Levels of key enzymes of methionine-homocysteine metabolism in preeclampsia.

OBJECTIVE: To evaluate the role of key enzymes in the methionine-homocysteine metabolism (MHM) in the physiopathology of preeclampsia (PE). METHODS: Plasma and placenta from pregnant women (32 controls and 16 PE patients) were analyzed after informed consent. Protein was quantified by western blot. RNA was obtained with RNA purification kit and was quantified by reverse transcritase followed by real-time PCR (RT-qPCR). Identification of the C677T and A1298C methylenetetrahydrofolate reductase (MTHFR) single-nucleotide polymorphisms (SNPs) and A2756G methionine synthase (MTR) SNP was performed using PCR followed by a high-resolution melting (HRM) analysis. S-adenosyl methionine (SAM) and S-adenosyl homocysteine (SAH) were measured in plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS). The SNP association analysis was carried out using Fisher's exact test. Statistical analysis was performed using a Mann-Whitney test. RESULTS: RNA expression of MTHFR and MTR was significantly higher in patients with PE as compared with controls. Protein, SAM, and SAH levels showed no significant difference between preeclamptic patients and controls. No statistical differences between controls and PE patients were observed with the different SNPs studied. CONCLUSION: The RNA expression of MTHFR and MTR is elevated in placentas of PE patients, highlighting a potential compensation mechanism of the methionine-homocysteine metabolism in the physiopathology of this disease.

Mesenchymal stem cells repress Th17 molecular program through the PD-1 pathway.

MSC display potent suppressive properties initially described a decade ago. More recently, MSC suppressive activities on T-cell effector pathways have been investigated. MSC modulate CD4 differentiation through different mechanisms depending on culture conditions and display disparate activities on T cells according to their differentiation status. A significant amount of evidence for MSC effects on Th17 cells revealed that MSC could be suppressive under diverse circumstances but also enhance Th17 cell activity under other conditions. In the present study, we investigated the suppressive effects of MSC on Th1 and Th17 subsets of T cells using T cells undergoing Th1 and Th17 polarization or mature Th1 and Th17 cells. MSC inhibited the proliferation of T cells during their differentiation toward Th1 cells and mature Th1 cells. This suppressive effect was maintained in a transwell cell culture insert demonstrating the major role played by soluble factors. Using the transwell cell separation barrier, we observed that MSC decrease the number of T cells undergoing Th17 differentiation whereas they did not affect IL-17 production by mature Th17, demonstrating the need for cell contact for suppressing Th17 cell function. Moreover, we reported that PD-L1 is highly expressed on MSC co-cultured with differentiating or polarized Th1 and Th17 cells. Using neutralizing antibodies specific for PD-L1 and PD-1 we showed that the mechanisms by which MSC mediate Th17 cell repolarization depend on PD-L1 expression on MSC. Taken together our results demonstrated a cell-to-cell contact depend mechanism in the selective immunosuppression of MSC on mature Th17 cells through up-regulation of PD-L1.

Functional characterization of UDP-glucose:undecaprenyl-phosphate glucose-1-phosphate transferases of Escherichia coli and Caulobacter crescentus.

Escherichia coli K-12 WcaJ and the Caulobacter crescentus HfsE, PssY, and PssZ enzymes are predicted to initiate the synthesis of colanic acid (CA) capsule and holdfast polysaccharide, respectively. These proteins belong to a prokaryotic family of membrane enzymes that catalyze the formation of a phosphoanhydride bond joining a hexose-1-phosphate with undecaprenyl phosphate (Und-P). In this study, in vivo complementation assays of an E. coli K-12 wcaJ mutant demonstrated that WcaJ and PssY can complement CA synthesis. Furthermore, WcaJ can restore holdfast production in C. crescentus. In vitro transferase assays demonstrated that both WcaJ and PssY utilize UDP-glucose but not UDP-galactose. However, in a strain of Salmonella enterica serovar Typhimurium deficient in the WbaP O antigen initiating galactosyltransferase, complementation with WcaJ or PssY resulted in O-antigen production. Gas chromatography-mass spectrometry (GC-MS) analysis of the lipopolysaccharide (LPS) revealed the attachment of both CA and O-antigen molecules to lipid A-core oligosaccharide (OS). Therefore, while UDP-glucose is the preferred substrate of WcaJ and PssY, these enzymes can also utilize UDP-galactose. This unexpected feature of WcaJ and PssY may help to map specific residues responsible for the nucleotide diphosphate specificity of these or similar enzymes. Also, the reconstitution of O-antigen synthesis in Salmonella, CA capsule synthesis in E. coli, and holdfast synthesis provide biological assays of high sensitivity to examine the sugar-1-phosphate transferase specificity of heterologous proteins.

