Efecto del embarazo y el parto en la musculatura perineal de mujeres con obesidad. Un estudio de cohorte longitudinal / Effect of pregnancy and delivery on perineal musculature in obese women: A longitudinal cohort study

Objetivo: Evaluar la evolución de la musculatura perineal en mujeres primigestas afectas de obesidad (índice de masa corporal [IMC]≥30) y comparar dicha evolución con mujeres primigestas con un IMC en rango de normopeso. Método: Se realizó un estudio cohorte longitudinal prospectivo donde se estudió a las gestantes en 3 momentos: en semana 12 de gestación, la semana 34 de gestación y a las 12 semanas tras el parto. En estas consultas se realizó medición mediante perinometría para determinar el tono basal (TB), la fuerza máxima contráctil (FMax); y la fuerza ejercida (FE). Además, se utilizó el test de Oxford modificado (MOS). Para el estudio estadístico se utilizaron modelos estadísticos generales mixtos. Resultados: Se reclutaron al inicio 50 gestantes nulíparas (25 con IMC≥30 y 25 con IMC<25), concluyendo el estudio completo 39. Al inicio del embarazo el tono perineal basal fue de 4,62±0,24Newton (Nw) y descendió a 4,18±0,26Nw tras el parto. La capacidad contráctil fue de 5,56±0,79Nw en la semana 12 y aumento a 6,34±1,24Nw tras el parto. Al comparar la FCMax en la semana 12 en gestantes obesas vs. Normopeso se observan valores de 5,51±87Nw vs. 5,61±0,71Nw (p=0,941). Se observaron valores posparto 6,72±1,17Nw vs. 5,95±1,21Nw (p=0,024), superiores en la población con obesidad. Conclusiones: Existe una disminución significativa del tono basal que es contrarrestada con un aumento de la fuerza contráctil objetivable a las 12 semanas posparto. El organismo dispone de mecanismos compensadores que permiten la recuperación a partir de los 3 meses posparto existiendo una mayor fuerza muscular dentro de las pacientes con obesidad.(AU)

Objective: Evaluating the changes of the perineal muscles in primigravid women with obesity (BMI≥30) and to compare the progress with primigravid women in normal BMI range. Methods: A prospective longitudinal cohort study was carried out. The pregnant women were studied at three moments: at 12 weeks’ gestation, at 34 weeks’ gestation and at 12 weeks after delivery. During the ckeck-up, perinometric measurements were taken to determine basal tone (BT), maximum contractile force (FMax) and applied forced (AF). In addition, the Oxford test (MOS) was used. General statistical mixed models were used for the statistical study. Results: Fifty nulliparous pregnant women (25 with BMI≥30 and 25 with BMI<25) were recruited at the beginning of the study and 39 completed the entire study. Basal tone (BT) was 4.62±0.24 Nw at the beginning of pregnancy and decreased to 4.18±0.26 Nw after delivery. Contractile capacity was 5.56±0.79 Nw at week 12 and increased to 6.34±1.24 Nw after delivery. When comparing the FCMax at week 12 in obese vs. normal weight pregnant women, values of 5.51±87 Nw vs. 5.61±0.71 Nw were observed (P=.941). Postpartum values were 6.72±1.17 Nw vs. 5.95±1.21 Nw (P=.024), higher in the obese population. Conclusions: There is an increase in contractile strength (Fmax) at 12 weeks postpartum in order to counteract the significant decrease in basal tone (BT). The body has compensatory mechanisms that allow recovery after 3 months postpartum, with greater muscle strength in obese patients.(AU)

Nucleoside Metabolism Is Induced in Common Bean During Early Seedling Development.

