biodiversity (REMIB), the Global Biodiversity and Information Facility (GBIF), and we also used data from live access to BUAP's vanilla germplasm bank, obtained between 2008 and 2014. Distribution maps were generated using a geographical information system. Bioclimatic profiles for each species were obtained considering 19 WorldClim variables and altitude at a spatial resolution of approximately 1 Km2. Variance, Mean, and standard deviation for each of the 19 variables were calculated at each of the registered points. Extreme environmental condition intervals (minimum, average, and maximum) were also obtained. In order to determine the most important distribution variables of the species, we performed a principal component analysis and carried out Kruskal-Wallis and Dunn's tests on the variables identified as significant. Results indicated records for V. planifolia, V. pompona, V. insignis, V. inodora V. odorata, V. cribbiana, and V. sprucei in Mexico, distributed throughout nine states in the country. V. planifolia presented wide intervals of temperature and rain precipitation, while V. pompona, V. odorata, V. insignis and V. inodora presented intermediate intervals. The amplitudes of extreme data for each species can be considered in locating areas where ex situ regional preservation strategies could be put in place, as well as in establishing areas for cultivation. The bioclimatic profile we found, allows for an indirect inference of each species' genetic condition, which could be used in genetic improvement programs; for instance, V. odorata grows at high altitudes and tolerates low temperatures, while V. inodora tolerates high temperatures, and V. odorata, V. pompona and V. planifolia tolerate low rain precipitation. Rev. Biol. Trop. 65 (3): 975-987. Epub 2017 September 01.

Leucorrea como signo de infecciones cérvicovaginales / Leucorrhea as a sign of cervico-vaginal infections

Las infecciones vaginales representan uno de los problemas ginecológicos más comunes en mujeres en edad reproductiva. Objetivo: Clasificar la leucorrea fisiológica y patológica de las pacientes investigadas, además de analizar si esta última obedece a infecciones cervico-vaginales. Método: La presente investigación se realizó a población abierta de todas las pacientes que acudieron al laboratorio de biología celular de la Benemérita Universidad Autónoma de Puebla al programa de detección oportuna de cáncer (Papanicolaou) desde enero del 2001 a Diciembre 2012, en las que se obtuvo un total de 1 679 muestras vaginales mismas que fueron teñidas con el tren de tinción de Papanicolaou modificado para su posterior diagnóstico microscópico. Resultados: Se encontró que 923 fueron positivas a leucorrea, de las cuales 489 corresponden a leucorrea fisiológica, 285 leucorrea patológica y las 149 no se ubican en ningún grupo. Discusión: Las infecciones cervico-vaginales se pueden presentar como respuesta a la presencia polimicrobiana que coloniza la cavidad vaginal y que esto conlleva un conjunto de signos y síntomas no específicos de los agentes que las causen, entre los que se encuentra la leucorrea como dato clínico que motiva en muchos casos a la consulta ginecológica. De las infecciones cervico-vaginales, la vaginitis es la causa más común de descarga vaginal patológica o leucorrea seguida de la cervicitis siendo ambas causadas por diversos agentes y de los que se mencionan los más frecuentes.

Vaginal infections are one of the most common gynecological problems in women of reproductive age. Objective: To classify physiological and pathological leucorrhoea in patients investigated, in addition to analyzing whether the latter is due to cervico-vaginal infections. Methods: This research was performed with an open population of all patients who came to the cancer screening program (Pap) at the Cell Biology Laboratory of the Benemérita Universidad Autónoma de Puebla from January 2001 to December 2012, where 1 679 vaginal samples were stained with the Papanicolaou stain set, modified for subsequent microscopic diagnosis. Results: Nine hundred twenty three were found to be positive for leucorrhea; of which 489 correspond to physiological leucorrhea and 285 to pathological leucorrhea; 149 are not located in any group. Discussion: Cervico-vaginal infections may occur in response to polymicrobial presence which colonizes the vaginal cavity; and this entails a set of signs and symptoms produced by nonspecific agents, among which is leucorrhoea as a clinical manifestation that motivates, in many case, a gynecological consultation. Among cervicovaginal infections, vaginitis is the most common cause of pathological vaginal discharge or leucorrhoea; followed by cervicitis; both of which are caused by various agents, of which we mention here those most frequently found.

