Aislamiento enzimático de protoplastos a partir del mesófilo de vitroplantas de Gynerium sagittatum Aubl. Beauv. cv. "Criolla"

Gynerium sagittatum es una gramínea ampliamente utilizada en la costa Caribe colombiana como fuente de fibra natural para la elaboración de artesanías, particularmente por la comunidad Zenú. En la presente investigación se evaluó el efecto de diferentes concentraciones de enzimas: celulasa y macerozima a diferentes tiempos de incubación y sus interacciones en el aislamiento de protoplastos. Los protoplastos se obtuvieron del mesófilo foliar de vitroplantas de G. sagittatum expuesto a combinaciones enzimáticas de celulasa (1.5 y 2.0%), con macerozima (0.3, 0.6 y 0.9%), durante 3, 6 y 9 horas de incubación, para un total de 18 tratamientos con 5 réplicas cada uno. Los mayores números de protoplastos aislados correspondieron a T18 (2.0% celulasa, 0.9% macerozima), T12 (2.0% de celulasa, 0.3% macerozima), T3 (1.5% de celulasa, 0.3% de macerozima) y T6 (1.5% de celulasa, 0.6% de macerozima) por 9 horas de incubación cada uno, con valores de 88.625, 83.000, 75.000 y 53.375 protoplastos/mL respectivamente. El tiempo de incubación fue significativo en el aislamiento de los protoplastos (p<0.05). Las predicciones entre factores mostraron que una interacción de 2.0% de celulasa y 0.9% de macerozima permite obtener 44.302 protoplastos/mL, mientras que las interaciciones tiempo de incubación-celulasa y tiempo de incubación-macerozima mostraron que es posible obtener 72.073 y 71.212 protoplastos/mL con 2.0% de celulasa y 0.9% macerozima por 9 horas de incubación cada una respectivamente. Los resultados indican que la aplicación de estas enzimas permite obtener cantidades considerables de protoplastos de G. sagittatum a partir de explantes cultivados in vitro.

Gynerium sagittatum is a graminaceous plant widely used in the Caribbean coast of Colombia as a natural fiber source for the elaboration of handicrafts, particularly by the Zenú community. In the present investigation, the effect of different concentrations of cellulase and macerozyme enzymes at different incubation times and their interaction in the isolation of protoplasts was evaluated. Protoplasts were obtained from leaf mesophyll of G. sagittatum vitroplants exposed to enzymatic combinations of cellulase (1.5 and 2.0%), with macerozyme (0.3, 0.6 and 0.9%), for 3, 6 and 9 hours of incubation, for a total of 18 treatments with 5 replicates each. The highest numbers of isolated protoplasts corresponded to T18 (2.0% cellulase, 0.9% macerozyme), T12 (2.0% cellulase, 0.3% macerozyme), T3 (1.5% cellulase, 0.3% macerozyme) and T6 (1.5% cellulase, 0.6% macerozyme); at 9 hours incubation. The protoplast number for these treatments were: 88.625, 83.000, 75.000 and 53.375 protoplasts/mL respectively. Incubation time was significant in the isolation of protoplasts (p<0.05). The predictions between the factors showed that with an interaction of 2.0% cellulase and 0.9% macerozyme it is possible to obtain 44.302 protoplasts/mL, likewise, the incubation time-cellulase and incubation time-macerozyme interactions showed that it is possible to obtain 72.073 and 71.212 protoplasts/mL with 2.0% cellulase and 0.9% macerozyme for 9 hours of incubation respectively. The results indicate that the use of these enzymes and time, allows the isolation of of protoplasts from G. sagittatum in vitro plants.

Knowledge and practices about taeniasis/cysticercosis complex within the framework of the national plan for its elimination in Colombia, 2019.

