Epigenetic variation in neonatal tissues in infants conceived using capacitation-in vitro maturation vs. in vitro fertilization.

OBJECTIVE: To investigate alterations of the global DNA methylation profile in placenta, cord blood, and neonatal buccal smears in infants conceived using in vitro maturation (IVM) with a prematuration step (capacitation-IVM [CAPA-IVM]) vs. in vitro fertilization (IVF). DESIGN: Analysis of data from the offspring of participants in a randomized controlled trial. SETTING: Private clinic. PATIENTS: Forty-six women with polycystic ovary syndrome and/or high antral follicle count and their offspring (58 newborns). INTERVENTION(S): Women with polycystic ovary syndrome and/or a high antral follicle count participating in the clinical trial were randomized to undergo CAPA-IVM or conventional IVF. MAIN OUTCOME MEASURE(S): At delivery, biological samples including cord blood, placental tissue, and a neonatal buccal smear were collected. Genome-wide DNA methylation was determined using the Illumina Infinium MethylationEPIC BeadChip. Variability in methylation was also considered, and mean variances for the two treatment categories were compared. RESULTS: In neonatal buccal smears, there were no significant differences between the CAPA-IVM and conventional IVF groups on the basis of the CpG probe after linear regression analysis using a significant cut-off of false-discovery rate <0.05 and |Δß|≥0.05. In cord blood, only one CpG site showed a significant gain of methylation in the CAPA-IVM group. In the placenta, CAPA-IVM was significantly associated with changes in methylation at five CpG sites. Significantly more variable DNA methylation was found in five probes in the placenta, 54 in cord blood, and two in buccal smears after IVM of oocytes. In cord blood samples, 20 CpG sites had more variable methylation in the conventional IVF vs. IVM group. Isolated CpG sites showing differences in methylation in cord blood were not associated with changes in gene expression of the overlapping genes. CONCLUSION(S): Capacitation-IVM appeared to be associated with only a small amount of epigenetic variation in cord blood, placental tissue, and neonate buccal smears. CLINICAL TRIAL REGISTRATION NUMBER: NCT03405701 (www. CLINICALTRIALS: gov).

Arbuscular Mycorrhizal Fungi Associated with Maize (Zea mays L.) in the Formation and Stability of Aggregates in Two Types of Soil.

Knowledge of native Arbuscular Mycorrhizal Fungi (AMF) and their relationship with the edaphic characteristics where they live is important to establish the influence of allochthonous AMF, which were inoculated, on the development and stability of soil aggregates. The objectives of this research were to know the composition of native AMF species from two contrasting soils, and to establish the development and stability of aggregates in those soils with corn plants after inoculating them with allochthonous AMF. The experiment had three factors: Soil (two levels [S1 and S2]), HMA (three levels: without application [A0], with the application of Claroideoglomus claroideum [A1] and with the application of a consortium [A2]) and Fertilization (two levels (without fertilization [f0] and with fertilization [f1])). Twelve treatments were generated, with five replicates (60 experimental units [EU]). The EU consisted of a pot with a corn plant and the distribution was completely random. The results demonstrated that the Typic Ustifluvent presented nine species of native AMF, while the Typic Dystrustert had three; the native AMF in each soil influenced the activity of allochthonous AMF, such as their colonization and sporulation. Likewise, differences were found in the stability of macro-sized aggregates (0.5 to 2.0 mm).

Silencing of the Laccase (lacc2) Gene from Pleurotus ostreatus Causes Important Effects on the Formation of Toxocyst-like Structures and Fruiting Body.

