Bacillus subtilis biofilm matrix components target seed oil bodies to promote growth and anti-fungal resistance in melon.

Beneficial microorganisms are used to stimulate the germination of seeds; however, their growth-promoting mechanisms remain largely unexplored. Bacillus subtilis is commonly found in association with different plant organs, providing protection against pathogens or stimulating plant growth. We report that application of B. subtilis to melon seeds results in genetic and physiological responses in seeds that alter the metabolic and developmental status in 5-d and 1-month-old plants upon germination. We analysed mutants in different components of the extracellular matrix of B. subtilis biofilms in interaction with seeds and found cooperation in bacterial colonization of seed storage tissues and growth promotion. Combining confocal microscopy with fluorogenic probes, we found that two specific components of the extracellular matrix, amyloid protein TasA and fengycin, differentially increased the concentrations of reactive oxygen species inside seeds. Further, using electron and fluorescence microscopy and metabolomics, we showed that both TasA and fengycin targeted the oil bodies in the seed endosperm, resulting in specific changes in lipid metabolism and accumulation of glutathione-related molecules. In turn, this results in two different plant growth developmental programmes: TasA and fengycin stimulate the development of radicles, and fengycin alone stimulate the growth of adult plants and resistance in the phylloplane to the fungus Botrytis cinerea. Understanding mechanisms of bacterial growth promotion will enable the design of bespoke growth promotion strains.

An Alternative and Conserved Cell Wall Enzyme That Can Substitute for the Lipid II Synthase MurG.

The cell wall is a stress-bearing structure and a unifying trait in bacteria. Without exception, synthesis of the cell wall involves formation of the precursor molecule lipid II by the activity of the essential biosynthetic enzyme MurG, which is encoded in the division and cell wall synthesis (dcw) gene cluster. Here, we present the discovery of a cell wall enzyme that can substitute for MurG. A mutant of Kitasatospora viridifaciens lacking a significant part of the dcw cluster, including murG, surprisingly produced lipid II and wild-type peptidoglycan. Genomic analysis identified a distant murG homologue, which encodes a putative enzyme that shares only around 31% amino acid sequence identity with MurG. We show that this enzyme can replace the canonical MurG, and we therefore designated it MglA. Orthologues of mglA are present in 38% of all genomes of Kitasatospora and members of the sister genus Streptomyces CRISPR interference experiments showed that K. viridifaciens mglA can also functionally replace murG in Streptomyces coelicolor, thus validating its bioactivity and demonstrating that it is active in multiple genera. All together, these results identify MglA as a bona fide lipid II synthase, thus demonstrating plasticity in cell wall synthesis.IMPORTANCE Almost all bacteria are surrounded by a cell wall, which protects cells from environmental harm. Formation of the cell wall requires the precursor molecule lipid II, which in bacteria is universally synthesized by the conserved and essential lipid II synthase MurG. We here exploit the unique ability of an actinobacterial strain capable of growing with or without its cell wall to discover an alternative lipid II synthase, MglA. Although this enzyme bears only weak sequence similarity to MurG, it can functionally replace MurG and can even do so in organisms that naturally have only a canonical MurG. The observation that MglA proteins are found in many actinobacteria highlights the plasticity in cell wall synthesis in these bacteria and demonstrates that important new cell wall biosynthetic enzymes remain to be discovered.

Functional neuroanatomy of interoceptive processing in children and adolescents: a pilot study.

In adults, interoception - the sense of the physiological condition of the body - appears to influence emotion processing, cognition, behavior and various somatic and mental health disorders. Adults demonstrate frontal-insula-parietal-anterior cingulate cortex activation during the heartbeat detection task, a common interoceptive measure. Little, however, is known about the functional neuroanatomy underlying interoception in children. The current pilot study examined interoceptive processing in children and adolescents with fMRI while using the heartbeat detection task. Our main findings demonstrate that children as young as the age of six activate the left insula, cuneus, inferior parietal lobule and prefrontal regions. These findings are similar to those in adults when comparing heartbeat and tone detection conditions. Age was associated with increased activation within the dACC, orbital frontal cortex and the mid-inferior frontal gyri. Thus, our pilot study may provide important information about the neurodevelopment of interoceptive processing abilities in children and a task for future interoception neuroimaging studies in children.

Critical congenital heart disease screening by pulse oximetry in a neonatal intensive care unit.

