Identification of a calcium-dependent matrix metalloproteinase complex in rat chorioallantoid membranes during labour.

The induction of the expression of matrix metalloproteinases (MMPs) and their extracellular activation are key processes in connective tissue degradation in the chorioallantoid membrane during rat labour. However, the regulatory mechanisms remain largely unknown. Here, we report the identification of a calcium-dependent high molecular weight complex composed of MMP-9, MMP-3, MMP-2, tissue inhibitor of metalloproteinase 1 (TIMP-1) and TIMP-2, identified by zymography and western blotting. Molecular sieve chromatography confirmed the presence of a complex of MMPs and TIMPs with an exclusion volume >670 kDa. Differential scanning calorimetry of the complex confirmed the existence of a macromolecular complex that unfolds with a broad transition; it is denatured over a wide range of temperatures and has a T(m) of 72 degrees C in the presence of Ca(2+). When denatured in the absence of Ca(2+), there were at least eight transitions with T(m)s that corresponded to pro-MMP-9, MMP-9, pro-MMP-3, MMP-3, pro-MMP-2, MMP-2, TIMP-1 and TIMP-2. Co-localization of the same molecular components was demonstrated by confocal microscopy using cell-depleted chorioallantoid membranes. The assembly and disassembly of the complex can be reproduced at physiological concentrations of Ca(2+). This complex provides a potential mechanism for the enzymatic regulation of MMPs, which may participate in connective tissue degradation leading to the rupture of the fetal membranes during labour.

El alga marina Sargassum (Sargassaceae): una alternativa tropical para la alimentación de ganado caprino / The seaweed Sargassum (Sargassaceae) as tropical alternative for goats’ feeding

The nutritive value of seaweed (Sargassum spp.) was studied in Baja California Sur, Mexico. Twenty female Nubian goats (43-weeks old) were randomly distributed into two groups of 10 goats each and were housed in individual pens. One group was fed with a control diet and the other with a diet supplemented with 25% of Sargassum spp. Feed and water intake were recorded daily and individually for 60 days. The weight of each goat was recorded every 15 days. The nutritional content of Sargassum spp. was 89% dry mater, 8% crude protein, 31% ash, 2% ether extract, and 39% carbohydrates. Fiber fractions, minerals, vitamins, fatty acids, and antinutritional factors were also determined. There were no significant differences in body weight (8.6 kg control and 9 kg experimental), feed intake (1.3 kg control and 1.6 kg experimental), and feed conversion rate (11.1 control and 12.6 experimental). Water consumption was greater in the goats that ate the Sargassum diet (5.3 l). From these results,Sargassum spp.can be considered as an alternative feedstuff for goats.

Se estudió el valor nutricional de varias algas del género Sargassum. Veinte cabras hembras (Nubia) en crecimiento (43 semanas)se distribuyeron aleatoriamente en dos grupos de 10 cabras y se alojaron en corraletas individuales. Un grupo fue alimentado con una dieta testigo elaborada con insumos regionales y el otro con una dieta que incluyó el 25% de Sargassum spp. Durante 60 días se midieron diariamente y en forma individual, el alimento y agua consumidos, el peso corporal se cuantificó cada 15 días. La composición química proximal de Sargassum fue de: 89% materia seca, 7.7% proteína, 31% cenizas, 2% extracto etéreo y 39% de carbohidratos. También se determinaron las fracciones de fibra, minerales, vitaminas, ácidos grasos y factores antinutricios. No se encontraron diferencias significativas en el peso corporal (8.6 kg grupo control y 9 kg experimental), alimento consumido (1.31 kg grupo control y 1.6 experimental)y tasa de conversión alimenticia (11.1 grupo control y 12.6 experimental) entre los tratamientos. El consumo de agua fue mayor en el grupo alimentado con la dieta que incluyó Sargassum spp (5.1 l). De los resultados se deduce que las algas marinas de este género pueden ser utilizadas como un forraje alternativo de mediana calidad para el ganado caprino.

[The seaweed Sargassum (Sargassaceae) as tropical alternative for goats' feeding]. / El alga marina Sargassum (Sargassaceae): una alternativa tropical para la alimentación de ganado caprino.

The seaweed Sargassum (Sargassaceae) as tropical alternative for goats' feeding. The nutritive value of seaweed (Sargassum spp.) was studied in Baja California Sur, Mexico. Twenty female Nubian goats (43-weeks old) were randomly distributed into two groups of 10 goats each and were housed in individual pens. One group was fed with a control diet and the other with a diet supplemented with 25% of Sargassum spp. Feed and water intake were recorded daily and individually for 60 days. The weight of each goat was recorded every 15 days. The nutritional content of Sargassum spp. was 89% dry mater, 8% crude protein, 31% ash, 2% ether extract, and 39% carbohydrates. Fiber fractions, minerals, vitamins, fatty acids, and antinutritional factors were also determined. There were no significant differences in body weight (8.6 kg control and 9 kg experimental), feed intake (1.3 kg control and 1.6 kg experimental), and feed conversion rate (11.1 control and 12.6 experimental). Water consumption was greater in the goats that ate the Sargassum diet (5.3 1). From these results, Sargassum spp. can be considered as an alternative feedstuff for goats.