A wide variety of biological functions, including those involved in the morphogenesis process of basidiomycete fungi, have been attributed to laccase enzymes. In this work, RNA interference (RNAi) was used to evaluate the role of the laccase (lacc2) gene of Pleurotus ostreatus PoB. Previously, transformant strains of P. ostreatus were obtained and according to their level of silencing they were classified as light (T7), medium (T21) or severe (T26 and T27). The attenuation of the lacc2 gene in these transformants was determined by RT-PCR. Silencing of lacc2 resulted in a decrease in laccase activity between 30 and 55%, which depended on the level of laccase expression achieved. The silenced strains (T21, T26, and T27) displayed a delay in the development of mycelium on potato dextrose agar (PDA) medium, whereas in the cultures grown on wheat straw, we found that these strains were incapable of producing aerial mycelium, primordia, and fruiting bodies. Scanning electron microscopy (SEM) showed the presence of toxocyst-like structures. The highest abundance of these structures was observed in the wild-type (PoB) and T7 strains. However, the abundance of toxocysts decreased in the T21 and T26 strains, and in T27 they were not detected. These results suggest that the presence and abundance of toxocyst-like structures are directly related to the development of fruiting bodies. Furthermore, our data confirm that lacc2 is involved in the morphogenesis process of P. ostreatus.

Clinical atrophy patterns of filiform papillae of the tongue and their relationship with the serum levels of iron, vitamin B12 and folic acid in patients with systemic diseases.

BACKGROUND: The purpose of the study was to evaluate the clinical patterns of atrophy of the filiform papillae (FP) of the tongue and their relationship with the serum levels of iron and vitamin B12 among patients with systemic diseases, in a tertiary care center. METHODS: A cross-sectional, analytical, research study was designed. A systematic tongue examination was performed to evaluate the presence and clinical patterns of FP atrophy. We collected epidemiologic, clinical, and laboratory data. Statistical analysis included χ2 test, Fisher's exact test, Kruskal-Wallis test, and a logistic regression analysis. RESULTS: A total of 87 patients (83.9% females) were included [median age = 55 (range 20-89) years]. Endocrinopathy (60.9%) was the most frequent comorbidity. We found atrophy of the FP in 90.8% of the patients; the atrophy was mild in 83.5% of the cases, and severe in 16.5%. The most common atrophic patterns were as follows: focalized in 64 (73.6%) cases, "U"-shaped pattern in 60 (69%), and generalized in 30 (34.5%). Geographic tongue and median rhomboid glossitis were observed in 12 (13.8%) and 11 (12.6%) subjects, respectively. Lower titers of serum iron were detected in cases with focal (median = 71 vs. 110 mcg/dl) and generalized (median = 55 vs. 78 mcg/dl) FP atrophy (P = 0.03 and P = 0.009, respectively), than their counterparts. The presence of symptomatology was related to the focal pattern of atrophy (P = 0.038). CONCLUSIONS: A high frequency of filiform papillary atrophy of the tongue was observed in patients with comorbidities. Some atrophic patterns of the tongue were significantly associated with certain medical conditions.

Trauma-Related Clinical Practice Variation in Dutch Emergency Departments.

Structural insights in the use of protocols and the extent of practice variation in EDs are lacking. The objective is to determine the extent of practice variation in EDs in The Netherlands, based on specified common practices. We performed a comparative study on Dutch EDs that employed emergency physicians to determine practice variation. Data on practices were collected via a questionnaire. Fifty-two EDs across The Netherlands were included. Thrombosis prophylaxis was prescribed for below-knee plaster immobilization in 27% of EDs. Vitamin C was prescribed in 50% of EDs after a wrist fracture. Splitting of applied casts to the upper or lower limb was performed in one-third of the EDs. Analysis of the cervical spine after trauma was performed by the NEXUS criteria (69%), the Canadian C-spine Rule (17%) or otherwise. The imaging modality for cervical spine trauma in adults was a CT scan (98%). The cast used for scaphoid fractures was divided between the short arm cast (46%) and the navicular cast (54%). Locoregional anaesthesia for femoral fractures was applied in 54% of the EDs. EDs in The Netherlands showed considerable practice variation in treatments among the subjects studied. Further research is warranted to gain a full understanding of the variation in practice in EDs and the potential to improve quality and efficiency.