OBJECTIVE: Critical congenital heart disease (CCHD) screening is effective in asymptomatic late preterm and term newborn infants with a low false-positive rate (0.035%). (1) To compare 2817 neonatal intensive care unit (NICU) discharges before and after implementation of CCHD screening; and (2) to evaluate CCHD screening at <35 weeks gestation. STUDY DESIGN: Collection of results of CCHD screening including pre- and postductal pulse oximetry oxygen saturation (SpO2) values. RESULT: During the pre-CCHD screen period, 1247 infants were discharged from the NICU and one case of CCHD was missed. After 1 March 2012, 1508 CCHD screens were performed among 1570 discharges and no CCHDs were missed. The pre- and postductal SpO2 values were 98.8 ± 1.4% and 99 ± 1.3%, respectively, in preterm and 98.9 ± 1.3% and 98.9 ± 1.4%, respectively, in term infants. Ten infants had false-positive screens (10/1508 = 0.66%). CONCLUSION: Performing universal screening in the NICU is feasible but is associated with a higher false-positive rate compared with asymptomatic newborn infants.

Is low docosahexaenoic acid associated with disturbed rhythms and neurodevelopment in offsprings of diabetic mothers?

BACKGROUND/OBJECTIVE: To evaluate the relation between docosahexaenoic acid (DHA) status and neurodevelopment in the offsprings of gestational diabetic mothers (ODMs). SUBJECTS/METHODS: A prospective cohort study was performed. The offspring of 63 pregnant women (23 controls, 21 diet-controlled gestational diabetes mellitus (GDM) and 19 insulin-treated GDM) were recruited. Maternal and venous cord plasma DHA percentages were analyzed. Skin temperature and activity in children were recorded for 72 h at 3 and 6 months of life. Neurodevelopment was assessed using the Bayley Scale of Infant Development II (BSID II) at 6 and 12 months of age. RESULTS: Cord plasma DHA percentage was significantly lower in the ODMs compared with that in the controls (Control 6.43 [5.04-7.82](a); GDM+diet 5.65 [4.44-6.86](ab); GDM+insulin 5.53 [4.45-6.61](b)). Both mental (Control 102.71 [97.61-107.81](a); GDM+diet 100.39 [91.43-109.35](a); GDM+insulin 93.94 [88.31-99.57](b)) and psychomotor (Control 91.52 [81.82-101.22](a); GDM+diet 81.67 [73.95-89.39](b); GDM+insulin 81.89 [71.96-91.85](b)) scores evaluated by the BSID II were significantly lower at 6 months in ODMs, even after adjusting for confounding factors such as breastfeeding, maternal educational level and gender. Cord plasma DHA percentage correlated with the psychomotor score from BSID II (r=0.27; P=0.049) and with the intra-daily variability in activity (r=-0.24; P=0.043) at 6 months. Maternal DHA was correlated with several sleep rhythm maturation parameters at 6 months. CONCLUSIONS: Lower DHA levels in cord plasma of ODMs could affect their neurodevelopment. Maternal DHA status was also associated with higher values in the sleep rhythm maturation parameters of children.

Use of CT angiography in the diagnosis of total anomalous venous return.

Total anomalous pulmonary venous return (TAPVR) is an uncommon cause of cyanotic congenital heart disease in the neonatal period. This diagnosis is often made by echocardiography. We present two cases where echocardiography was not confirmatory. Computerized tomography (CT) scan of the chest and abdomen with contrast was performed instead of cardiac catheterization to establish the diagnosis. We suggest that CT with contrast is a noninvasive modality to obtain anatomic details of pulmonary venous drainage in TAPVR when echocardiography is inconclusive.

Hábito tabáquico en médicos de las provincias de Talca y Linares / Smoking in physicians of the provinces of Talca and Linares

Objetivos: conocer aspectos descriptivos epidemiológicos del hábito tabáquico en médicos de las provincias de Talca y Linares. Método: Descriptivo transversal, aplicando como instrumento una encuesta a un N de 122 médicos cuya residencia o lugar de trabajo se encontraban en las provincias de Talca y Linares. Resultados: De la muestra compuesta por 42 mujeres y 80 hombres, en su mayoría médicos generales, un 36 por ciento se declaró actualmente fumador y un 64 por ciento no fumador. La prevalencia histórica del hábito en la muestra estudiada, es decir, la suma de los médicos que actualmente fuman y los que antes fumaban resultó ser del 59 por ciento. La percepción de los fumadores de que el hábito es dañino para su salud es de un 95 por ciento y han intentado suspender el hábito un 72 por ciento de ellos. De los 78 no fumadores, 28 (36 por ciento) eran fumadores y la razón más frecuente para suspender el hábito fue la salud.

