Post-transcriptional silencing of the SGE1 gene induced by a dsRNA hairpin in Fusarium oxysporum f. sp cubense, the causal agent of Panama disease.

Fusarium oxysporum f. sp cubense (Foc), the causal agent of Panama disease, is responsible for economic losses in banana crops worldwide. The identification of genes that effectively act on pathogenicity and/or virulence may contribute to the development of different strategies for disease control and the production of resistant plants. The objective of the current study was to analyze the importance of SGE1 gene expression in Foc virulence through post-transcriptional silencing using a double-stranded RNA hairpin. Thirteen transformants were selected based on different morphological characteristics, and sporulation in these transformants was significantly reduced by approximately 95% (P < 0.05) compared to that of the wild-type strain. The relative SGE1 expression levels in the transformant strains were reduced by 27 to 47% compared to those in the wild-type strain. A pathogenicity analysis revealed that the transformants were able to reach the rhizomes and pseudostems of the inoculated banana plants. However, the transformants induced initial disease symptoms in the banana plants approximately 10 days later than that by the wild-type Foc, and initial disease symptoms persisted even at 45 days after inoculation. These results indicate that the SGE1 gene is directly involved in the virulence of Foc. Therefore, SGE1 may be a potential candidate for host-induced gene silencing in banana plants.

[Treatment of postpartum hemorrhage with intrauterine hydrostatic condom]. / Tratamiento de la hemorragia posparto con condón hidrostático intrauterine.

OBJECTIVE: To report of experience of intrauterine hydrostatic condom to control obstetric hemorrhage. MATERIAL AND METHOD: Descriptive, retro-prospective study. The method was use in patients who had obstetric hemorrhage and do not responded to medical management during the period from March 1st to August27, 2015 in a rural facility. RESULTS: 955 patients that had a vaginal delivery were identified, 40 (4.1%) of which needed the application of the method. The method was unsuccessful in 2 of 40 patients (5%), one requiring emergency obstetric hysterectomy and other exploratory laparotomy with conservative measures. 11 of 40 patients (2 7.5%) required at least 1-3 globular packs transfusion. None of the 38 patients (95%) who responded to the method presented endometritis in the postpartum follow up or complications associated with the use of hydrostatic condom. CONCLUSION: The use of intrauterine hydrostatic condom is an effective method to control postpartum obstetric hemorrhage secondary to uterine atony unresponsive to medical management.

Serial determinations of asymmetric dimethylarginine and homocysteine during pregnancy to predict pre-eclampsia: a longitudinal study.

OBJECTIVE: To evaluate the usefulness of serial determinations of asymmetric dimethylarginine (ADMA) and homocysteine (Hcy) concentrations during pregnancy to predict pre-eclampsia, taking into account maternal obesity and B vitamin status. DESIGN: Longitudinal study. SETTING: Two obstetric referral hospitals. SAMPLE: Two hundred and fifty-two of 411 women invited to participate in the study. METHODS: The women made monthly visits from ≤20 weeks of gestation until delivery for measurements of plasma ADMA, Hcy, and vitamins B6 , B12, and folic acid, and for the recording of clinical information. MAIN OUTCOME MEASURE: Early elevations in plasma ADMA and Hcy related to the development of pre-eclampsia. RESULTS: Of the 252 women who completed the study, 179 had no complications, 49 developed pre-eclampsia, and 24 presented with complications other than pre-eclampsia. ADMA and Hcy increased gradually throughout pregnancy in the pre-eclampsia group, independent of maternal B-vitamin status and obesity, but remained constant in women with no complications. Relative to the preceding month, ADMA and Hcy levels increased 1 month prior to the onset of pre-eclampsia: 124 ± 27 nmol (P < 0.001) and 1177 ± 278 nmol (P = 0.001), respectively, in the pre-eclampsia group. The group of women with no complications did not show any significant changes. Increases of 80 nmol ADMA and 1000 nmol Hcy at 1 month prior to the onset of pre-eclampsia demonstrated the best potential for prediction. CONCLUSIONS: Increased ADMA and Hcy levels precede clinical manifestations of pre-eclampsia. Therefore, serial determinations of their concentrations may be helpful in identifying women at risk. TWEETABLE ABSTRACT: Increased ADMA and Hcy precede clinical pre-eclampsia and may identify women at risk.