Physiological Responses of Agave maximiliana to Inoculation with Autochthonous and Allochthonous Arbuscular Mycorrhizal Fungi.

The benefits of mycorrhizal interactions are only known in 8 of 210 recognized Agave taxa. We evaluated the effects of autochthonous and allochthonous arbuscular mycorrhizal fungi (AMF) on growth and nutrient assimilation in Agave maximiliana. The autochthonous consortium (Cn) of eight species was propagated from the rhizospheric soil of A. maximiliana, while Claroideoglomus claroideum (Cc) and Claroideoglomus etunicatum (Ce) were employed as allochthonous AMF. Six treatments were included in the study: Cn, Ce, Cc, Ce + Cc, Tf (fertilized control), and Tn (non-fertilized control, not inoculated). Mycorrhizal colonization increased over time, and the colonization percentages produced by Cn and the allochthonous AMF, both alone and mixed together, were equal at 6, 12, and 18 months. Height increased steadily and was higher in AMF-treated plants from seven months onward. Growth indicators of AMF-treated and AMF-free plants were equal at 6 months, but the beneficial effects of allochthonous and autochthonous AMF were evident in all growth indicators at 18 months and in sugar and mineral (P, K, Ca, Mg, and Fe) content. Arbuscular mycorrhizal fungi significantly improved all growth parameters of A. maximiliana regardless of the origin of the inoculums. This is the first study to report the positive effects of AMF colonization in A. maximiliana.

Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts.

BACKGROUND: In their attempt to fulfill the wish of having children, women who suffer from fertility issues often undergo assisted reproductive technologies such as ovarian stimulation, which has been associated with adverse health outcomes and imprinting disorders in children. However, given the crucial role of exogenous hormone stimulation in improving human infertility treatments, a more comprehensive analysis of the potential impacts on DNA methylation in embryos following ovarian stimulation is needed. Here, we provide genome-wide DNA methylation profiles of blastocysts generated after superovulation of prepubertal or adult mice, compared with blastocysts derived from non-stimulated adult mice. Additionally, we assessed the impact of the in vitro growth and maturation of oocytes on methylation in blastocysts. RESULTS: Neither hormone stimulation nor sexual maturity had an impact on the low global methylation levels characteristic of the blastocyst stage or was associated with extensive DNA methylation alterations. However, we found hormone- and age-associated changes at specific positions but dispersed throughout the genome. In particular, we detected anomalous methylation at a limited number of CpG islands. Additionally, superovulation in adult mice was associated with alterations at the Sgce and Zfp777 imprinted genes. On the other hand, in vitro culture of follicles from the early pre-antral stage was associated with globally reduced methylation and increased variability at imprinted loci in blastocysts. CONCLUSIONS: Our results indicate a minimal effect of ovarian stimulation of adult and prepubertal mice on the DNA methylation landscape attained at the blastocyst stage, but potentially greater impacts of in vitro growth and maturation of oocytes. These findings have potential significance for the improvement of assisted reproductive techniques, in particular for those related to treatments in prepubertal females, which could be crucial for improving human fertility preservation strategies.

Prehospital transdermal glyceryl trinitrate in patients with presumed acute stroke (MR ASAP): an ambulance-based, multicentre, randomised, open-label, blinded endpoint, phase 3 trial.