The taeniasis/cysticercosis complex (TCC) belongs to the group of neglected infectious diseases with a multifactorial transmission that includes hosts such as humans and pigs of the parasitic tapeworm (Taenia solium) and environmental factors. This study represents the first phase of the National and Cross-sectoral Plan for TCC Elimination. OBJECTIVES: To obtain data about knowledge and practices in relation to the TCC from knowledge, attitudes, and practices (KAP) surveys applied in two pilot localities for building a baseline of the potential risk factors related to identification, prevention, and control of TCC in Colombia. METHODS: A descriptive cross-sectional survey from October to November in 2019 was carried out using a structured questionnaire and random sampling by single-stage conglomerates in the municipalities of Mahates, Bolívar (n = 152) (Atlantic Region) and Mercaderes, Cauca (n = 152) (Andean Region). RESULTS: Respondents in Mahates showed moderate knowledge about TCC compared to those in Mercaderes, however, risky practices were identified in both places (consumption of pork with cysts and defecation in places other than the bathroom or latrine). Deficiency in infrastructure and failure in basic services were observed as potential risk factors for TCC transmission in both municipalities. CONCLUSION: Based on the framework of the National and Cross-sectoral Plan for TCC Elimination, adequate knowledge on identification, prevention, and control of the disease must be reinforced; cultural and ecological differences should be considered when designing communication and knowledge transmission tools. We consider that major investment should be made in improving basic services and creating sustainable modernized pig farming in Mahates and Mercaderes.

3'5-Dimaleamylbenzoic Acid Attenuates Bleomycin-Induced Pulmonary Fibrosis in Mice.

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by parenchymal scarring, leading progressively to alveolar architecture distortion, respiratory failure, and eventually death. Currently, there is no effective treatment for IPF. Previously, 3'5-dimaleamylbenzoic acid (3'5-DMBA), a maleimide, demonstrated pro-apoptotic, anti-inflammatory, and anti-cancer properties; however, its potential therapeutic effects on IPF have not been addressed. Bleomycin (BLM) 100 U/kg was administered to CD1 mice through an osmotic minipump. After fourteen days of BLM administration, 3'5-DMBA (6 mg/kg or 10 mg/kg) and its vehicle carboxymethylcellulose (CMC) were administered intragastrically every two days until day 26. On day 28, all mice were euthanized. The 3'5-DMBA effect was assessed by histological and immunohistochemical staining, as well as by RT-qPCR. The redox status on lung tissue was evaluated by determining the glutathione content and the GSH/GSSG ratio. 3'5-DMBA treatment re-established typical lung histological features and decreased the expression of BLM-induced fibrotic markers: collagen, α-SMA, and TGF-ß1. Furthermore, 3'5-DMBA significantly reduced the expression of genes involved in fibrogenesis. In addition, it decreased reduced glutathione and increased oxidized glutathione content without promoting oxidative damage to lipids, as evidenced by the decrease in the lipid peroxidation marker 4-HNE. Therefore, 3'5-DMBA may be a promising candidate for IPF treatment.

Proteomic Analysis Reveals Differential Expression Profiles in Idiopathic Pulmonary Fibrosis Cell Lines.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, irreversible lung disorder of unknown cause. This disease is characterized by profibrotic activation of resident pulmonary fibroblasts resulting in aberrant deposition of extracellular matrix (ECM) proteins. However, although much is known about the pathophysiology of IPF, the cellular and molecular processes that occur and allow aberrant fibroblast activation remain an unmet need. To explore the differentially expressed proteins (DEPs) associated with aberrant activation of these fibroblasts, we used the IPF lung fibroblast cell lines LL97A (IPF-1) and LL29 (IPF-2), compared to the normal lung fibroblast cell line CCD19Lu (NL-1). Protein samples were quantified and identified using a label-free quantitative proteomic analysis approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS). DEPs were identified after pairwise comparison, including all experimental groups. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Protein-Protein Interaction (PPI) network construction were used to interpret the proteomic data. Eighty proteins expressed exclusively in the IPF-1 and IPF-2 clusters were identified. In addition, 19 proteins were identified up-regulated in IPF-1 and 10 in IPF-2; 10 proteins were down-regulated in IPF-1 and 2 in IPF-2 when compared to the NL-1 proteome. Using the search tool for retrieval of interacting genes/proteins (STRING) software, a PPI network was constructed between the DEPs and the 80 proteins expressed exclusively in the IPF-2 and IPF-1 clusters, containing 115 nodes and 136 edges. The 10 hub proteins present in the IPP network were identified using the CytoHubba plugin of the Cytoscape software. GO and KEGG pathway analyses showed that the hub proteins were mainly related to cell adhesion, integrin binding, and hematopoietic cell lineage. Our results provide relevant information on DEPs present in IPF lung fibroblast cell lines when compared to the normal lung fibroblast cell line that could play a key role during IPF pathogenesis.