Guía para la prevención de úlceras en pacientes encamados / No disponible

Se presenta la elaboración que han efectuado los autores de una rueda para prevenir úlceras en pacientes encamados. Diseñada en un formato práctico y de fácil comprensión para todos aquellos cuidadores principales que están a cargo de algún paciente encamado, es un instrumento de gran utilidad para enfermería

The authors present a report they produced dealing with how toprevent decubitus ulcers in bedridden patients. This report, designed in a practical use format, can be easily understood by all those nursing care professionals who have to care for bedridden patients; it is a highly useful tool for the nursing field

Attenuation of frontal asymmetry in pediatric posttraumatic stress disorder.

BACKGROUND: Volumetric imaging research has shown abnormal brain morphology in adults with posttraumatic stress disorder (PTSD) when compared with matched control subjects. In this article, we present brain imaging findings from a study of children with PTSD symptoms. METHODS: Twenty-four children between the ages of 7 and 14 with a history of trauma and PTSD symptoms were assessed with the Clinician-Administered PTSD Scale for Children and Adolescents (CAPS-CA). The sample underwent magnetic resonance imaging in a 1.5 T scanner. Brain images were analyzed by raters blind to diagnostic status using well-standardized methods, and images were compared with age- and gender-matched healthy control subjects. RESULTS: The clinical group demonstrated attenuation of frontal lobe asymmetry and smaller total brain and cerebral volumes when compared with the control group. There were no statistically significant differences in hippocampal volume between clinical and control subjects. CONCLUSIONS: Frontal lobe abnormalities may occur as a result of PTSD in children or, alternatively, be a risk factor for the development of the syndrome in this age group. The implications of the findings and their consistency with previous research are discussed.

Estudio ecuatoriano abierto multicÚntrico con etofibrato en el tratamiento de las dislipidemias / Multicentric opened Ecuadorian study with etofibrato in the treatment of Dislipidemias

Se realizó un estudio abierto multicÚntrico en 75 pacientes que presentaban dislipidemias con colesterol o triglicÚridos elevados, asÝ como pacientes con dislipidemias mixtas (de acuerdo a la nueva clasificación europea de las dislipidemias) fueron incluidos en el estudio pacientes que presentaban por lo menos un factor de riesgo cardiovascular (tabaquismo, diabetes, sedentarismo, hipertensión). Para evaluar la eficacia y tolerancia de etofibrato retard, fueron administrados 400 mg al dÝa previo un perÝodo de dieta de 4 a 6 semanas. Al finalizar el tratamiento se observó una reducción promedio del 17 por ciento en los valores de colesterol, de un 41 por ciento en los triglicÚridos y un aumento de un 16 por ciento de las HDL...

Bacterial toxins and enteral feeding of premature infants at risk for necrotizing enterocolitis.

Concordance between gram-negative enteric and other toxin-producing bacteria in blood and stool culture, endotoxin (lipopolysaccharide), and interleukin 6 (IL-6) was measured in 60 preterm infants (600-1600 g) as a clinical index in neonatal necrotizing enterocolitis (NEC). Escherichia coli, Klebsiella, Enterobacter, and Clostridium spp, identified by routine bacteriology, were each strongly associated with elevated concentrations of endotoxin (P < 0.01) in stool filtrates with Clostridium spp most strongly associated with NEC disease. Stool filtrate endotoxin (endotoxin units [EU] per gram) measured by a Limulus amebocyte lysate assay was age-dependent. Samples from stage I NEC (61%) and infants with advanced disease (67%) had notably elevated levels of stool endotoxin (>10 ln EU/g) compared with NEC-negative (47%) samples tested. Plasma and stool IL-6 generally tested at the low, nonmeasurable limit of the enzyme-linked immunosorbent assay (ELISA) for NEC-negative (88%) and stage I NEC (93%), although a small proportion of samples (25%) from infants with stage II or II NEC had elevated stool concentrations of IL-6. We conclude that identification of toxin-producing organisms and endotoxin elevations in stool filtrates are more useful than circulating levels of endotoxin in plasma in predicting mucosally limited disease in the gastrointestinal tract. The prognostic value of monitoring stool endotoxin in infants with overgrowth of gram-negative bacteria has implications for therapeutic strategies for patients with early and advanced stages of disease. Monitoring inflammatory cytokines (IL-6) in relation to endotoxin values in stool appears of limited clinical value in controlling this devastating disease in preterm neonates.