Oclusión tubárica bilateral relacionada a dispareunia / Fallopian tube occlusion and dispareunia

La dispareunia (dolor durante o después de la relación sexual) tiene una etiología multifactorial y multisistémica; se presenta hasta en un 45% de las mujeres que han dado a luz. La oclusión tubárica bilateral (OTB) es uno de los métodos más utilizados en planificación familiar en México y en el mundo; debido a que toda cirugía implica la formación de una cicatriz -y por tanto dolor-, es lógico pensar en problemas ginecológicos en general como efectos colaterales de este procedimiento, aunado al hecho de que la OTB puede afectar las ramas terminales de las arterias uterina y ovárica, además de la sección de fibras nerviosas. El objetivo de esta investigación es evaluar la existencia de dispareunia en mujeres como posible consecuencia de la OTB. Se analizaron los resultados obtenidos del interrogatorio a todas las pacientes que acudieron al Laboratorio de Biología Celular del programa de Detección Oportuna de Cáncer (DOC) del 2001 al 2008. De un total de 921 pacientes, 287 fueron sometidas a OTB; de ellas, 45 mujeres (15.7%) refirieron presentar dispareunia. La dispareunia de acuerdo con nuestro estudio tiene una incidencia muy baja en pacientes con OTB, por lo que es posible que no represente un efecto colateral de este procedimiento.

Dyspareunia (pain during or after sexual intercourse) has a multifactorial and multisystemic etiology, occurring in up to 45% of the women who have given birth. Bilateral tubal occlusion is one of the top methods in family planning in Mexico and in the world; because all surgery involves the formation of a scar and therefore pain and so the fact that Bilateral tubal occlusion can affect the terminal branches of the uterine and ovarian arteries in addition to the section of nerve fibers, it's possible to think of general gynecological problems as side effects of this procedure. The objective of this research is to evaluate the existence of dyspareunia in women as a possible consequence of the Bilateral tubal occlusion. We analyzed the results of the questioning of all patients who attended the Laboratorio de Biología Celular to the program Early Detection of Cancer from 2001 to 2008. From a total of 921 patients, 287 were submitted to bilateral tubal occlusion, of whom 45 women (15.7%) reported dyspareunia. According to our study dyspareunia has a very low incidence in patients with bilateral tubal occlusion and therefore may not represent a side effect of this procedure.

Formación de biopelículas por micoplasmas de importancia médica / Biofilm formation by mycoplasmas of medical importance

El objetivo de este estudio fue evaluar la formación de biopelículas por micoplasmas de interés médico.Métodos. La formación de las biopelículas se realizó en microplacas, fueron enjuagadas con solución PBS para remover las células que no se adhirieron y se tiñeron con soluciónde cristal violeta al 0,5% durante 30 minutos. La formación de biopelículas por parte de Mycoplasma pneumoniae, Mycoplasma fermentans y Mycoplasma penetrans se analizó por medio de microscopía óptica y microscopía electrónica de barrido. Resultados. Los micoplasmas evaluados mostraron la capacidad para formar biopelículas,lo cual se evidenció por medio de la tinción de cristal violeta. Las biopelículas formadas en las microplacas y analizadas por microscopía mostraron agregados de microcolonias. La formación de biopelículas por parte de M. fermentans, M. pneumoniae y M. penetrans se presentó a las 72 horas de incubación. Se comparó la formación de biopelícula y la cuantificación de plancton, y se encontró correlación. Conclusión. Se debe considerar que este estudio se hizo bajo condiciones de laboratorioy que, para trabajos futuros, se recomienda utilizar modelos animales para definir cómo contribuyen estos agregados en la persistencia en los huéspedes. Estos resultados sugierenque la capacidad de M. fermentans, M. pneumoniae y M. penetrans para formar biopelículas puede considerarse un factor de virulencia, y un evento importante en la patogénesis yevolución en infecciones asociadas con el uso de dispositivos médicos.