Human Platelets Contain, Translate, and Secrete Azurocidin; A Novel Effect on Hemostasis.

Platelets play a significant role in hemostasis and perform essential immune functions, evidenced by the extensive repertoire of antimicrobial molecules. Currently, there is no clear description of the presence of azurocidin in human platelets. Azurocidin is a 37 kDa cationic protein abundant in neutrophils, with microbicidal, opsonizing, and vascular permeability-inducing activity. Therefore, this work aimed to characterize the content, secretion, translation, and functions of azurocidin in platelets. Our results show the presence of azurocidin mRNA and protein in α-granules of platelet and megakaryoblasts, and stimulation with thrombin, ADP, and LPS leads to the secretion of free azurocidin as well as within extracellular vesicles. In addition, platelets can translate azurocidin in a basal or thrombin-induced manner. Finally, we found that the addition of low concentrations of azurocidin prevents platelet aggregation and activation. In conclusion, we demonstrate that platelets contain, secrete, and translate azurocidin, and this protein may have important implications for hemostasis.

Características clinicoepidemiológicas de pacientes con infecciones intrahospitalarias en un servicio de medicina interna de Santiago de Cuba / Clinical-epidemiological characteristics of patients with hospital acquired infections in a service of internal medicine from Santiago de Cuba

Introducción: La infección nosocomial o intrahospitalaria constituye un importante problema de salud en todos los hospitales del orbe. Objetivo: Describir las características clínicas y epidemiológicas de pacientes con infecciones intrahospitalarias. Métodos: Se realizó un estudio descriptivo y transversal de 57 pacientes con infecciones intrahospitalarias, ingresados en el Servicio de Medicina Interna del Hospital Provincial Docente Clínico-Quirúrgico Saturnino Lora de Santiago de Cuba, de octubre a diciembre de 2019, para lo cual se analizaron las variables edad, enfermedades asociadas, factores predisponentes, tipo de infección y gérmenes aislados. Resultados: En la serie sobresalieron el grupo etario de 60-69 años y la hipertensión arterial como enfermedad crónica concomitante (26,0 %). Entre los factores predisponentes resultó más frecuente el tabaquismo (32,8 %) y el tipo de infección preponderante fue la bronconeumonía bacteriana (47,0 %), cuyo germen causal en la mayoría de los casos (35,1 %) fue la Klebsiella pneumoniae. Conclusiones: Las infecciones intrahospitalarias aquejaron principalmente a pacientes de edad avanzada con enfermedades crónicas asociadas, como la diabetes mellitus y la hipertensión arterial. Cabe destacar la importancia de conocer la flora microbiana existente en el servicio donde se adquiere la infección, a fin de lograr tanto la prevención como el diagnóstico oportuno y el tratamiento adecuado.

Introduction: The cross or hospital infections constitute an important health problem in all the hospitals of the world. Objective: To describe the clinical and epidemiological characteristics of patients with hospital infections. Methods: A descriptive and cross-sectional study of 57 patients with hospital acquired infections was carried out. They were admitted to the Internal Medicine Service of Saturnino Lora Clinical-surgical Teaching Provincial Hospital of Santiago de Cuba, from October to December, 2019, for which the variables age, associated diseases, predisposing factors, type of infection and isolated germs were analyzed. Results: In the series the 60-69 years age group and hypertension as concomitant chronic disease (26.0 %) were notable. Among the predisposing factors nicotine addiction (32.8 %) was more frequent and the preponderant type of infection was the bacterial bronchopneumonia (47.0 %) whose causal germ in most of the cases (35.1 %) was the Klebsiella pneumoniae. Conclusions: The hospital acquired infections mainly afflicted patients of advanced age with associated chronic diseases, as diabetes mellitus and hypertension. It is necessary to highlight the importance of knowing the existent microbial flora in the service where the infection is acquired, in order to achieve boththe prevention, the opportune diagnosis and the appropriate treatment.