Operational taxonomic units (OTUs) of endophytic bacteria isolated from banana cultivars in the Amazon.

Endophytic microorganisms colonize plants, inhibit the growth of pathogens (by competing for nutrients and/or space), or produce antagonistic substances. Fifty-five endophytic bacteria were isolated from the leaf tissue of the FHIA 18 banana cultivar. Genetic diversity analyses were performed using the enterobacterial repetitive intergenic consensus sequence polymerase chain reaction method and BOX molecular markers. These analyses resulted in 33 and 21 polymorphic bands, respectively. The similarity data, obtained using the Dice coefficient based on the polyphasic analysis method, ranged from 22 to 100%. This indicated a high genetic diversity among the analyzed isolates. Sixty percent similarity was utilized as the cut-off criterion for the formation of operational taxonomic units (OTUs); this resulted in the identification of 32 possible OTUs, indicating a high number of potential species.

Endophytic bacteria from banana cultivars and their antifungal activity.

Endophytic microorganisms consist of fungi, bacteria, and actinomycetes that play important roles in the process of plant adaptation to the environment. Currently, the natural associations between microorganisms and plant species are being explored for a large number of biotechnological applications. In this study, 122 endophytic bacteria were isolated from 5 cultivars of Musa spp from the state of Amazonas (Brazil). Four strains were selected because they exhibited antagonistic activities against Fusarium oxysporum f. sp cubense and Colletotrichum guaranicola, with inhibitions ranging from 19 to 30% and 27 to 35%, respectively. Phylogenetic analysis of the 16S rDNA regions of these bacteria with antifungal activity showed that they are phylogenetically related to 3 different species of Bacillus - B. amyloliquefaciens, B. subtilis subsp subtilis, and B. thuringiensis.

Application of nanosilica in the construction industry: A bibliometric analysis using Methodi Ordinatio.

The number of publications related to the implementation of nanotechnology in the construction industry, and specifically to the application of nanosilica (SiO2), has had a constant increase in recent years. Based on this, in the present work, an analysis was carried out using bibliometric techniques, with the aim at characterizing the development of specialized literature and identifying the largest areas of growth in the field, maintaining hydrophobic nanosilica as the research guideline. This analysis acquired information from the Scopus and Web of Science (WoS) databases to compare bibliometric indicators of the publications. It should be noted that, even though bibliometric analysis is useful to identify the study areas of greatest interest, to complement this work, the implementation of a method that helped in the research process to obtain the most important bibliography was required. This study implemented Methodi Ordinatio, which helped to take a new direction. Therefore, based on this method, a list of articles cataloged and ranked is obtained, which is the basis for integrating the final bibliographic portfolio. •The study applies the Methodi Ordinatio to obtain a portfolio of the most relevant articles to guide the researchers' work.•Insightful information can be obtained using VOSviewer to analyze and visualize metadata of the bibliographic portfolio.•The study demonstrates how the alpha value in the InOrdinatio formula modifies the resulting portfolio.

Does chlorhexidine reduce the incidence of ventilator-associated pneumonia in ICU patients? A systematic review and meta-analysis.

OBJECTIVE: This study aimed to investigate chlorhexidine's efficacy in preventing ventilator-associated pneumonia (VAP). DESIGN: A systematic review and meta-analysis were conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses. SETTINGS: The data were obtained from Pubmed, Cochrane Library, and EMBASE. PATIENTS OR PARTICIPANTS: Only mechanically ventilated patients for at least 48h were included. INTERVENTIONS: Randomized clinical trials applying any dosage form of chlorhexidine were eligible. MAIN VARIABLES OF INTEREST: The relative risk (RR) of the VAP incidence and all-cause mortality was assessed using the random-effects model. The mean difference in days of mechanical ventilation duration and intensive care unit (ICU) length of stay were also appraised. RESULTS: Ten studies involving 1233 patients were included in the meta-analysis. The oral application of CHX reduced the incidence of VAP (RR, 0.73 [95% CI, 0.55, 0.97]) and did not show an increase in all-cause mortality (RR, 1.13 [95% CI, 0.96, 1.32]). CONCLUSIONS: CHX proved effective to prevent VAP. However, a conclusion on mortality rates could not be drawn because the quality of the evidence was very low for this outcome.