In-vitro secretion of proinflammatory cytokines by human amniochorion carrying hyper-responsive gene polymorphisms of tumour necrosis factor-alpha and interleukin-1beta.

The identification of polymorphisms in genes encoding proinflammatory cytokines that affect transcription or the secretion rate has opened new ways to understand the variation in responses to infection during pregnancy. In this study, human amniochorion carrying hyper-responsive alleles of tumour necrosis factor-alpha (TNF-alpha: TNF*2 at -308) and interleukin-1beta (IL-1beta: IL-1*2 at +3953) were stimulated in vitro with bacterial lipopolysaccharide (LPS) and compared with tissues carrying the common alleles (TNF*1 and IL-1*1). Fetal membranes carrying the TNF*1 allele displayed an identical dose-response pattern to tissues carrying a TNF*2 allele, except at the highest dose of LPS tested (50 ng/ml) there was a significantly greater production of TNF-alpha in the presence of a TNF*2 allele. Membranes carrying the IL-1*2 polymorphism secreted IL-1beta in a dose-response curve that was different from IL-1* tissues when challenged with 5, 10 and 50 ng/ml LPS. These observations support the hypothesis that reproductive tissues carrying hyper-responsive proinflammatory cytokine genes may over-respond to intrauterine infection secreting higher amounts of cytokines, which in turn, may lead to adverse pregnancy outcomes.

[Effect of Palomo procedure on the quality of the semen in infertile patients with varicocele and oligoastenospermia]. / Efecto de la operación de Palomo en la calidad del semen de pacientes infértiles con varicocele y oligoastenospermia.

OBJECTIVE: To evaluate the effect of Palomo's surgery on the semen quality from oligospermic infertile men with varicocele. STUDY DESIGN: Retrolective cohort. SETTING: Hospital General Regional No. 1, IMSS, Culiacán, Sinaloa. PATIENTS: 39 oligospermic infertile men with varicocele. INTERVENTIONS: Palomo's surgery. MEASUREMENTS: A spermatobioscopy before the Palomo's surgery was performed, and after the surgery two spermatobioscopies in different times of six months. RESULTS: Spermatobioscopy before the surgery: Concentration/mL = 8.8 +/- 4.1 millions, Motility = 19.5 +/- 10.0%, Vitality = 66.2 +/- 14.4%. First spermatobioscopy after surgery: Concentration/mL = 29 3 +/- 18 3 Motility = 41 5 +/- 19 1, Vitality = 74.7 +/- 14.5. Second spermatobioscopy after surgery: Concentration/mL = 14.6 +/- 10.3, Motility = 28.0 +/- 14.8, Vitality = 78.7 +/- 16.8. All differences from the spermatobioscopy before the surgery were significant. CONCLUSION: Palomo's surgery has a favorable effect on the quality of the semen, with a diminishing trend in concentration and motility in the second semester after the surgery. The pregnancy rate was 15%.

[Tumor necrosis factor-alpha and interleukin-1 beta in maternal, fetal and retroplacental intravascular compartments at term and preterm labor]. / Factor de necrosis tumoral-alpha e interleucina-1 beta en los compartimentos intravascular materno, fetal y retroplacentario en parto a término y pretérmino.

Neonato preterm birth (before 37 pregnancy weeks) account more than 80% perinatal deaths not attributable to congenital malformations. Preterm and term labor full mechanisms are unknown at present. Proinflammatory cytokinesis direct participation have been involved in the phenomena by several experimental evidence. The study's aim was to determine TNF-alpha and IL-1 beta concentration at maternal, fetal and fetal-maternal vascular compartments in women with term and preterm delivery and in women at term childbirth without labor. TNF-alpha and IL-1 beta concentration were determinated by commercial immunoassay. TNF-alpha concentration showed a tendency to be in more proportion at fetal and fetal-maternal compartments in preterm and term childbirth groups versus TNF-alpha concentration in term group without labor at same places. IL-1 beta concentration showed same tendency of increase than TNF-alpha in preterm and term childbirth groups, but alone at fetal-maternal compartment. Statistical difference were not documented at any compartment or group compared. Data allow to identify fetal-maternal compartments as target places where TNF-alpha and IL-1 beta were synthesized. Gradient concentration synthesis of cytokinesis allows to intend fetus as TNF-alpha initial producer.

[Apoptosis in trophoblast of patients with recurrent spontaneous abortion of unidentified cause]. / Apoptosis en trofoblasto de pacientes con aborto espontáneo recurrente de causa no identificada.

Recurrent spontaneous abortion of unidentified cause or idiopatic is an important problem of reproduction health. In this study, the study has been started of one of the mechanisms that could act in the pathology of human pregnancy. The general hypothesis proposed, is that in the case of patients with recurrent spontaneous abortion of non identified cause (RSA) there are phenomenons in the materno/fetal/placentary means that manifest themselves in harm to the functionality and/or vitality of placentary tissue. The hypothesis was consistent with the finding of massive activation of cellular death in all the cases with RSA studied in this work.