Nucleoside hydrolases (NSH; nucleosidases) catalyze the cleavage of nucleosides into ribose and free nucleobases. These enzymes have been postulated as key elements controlling the ratio between nucleotide salvage and degradation. Moreover, they play a pivotal role in ureidic legumes by providing the substrate for the synthesis of ureides. Furthermore, nucleotide metabolism has a crucial role during germination and early seedling development, since the developing seedlings require high amount of nucleotide simultaneously to the mobilization of nutrient in cotyledons. In this study, we have cloned two nucleosidases genes from Phaseolus vulgaris, PvNSH1 and PvNSH2, expressed them as recombinant proteins, and characterized their catalytic activities. Both enzymes showed a broad range of substrate affinity; however, PvNSH1 exhibited the highest activity with uridine, followed by xanthosine, whereas PvNSH2 hydrolyses preferentially xanthosine and shows low activity with uridine. The study of the regulation of nucleosidases during germination and early postgerminative development indicated that nucleosidases are induced in cotyledons and embryonic axes just after the radicle emergence, coincident with the induction of nucleases activity and the synthesis of ureides in the embryonic axes, with no remarkable differences in the level of expression of both nucleosidase genes. In addition, nucleosides and nucleobase levels were determined as well in cotyledons and embryonic axes. Our results suggest that PvNSH1 and PvNSH2 play an important role in the mobilization of nutrients during this crucial stage of plant development.

Differential Regulation of Drought Responses in Two Phaseolus vulgaris Genotypes.

Drought is probably the most harmful stress affecting common bean crops. Domestication, worldwide spread and local farming practices has entailed the development of a wide variety of common bean genotypes with different degrees of resistance to water stress. In this work, physiological and molecular responses to water stress have been compared in two common bean accessions, PHA-0683 and PMB-0220, previously identified as highly and moderately resistant to water stress, respectively. Our hypothesis was that only quantitative differences in the expression patterns of key genes should be found if molecular mechanisms regulating drought resistance are similar in the two accessions. However, results presented here indicate that the resistance to drought in PMB-0220 and PHA-0683 common bean accessions is regulated by different molecular mechanisms. Differential regulation of ABA synthesis and ABA signaling related genes among the two genotypes, and the control of the drought-induced senescence have a relevant contribution to the higher resistance level of PHA-0683 accession. Our results also suggest that expression patterns of key senescence-related transcription factors could be considered in the screening for drought resistance in common bean germplasm collections.

Transcriptomic Response to Water Deficit Reveals a Crucial Role of Phosphate Acquisition in a Drought-Tolerant Common Bean Landrace.

Drought is one of the most critical factors limiting legume crop productivity. Understanding the molecular mechanisms of drought tolerance in the common bean is required to improve the yields of this important crop under adverse conditions. In this work, RNA-seq analysis was performed to compare the transcriptome profiles of drought-stressed and well-irrigated plants of a previously characterized drought-tolerant common bean landrace. The analysis revealed responses related with the abscisic acid signaling, including downregulation of a phosphatase 2C (PP2C) and an abscisic acid-8' hydroxylase, and upregulation of several key transcription factors and genes involved in cell wall remodeling, synthesis of osmoprotectants, protection of photosynthetic apparatus, and downregulation of genes involved in cell expansion. The results also highlighted a significant proportion of differentially expressed genes related to phosphate starvation response. In addition, the moderate detrimental effects of drought in the biomass of these tolerant plants were abolished by the addition of phosphate, thus indicating that, besides the ABA-mediated response, acquisition of phosphate could be crucial for the drought tolerance of this common bean genotype. These results provided information about the mechanisms involved in drought response of common bean response that could be useful for enhancing the drought tolerance of this important crop legume.

Biochemical and Molecular Characterization of PvNTD2, a Nucleotidase Highly Expressed in Nodules from Phaseolus vulgaris.

Nucleotides are molecules of great importance in plant physiology. In addition to being elementary units of the genetic material, nucleotides are involved in bio-energetic processes, play a role as cofactors, and are also components of secondary metabolites and the hormone cytokinin. The common bean (Phaseolus vulgaris) is a legume that transports the nitrogen fixed in nodules as ureides, compounds synthetized from purine nucleotides. The first step in this pathway is the removal of the 5'-phosphate group by a phosphatase. In this study, a gene that codes for a putative nucleotidase (PvNTD2) has been identified in P. vulgaris. The predicted peptide contains the conserved domains for haloacid dehalogenase-like hydrolase superfamily. The protein has been overexpressed in Escherichia coli, and the purified protein showed molybdate-resistant phosphatase activity with nucleoside monophosphates as substrates, confirming that the identified gene codes for a nucleotidase. The optimum pH for the activity was 7-7.5. The recombinant enzyme did not show special affinity for any particular nucleotide, although the behaviour with AMP was different from that with the other nucleotides. The activity was inhibited by adenosine, and a regulatory role for this nucleoside was proposed. The expression pattern of PvNTD2 shows that it is ubiquitously expressed in all the tissues analysed, with higher expression in nodules of adult plants. The expression was maintained during leaf ontogeny, and it was induced during seedling development. Unlike PvNTD1, another NTD previously described in common bean, the high expression of PvNTD2 was maintained during nodule development, and its possible role in this organ is discussed.

