Trichomonas vaginalis acidic phospholipase A2: isolation and partial amino acid sequence.

Sexually transmitted diseases are a major cause of acute disease worldwide, and trichomoniasis is the most common and curable disease, generating more than 170 million cases annually worldwide. Trichomonas vaginalis is the causal agent of trichomoniasis and has the ability to destroy in vitro cell monolayers of the vaginal mucosa, where the phospholipases A2 (PLA2) have been reported as potential virulence factors. These enzymes have been partially characterized from the subcellular fraction S30 of pathogenic T. vaginalis strains. The main objective of this study was to purify a phospholipase A2 from T. vaginalis, make a partial characterization, obtain a partial amino acid sequence, and determine its enzymatic participation as hemolytic factor causing lysis of erythrocytes. Trichomonas S30, RF30 and UFF30 sub-fractions from GT-15 strain have the capacity to hydrolyze [2-(14)C-PA]-PC at pH 6.0. Proteins from the UFF30 sub-fraction were separated by affinity chromatography into two eluted fractions with detectable PLA A2 activity. The EDTA-eluted fraction was analyzed by HPLC using on-line HPLC-tandem mass spectrometry and two protein peaks were observed at 8.2 and 13 kDa. Peptide sequences were identified from the proteins present in the eluted EDTA UFF30 fraction; bioinformatic analysis using Protein Link Global Server charged with T. vaginalis protein database suggests that eluted peptides correspond a putative ubiquitin protein in the 8.2 kDa fraction and a phospholipase preserved in the 13 kDa fraction. The EDTA-eluted fraction hydrolyzed [2-(14)C-PA]-PC lyses erythrocytes from Sprague-Dawley in a time and dose-dependent manner. The acidic hemolytic activity decreased by 84% with the addition of 100 µM of Rosenthal's inhibitor.

Identification and characterization of microRNAs from Entamoeba histolytica HM1-IMSS.

BACKGROUND: Entamoeba histolytica is the causative agent of amebiasis, a disease that is a major source of morbidity and mortality in the developing world. MicroRNAs (miRNAs) are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and post-transcriptional regulation. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced a small RNA library prepared from a culture of trophozoites of Entamoeba histolytica Strain HM1-IMSS using a deep DNA sequencing approach. Deep sequencing yielded 16 million high-quality short sequence reads containing a total of 5 million non-redundant sequence reads. Based on a bioinformatics pipeline, we found that only 0.5% of these non-redundant small RNA reads were a perfect match with the drafted E. histolytica genome. We did not find miRNA homologs in plant or animal miRNAs. We discovered 199 new potential Entamoeba histolytica miRNAs. The expression and sequence of these Ehi-miRNAs were further validated through microarray by µParaflo Microfluidic Biochip Technology. Ten potential miRNAs were additionally confirmed by real time RT-PCR analysis. Prediction of target genes matched 32 known genes and 34 hypothetical genes. CONCLUSIONS/SIGNIFICANCE: These results show that there is a number of regulatory miRNAs in Entamoeba histolytica. The collection of miRNAs in this parasite could be used as a new platform to study genomic structure, gene regulation and networks, development, and host-parasite interactions.

Chronic environmental exposure to lead affects semen quality in a Mexican men population.

BACKGROUND: Male infertility is affected by several factors. Lead is one of the heavy metals more bioavailable than usually modifies the sperm quality in humans. OBJECTIVE: The aim of this study was to establish the role of lead in semen quality in environmentally exposed men. MATERIALS AND METHODS: Semen and blood samples were obtained from two groups: the exposed group (EG=20) and the non-exposed group (NEG=27). Two semen aliquots were used, one to evaluate spermatic quality and the other for lead determination. Blood (PbB) and semen lead (PbS) determination was performed by atomic absorption spectrophotometry. RESULTS: The PbB concentration was significantly greater in the EG, 10.10±0.97 µgdL-1 than in the NEG, 6.42±0.38 µgdL-1 (p<0.01), as well as the PbS concentration, with 3.28±0.35 and 1.76±0.14µgdL-1 in the EG and NEG respectively (p=0.043). A significant correlation between PbS and PbB concentration in the EG was found (r=0.573, p=0.038). Overall, the spermatic quality was lower in the EG than in the NEG. Specifically, there were significant differences in the spermatic concentration [EG=43.98±6.26 and NEG=68.78±8.51X10(6) cellmL-1 (p<0.01)], motility [EG=49±7 and NEG=67±4% (p=0.029)], viability [EG=36.32±3.59 and NEG=72.12±1.91% (p<0.01)] and abnormal morphology [EG=67±18 and NEG=32±12% (p<0.01)]. In the immature germ cells (IGC) concentration differences were found only for A cells (EG=8.1±1.1x100 and NEG=3.2±1.9X100 spermatozoa) (p<0.01) and for Sab cells (EG=3.4±2.2x100 and NEG=1.1±1.0X100 spermatozoa) (p=0.041). CONCLUSION: These results suggest that chronic environmental exposure to low levels of lead adversely affect the spermatic quality.