[Trans-obturator-tape (TOT) for the surgical repair of stress urinary incontinence: our experience]. / Experiencia en la corrección quirúrgica de la incontinencia de orina de esfuerzo con técnica TOT (trans-obturator-tape).

OBJECTIVES: To evaluate the safety and efficacy of the TOT surgical technique in the treatment of female stress urinary incontinence (SUI). METHODS: Prospective study of 49 patients undergoing TOT at the Gynecology Unit of the Obstetrics and Gynecology Department in Barros Luco-Trudeau Hospital between November 2003 and January 2005. The mesh used was polypropylene, macropore, monofilament (Prolene). Median age was 52 years. The observational phase was completed in November 2005. RESULTS: Median operative time was 15 minutes. Thirty-two cases (65%) underwent a second gynecologic surgery. One patient had an intraoperative bladder lesion. In the immediate postoperative period there was one case of lower urinary tract infection and another acute pyelonephritis. At the end of the observation phase 47 (96%) patients were cured, 1 (2%) improved, and 1 (2%) had a failure. The failure appeared after a trauma three months after surgery. CONCLUSIONS: The TOT is an effective technique for the correction of SUI. It is easier than TVT. Although rare, it is not free of complications.

Experiencia en la corrección quirúrgica de la incontinencia de orina de esfuerzo con la técnica TOT (Trans Obturator Tape) / Trans-Obturator-Tape (TOT) for the surgical repair of stress urinary incontinence: our experience

OBJETIVO : Evaluar la seguridad y eficacia de la técnica quirúrgica de TOT en el tratamiento de la incontinencia de orina de esfuerzo femenina. MÉTODO: Seguimiento prospectivo de 49 pacientes sometidas a TOT, en la Unidad de Ginecología del Departamento de Ginecología y Obstetricia del Hospital Barros Luco-Trudeau, entre noviembre 2003 y enero 2005. Se utilizó malla de polipropileno, macroporo, monofilamento (Prolene). Mediana de edad 52 años, paridad 3. Se completó un período de observación hasta noviembre de 2005. RESULTADO: Mediana de tiempo operatorio de TOT, 15 minutos. En 32 (65%) se asoció otra cirugía ginecológica. En el intraoperatorio se presentó una lesión de vejiga. En el postoperatorio inmediato un caso de infección del tracto urinario bajo, y otro de pielonefritis aguda. Al finalizar el período de observación, se obtiene cura en 47(96%) casos, mejoría 1(2%) y falla 1(2%). El caso de falla resulta después de un traumatismo a los tres meses. CONCLUSIONES: El TOT se presenta como una técnica eficaz en la corrección de la IOE. Además es de mayor facilidad que el TVT. Aunque escasas, no está exento de complicaciones

OBJECTIVES: To evaluate the safety and efficacy of the TOT surgical technique in the treatment of female stress urinary incontinence (SUI). METHODS: Prospective study of 49 patients undergoing TOT at the Gynecology Unit of the Obstetrics and Gynecology Department in Barros Luco-Trudeau Hospital between November 2003 and January 2005. The mesh used was polypropylene, macropore, monofilament (Prolene). Median age was 52 years. The observational phase was completed in November 2005. RESULTS: Median operative time was 15 minutes. Thirty-two cases (65%) underwent a second gynecologic surgery. One patient had an intraoperative bladder lesion. In the immediate postoperative period there was one case of lower urinary tract infection and another acute pyelonephritis. At the end of the observation phase 47 (96%) patients were cured, 1 (2%) improved, and 1 (2%) had a failure. The failure appeared after a trauma three months after surgery. CONCLUSIONS: The TOT is an effective technique for the correction of SUI. It is easier than TVT. Although rare, it is not free of complications

Efecto de beclometasona en el eje hipotálamo-hipófisis suprarrenal en niños / Effect of beclomethasone on the hypothalamic-pituitary adrenal axis in children

Se estudió el efecto del dipropionato de beclometasona en 10 niños catalogados como asmáticos con prueba de hiperrectividad bronquial positiva, usando dosis de 400 y 1.000ug/día durante un período de tres meses. Se demostró formación del eje hipotálamo-hipófisis-suprarrenal en dos de los 6 pacientes que recibieron la dosis mayor. No hubo efecto sobre el eje en los pacientes que usaron de 400 ug/día