Genetic and antigenic characterization of Brazilian SRLV strains: Natural small ruminant interspecies transmission from mixed herds.

Cross-species transmission events and mixed infection of small ruminant lentiviruses (SRLVs) were studied in seven goats and two sheep from three small ruminant mixed flocks from Northeast and Southeast Brazil. Genetic and antigenic analyses with gag/env genes and ELISA multiepitope SU1/SU5 recombinant antigens were carried out, respectively. The genetic analysis of gag and env sequences showed high viral diversity in both species, MVV-like (subtype A1) and CAEV-like B1 in goats, and CAEV-like (subtype B1) in sheep, revealing SRLV interspecies transmission from sheep to goats and vice versa in Brazilian farms. Two Brazilian caprine lentiviruses were segregated in two new genetic clades based on gag analyses, which suggests a new classification into heterogenic genotype A. Furthermore, goat isolates were grouped into subtype A1 and B1 clusters. Cross-reactive antibodies were detected in goats using ELISA with a recombinant antigen carrying SU1 and SU5 immunodominant epitopes; the results showed anti-CAEV and MVV antibodies in goats and anti-CAEV antibodies in sheep. This result can be associated with the high divergence in the V4 region due to SRLV variability. All results confirm cross-species infection of SRLV in Brazilian mixed herds.

Effects of Hyperosmolality on Hypothalamic Astrocytic Area, mRNA Expression and Glutamate Balance In Vitro.

During prolonged dehydration, body fluid homeostasis is challenged by extracellular fluid (ECF) hyperosmolality, which induce important functional changes in the hypothalamus, in parallel with other effector responses, such as the activation of the local renin-angiotensin system (RAS). Therefore, in the present study we investigated the role of sodium-driven ECF hyperosmolality on glial fibrillary acid protein (GFAP) immunoreactivity and protein expression, membrane capacitance, mRNA expression of RAS components and glutamate balance in cultured hypothalamic astrocytes. Our data show that hypothalamic astrocytes respond to increased hyperosmolality with a similar decrease in GFAP expression and membrane capacitance, indicative of reduced cellular area. Hyperosmolality also downregulates the transcript levels of angiotensinogen and both angiotensin-converting enzymes, whereas upregulates type 1a angiotensin II receptor mRNA. Incubation with hypertonic solution also decreases the immunoreactivity to the membrane glutamate/aspartate transporter (GLAST) as well as tritiated-aspartate uptake by astrocytes. This latter effect is completely restored to basal levels when astrocytes previously exposed to hypertonicity are incubated under isotonic conditions. Together with a direct effect on two important local signaling systems (glutamate and RAS), these synaptic rearrangements driven by astrocytes may accomplish for a coordinated increase in the excitatory drive onto the hypothalamic neurosecretory system, ultimately culminating with increased AVP release in response to hyperosmolality.

Intermittent activation of peripheral chemoreceptors in awake rats induces Fos expression in rostral ventrolateral medulla-projecting neurons in the paraventricular nucleus of the hypothalamus.