Nuclease and ribonuclease activities in response to salt stress: Identification of PvRNS3, a T2/S-like ribonuclease induced in common bean radicles by salt stress.

The increase in soil salinization due to global climate change could cause large losses in crop productivity affecting, among other biological processes, to germination and seedling development. We have studied how salt stress affects nucleic acid degrading activities in radicles of common bean during seedling development. In radicles of common bean, a main nuclease of 37 kDa and two ribonucleases of 17 and 19 kDa were detected. Saline stress did not alter these three activities but induced a new ribonuclease of 16 kDa. All three ribonucleases are acidic enzymes that were inhibited by Zn. The 16 and 17 kDa ribonucleases are inhibited by guanilates. In the genome of common bean, we have identified 13 genes belonging to the T2 ribonuclease family and that are grouped in the 3 classes of T2 ribonucleases. The analysis of the expression of the 3 genes belonging to Class I (PvRNS1 to 3) and the unique gene from Class II (PvRNS4) in radicles showed that PvRNS3 is highly induced under salt stress.

Molecular and biochemical analysis of XDH from Phaseolus vulgaris suggest that uric acid protects the enzyme against the inhibitory effects of nitric oxide in nodules.

Xanthine dehydrogenase (XDH) is essential for the assimilation of symbiotically fixed nitrogen in ureidic legumes. Uric acid, produced in the reaction catalyzed by XDH, is the precursor of the ureides, allantoin and allantoate, which are the main N-transporting molecules in these plants. XDH and uric acid have been reported to be involved in the response to stress, both in plants and animals. However, the physiological role of XDH under stressful conditions in ureidic legumes remains largely unexplored. In vitro assays showed that Phaseolus vulgaris XDH (PvXDH) can behave as a dehydrogenase or as an oxidase. Therefore, it could potentially protect against oxidative radicals or, in contrast, it could increase their production. In silico analysis of the upstream genomic region of XDH coding gene from P. vulgaris revealed the presence of several stress-related cis-regulatory elements. PvXDH mRNA and enzymatic activity in plants treated with stress-related phytohormones or subjected to dehydration and stressful temperatures showed several fold induction. However, PvXDH activity was in vivo and in vitro inhibited by nitric oxide in leaves but not in nodules. In extracts from RNAi PvXDH silenced nodules, with lower levels of uric acid, XDH activity was inhibited by SNP which indicates that uric acid produced by XDH in the nodules of this ureidic legume could help to protect XDH against the inhibitory effects of nitric oxide.

Potential impacts of horizontal gene transfer on human health and physiology and how anthropogenic activity can affect it.

Horizontal gene transfer (HGT) is widespread among prokaryotes driving their evolution. In this paper, we review the potential impact in humans of the HGT between prokaryotes living in close association with humans in two scenarios: horizontal transfer in human microbiomes and transfer between microbes living in human managed environments. Although our vision is focused on the possible impact of these transfers in the propagation of antibiotic resistance genes or pathogenicity determinants, we also discuss possible human physiological adaptations via gene transfer between resident and occasional bacteria in the human microbiome.

Relationship between ureidic/amidic metabolism and antioxidant enzymatic activities in legume seedlings.