Case Report of a Metastatic Biliary Tract Neoplasm as an Unusual Cause of Appendicitis / Reporte de caso de neoplasia biliar metastásica como causa inusual de apendicitis

Abstract Introduction: We describe the case of a patient with appendiceal metastasis as the first manifestation of a cholangiocarcinoma. Main symptoms: Abdominal pain, jaundice, hyporexia, and choluria. Methods and results: We documented an appendiceal plastron histologically compatible with metastatic appendiceal adenocarcinoma, common hepatic duct stricture, and a suspected cholangiocarcinoma, later corroborated by endoscopic retrograde cholangiopancreatography. Conclusions: Metastatic appendiceal tumors are an infrequent and poorly studied manifestation, whereas those secondary to bile duct neoplasia have rarely been reported in the literature.

Resumen Introducción: se describe el caso de una paciente con una metástasis apendicular como primera manifestación encontrada de un colangiocarcinoma. Síntomas principales: expresado con dolor abdominal, ictericia, hiporexia y coluria. Métodos y resultados: se documentó un plastrón apendicular histológicamente compatible con adenocarcinoma apendicular metastásico, estrechez del conducto hepático común, con alta sospecha de colangiocarcinoma, corroborado luego con la realización de una colangiopancreatografía retrógrada endoscópica. Conclusiones: los tumores apendiculares metastásicos son una presentación infrecuente y poco estudiada, donde los secundarios a neoplasia de vía biliar se han reportados de forma muy escasa en la literatura.

Mechanisms of Immunothrombosis by SARS-CoV-2.

SARS-CoV-2 contains certain molecules that are related to the presence of immunothrombosis. Here, we review the pathogen and damage-associated molecular patterns. We also study the imbalance of different molecules participating in immunothrombosis, such as tissue factor, factors of the contact system, histones, and the role of cells, such as endothelial cells, platelets, and neutrophil extracellular traps. Regarding the pathogenetic mechanism, we discuss clinical trials, case-control studies, comparative and translational studies, and observational studies of regulatory or inhibitory molecules, more specifically, extracellular DNA and RNA, histones, sensors for RNA and DNA, as well as heparin and heparinoids. Overall, it appears that a network of cells and molecules identified in this axis is simultaneously but differentially affecting patients at different stages of COVID-19, and this is characterized by endothelial damage, microthrombosis, and inflammation.

Interaction of Spike protein and lipid membrane of SARS-CoV-2 with Ursodeoxycholic acid, an in-silico analysis.

Numerous repositioned drugs have been sought to decrease the severity of SARS-CoV-2 infection. It is known that among its physicochemical properties, Ursodeoxycholic Acid (UDCA) has a reduction in surface tension and cholesterol solubilization, it has also been used to treat cholesterol gallstones and viral hepatitis. In this study, molecular docking was performed with the SARS-CoV-2 Spike protein and UDCA. In order to confirm this interaction, we used Molecular Dynamics (MD) in "SARS-CoV-2 Spike protein-UDCA". Using another system, we also simulated MD with six UDCA residues around the Spike protein at random, naming this "SARS-CoV-2 Spike protein-6UDCA". Finally, we evaluated the possible interaction between UDCA and different types of membranes, considering the possible membrane conformation of SARS-CoV-2, this was named "SARS-CoV-2 membrane-UDCA". In the "SARS-CoV-2 Spike protein-UDCA", we found that UDCA exhibits affinity towards the central region of the Spike protein structure of - 386.35 kcal/mol, in a region with 3 alpha helices, which comprises residues from K986 to C1032 of each monomer. MD confirmed that UDCA remains attached and occasionally forms hydrogen bonds with residues R995 and T998. In the presence of UDCA, we observed that the distances between residues atoms OG1 and CG2 of T998 in the monomers A, B, and C in the prefusion state do not change and remain at 5.93 ± 0.62 and 7.78 ± 0.51 Å, respectively, compared to the post-fusion state. Next, in "SARS-CoV-2 Spike protein-6UDCA", the three UDCA showed affinity towards different regions of the Spike protein, but only one of them remained bound to the region between the region's heptad repeat 1 and heptad repeat 2 (HR1 and HR2) for 375 ps of the trajectory. The RMSD of monomer C was the smallest of the three monomers with a value of 2.89 ± 0.32, likewise, the smallest RMSF was also of the monomer C (2.25 ± 056). In addition, in the simulation of "SARS-CoV-2 membrane-UDCA", UDCA had a higher affinity toward the virion-like membrane; where three of the four residues remained attached once they were close (5 Å, to the centre of mass) to the membrane by 30 ns. However, only one of them remained attached to the plasma-like membrane and this was in a cluster of cholesterol molecules. We have shown that UDCA interacts in two distinct regions of Spike protein sequences. In addition, UDCA tends to stay bound to the membrane, which could potentially reduce the internalization of SARS-CoV-2 in the host cell.