BACKGROUND: Pooled analyses of previous randomised studies have suggested that very early treatment with glyceryl trinitrate (also known as nitroglycerin) improves functional outcome in patients with acute ischaemic stroke or intracerebral haemorrhage, but this finding was not confirmed in a more recent trial (RIGHT-2). We aimed to assess whether patients with presumed acute stroke benefit from glyceryl tr initrate started within 3 h after symptom onset. METHODS: MR ASAP was a phase 3, randomised, open-label, blinded endpoint trial done at six ambulance services serving 18 hospitals in the Netherlands. Eligible participants (aged ≥18 years) had a probable diagnosis of acute stroke (as assessed by a paramedic), a face-arm-speech-time test score of 2 or 3, systolic blood pressure of at least 140 mm Hg, and could start treatment within 3 h of symptom onset. Participants were randomly assigned (1:1) by ambulance personnel, using a secure web-based electronic application with random block sizes stratified by ambulance service, to receive either transdermal glyceryl trinitrate 5 mg/day for 24 h plus standard care (glyceryl trinitrate group) or to standard care alone (control group) in the prehospital setting. Informed consent was deferred until after arrival at the hospital. The primary outcome was functional outcome assessed with the modified Rankin Scale (mRS) at 90 days. Safety outcomes included death within 7 days, death within 90 days, and serious adverse events. Analyses were based on modified intention to treat, and treatment effects were expressed as odds ratios (ORs) or common ORs, with adjustment for baseline prognostic factors. We separately analysed the total population and the target population (ie, patients with intracerebral haemorrhage, ischaemic stroke, or transient ischaemic attack). The target sample size was 1400 patients. The trial is registered as ISRCTN99503308. FINDINGS: On June 24, 2021, the MR ASAP trial was prematurely terminated on the advice of the data and safety monitoring board, with recruitment stopped because of safety concerns in patients with intracerebral haemorrhage. Between April 4, 2018, and Feb 12, 2021, 380 patients were randomly allocated to a study group. 325 provided informed consent or died before consent could be obtained, of whom 170 were assigned to the glyceryl trinitrate group and 155 to the control group. These patients were included in the total population. 201 patients (62%) had ischaemic stroke, 34 (10%) transient ischaemic attack, 56 (17%) intracerebral haemorrhage, and 34 (10%) a stroke-mimicking condition. In the total population (n=325), the median mRS score at 90 days was 2 (IQR 1-4) in both the glyceryl trinitrate and control groups (adjusted common OR 0·97 [95% CI 0·65-1·47]). In the target population (n=291), the 90-day mRS score was 2 (2-4) in the glyceryl trinitrate group and 3 (1-4) in the control group (0·92 [0·59-1·43]). In the total population, there were no differences between the two study groups with respect to death within 90 days (adjusted OR 1·07 [0·53-2·14]) or serious adverse events (unadjusted OR 1·23 [0·76-1·99]). In patients with intracerebral haemorrhage, 12 (34%) of 35 patients allocated to glyceryl trinitrate versus two (10%) of 21 allocated to the control group died within 7 days (adjusted OR 5·91 [0·78-44·81]); death within 90 days occurred in 16 (46%) of 35 in the glyceryl trinitrate group and 11 (55%) of 20 in the control group (adjusted OR 0·87 [0·18-4·17]). INTERPRETATION: We found no sign of benefit of transdermal glyceryl trinitrate started within 3 h of symptom onset in the prehospital setting in patients with presumed acute stroke. The signal of potential early harm of glyceryl trinitrate in patients with intracerebral haemorrhage suggests that glyceryl trinitrate should be avoided in this setting. FUNDING: The Collaboration for New Treatments of Acute Stroke consortium, the Brain Foundation Netherlands, the Ministry of Economic Affairs, Stryker, Medtronic, Cerenovus, and the Dutch Heart Foundation.

Characterization of carbohydrate metabolism in in vivo- and in vitro-grown and matured mouse antral follicles†.

Establishing an ideal human follicle culture system for oncofertility patients relies mainly on animal models since donor tissue is scarce and often of suboptimal quality. The in vitro system developed in our laboratory supports the growth of prepubertal mouse secondary follicles up to mature oocytes. Given the importance of glucose in preparing the oocyte for proper maturation, a baseline characterization of follicle metabolism both in the culture system and in vivo was carried out. Markers of glucose-related pathways (glycolysis, tricarboxylic acid [TCA] cycle, pentose phosphate pathway [PPP], polyol pathway, and hexosamine biosynthetic pathway), as well as the antioxidant capacity, were measured in the different follicle cell types by both enzymatic activities (spectrophotometric detection) and gene expression (qPCR). This study confirmed that in vivo the somatic cells, mainly granulosa, exhibit intense glycolytic activity, while oocytes perform PPP. Throughout the final maturation step, oocytes in vivo and in vitro showed steady levels for all the key enzymes and metabolites. On the other hand, ovulation triggers a boost of pyruvate and lactate uptake in cumulus cells in vivo, consumes reduced nicotinamide adenine dinucleotide phosphate, and increases TCA cycle and small molecules antioxidant capacity activities, while in vitro, the metabolic upregulation in all the studied pathways is limited. This altered metabolic pattern might be a consequence of cell exhaustion because of culture conditions, impeding cumulus cells to fulfill their role in providing proper support for acquiring oocyte competence.