Trauma and dissociation in delinquent adolescents.

OBJECTIVES: To assess history of trauma and dissociation in a group of juvenile delinquents and to assess how adolescents would respond to a structured interview for dissociative symptoms. METHOD: Sixty-four adolescents in juvenile probation hall participated in 2 investigational sessions in 1996-1997. For session 1 they answered the Childhood Trauma Questionnaire (CTQ), the Response Evaluation Measure for Youth-71 (REMY-71), and the Weinberger Adjustment Inventory. For session 2 they were given the Childhood Trauma Interview (CTI) and the Structured Clinical Interview for DSM-IV Dissociative Disorders (SCID-D). RESULTS: In this sample 28.3% met criteria for a dissociative disorder and 96.8% endorsed a history of traumatic events. There were significant positive correlations between CTI and CTQ trauma scores and SCID-D and REMY-71 dissociative symptoms. All dissociative symptoms were endorsed, but depersonalization was the most common experience. There was a lack of congruence between the different methods of assessing dissociation. CONCLUSIONS: This study provides support for an early link between history of trauma and dissociation. Adolescents were able to answer questions from a structured interview assessing dissociation.

Concordance of bacterial cultures with endotoxin and interleukin-6 in necrotizing enterocolitis.

Concordance between gram-negative enteric and other toxin-producing bacteria in blood and stool culture, endotoxin (lipopolysaccharide), and interleukin-6 (IL-6) was measured in 60 preterm infants (600-1600 g) as a clinical index in neonatal necrotizing enterocolitis (NEC). E. coli, Klebsiella, Enterobacter, and Clostridium spp., identified by routine bacteriology, were each strongly associated with elevated concentrations of endotoxin (P < 0.01) in stool filtrates, with Clostridium spp. most strongly associated with NEC disease. Stool filtrate endotoxin (EU/g) measured by a Limulus amebocyte lysate assay was age dependent. Samples from stage I NEC (61%) and infants with advanced disease (67%) had notably elevated levels of stool endotoxin (> 10 ln EU/g) compared to NEC-negative (47%) samples tested. Plasma and stool IL-6 generally tested at the low, nonmeasurable limit of the ELISA for NEC-negative (88%) and stage I NEC (93%), although a small proportion of samples (25%) from infants with stage II or III NEC had elevated stool concentrations of IL-6. We conclude that identification of toxin-producing organisms and endotoxin elevations in stool filtrates are more useful than circulating levels of endotoxin in plasma in predicting mucosally limited disease in the gastrointestinal tract. The prognostic value of monitoring stool endotoxin in infants with overgrowth of gram-negative bacteria has implications for therapeutic strategies in patients with early and advanced stages of disease. Monitoring inflammatory cytokines (IL-6) in relation to endotoxin values in stool appears of limited clinical value in controlling this devastating disease in preterm neonates.

Effect of methylene blue on refractory neonatal hypotension.

Excess nitric oxide is a mediator of the hypotension in septic shock. Nitric oxide dilates vascular smooth muscle through activation of soluble guanylate cyclase. We report the increase in blood pressure caused by methylene blue (MB), a soluble guanylate cyclase inhibitor, in five neonates with presumed septic shock unresponsive to colloids, inotropic agents, and corticosteroids. MB was given intravenously at a dose of 1 mg/kg during a 1-hour period. MB increased blood pressure in each patient (average, 33% +/- 20%). Blood pressure subsequently decreased to near baseline values in three patients, who then received a second infusion of MB. Blood pressure again increased in these patients. Three of five patients were weaned from inotropic support within 72 hours. Three of five patients survived and were discharged home. We suggest that MB increased blood pressure in these neonates with refractory hypotension.

A multicenter randomized, masked comparison trial of natural versus synthetic surfactant for the treatment of respiratory distress syndrome.