[Mental health and human rights: The experience of professionals in training with the use of mechanical restraints in Madrid, Spain]. / Salud mental y derechos humanos: La experiencia de los profesionales en formación en el uso de sujeciones mecánicas en Madrid, España.

Mechanical restraint is a coercive procedure in psychiatry, which despite being permitted in Spain, raises significant ethical conflicts. Several studies argue that non-clinical factors - such as professionals' experiences and contextual influences - may play a more important role than clinical factors (diagnosis or symptoms) in determining how these measures are employed. The aim of this study is to understand how the experiences of mental health professionals in training relate to the use of mechanical restraints in Madrid's mental health network. Qualitative phenomenological research was conducted through focus groups in 2017. Interviews were transcribed for discussion and thematic analysis with Atlas.ti. Descriptive results suggest that these measures generate emotional distress and conflict with their role as caregivers. Our findings shed light on different factors related to their experiences and contexts that are important in understanding the use of mechanical restraint, as well as the contradictions of care in clinical practice.

La sujeción mecánica en psiquiatría es un procedimiento permitido en España que despierta importantes conflictos éticos. Diversos estudios sostienen que su uso depende de factores no clínicos, como las experiencias de los profesionales y las influencias del contexto, más que de factores clínicos (diagnósticos o síntomas). El objetivo del estudio es comprender las experiencias de profesionales de salud mental en formación en relación con el uso de sujeciones en la red de salud mental de Madrid. Es un estudio cualitativo de tipo fenomenológico mediante grupos focales, realizados en 2017. Las entrevistas fueron transcritas para su discusión y análisis temático mediante Atlas.ti. Los resultados descriptivos sugieren que estas medidas producen malestar y conflicto con relación al rol de cuidador y se observan estrategias de adaptación a los mismos. A partir de los hallazgos, se reflexiona acerca de aspectos de sus experiencias y del contexto que influyen en su uso, así como de las contradicciones del cuidado en la práctica clínica.

Analysis of SARS-CoV-2 mutations in Mexico, Belize, and isolated regions of Guatemala and its implication in the diagnosis.

The genomic sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide are publicly available and are derived from studies due to the increase in the number of cases. The importance of study of mutations is related to the possible virulence and diagnosis of SARS-CoV-2. To identify circulating mutations present in SARS-CoV-2 genomic sequences in Mexico, Belize, and Guatemala to find out if the same strain spread to the south, and analyze the specificity of the primers used for diagnosis in these samples. Twenty three complete SARS-CoV-2 genomic sequences, available in the GISAID database from May 8 to September 11, 2020 were analyzed and aligned versus the genomic sequence reported in Wuhan, China (NC_045512.2), using Clustal Omega. Open reading frames were translated using the ExPASy Translate Tool and UCSF Chimera (v.1.12) for amino acid substitutions analysis. Finally, the sequences were aligned versus primers used in the diagnosis of COVID-19. One hundred and eighty seven distinct variants were identified, of which 102 are missense, 66 synonymous and 19 noncoding. P4715L and P5828L substitutions in replicase polyprotein were found, as well as D614G in spike protein and L84S in ORF8 in Mexico, Belize, and Guatemala. The primers design by CDC of United States showed a positive E value. The genomic sequences of SARS-CoV-2 in Mexico, Belize, and Guatemala present similar mutations related to a virulent strain of greater infectivity, which could mean a greater capacity for inclusion in the host genome and be related to an increased spread of the virus in these countries, furthermore, its diagnosis would be affected.