Complicated placenta accreta spectrum: identifying a high-risk cohort.

OBJECTIVE: To assess differences in the perioperative complication rate between patients with placenta accreta spectrum (PAS) with and without complicating factors. METHODS: This retrospective cohort study included subjects who underwent cesarean hysterectomy with histology-proven PAS between 23 0/7 and 42 0/7 weeks gestational age (GA) from 1 July 2008 to 11 April 2017. Perioperative outcomes were compared between those with uncomplicated PAS and "complicated PAS," defined as PAS subjects who experienced ≥2 bleeding episodes, preterm premature rupture of membranes (PPROM), or premature contractions requiring tocolysis. RESULTS: Overall, 26 complicated PAS and 27 uncomplicated PAS cases were compared; no difference in the rate of perioperative complications was identified. An increased proportion of complicated PAS cases required blood product transfusion before delivery: 2 (40%), 3 (27.3%), and 2 patients (20%) for those with PPROM, preterm contractions, and ≥2 bleeding episodes respectively, compared to patients with uncomplicated PAS, having no transfusions (p = .001). Time of delivery was earlier for patients with complicated compared to uncomplicated PAS (median GA 30.9 [Q1 = 27.9; Q3 = 31.9] and 34.9 [Q1 = 32.1; Q3 = 35.7], p < .001). Median birthweights were lower (p < .0144) and maternal length of stay longer (p < .0012) for complicated PAS. CONCLUSION: Patients with complicated PAS were not at higher risk for perioperative complications but were associated with earlier delivery, required more antenatal blood transfusions, and had a longer LOS.

Reversing complete mechanical transzonal projections disruption during mouse in vitro follicle culture with unaltered oocyte competence†.

In vitro oocyte growth is widely studied as an alternative fertility preservation approach. Several animal models are used to generate extensive information on this complex process regulated by the constant and dynamic interaction between the oocyte and its somatic compartment throughout follicle growth and maturation. A two-dimensional attachment mouse secondary follicle culture system was used to assess the oocyte's capacity to overcome disconnection from its somatic companions at different developmental stages for final competence acquisition. To test this, complete mechanical denudation of oocytes from preantral (PA) and early antral (EA) follicles was performed. Established endpoints were the oocyte's potential to reconnect with somatic cells and the impact of connectivity disruption on mature oocyte quality. This study proves that oocytes from PA and EA cultured mouse follicles can overcome complete denudation, restoring likely functional transzonal projections with no significant differences in meiotic and developmental competence compared with those from intact cultured follicles. These novel findings constitute good premises for developing successful strategies to rescue human oocyte competence in the context of in vitro culture approaches such as nonhuman chorionic gonadotropin triggered in vitro maturation.

Glucose metabolism characterization during mouse in vitro maturation identifies alterations in cumulus cells†.