Ureides are nitrogenous compounds with a special function in some legume under nitrogen fixing conditions, the ureidic legumes. In this group, ureides are the predominant nitrogen transport molecule from nodules to the upper part, whereas amidic legumes use amides as nitrogen transport compounds. In this study, the ureide levels have been analysed in seedlings from four ureidic and four amidic legume plants. It has been found that the differentiation among ureide and amide plants already exists in seedlings during early seedling development, with high levels of ureide and allantoinase activity in cotyledons and embryonic axes from ureide plants. Since ureides have been implicated in the response of plant to several stress, total hydrosoluble antioxidant capacity and the levels of several antioxidant activities have been determined and compared among these two legume groups. The total antioxidant capacity did not follow any differential pattern in cotyledons or embryonic axes for the analysed plants. The levels of superoxide dismutase, guaiacol peroxidase and ascorbate peroxidase in both embryonic axes and cotyledons are statistical different between amide and ureide seedlings, whereas the catalase activity was similar among these groups of plants. We discuss than amides and ureides could follow different strategies to protect against oxidation.

The origin of aliphatic hydrocarbons in olive oil.

BACKGROUND: There are many substances that can interfere with olive oil quality. Some of them are well characterized, but many others have an unknown origin. Saturated hydrocarbons make an extraordinary complex family of numerous molecules, some of them present naturally in vegetable oils. When major natural saturated hydrocarbons are analyzed by standard chromatographic methods, this complex mixture of saturated hydrocarbons appears as a hump in the chromatogram and is commonly named as unresolved complex mixture (UCM), whose origin remains unknown. RESULTS: In this work we studied the occurrence and the origin of aliphatic saturated hydrocarbons in olive oil. Hydrocarbons were analyzed in olive oil and along the industrial process of oil extraction. We also analyzed n-alkanes and the UCM fraction of hydrocarbons in leaf, fruit and oil from different varieties and different locations, and we also analyzed the soils at these locations. CONCLUSIONS: We conclude that the hydrocarbons present in olive oil do not necessarily have their origin in a contamination during olive oil elaboration; they seem to have a natural origin, as a result of olive tree metabolism and/or as the result of an intake and accumulation by the olive tree directly from the environment during its entire life cycle. © 2017 Society of Chemical Industry.

Functional specialization of one copy of glutamine phosphoribosyl pyrophosphate amidotransferase in ureide production from symbiotically fixed nitrogen in Phaseolus vulgaris.

Purines are essential molecules formed in a highly regulated pathway in all organisms. In tropical legumes, the nitrogen fixed in the nodules is used to generate ureides through the oxidation of de novo synthesized purines. Glutamine phosphoribosyl pyrophosphate amidotransferase (PRAT) catalyses the first committed step of de novo purine synthesis. In Phaseolus vulgaris there are three genes coding for PRAT. The three full-length sequences, which are intron-less genes, were cloned, and their expression levels were determined under conditions that affect the synthesis of purines. One of the three genes, PvPRAT3, is highly expressed in nodules and protein amount and enzymatic activity in these tissues correlate with nitrogen fixation activity. Inhibition of PvPRAT3 gene expression by RNAi-silencing and subsequent metabolomic analysis of the transformed roots shows that PvPRAT3 is essential for the synthesis of ureides in P. vulgaris nodules.

Histological Analysis of Intra-Abdominal Adhesions Treated with Sodium Hyaluronate and Carboxymethylcellulose Gel.

PURPOSE: To evaluate macro and microscopically the adhesions developed after using the anti-adherence compound sodium hyaluronate and carboxymethylcellulose (SH-CBMC) gel and to determine the volume of the adhesions using a stereological estimation. METHODS: The study was experimental, random, comparative, and prospective. The subjects of the study were male Wistar rats divided in three groups (n = 10). Group I (control) included rats with no peritoneal injury. Group II rats had a 2 cm diameter injury created bilaterally in the parietal peritoneum at 3 cm from the abdominal midline with electrocautery coated with physiological solution. Group III rats were given the same injuries and coated with SH-CBMC gel. All groups were followed up postoperatively for 30 days, after which a laparotomy was performed to macroscopically determine the presence and type of adhesions. Experimental models were euthanized with anesthetic overdose and biopsies were taken for histopathological examination and stereological estimate of the volume of adhesions. RESULTS: Macroscopic adhesions were 20% less prevalent in Group III compared to Group II, which presented 40% more multiple and firm adhesions, unlike in Group III, in which they were unique and lax. There was a statistically significant decrease in the presence and number of adhesions in rats treated with SH-CBMC gel. Inflammatory infiltrate was significantly lower in rats treated with SH-CBMC gel, but there were no differences in connective tissue, fibrosis, and angiogenesis among groups. There was no statistical difference in the overall volume of adhesions among the treatment groups. CONCLUSIONS: SH-CBMC gel reduces macroscopic presence and number of adhesions and the severity of the inflammatory infiltrate.