Despite the well-established sympathoexcitation evoked by chemoreflex activation, the specific sub-regions of the CNS underlying such sympathetic responses remain to be fully characterized. In the present study we examined the effects of intermittent chemoreflex activation in awake rats on Fos-immunoreactivity (Fos-ir) in various subnuclei of the paraventricular nucleus of the hypothalamus (PVN), as well as in identified neurosecretory preautonomic PVN neurons. In response to intermittent chemoreflex activation, a significant increase in the number of Fos-ir cells was found in autonomic-related PVN subnuclei, including the posterior parvocellular, ventromedial parvocellular and dorsal-cap, but not in the neurosecretory magnocellular-containing lateral magnocellular subnucleus. No changes in Fos-ir following chemoreflex activation were observed in the anterior PVN subnucleus. Experiments combining Fos immunohistochemistry and neuronal tract tracing techniques showed a significant increase in Fos-ir in rostral ventrolateral medulla (RVLM)-projecting (PVN-RVLM), but not in nucleus of solitarii tract (NTS)-projecting PVN neurons. In summary, our results support the involvement of the PVN in the central neuronal circuitry activated in response to chemoreflex activation, and indicate that PVN-RVLM neurons constitute a neuronal substrate contributing to the sympathoexcitatory component of the chemoreflex.

BPA and NP removal from municipal wastewater by tropical horizontal subsurface constructed wetlands.

It has been recognized that numerous synthetic compounds like Bisphenol A (BPA) and nonylphenols (NP) are present in effluents from wastewater treatment plants (WWTP) at levels of parts per billion (µg L(-1)) or even parts per trillion (ng L(-1)) with a high potential to cause endocrine disruption in the aquatic environment. Constructed wetlands (CW) are a cost-effective wastewater treatment alternative with promising performance to treat these afore mentioned compounds. This research was aimed to evaluate the efficacy of CW treatment of WWTP effluent for mitigating the effects endocrine disrupting compounds (EDCs). This research goal was accomplished by (1) quantifying the removal of BPA and NP in CWs; (2) isolating CW fungal strains and testing for laccase production; and (3) performing endocrine disruption (reproduction) bioassays using the fruit fly Drosophila melanogaster. Three pilot scale horizontal subsurface flow constructed wetlands (HSSF-CW) were operated for eight weeks: one planted with Phragmites australis; one planted with Heliconia psitacorum; and one unplanted. The Heliconia CW showed a removal efficiency of 73.3(± 19%) and 62.8(± 20.1%) for BPA and NP, respectively; while the Phragmites CW demonstrated a similar removal for BPA (70.2 ± 27%) and lower removal efficiency for NP 52.1(± 37.1%).The unplanted CW achieved 62.2 (± 33%) removal for BPA and 25.3(± 37%) removal for NP. Four of the eleven fungal strains isolated from the Heliconia-CW showed the capacity to produce laccase. Even though complete removal of EDCs was not achieved by the CWs, the bioassay confirmed a significant improvement (p < 0.05) in fly viability for all CWs, with Heliconia sp. being the most effective at mitigating adverse effects on first and second generational reproduction. This study showed that a CW planted with a native Heliconia sp. CW demonstrated a higher removal of endocrine disrupting compounds and better mitigation of reproductive disruption in the bioassay.

Ibandronate decreases bone disease development and osteoclast stimulatory activity in an in vivo model of human myeloma.

The benefits of bisphosphonate therapy for multiple myeloma bone disease have been clearly documented. However, the effects of bisphosphonates on the osteoclast stimulatory activity (OSA) that is present in the marrow of patients with multiple myeloma, even before the bone disease is detectable, are unknown. Therefore, we examined the effects of ibandronate (IB) treatment prior to the development of bone disease in a murine model of human myeloma. Sublethally irradiated severe combined immunodeficient (SCID) mice were transplanted with ARH-77 cells on day 0. These ARH-77 mice were treated daily with subcutaneous injections of IB started before or at different times after tumor injection as follows: group 1 was started on day -7; group 2 on day 0; group 3 on day +7; group 4 on day +14 after IB administration; and group 5 (control) received no IB. Mice were sacrificed after they developed paraplegia. The onset of paraplegia was delayed in group 1 vs all other groups (mean day 27 vs day 32; p = 0.0098). The number of lytic lesions and the bone surface area of resorption (mm(2)) were significantly decreased in groups 1, 2, and 3, which were treated early with IB, when compared with groups 4 and 5 (p = 0.003 and 0.002, respectively). OSA, as measured by the capacity of bone marrow plasma from ARH-77 mice to induce osteoclast (OCL) formation in human bone marrow cultures, was decreased proportionally to the length of IB treatment. Group 1 had the lowest OSA compared with the other groups (p = 0.003). However, all mice eventually developed paraplegia, and at time of sacrifice, tumor burden was not grossly different among the groups. Interestingly, macroscopic abdominal tumors were more frequent in mice treated with IB. These data demonstrate that early treatment of ARH-77 mice with IB prior to development of myeloma bone disease decreases OSA and possibly retards the development of lytic lesions, but not eventual tumor burden.