Salud mental y derechos humanos: La experiencia de los profesionales en formación en el uso de sujeciones mecánicas en Madrid, España / Mental health and human rights: The experience of professionals in training with the use of mechanical restraints in Madrid, Spain

RESUMEN La sujeción mecánica en psiquiatría es un procedimiento permitido en España que despierta importantes conflictos éticos. Diversos estudios sostienen que su uso depende de factores no clínicos, como las experiencias de los profesionales y las influencias del contexto, más que de factores clínicos (diagnósticos o síntomas). El objetivo del estudio es comprender las experiencias de profesionales de salud mental en formación en relación con el uso de sujeciones en la red de salud mental de Madrid. Es un estudio cualitativo de tipo fenomenológico mediante grupos focales, realizados en 2017. Las entrevistas fueron transcritas para su discusión y análisis temático mediante Atlas.ti. Los resultados descriptivos sugieren que estas medidas producen malestar y conflicto con relación al rol de cuidador y se observan estrategias de adaptación a los mismos. A partir de los hallazgos, se reflexiona acerca de aspectos de sus experiencias y del contexto que influyen en su uso, así como de las contradicciones del cuidado en la práctica clínica

ABSTRACT Mechanical restraint is a coercive procedure in psychiatry, which despite being permitted in Spain, raises significant ethical conflicts. Several studies argue that non-clinical factors - such as professionals' experiences and contextual influences - may play a more important role than clinical factors (diagnosis or symptoms) in determining how these measures are employed. The aim of this study is to understand how the experiences of mental health professionals in training relate to the use of mechanical restraints in Madrid's mental health network. Qualitative phenomenological research was conducted through focus groups in 2017. Interviews were transcribed for discussion and thematic analysis with Atlas.ti. Descriptive results suggest that these measures generate emotional distress and conflict with their role as caregivers. Our findings shed light on different factors related to their experiences and contexts that are important in understanding the use of mechanical restraint, as well as the contradictions of care in clinical practice

A preliminary study of platelet hyperactivity in the chronic indeterminate phase of Chagas' disease.

The chronic indeterminate phase of Chagas' disease is asymptomatic despite positive test results for antibodies specific to Trypanosoma cruzi. CD62P-APC (P-selectin) and PAC-1 FITC (GpIIb/IIIa) may improve diagnosis as biomarkers of platelet activity. Nine asymptomatic seropositive subjects, previously untreated, were selected from a blood bank within a year of Chagas' disease detection, in addition to a control group of four. All subjects were evaluated by flow cytometry for CD62P, PAC-1 and CD41, and in a complementary study, by Tissue Doppler Echocardiography for isovolumic relaxation times (IVRT) and E/A ratios. The subjects were classified as positive or negative for CD62P and PAC-1 by a cut off obtained from their mean±2SD. For IVRT and E/A ratios, cut offs were obtained from the American Society of Echocardiography and the European Association of Cardiovascular Imaging recommendations. Fisher's exact test was used for associated findings. Pre-test and post-test probability, sensitivity, specificity, positive and negative predictive values and likelihood ratios were calculated. Abnormalities were expressed as platelet hyperactivity and ventricular dysfunction in CD62P, PAC-1, IVRT and E/A ratios. CD62P appears to have greater sensitivity (0.75) and PAC-1, more accurate specificity (0.75), which may explain thrombotic events in Chagas' disease. We recommend the use of CD62P and PAC-1 as biomarkers of platelet hyperactivity in patients in the chronic indeterminate phase of Chagas' disease.