Homogentisate phytyltransferase from the unicellular green alga Chlamydomonas reinhardtii.

Homogentisate phytyltransferase (HPT) (EC 2.5.1.-) catalyzes the first committed step of tocopherol biosynthesis in all photosynthetic organisms. This paper presents the molecular characterization and expression analysis of HPT1 gene, and a study on the accumulation of tocopherols under different environmental conditions in the unicellular green alga Chlamydomonas reinhardtii. The Chlamydomonas HPT1 protein conserves all the prenylphosphate- and divalent cation-binding sites that are found in polyprenyltransferases and all the amino acids that are essential for its catalytic activity. Its hydrophobicity profile confirms that HPT is a membrane-bound protein. Chlamydomonas genomic DNA analysis suggests that HPT is encoded by a single gene, HPT1, whose promoter region contains multiple motifs related to regulation by jasmonate, abscisic acid, low temperature and light, and an ATCTA motif presents in genes involved in tocopherol biosynthesis and some photosynthesis-related genes. Expression analysis revealed that HPT1 is strongly regulated by dark and low-temperature. Under the same treatments, α-tocopherol increased in cultures exposed to darkness or heat, whereas γ-tocopherol did it in low temperature. The regulatory expression pattern of HPT1 and the changes of tocopherol abundance support the idea that different tocopherols play specific functions, and suggest a role for γ-tocopherol in the adaptation to growth under low-temperature.

Identification and characterization of a gene encoding for a nucleotidase from Phaseolus vulgaris.

Nucleotidases are phosphatases that catalyze the removal of phosphate from nucleotides, compounds with an important role in plant metabolism. A phosphatase enzyme, with high affinity for nucleotides monophosphate previously identified and purified in embryonic axes from French bean, has been analyzed by MALDI TOF/TOF and two internal peptides have been obtained. The information of these peptide sequences has been used to search in the genome database and only a candidate gene that encodes for the phosphatase was identified (PvNTD1). The putative protein contains the conserved domains (motif I-IV) for haloacid dehalogenase-like hydrolases superfamily. The residues involved in the catalytic activity are also conserved. A recombinant protein overexpressed in Escherichia coli has shown molybdate resistant phosphatase activity with nucleosides monophosphate as substrate, confirming that the identified gene encodes for the phosphatase with high affinity for nucleotides purified in French bean embryonic axes. The activity of the purified protein was inhibited by adenosine. The expression of PvNTD1 gene was induced at the specific moment of radicle protrusion in embryonic axes. The gene was also highly expressed in young leaves whereas the level of expression in mature tissues was minimal.

Absorbable bioprosthesis for the treatment of bile duct injury in an experimental model.

INTRODUCTION: Cholecystectomy is a common surgical procedure in which complications may occur, such as injury to the biliary tract, which are associated with high morbidity. The aim of this study was to demonstrate the efficacy of a polymer-based absorbable bioprosthesis with bone scaffold for the treatment of bile duct injury in an animal model. MATERIALS AND METHODS: An absorbable bioprosthesis was used to replace the common bile duct in 15 pigs which were divided into 3 groups with different follow-ups at 1, 3 and 6 months. The animals were anesthetized at these time points and laboratory tests, Magnetic Resonance Cholangiopancreatogram [MRCP], Choledochoscopy using Spyglass and Endoscopic retrograde Cholangiopancreatogram [ERCP] were performed. After radiological evaluation was complete, the animals were euthanized and histological and immunohistochemical analyses were performed. RESULTS: Liver function tests at different time points demonstrated no significant changes. No mortality or postoperative complications were found in any of the experimental models. Imaging studies ([MRCP], [ERCP] and Choledochoscopy with SpyGlass(™)) showed absence of stenosis or obstruction in all the experimental models. DISCUSSION: Histological and immunohistochemical staining (CK19 and MUC5+) revealed the presence of biliary epithelium with intramural biliary glands in all the experimental models. There was no stenosis or obstruction in the bile duct. CONCLUSIONS: The bioprosthesis served as scaffolding for tissue regeneration. There was no postoperative complication at 6 months follow-up. This bioprosthesis could be used to replace the bile duct in cancer or bile duct injury. The bioprosthesis may allow different modeling depending on the type of bile duct injury.