OBJECTIVE: To compare the efficacy and safety of two surfactant preparations in the treatment of respiratory distress syndrome (RDS). METHODS: We conducted a randomized, masked comparison trial at 21 centers. Infants with RDS who were undergoing mechanical ventilation were eligible for treatment with two doses of either a synthetic (Exosurf) or natural (Infasurf) surfactant if the ratio of arterial to alveolar partial pressure of oxygen was less than or equal to 0.22. Crossover treatment was allowed within 96 hours of age if severe respiratory failure (defined as two consecutive arterial/alveolar oxygen tension ratios < or = 0.10) persisted after two doses of the randomly assigned surfactant. Four primary outcome measures of efficacy (the incidence of pulmonary air leak (< or = 7 days); the severity of RDS; the incidence of death from RDS; and the incidence of survival without bronchopulmonary dysplasia (BPD) at 28 days after birth) were compared by means of linear regression techniques. RESULTS: The primary analysis of efficacy was performed in 1033 eligible infants and an analysis of safety outcomes in the 1126 infants who received study surfactant. Preentry demographic characteristics and respiratory status were similar for the two treatment groups, except for a small but significant difference in mean gestational age (0.5 week) that favored the infasurf treatment group. Pulmonary air leak (< or = 7 days) occurred in 21% of Exosurf- and 11% of infasurf-treated infants (adjusted relative risk, 0.53; 95% confidence interval, 0.40 to 0.71; p < or = 0.0001). During the 72 hours after the initial surfactant treatment, the average fraction of inspired oxygen (+/-SEM) was 0.47 +/- 0.01 for Exosurf- and 0.39 +/- 0.01 for infasurf-treated infants (difference, 0.08; 95% confidence interval, 0.06 to 0.10; p < 0.0001); the average mean airway pressure (+/-SEM) was 8.6 +/- 0.1 cm H2O; for Exosurf- and 7.2 +/- 0.1 cm H2O for Infasurf-treated infants (difference, 1.4 cm H2O; 95% confidence interval, 1.0 to 1.8 cm H2O; p < 0.0001). The incidences of RDS-related death, total respiratory death, death to discharge, and survival without bronchopulmonary dysplasia at 28 days after birth did not differ. The number of days of more than 30% inspired oxygen and of assisted ventilation, but not the duration of hospitalization, were significantly lower in Infasurf-treated infants. CONCLUSION: Compared with Exosurf, Infasurf provided more effective therapy for RDS as assessed by significant reductions in the severity of respiratory disease and in the incidence of air leak complications.

Roadblocks to do-not-resuscitate orders. A study in policy implementation.

BACKGROUND: Cardiopulmonary resuscitation, a potentially lifesaving procedure, is initiated on hospitalized patients who have an arrest in the absence of a written do-not-resuscitate (DNR) order. New York State Law specifies that attending physicians may write a DNR order on an adult patient either with his/her consent or that of a surrogate. Under specified circumstances, concurring physician and witness signatures are also required. This study examines potential obstacles physicians may encounter when implementing a DNR order for a hospitalized patient. METHODS: Sixty house staff officers and 45 attending physicians at two New York City medical centers responded to a questionnaire listing 18 potential problems in obtaining a DNR order. Using a Likert scale, respondents rated the prevalence of each problem. RESULTS: Analysis of the data indicates that attending physician's failure to discuss DNR issues with patients and situations involving surrogate decision making are considered major obstacles to obtaining a DNR order. Procedural regulations, including abundant paperwork and witnessed signatures, are not identified as major obstacles. CONCLUSIONS: This study suggests a need for improved communication among physicians, patients, and surrogates about advance directives, when feasible, either prior to hospitalization or early in its course, in an effort to comply with DNR legislation in a manner that reflects the patient's wishes and best interests.

Prevention of neonatal necrotizing enterocolitis.

Small premature infants are often hypochlorhydric, and frequently their stomachs are colonized by enteric, gram-negative bacteria. We tested a hypothesis that gastric pH affected the colonization of the stomach with enteric bacteria and that this colonization was causally related to the risk or severity of necrotizing enterocolitis. A prospective, double-blind study was conducted that compared a group of infants supplemented with 0.01-0.02 ml of 1 N HCl/ml of milk to a group with a similar supplement of water. Gastric pH, gastric enteric bacteria counts, and the incidence and severity of necrotizing enterocolitis were monitored. The median gastric pH of the HCl-supplemented group was lower (3.0) than controls (4.0) throughout the study (p less than 0.001). The gastric enteric bacterial colonization rate and the quantitative bacterial counts were strongly correlated with gastric pH over 4 (p less than 0.001). Somatic growth rates in infants in the HCl-supplemented group were equal to, or exceeded, those in the control group. There was 1 case of necrotizing enterocolitis among the 34 infants in the HCl-supplemented group and 8 cases among the 34 in the control group (p = 0.02). It appears that acidifying the feedings of small premature infants to a pH low enough to inhibit bacterial proliferation in the stomach significantly lowers the risk of necrotizing enterocolitis.