In vitro maturation (IVM) is an assisted reproduction technique with reduced hormone-related side-effects. Several attempts to implement IVM in routine practice have failed, primarily due to its relatively low efficiency compared with conventional in vitro fertilization (IVF). Recently, capacitation (CAPA)-IVM-a novel two-step IVM method-has improved the embryology outcomes through synchronizing the oocyte nuclear and cytoplasmic maturation. However, the efficiency gap between CAPA-IVM and conventional IVF is still noticeable especially in the numerical production of good quality embryos. Considering the importance of glucose for oocyte competence, its metabolization is studied within both in vivo and CAPA-IVM matured mouse cumulus-oocyte-complexes (COCs) through direct measurements in both cellular compartments, from transcriptional and translational perspectives, to reveal metabolic shortcomings within the CAPA-IVM COCs. These results confirmed that within in vivo COC, cumulus cells (CCs) are highly glycolytic, whereas oocytes, with low glycolytic activity, are deviating their glucose towards pentose phosphate pathway. No significant differences were observed in the CAPA-IVM oocytes compared with their in vivo counterparts. However, their CCs exhibited a precocious increase of glycolytic activity during the pre-maturation culture step and activity was decreased during the IVM step. Here, specific alterations in mouse COC glucose metabolism due to CAPA-IVM culture were characterized using direct measurements for the first time. Present data show that, while CAPA-IVM CCs are able to utilize glucose, their ability to support oocytes during final maturation is impaired. Future CAPA-IVM optimization strategies could focus on adjusting culture media energy substrate concentrations and/or implementing co-culture strategies.

Evaluación del Plan de Alta por enfermería en el hospital San José, Popayán 2017 / Evaluation of the effectiveness of hospital discharge planning at San José hospital, Popayán 2017

Resumen Introducción: Durante el alta hospitalaria es importante que el enfermero identifique los cuidados que requiere el paciente, permitiéndole guiar intervenciones mediante el plan de alta que fomenta el autocuidado, enfocado a necesidades específicas de cada persona, para prevenir complicaciones y disminuir reingresos. Objetivo: Evaluar la ejecución del plan de alta por los profesionales de enfermería en los servicios de medicina interna y quirúrgicas del Hospital San José, Popayán durante el año 2017. Materiales y métodos : Estudio de tipo cuantitativo, diseño descriptivo y transversal. La población conformada por los enfermeros de los servicios de medicina interna y quirúrgicas del Hospital San José y pacientes con alta programada de 24 horas o menos antes del egreso hospitalario. Resultados: La información se recolectó por medio de un instrumento de valoración realizado a participantes que cumplen con criterios de inclusión. El 94,7% de los pacientes tenían egreso programado, de estos 58,6% recibieron indicaciones en las últimas 48 horas sobre los cuidados en casa y el 100% manifestaron entender las recomendaciones brindadas por el enfermero. Respecto al profesional de enfermería el 100% refiere no conocer alguna metodología para elaborar el plan de alta y el 80% no conoce si en la institución existe un procedimiento de egreso y seguimiento al paciente. Conclusiones: El plan de alta es un proceso que no se encuentra estandarizado y por lo tanto no es evaluado en la institución, el personal de Enfermería se limita a brindar educación sobre las indicaciones médicas de egreso y no sobre un plan de alta propio de Enfermería.

Abstract: Introduction: When the patients are going to be discharged from the hospital, the nurses must execute the discharge plan based on their specific needs. It is essential to promote self-care in patients to prevent future medical complications and readmissions. Objective: To evaluate the implementation of the discharge plan by the nursing professionals in the internal medicine and surgical services at the San José hospital in Popayán, Colombia, in 2017. Materials and methods: This quantitative study has a descriptive and transversal design. The research population is composed of nurses from the internal medicine and surgical services from the San José Hospital and patients that are going to be discharged from the clinic in 24 hours or less. Results: The information was collected by using an instrument that was designed for the participants to confirm if they fit the inclusion criteria. According to collected data, 94.7% of the patients had scheduled discharges, 58.6% of them had received indications in the last 48 hours before the discharge on how the self-care routines must be followed at home. Additionally, 100% of people said they understood the recommendations given by the nurses. Finally, 100% of the nursing professionals claimed that they did not know any methodology to prepare the discharge plan, and about 80% of them did not know about the existence of a patient's exit procedure and patient's medical follow-up in the institution. Conclusions: The patient discharge plan is not standardized, so it is not evaluated in the institution. The nursing staff is focused only on explaining the medical indications, but they are not focused on creating their discharge plans.