In vitro amoebicidal activity of borage (Borago officinalis) extract on Entamoeba histolytica.

Borage (Borago officinalis) is a plant with nutritional value that is also used in traditional medicine to treat gastrointestinal disease. This study investigated the amoebicidal activity of a methanol extract of borage. The 50% inhibitory concentration (IC50) of the extract for Entamoeba histolytica was 33 µg/mL. The 50% lethal dose of the extract for brine shrimp was greater than 1,000 µg/mL. The IC50 of the extract for Vero cells was 203.9 µg/mL. These results support the use of borage to prevent diseases associated with E. histolytica infection.

Efecto del albendazol en ratas long evans gestantes e infectadas con T. spiralis / Albendazol effect in Long Evans Rats pregnant and infected with T. spiralis

El Albendazol es el antihelmíntico de primera elección para varias parasitosis, entre ellas la Trichinellosis, sin embargo, el tiempo de tratamiento debe ser por periodos prolongados. El objetivo del trabajo fue determinar el efecto de este medicamento sobre ratas Long Evans gestantes e infectadas con T. spiralis. Se sincronizaron las ratas para gestarlas e infectarlas, el tratamiento fue de uno a catorce días, se utilizó la técnica de Hematoxilina-Eosina para evaluar el efecto sobre el hígado y el aparato reproductor, se determino el número de crías por tratamiento y las características de las crías obtenidas, los resultados indican una degeneración de los tejidos conforme aumenta el tiempo de tratamiento además se observa que el numero de crías es afectado directamente al aumentar el tiempo de tratamiento

The Albendazol is the antihelmintic of first election for several parasitism, among them trichinellosis, nevertheless, the time of treatment must be per prolonged periods. The aim of this work was to determine the effect of this medicine on Long rats Evans pregnant and infected with T. spiralis. We synchronize the rats to develop them and to infect them, the treatment went of one to fourteen days, used the Hematoxilina-Eosin technique to verify the effect on the liver and the reproductive apparatus and verified the number of young for each treatment and if correspondence between the obtained characteristics of the reproductive apparatus and young existed, the results indicate a degeneration of weaves as increases the time of treatment in addition is observed that I number of young is directly affected when increasing the treatment

Detección de los cambios fenotípicos en productos de ratas long evans infectadas con trichinella spiralis y tratadas con albendazol / Phenotypic changes detection in new born Long Evans Rat infected with T. spiralis and Albendazol treated

Trichinella spiralis es uno de los agentes causantes de la Trichinellosis, es una zoonosis de distribución mundial, afecta a gran número de mamíferos así como al humano. En México la causa principal de infección se asocia con el consumo de carne de cerdo infectada con T. spiralis. En este trabajo se detectaron los cambios fenotípicos en productos de ratas Long Evans infectas con T. spiralis y tratadas con Albendazol. Utilizamos ratas cepa Long Evans en edad reproductiva. Divididas en grupos, incluidos controles y con tratamiento de uno a catorce días de tratamiento, para sincronizar la gestación en las ratas se realizaron frotis vaginales. Para la detección del parásito se utilizaron técnicas directas de compresión en placa y digestión artificial y la técnica indirecta de MIDD, para los cambios fenotípicos de crías obtenidas se realizo análisis detallado con microscopio estereoscópico

Trichinella spiralis is one of the causative agents of the Trichinellosis, is a zoonosis of world-wide distribution, affects to big number of mammals like this as to the human. In Mexico the main cause of infection associate with the consumption of pig infected meat with T. spiralis. In this work detected the phenotypical changes in long Evans rats products infected with Trichinells spiralis and threated with albendazol medication. Use rats cepa long evans ina reproductive age. Divided in groups, included controls and with treatment varying of one to fourteen days of treatment, to synchronize the management in the rats realized vaginal.smears For the detection of the parasites used direct techniques of understanding in plate and artificial digestion and the indirect technician of MIID, for the phenotypical changes of baby rats obtained realized a detailed analysis with stereoscopic microscope

Activity of intracellular phospholipase A1 and A2 in Giardia lamblia.

Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have been identified in Giardia lamblia, even though they are essential for lipid metabolism in this parasite. A method to identify, isolate, and characterize these enzymes is needed. The activities of PLA A1 and PLA A2 were analyzed in a total extract (TE) and in vesicular (P30) and soluble (S30) subcellular fractions of G. lamblia trophozoites; the effects of several chemical and physicochemical factors on their activities were investigated. The assays were performed using substrate labeled with 14C, and the mass of the 14C-product was quantified. PLA A1 and PLA A2 activity was present in the TE and the P30 and S30 fractions, and it was dependent on pH and the concentrations of protein and Ca2+. In all trophozoite preparations, PLA A1 and PLA A2 activities were inhibited by ethylenediaminetetraacetic acid and Rosenthal's inhibitor. These results suggest that G. lamblia possesses several PLA A1 and PLA A2 isoforms that may be soluble or associated with membranes. In addition to participating in G. lamblia phospholipid metabolism, PLA A1 and PLA A2 could play important roles in the cytopathogenicity of this parasite.

Evaluación del efecto protector de 2 inmunógenos de Trchinella spiralis en ratas Long Evans con modificación nutricional e infectado con Trichinella spiralis / Evaluation of the protective effect of 2 inmunogenos of Trchinella spiralis in rats Long Evans with nutritional modification and infected with Trichinella spiralis

La Trichinellosis es una zoonosis endémica, cosmopolita, sus huéspedes son, ratas, cerdo y otros mamíferos entre ellos el hombre, la presencia de Trichinellosis se debe a la ingestión de carne de cerdo insuficientemente cocinada, afecta a países con bajos recursos económicos. Se han caracterizado Inmunógenos, siendo el inmunodominante el de 45 kDa, efectivo contra T. spiralis, desafortunadamente no ha cristalizado en una vacuna. Se han descrito efectos de desnutrición (DN) sobre órganos linfáticos. Los mecanismos de defensa del huésped son alterados con DN proteico-calórico (DPC). Evaluar el efecto protector de 2 inmunógenos de T. spiralis en ratas Long Evans con modificación Nutricional e infectado con T. spiralis. 80 ratas Long Evans de 30 días de edad, divididas en 2 grupos: 40 Nut con 24 de proteína y 40 DN con 12 de proteína de los cuales se subdividieron en 8 sub grupos con tratamiento: a) 10 animales control b)10 animales infectados con T. spiralis, c) 10 animales inmunizados con antígeno soluble total (AST), d) 10 animales inmunizados con inmunógeno de 45 kDa de T. spiralis (esquema de inmunización una aplicación cada semana por 4 ocasiones), retados a la 1era semana de la culminación de inmunización, sacrificadas a la 6 sexta semana post-infección. Parámetros a evaluar: Determinación de la carga parasitaria mediante la Digestión Artificial (D/A), Determinación de las características morfológicas de la célula nodriza por la técnica de compresión de tejidos, detección de la respuesta inmune por WB. Con la técnica de D/A, se obtuvieron en los diferentes tratamientos, de las ratas Nut, DN, Inmunizadas e Infectadas. Mostró diferencia significativa en la cantidad de Ll que se presentaron en 30gr de músculos estriado, los grupos controles de ratas Nut y DN sin infección, ausencia de Ll, y en los grupos de ratas Nut infectadas, se recolectaron 200 µl de Ll, para el grupo de DN infectado se recolecto 400 µl de Ll, en el grupo de Nut inmunizadas con AST.

Evaluación del albendazol, ivermectina y nitazoxanida en infección causada por trichinella spiralis en modelo suino / Evaluation of the abendazol, ivermectina and nitazoxanida in infecion caused by trichinella spiralis in model suino

La Trichinellosis es una enfermedad parasitaria zoonótica que se trasmite de modo accidental al hombre por ingestión de productos cárnicos insuficientemente cocidos, procedentes de animales infectados. En México el cerdo actúa como principal fuente de contaminación, los tratamientos aplicados han sido inespecíficos. Desconocemos a ciencia cierta el tratamiento efectivo para esta enfermedad. Este estudio esta dirigido a conocer si el Albendazol (ABZ), la Nitazoxanida (NZX) y la Ivermectina (IVM) son efectivos en el tratamiento de la fase intestinal (FI) y fase muscular (FM) de la Trichinellosis. Así mismo consideramos conveniente conocer, cual de los tres fármacos es más efectivo contra el parásito T. spiralis en ambas fases. Se utilizaron 18 cerdos raza York, se formaron 10 grupos: 1- Control sano en FI, 2-Control infectado en FI, 3- Infectado tratado con ABZ en FI, 4- Infectado y tratado con NZX en FI, 5- Infectado y tratado con IVM en FI, 6-Control sano en FM, 7-Control infectado en FM, 8- Infectado y tratado con ABZ en FM, 9- Infectado y tratado con NZX en FM, 10- Infectado y tratado con IVM en FM. El grupo 1 y 6 fueron las mismas cifras y de igual forma el 2 y 7 para la realización del análisis estadístico que fue, un diseño experimental completamente al azar y un diseño de tratamientos factorial. Concluyendo que el ABZ además de ser más efectivo en fase intestinal, en fase muscular presenta una eficacia del 100 por ciento contra la infección.