OSL and TL response characterization of LiF:MG,Ti microdosimeters to be applied to VMAT quality assurance.

This paper aims to evaluate the feasibility of applying LiF:Mg,Ti microdosimeters as a new method of dosimetry to volumetric-modulated arc therapy (VMAT) RapidArc. The response of microdosimeters presented a maximum variation of ±3.18% and ±0.510% using optically stimulated luminescence (OSL) and thermoluminescence (TL) techniques, respectively. Although studies were conducted on LiF:Mg,Ti microdosimeters previously, the microdosimeters in this study showed precision and high potential of application in VMAT dosimetry and in the verification of treatment planning using the VMAT technique.

Radiosurgery for brain metastases: who may not benefit?

PURPOSE: To select a group of patients with brain metastases for whom stereotactic radiosurgery (SRS) may not be beneficial. PATIENTS, MATERIALS, AND METHODS: Actuarial survival of 87 patients with brain metastases treated with SRS between July 1993 and May 1999 was retrospectively analyzed under stratification by the Score Index for Stereotactic Radiosurgery for Brain Metastases (SIR). To identify the group of patients most likely to survive less than 6 months after SRS, Cox model survival curves were calculated for all SIR values, and Kaplan-Meier survival curves were calculated for two SIR subsets (0-5 and 6-10) and were compared by log-rank test. RESULTS: Overall median survival after SRS was 6.88 months. The stratification of patients into two SIR subsets (0-5 and 6-10) sustained statistical significance regarding survival with p = 0.0001. The median survival time for the group of patients with SIR between 0 and 5 was 4.52 months (95% confidence interval of 2.82 to 5.84 months). Survival probability at 6 months for this group of patients with poor prognosis was 35.6%. CONCLUSION: Patients with brain metastases and SIR of 5 or lower have an expected median survival of less than 6 months after treatment with radiosurgery. Thus, radiosurgery may not be beneficial for this group of patients.

Radiosurgery for brain metastases: a score index for predicting prognosis.

PURPOSE: To analyze a prognostic score index for patients with brain metastases submitted to stereotactic radiosurgery (the Score Index for Radiosurgery in Brain Metastases [SIR]). METHODS AND MATERIALS: Actuarial survival of 65 brain metastases patients treated with radiosurgery between July 1993 and December 1997 was retrospectively analyzed. Prognostic factors included age, Karnofsky performance status (KPS), extracranial disease status, number of brain lesions, largest brain lesion volume, lesions site, and receiving or not whole brain irradiation. The SIR was obtained through summation of the previously noted first five prognostic factors. Kaplan-Meier actuarial survival curves for all prognostic factors, SIR, and recursive partitioning analysis (RPA) (RTOG prognostic score) were calculated. Survival curves of subsets were compared by log-rank test. Application of the Cox model was utilized to identify any correlation between prognostic factors, prognostic scores, and survival. RESULTS: Median overall survival from radiosurgery was 6.8 months. Utilizing univariate analysis, extracranial disease status, KPS, number of brain lesions, largest brain lesion volume, RPA, and SIR were significantly correlated with prognosis. Median survival for the RPA classes 1, 2, and 3 was 20.19 months, 7.75 months, and 3. 38 months respectively (p = 0.0131). Median survival for patients, grouped under SIR from 1 to 3, 4 to 7, and 8 to 10, was 2.91 months, 7.00 months, and 31.38 months respectively (p = 0.0001). Using the Cox model, extracranial disease status and KPS demonstrated significant correlation with prognosis (p = 0.0001 and 0.0004 respectively). Multivariate analysis also demonstrated significance for SIR and RPA when tested individually (p = 0.0001 and 0.0040 respectively). Applying the Cox Model to both SIR and RPA, only SIR reached independent significance (p = 0.0004). CONCLUSIONS: Systemic disease status, KPS, SIR, and RPA are reliable prognostic factors for patients with brain metastases submitted to radiosurgery. Applying SIR and RPA classifications to our patients' data, SIR demonstrated better accuracy in predicting prognosis. SIR should be further tested with larger patient accrual and for all patients with brain metastases subjected or not to stereotactic radiosurgery.