A competent catalytic active site is necessary for substrate induced dimer assembly in triosephosphate isomerase.

The protozoan parasite Trichomonas vaginalis contains two nearly identical triosephosphate isomerases (TvTIMs) that dissociate into stable monomers and dimerize upon substrate binding. Herein, we compare the role of the "ball and socket" and loop 3 interactions in substrate assisted dimer assembly in both TvTIMs. We found that point mutants at the "ball" are only 39 and 29-fold less catalytically active than their corresponding wild-type counterparts, whereas Δloop 3 deletions are 1502 and 9400-fold less active. Point and deletion mutants dissociate into stable monomers. However, point mutants assemble as catalytic competent dimers upon binding of the transition state substrate analog PGH, whereas loop 3 deletions remain monomeric. A comparison between crystal structures of point and loop 3 deletion monomeric mutants illustrates that the catalytic residues in point mutants and wild-type TvTIMs are maintained in the same orientation, whereas the catalytic residues in deletion mutants show an increase in thermal mobility and present structural disorder that may hamper their catalytic role. The high enzymatic activity present in monomeric point mutants correlates with the formation of dimeric TvTIMs upon substrate binding. In contrast, the low activity and lack of dimer assembly in deletion mutants suggests a role of loop 3 in promoting the formation of the active site as well as dimer assembly. Our results suggest that in TvTIMs the active site is assembled during dimerization and that the integrity of loop 3 and ball and socket residues is crucial to stabilize the dimer.

Does Plant Origin Influence the Fitness Impact of Flower Damage? A Meta-Analysis.

Herbivory has been long considered an important component of plant-animal interactions that influences the success of invasive species in novel habitats. One of the most important hypotheses linking herbivory and invasion processes is the enemy-release hypothesis, in which exotic plants are hypothesized to suffer less herbivory and fitness-costs in their novel ranges as they leave behind their enemies in the original range. Most evidence, however, comes from studies on leaf herbivory, and the importance of flower herbivory for the invasion process remains largely unknown. Here we present the results of a meta-analysis of the impact of flower herbivory on plant reproductive success, using as moderators the type of damage caused by floral herbivores and the residence status of the plant species. We found 51 papers that fulfilled our criteria. We also included 60 records from unpublished data of the laboratory, gathering a total of 143 case studies. The effects of florivory and nectar robbing were both negative on plant fitness. The methodology employed in studies of flower herbivory influenced substantially the outcome of flower damage. Experiments using natural herbivory imposed a higher fitness cost than simulated herbivory, such as clipping and petal removal, indicating that studies using artificial herbivory as surrogates of natural herbivory underestimate the real fitness impact of flower herbivory. Although the fitness cost of floral herbivory was high both in native and exotic plant species, floral herbivores had a three-fold stronger fitness impact on exotic than native plants, contravening a critical element of the enemy-release hypothesis. Our results suggest a critical but largely unrecognized role of floral herbivores in preventing the spread of introduced species into newly colonized areas.

One-Year Outcome Comparison of Laparoscopic, Robotic, and Robotic Intrafascial Simple Prostatectomy for Benign Prostatic Hyperplasia.

OBJECTIVE: To compare preoperative, intraoperative, and postoperative variables at 1, 6, and 12 months after laparoscopic simple prostatectomy (LSP), robotic simple prostatectomy (RSP), and intrafascial robotic simple prostatectomy (IF-RSP). PATIENTS AND METHODS: From January 2003 to November 2014, 315 simple prostatectomies were performed using three techniques, LSP, RSP, and IF-RSP; of the patients who underwent these procedures, 236 met the inclusion criteria for this study. RESULTS: No statistically significant difference (SSD) was found in preoperative or perioperative variables. Of the postoperative variables that were analyzed, an SSD (p > 0.01) in prostate-specific antigen levels was found, with levels of 0.07 ± 1.1 ng/mL following IF-RSP, and the detection rate of prostate adenocarcinoma (26%) and high-grade prostatic intraepithelial neoplasia (HG-PIN; 12%) was higher for IF-RSP. We also found that lower International Prostate Symptom Scores (IPSS) were associated with LSP, at 4.8 ± 3.2. Erectile function was reduced in IF-RSP patients in the first 6 months after surgery but was similar in all patient groups at 12 months after surgery; continence and other measured parameters were also similar at 12 months for all three techniques. CONCLUSION: The IF-RSP technique is safe and effective, with results at 1-year follow-up for continence, IPSS, and Sexual Health Inventory for Men scores similar to those for the LSP and RSP techniques. IF-RSP also offers the advantages that it does not require postoperative irrigation, has an increased ability to detect prostate cancer (CA) and HG-PIN, and avoids the risk of future cancer and subsequent reintervention for possible new prostate growth.