The purpose of this study was to evaluate the development of biofilms by mycoplasmas of medical importance.Methods: Biofilms grown in microtiter plates were rinsed briefly in PBS to remove non-adherent cells and stained with 0,5% crystal violet solution for 30 minutes. The biofilm formation bymycoplasma species were analyzed by optical and scanning electron microscopy. Results: Three mycoplasma species were assessed for their ability to form biofilms. Crystal violet staining of biofilms in microtiter plates revealed the ability of mycoplasma to form a biofilm. Microscopic analysis of crystalviolet stained biofilms on microtiters indicated aggregation to form microcolonies. Biofilm growth by Mycoplasma fermentans, Mycoplasmapneumoniae and Mycoplasmapenetrans was followed over a time course of 72 hours. Mycoplasma were also analyzed quantitatively for biofilm formation and cell counts compared for both biofilm and plankton cells. Cell counts for biofilms showed a goodcorrelation with results obtained using crystal violet staining. Conclusion: This study has examined biofilm formation under in vitro laboratory conditions.Further studies on animal models will be crucial to determine if biofilms form in vivo and whether they contribute to mycoplasma persistence in thehost. These results suggest that the ability of M. fermentans, M. pneumoniae and M. penetrans to form a biofilm may be a virulence factor, and an important event in the pathogenesis and evolutionin infections associated with the use of medical devices.

Some biological features of mollicutes.

Mycoplasmas are a bacterial group that is classified in the Mollicute class which includes Mycoplasmas, Spiroplasmas and Acholeplasmas. One hundred and seventy six species have been described in this group. Mycoplasmas are the smallest self living prokaryotes, they do not have a bacterial wall, their genomic size ranges from 577 to 2220 bpk, they are nutritional exigent so it is hard to culture them, but the development of molecular biology techniques has let us detect more mycoplasmas in different hosts. Mycoplasmas have been associated to acute and chronic diseases mainly in animals and humans while spiroplasmas have been found in arthropods, plants and flowers producing or not damage. Some recent studies have shown the role of some structural components of Mycoplasmas in pathogenesis, such as cytoskeleton proteins and adhesins, and the influence of some genetic characteristics on the development of an infectious disease.

Incidenciadel bacilo de Dõderlein y su influencia en la presencia de otros microorganismos en el canal vaginal / Dõderlein bacillus incidence and its influence on the presence of other microorganisms in the vaginal tract

Objetivo. Investigar la presencia en cantidad normal o exagerada, o la ausencia de bacilos de Dõderlein en la muestra de población de estudio y determinar su influencia en asociación con otros microorganismos en el canal vaginal. Métodos. Se realizó estudio de campo, retrospectivo, de los frotis y la citología exfoliativacérvico-vaginal, o Papanicolaou, realizados a 740 mujeres en el Departamento de Biología Celular de la Facultad de Medicina de la Benemérita Universidad Autónoma de Puebla, de enero de 2001 a diciembre de 2006, con fines de prevención y detección temprana decáncer de cuello uterino. Resultados. De las 740 pacientes estudiadas, se identificaron lactobacilos en cantidades normales en 102 pacientes (14), no se encontraron lactobacilos en 564 pacientes (76

Interacción sinérgica entre Mycoplasma fermentans y la exposición a ceniza volcánica / Synergistic interaction between Mycoplasma fermentans and exposure to volcanic ash

Introducción: Mycoplasma fermentans presenta propiedades inmunorreguladoras y puede desencadenar infecciones crónicas con signos leves de la enfermedad. Por su parte, la materia partículada atmosférica es un componente diverso y complejo de la contaminación del aire y que se asocia a efectos adversos para la salud. Objetivo: Estudiar el daño producido por la inoculación de Mycoplasma fermentans y la exposición a ceniza volcánica. Materiales y métodos: Las muestras de ceniza se colectaron en zonas cercanas al volcán Popocatépetl. Diez hamsters se expusieron a la inhalación de ceniza volcánica, tros diez ejemplares fueron inoculados intratraquealmente con Mycoplasma fermentans y expuestos a inhalación de ceniza volcánica. Diez hamsters no fueron inoculados ni expuestos a la inhalación de ceniza, considerándose como grupo control. A partir del día 60 los ejemplares se sacrificaron, obteniéndose muestras de sangre para determinar el hematocrito, la cuenta diferencial y detección de anticuerpos. Se realizó estudio histopatológico al tejido pulmonar de los ejemplares. Resultados: El grupo de hamsters expuestos a la ceniza volcánica mostraron perdida en peso y decremento en el conteo de neutrófilos. No se detectaron anticuerpos contra Mycoplasma fermentans. El tejido pulmonar de los ejemplares expuestos a la ceniza mostró detritus celulares, reacción inflamatoria aguda e infiltrado linfocitario. Los hamsters previamente infectados y expuestos a la ceniza volcánica presentaronengrosamiento de la pared alveolar, infiltrado de células plasmáticas, neutrófilos y foco hemorrágico. Estos resultados indican que Mycoplasma fermentans puede interaccionar con estímulos ambientales, como es el caso de la ceniza volcánica, pontecializando efectos adversos en la salud.