Single-breath, room-air method for measuring closing volume (phase 4) in the normal human lung.

The purpose of this study was to evaluate a new method to measure closing volume (CV). This new method does not require oxygen or inert gases to be inhaled to obtain the onset of phase 4. Because there are regional differences in the concentrations of the resident alveolar gases (O2, CO2, and N2), there should be an abrupt change in the concentration of these gases at the terminal portion of a prolonged expired vital capacity (VC) that marks the onset of phase 4. Nine normal healthy subjects, 30 to 65 years of age, inspired room air from residual volume (to mimic the maneuver of the standard single breath N2 (SBN2) washout test) to total lung capacity. During the expiration (flow constant at 250 ml.s-1) following a 10-s breath hold at total lung capacity, the exhaled gas was analyzed with a mass spectrometer for fractions of O2, CO2, and N2. Although the onset of phase 4 can be shown as the change in concentration of any of the three alveolar resident gases, oxygen was selected because (1) it demonstrates a greater apex to base concentration gradient than that found with CO2 and N2, and (2) a clear identification of the onset of phase 4 (minimum value of O2 fraction). With this method, the mean +/- SEM of CV was 16.8 +/- 1.52 percent (CV x 100/VC). No significant difference was found among the room air method, SBN2 method, and the helium bolus technique.

Intercomparison of calibration procedures for 192Ir HDR sources in Brazil.

The lack of well established dosimetry protocols for HDR sources is a point of great concern regarding the uniformity of procedures within a particular country and worldwide. The main objective of this paper is to report the results from ten institutions of an intercomparison of calibration procedures for 192Ir HDR sources currently in use in Brazil. The treatment irradiator of one institution was calibrated by a reference system and used by all participants with their own measuring electrometers and ionization chambers under the same experimental conditions. Two methods were used: the calibration jig and the well-type ionization chamber. Each participant was allowed to use their own method and formalism. The results of this exercise were very positive since this was the first time in Brazil that a group of users gathered to share their experience and openly discuss the physical concepts behind the calibration procedures. The results were all within +/-3.0%, except one case where -4.6% was observed and later identified as a problem with the Nk value for x-rays. Though the magnitude of the deviations found was generally acceptable considering the diversity of formalisms currently in use, a proposal is now being prepared to be adopted as a national protocol. The identification of the institutions was left out for the sake of confidentiality.

Orbital hydatid cyst: treatment and prevention of recurrences with albendazole plus praziquantel.

A case of successful treatment of orbital echinococcosis without evidence of recurrence on prolonged follow-up is presented. The management of orbital hydatid cyst is discussed. ¿ 2000 The British Infection Society

A paradigm for computer-assisted radiology resident rotation scheduling.

RATIONALE AND OBJECTIVES: The scheduling of radiology residents remains a major annual undertaking of chief residents. In this article, we describe a paradigm to implement interactive computer programs to reduce the inefficiencies and inequities of planning the yearly schedule. METHODS: We used the programming language, Prolog, to develop a compact program that provides faster and more flexible performance than those reported in the literature. This interactive program stores scheduling requirements in data files separated from the control program and runs on a Macintosh computer. RESULTS: The schedule of any residency year is generated within 3-7 sec. The fast computation and query capabilities of this scheduling program have helped chief residents to identify conflicting requirements that were previously overlooked. CONCLUSION: Using our programming paradigm, we have developed a portable Prolog-based scheduling program that is quick and easy to use.