Fibrosis pulmonar idiopática, tratamiento con pirfenidona / Idiopathic pulmonary fibrosis, treatment with pirfenidone

La fibrosis pulmonar idiopática es el tipo más frecuente de neumonía intersticial se caracteriza por un proceso de cicatrización anormal, con exceso de tejido fibroso. Es una patología crónica, progresiva, limitada a los pulmones de etiología desconocida que afecta a adultos mayores de 50 años con un deterioro progresivo de la función pulmonar y del intercambio gaseoso con un pronóstico fatal en un periodo relativamente corto. Se ha publicado un nuevo documento de consenso de la ATS/ERS/JRS/ALAT, sobre el diagnóstico y tratamiento de esta patología y se ha recomendado como tratamiento un fármaco nuevo que ha generado grandes expectativas, la pirfenidona, el cual posee propiedades antiinflamatorias y antifibróticas con resultados prometedores. Presentamos el caso clínico, de una paciente de 60 años de edad que ingresa al Hospital Seguro Social Universitario con un cuadro respiratorio insidioso; durante su internación fue diagnosticada de fibrosis pulmonar idiopática con criterios clínicos, imagenológicos e histológico. Se inició tratamiento con Pirfenidona según la recomendación del consenso; la evolución y seguimiento al quinto mes de tratamiento fue favorable.

Idiopathic pulmonary fibrosis is the most frequent type of interstitial pneumonia. It is characterized by an abnormal healing process, with excess fibrous tissue. It is a chronic, progressive illness, limited to the lungs with unknown etiology; it affects adults over 50 years old, with a progressive deterioration of lung function and gas exchange and a fatal outcome in a relatively short period. It has been published a new consensus document of the ATS/ERS/JRS/ALAT, about the diagnostic and treatment of these disease, and it is been suggested as a treatment a new drug that has generated great expectations, pirfenidone. It has anti-inflammatory and anti-fibrotic properties with promising results. we report the case of a patient of 60 years of age admitted to university hospital social security with an insidious respiratory symptoms, during her hospitalization she was diagnosed with idiopathic pulmonary fibrosis with clinical imaging and histopathological criteria. Treatment with Pirfenidone was initiated as recommended by consensus; monitoring the evolution and the fifth month of treatment was favorable.

Yeast mitochondrial RNA polymerase primes mitochondrial DNA polymerase at origins of replication and promoter sequences.

Three proteins phylogenetically grouped with proteins from the T7 replisome localize to yeast mitochondria: DNA polymerase γ (Mip1), mitochondrial RNA polymerase (Rpo41), and a single-stranded binding protein (Rim1). Human and T7 bacteriophage RNA polymerases synthesize primers for their corresponding DNA polymerases. In contrast, DNA replication in yeast mitochondria is explained by two models: a transcription-dependent model in which Rpo41 primes Mip1 and a model in which double stranded breaks create free 3' OHs that are extended by Mip1. Herein we found that Rpo41 transcribes RNAs that can be extended by Mip1 on single and double-stranded DNA. In contrast to human mitochondrial RNA polymerase, which primes DNA polymerase γ using transcripts from the light-strand and heavy-strand origins of replication, Rpo41 primes Mip1 at replication origins and promoter sequences in vitro. Our results suggest that in ori1, short transcripts serve as primers, whereas in ori5 an RNA transcript longer than 29 nucleotides is used as primer.