Diagnóstico de Mycoplasma genitalium por amplificación de los genes MgPa y ARN ribosomal 16S / Diagnosis of Mycoplasma genitalium by MgPa and rRNA 16S gene amplification

OBJETIVO: El microorganismo Mycoplasma genitalium se ha relacionado con la uretritis no gonocócica (UNG). La técnica de PCR se ha convertido en el principal método de detección de este patógeno. En consecuencia, debe aplicarse un método de diagnóstico mediante la amplificación de fragmentos de ADN por la técnica PCR. MATERIAL Y MÉTODOS: Se seleccionaron los cebadores MGF-MGR y MgPaF-MgPaR, complementarios de los genes de ARNr 16S y MgPa de M. genitalium, respectivamente. Se efectuaron ensayos de especificidad y sensibilidad y se estudiaron muestras clínicas. RESULTADOS: La PCR con cada grupo de cebadores utilizado fue específica sólo para M. genitalium y la sensibilidad fue mayor con el grupo de cebadores MGF-MGR. En el estudio de 34 muestras clínicas, 18.5 por ciento fue positivo a M. genitalium y se encontró un mayor número de muestras positivas al utilizar los cebadores MgPaF-MgPaR. CONCLUSIONES: Debe aplicarse en la práctica clínica el diagnóstico de M. genitalium mediante la amplificación del ADN por PCR en los pacientes con UNG.

OBJECTIVE: Mycoplasma genitalium has been associated with nongonococcal urethritis (NGU). Diagnosis by PCR has become the primary detection method for this organism. Thus, diagnosis by DNA amplification using the PCR technique should be utilized. MATERIAL AND METHODS: GMF/GMR and MgpF/MgpR primer pairs, complementary to the M. genitalium 16S rRNA and MgPa genes, respectively, were selected. Specificity and sensibility assays were conducted and clinical samples were studied. RESULTS: The PCR with each primer pair was specific only for M. genitalium, and the sensibility was higher with the GMF/GMR primers. In the study of 34 clinical samples, 18,5 percent were positive for M. genitalium, with more positive samples when the MgpF/MgpR primers were used. CONCLUSIONS: DNA amplification by PCR should be applied in clinical practice to the diagnosis of M. genitalium in patients with NGU should using.

Diagnóstico clínico, de laboratorio y tratamiento de la vaginosis por Gardnerella vaginalis / Clinical and laboratory diagnosis of vaginosis produced by Gardnerella vaginalis and treatment

La vaginosis bacteriana es la infección de tejidos vaginales, generalmente por transmisión sexual, en cambio la vaginitis es la inflamación de la mucosa de la vagina y obedece a diversas etiologías. La vaginosis es más frecuente durante el periodo fértil y aunque su etiología es polimicrobiana se ha visto que uno de los agentes infecciosos más importante es Gardnerella vaginalis; identificada en la década de 1950 por Leopod, Gardner y Dukes, los que después de varios estudios la encontraron en un 98% de las mujeres con vaginosis y hasta un 50% en mujeres aparentemente sanas. Gardnerella vaginalis fue clasificada como una sola especie y fue establecida como agente causal de la vaginosis (antes conocida como vaginitis inespecífica). El cuadro clínico que presenta es caracterizado por una secreción blanca o blanco-grisácea que se percibe generalmente después de la relación sexual con olor fétido aminado (pescado). El diagnóstico certero es la base para evitar posibles complicaciones como la enfermedad inflamatoria pelviana y las complicaciones del embarazo. El tratamiento se basa principalmente en los fármacos como: metronidazol y clindamicina, debido a su efectividad y espectro, pero como todos se deben emplear con adecuada prudencia debido a su toxicidad. Además de que se deben corregir o modificar los factores predisponentes, ya que esta patología va en aumento convirtiéndose por su frecuencia en un problema de salud pública.