Molecular and functional characterization of allantoate amidohydrolase from Phaseolus vulgaris.

Allantoate degradation is an essential step for recycling purine-ring nitrogen in all plants, but especially in tropical legumes where the ureides allantoate and allantoin are the main compounds used to store and transport the nitrogen fixed in nodules. Two enzymes, allantoate amidohydrolase (AAH) and allantoate amidinohydrolase (allantoicase), could catalyze allantoate breakdown, although only AAH-coding sequences have been found in plant genomes, whereas allantoicase-related sequences are restricted to animals and some microorganisms. A cDNA for AAH was cloned from Phaseolus vulgaris leaves. PvAAH is a single-copy gene encoding a polypeptide of 483 amino acids that conserves all putative AAH active-site domains. Expression and purification of the cDNA in Nicotiana benthamiana showed that the cloned sequence is a true AAH protein that yields ureidoglycine and ammonia, with a Km of 0.46 mM for allantoate. Optimized in vitro assay, quantitative RT-PCR and antibodies raised to the PvAAH protein were used to study AAH under physiological conditions. PvAAH is ubiquitously expressed in common bean tissues, although the highest transcript levels were found in leaves. In accordance with the mRNA expression levels, the highest PvAAH activity and allantoate concentration also occurred in the leaves. Comparison of transcript levels, protein amounts and enzymatic activity in plants grown with different nitrogen sources and upon drought stress conditions showed that PvAAH is regulated at posttranscriptional level. Moreover, RNAi silencing of AAH expression increases allantoate levels in the transgenic hairy roots, indicating that AAH should be the main enzyme involved in allantoate degradation in common bean.

Molecular characterization of PVAS3: an asparagine synthetase gene from common bean prevailing in developing organs.

In common bean, asparagine synthetase (AS; EC 6.3.5.4) is encoded by three members of a multigene family called PVAS1, PVAS2 and PVAS3. Two of these genes, PVAS1 and PVAS2, have been extensively studied, but little is known about PVAS3, remaining unclear whether PVAS3 function is redundant to the other AS or if it plays a specific role in Phaseolus vulgaris metabolism. In this work, we used a molecular approach to characterize PVAS3 expression and to gain some knowledge about its physiological function. We showed that, in contrast to PVAS1 and PVAS2, PVAS3 was expressed in all organs analyzed. Interestingly, PVAS3 was the AS gene most highly expressed in nodules, leaves and pods at the earliest stages of development, and its expression decreased as these organs developed. Expression of PVAS3 parallels the accumulation of AS protein and the asparagine content during the earliest stages of nodule, leaf and pod development, suggesting an important role for PVAS3 in the synthesis of asparagine in that period. Furthermore, PVAS3 was not repressed by light, as most class-II AS genes. Surprisingly, fertilization of nodulated plants with nitrate or ammonium, conditions that induce PVAS1 and PVAS2 and the shift from ureides to amide synthesis, repressed the expression of PVAS3 in nodules and roots. The possible implications of this regulation are discussed.

Local inhibition of nitrogen fixation and nodule metabolism in drought-stressed soybean.

Drought stress is a major factor limiting symbiotic nitrogen fixation (NF) in soybean crop production. However, the regulatory mechanisms involved in this inhibition are still controversial. Soybean plants were symbiotically grown in a split-root system (SRS), which allowed for half of the root system to be irrigated at field capacity while the other half remained water deprived. NF declined in the water-deprived root system while nitrogenase activity was maintained at control values in the well-watered half. Concomitantly, amino acids and ureides accumulated in the water-deprived belowground organs regardless of transpiration rates. Ureide accumulation was found to be related to the decline in their degradation activities rather than increased biosynthesis. Finally, proteomic analysis suggests that plant carbon metabolism, protein synthesis, amino acid metabolism, and cell growth are among the processes most altered in soybean nodules under drought stress. Results presented here support the hypothesis of a local regulation of NF taking place in soybean and downplay the role of ureides in the inhibition of NF.