Evidence of endothelial cytotoxic compounds in placental extracts from preeclamptic women.

OBJECTIVE: To determine whether placenta and plasma of preeclamptic women contain factors that cause endothelial cell damage. METHODS: Placental extracts and plasma from preeclamptic and normotensive women were added to cultures of normal human umbilical vein endothelial cells and their effect on their viability, was determined by MTT reduction and 51chromium release. RESULTS: Placental extracts from normotensive and preeclamptic women were cytotoxic to endothelial cells, but not the plasma from both groups. Mean +/- standard deviation values of cytotoxicity index in preeclamptic and normotensive placental extracts using the MTT reduction were 70.3 +/- 6.76% and 51.4 +/- 8.81%, respectively, showing a significant difference (P < .0001). Using the 51chromium-release assay, preeclamptic placental extracts showed cytotoxic effects of 87.6 +/- 13.47% compared with 17 +/- 20.60% in control patients. The cytotoxic activity decreased after trypsin digestion and heat treatment in both groups. CONCLUSIONS: A cytotoxic factor to endothelial cells in placental extracts of preeclamptic women was identified. This compound is thermolabile and sensitive to trypsin digestion.

[Balance of the TH1/TH2 immune response in the materno-fetal interphase in normal and preterm pregnancy. Preliminary results]. / Balance de la respuesta inmune TH1/TH2 en la interfase materno-fetal de pacientes con embarazo normal y pretérmino. Resultados preliminares.

The subpopulations don's speak directly of the answers Th1 or Th2 in the compartment, as the are only reflexion of prevalent citocines in the microenvironment: However. It is important its exploration to the taken as starting point of this project.

[Peritoneal immune response in endometriosis]. / Respuesta inmune peritoneal en endometriosis.

The systematic analysis of immunity status in the peritoneum of women with endometriosis, is presented. Twenty five patients were studied, distributed as follows: A. Endometriosis, B. Sterility of non identified cause. In all the patients, peritoneal fluid was obtained (PF) during diagnostic laparoscopy, and a sample of peripheral blood was taken simultaneously. Both samples were processed analysis of lymphocytes subpopulations (ALS) by flux cytometry using monoclonal antibodies with reactivity to lymphocytes T total (CD+), lymphocytes B (CD19+), auxiliatory/inductory (CD3+CD4+), suppressor/cytotoxic (CD3+CD8+) and NK (CD3+CD16+ + 56+). The peritoneal ALS of both groups were characterized by a clear lymphocyte B diminution, moderate diminution of activators; a considerable increase of suppressors; altered CD4+/CD8+ relation in favour of suppression and NK diminution when comparing with its own blood compartment. All of these values were identical for both groups, and the only significant difference was found when comparing activated lymphocytes T subpopulation that was lesser in PF of women with endometriosis. This is the only fact that backs up the existence of altered immunity in the peritoneum of these patients, that affects the expression of activated lymphocytes. However, taking as a criterium the rest of subpopulations, in endometriosis as well as in ECNI the microenvironment goes for suppression and they are not distinguishable among them. On other side, it started signal characterization that conditions suppression induction for lymphocytes activation in peritoneal environment of women with endometriosis. Ten samples of PF of patients using peripheral purified lymphocytes by density gradient of voluntary donnors, normal, fertile, were analyzed. Cells were cultivated and stimulated in vitro with concanavaline A in presence of timidine 3H. In experimental holes PF was added at dose-response concentration, and it was evaluated as the difference in synthesis of radioactive ADN against the incubated cells in absence of PF. It was possible to demonstrate the presence of associated factors in PF of patients with endometriosis that inhibit proliferation of lymphocytes stimulated by Con A, and not so in samples of ECNI. With these factors one supposes the presence of soluble signals that condition local immunological suppression.

[Identification and initial characterization of a protease of immunoglobulin A associated with human spermatozoa]. / Identificación y caracterización inicial de una proteasa de inmunoglobulina A asociada a espermatozoides humanos.

The factors that the human semen use to block the humoral immune response at the uterine cervix level are not known. This response is mainly represented by immunoglobulin A, secretory type (IgAs). A possible mechanism would be hydrolisis od IgAs in an analogue ways as it has been described for some pathogenous bacteria that colonized human epitheliums and hydrolize IgAs. The objective of this study was to search for the presence of a protease for IgAs in human semen, seminal plasma and espermatozoons, using immunoelectrophoresis in agarose and electrophoresis in polyachrilamide, doing after immunotransference in nitrocellulose membrane. It was possible to identify a protease in sepermatic fraction, that hydrolizes IgAs, producing two fragments, the first one with molecular weight 66 to 70 kD, the degrees C. It is a metaloprotease (inhibited by EDTA). Its acti-enzymatic activity was optimal at 24 hours, pH 7.5 at 37 C. It is a metaloprotease (inhibited by EDTA). Its activity was specific for IgAs, and do not affected other immunoglobulins (IgG and IgM). The enzymatic activity of spermatozoon demonstrated in this study, may be on of the necessary factors used for blocking humoral immune response in femenine genital tract, and to permit that spermatozoon achieve the fecundation of human ovocyte.