Using a Macro Lens for Anterior Segment Imaging in Rural Panama.

Background: Visual impairment, specifically anterior segment pathology, presents a significant burden of disease in the world. Introduction: Inexpensive tools are necessary to improve eye health of residents in developing countries where care is difficult to access. Our study aimed at determining whether a $5 macro lens attached to a smartphone camera is an effective anterior segment imaging method for screening diseases. Materials and Methods: Fifty four (n = 54) patients had anterior segment imaging performed by using an Easy Macro lens and an iPhone. Imaging was performed at the Floating Doctors' mobile clinic sites in Panama. Images were sent back and graded by two board-certified ophthalmologists using a modified version of the FOTO-ED scale. Statistical analysis was performed by using a Wilcoxon signed-rank test to compare grades between the two imaging modalities. Results: There was no significant difference in overall clinical utility of images obtained by the iPhone versus Easy Macro lens. The iPhone was significantly superior in imaging of the lens and conjunctiva, whereas the Easy Macro lens was superior in regards to the anterior chamber, iris, and lens. Discussion: The imaging modality that best captures pathology is dependent on what part of the anterior segment is being examined. An imaging protocol with a pair of images, one from a smartphone and one from a macro lens, would have significant clinical utility. Conclusion: Our study demonstrates how minimally trained users can deliver effective eye screening via a telemedicine-based approach in a resource-deprived setting. Future directions would be to develop a telemedicine protocol and determine whether it improves clinically measurable outcomes in patients.

IMPACT OF CRUDE OIL ON FUNCTIONAL GROUPS OF CULTURABLE BACTERIA AND COLONIZATION OF SYMBIOTIC MICROORGANISMS IN THE Clitoria-Brachiaria RHIZOSPHERE GROWN IN MESOCOSMS / Impacto del petróleo crudo sobre grupos funcionales de bacterias cultivables y colonización de microorganismos simbióticos en la rizosfera de Clitoria-Brachiaria crecidas en mesocosmos

ABSTRACT This research evaluated the changes on populations of culturable N-fixing free bacteria (NFFB) and P-solubilizing bacteria (PSB), as well as on the root nodulation by native rhizobia, the root colonization and spore number of arbuscular mycorrhizal fungi (AMF), in the rhizosphere of Clitoria ternatea and Brachiaria brizantha grown in mesocosms contaminated with crude oil (0, 3000, 6000, 9000, and 12000 mg kg-1), for 240 days. After 24 h of soil contamination, the highest populations of NFFB and PSB (5.5 and 4.9 LogUFC, respectively) were found in control, and the lowest populations were obtained at 12000 mg kg-1 (5.1 and 4.2 LogUFC, respectively). In contrast, at 60 and 240 days, the control showed lower populations of NFFB and PSB (5.4 and 4.8 LogUFC, respectively) than contaminated treatments. The highest number of root nodules in C. ternatea was quantified in control at 60 and 240 days (25 and 27 nodules, respectively) in comparison to those observed at the treatment with 12000 mg kg-1 (7 and 1 nodule, respectively). At 60 days, AMF colonization in both plant species, and the number of spores significantly decreased as the crude oil concentration increased; however, at 240 days, the highest number of AMF spores was recorded at treatments with 6000 and 12000 mg kg-1. The dry weight of both plant species significantly decreased as crude oil concentrations increased. Although C. ternatea was more susceptible to the toxic effects of crude oil, this plant species showed greater content of total chlorophyll than B. brizantha.

RESUMEN Esta investigación evaluó los cambios en la población cultivable de bacterias de vida libre fijadoras de nitrógeno (BVLFN) y de bacterias solubilizadoras de fósforo (BSP), así como en la nodulación de raíces por rizobios nativos, y en la colonización y número de esporas de hongos micorrízicos arbusculares (HMA) en la rizósfera de Clitoria ternatea y Brachiaria brizantha cultivadas en mesocosmos contaminados con petróleo crudo (0, 3000, 6000, 9000 y 12000 mg kg-1), durante 240 días. A las 24 h de la contaminación del suelo, las poblaciones más altas de BVLFN y BSP (5,5 y 4,9 LogUFC, respectivamente) se encontraron en el control, mientras que las poblaciones más bajas se obtuvieron a 12000 mg kg-1 (5,1 y 4,2 LogUFC, respectivamente). En contraste, a los 60 y 240 días, el control mostró bajas poblaciones de BVLFN y BSP (5,4 y 4,8 LogUFC, respectivamente) que los tratamientos contaminados. El mayor número de nódulos en raíz de C. ternatea se cuantificó en el control a los 60 y 240 días (25 y 27 nódulos, respectivamente) en comparación con el tratamiento con 12000 mg kg-1 (7 y 1 nódulos, respectivamente). A los 60 días, la colonización de HMA en ambas especies vegetales y el número de esporas disminuyeron significativamente al aumentar la concentración de petróleo crudo; sin embargo, a los 240 días, se registró el mayor número de esporas de HMA en los tratamientos con 6000 y 12000 mg kg-1. El peso seco vegetal disminuyó significativamente al aumentar las concentraciones de petróleo crudo. Clitoria ternatea fue más susceptible a la toxicidad del petróleo, aunque esta especie vegetal mostró mayor contenido de clorofila total que B. brizantha.

The association between intravenous fluid resuscitation and mortality in older emergency department patients with suspected infection.

OBJECTIVE: Recent studies suggest that hypotension thresholds in current guidelines might be too low for older patients due to arterial stiffening, possibly leading to insufficient fluid resuscitation. We compared intravenous (IV) fluid volumes that older (≥ 70 years) and younger (< 70 years) patients with suspected infection with similar initial systolic blood pressure (SBP) received in the emergency department (ED) and investigated whether this was associated with in-hospital mortality in older patients. METHODS: This was an observational multicenter study using an existing database in which consecutive ED patients hospitalized with suspected infection were prospectively included. We first compared the fluid volumes older and younger ED patients received per initial SBP category. Patients were then stratified into two SBP categories (≤ or > 120 mmHg; 120 has been suggested to be a better threshold) and thereafter into three fluid volume categories: 0-1 L, 1-2 L, or > 2 L. In each SBP and fluid category, case-mix-adjusted in-hospital mortality was compared between older and younger patients, using multivariable logistic regression analysis. RESULTS: The included 981 (37%) older and 1678 (63%) younger ED patients received similar IV fluid volumes per initial SBP category. Older patients with an initial SBP > 120 mmHg had a higher adjusted OR of 2.06 (95% CI 1.02-4.16), in the 0-1 L category, while this association was not found in the higher fluid categories of 1-2 L or > 2 L. In the SBP ≤ 120 mmHg category, this association was also absent. CONCLUSION: This hypothesis-generating study suggests that older patients with suspected infection may need higher fluid volumes than younger patients, when having a seemingly normal initial SBP.

Zika Virus Protease Cleavage of Host Protein Septin-2 Mediates Mitotic Defects in Neural Progenitors.

Zika virus (ZIKV) targets neural progenitor cells in the brain, attenuates cell proliferation, and leads to cell death. Here, we describe a role for the ZIKV protease NS2B-NS3 heterodimer in mediating neurotoxicity through cleavage of a host protein required for neurogenesis. Similar to ZIKV infection, NS2B-NS3 expression led to cytokinesis defects and cell death in a protease activity-dependent fashion. Among binding partners, NS2B-NS3 cleaved Septin-2, a cytoskeletal factor involved in cytokinesis. Cleavage of Septin-2 occurred at residue 306 and forced expression of a non-cleavable Septin-2 restored cytokinesis, suggesting a direct mechanism of ZIKV-induced neural toxicity. VIDEO ABSTRACT.