Deciphering unusual uncultured magnetotactic multicellular prokaryotes through genomics

Candidatus Magnetoglobus multicellularis (Ca. M. multicellularis) is a member of a group of uncultured magnetotactic prokaryotes that possesses a unique multicellular morphology. To better understand this organism''s physiology, we used a genomic approach through pyrosequencing. Genomic data analysis corroborates previous structural studies and reveals the proteins that are likely involved in multicellular morphogenesis of this microorganism. Interestingly, some detected protein sequences that might be involved in cell adhesion are homologues to phylogenetically unrelated filamentous multicellular bacteria proteins, suggesting their contribution in the early development of multicellular organization in Bacteria. Genes related to the behavior of Ca. M. multicellularis (chemo-, photo- and magnetotaxis) and its metabolic capabilities were analyzed. On the basis of the genomic–physiologic information, enrichment media were tested. One medium supported chemoorganoheterotrophic growth of Ca. M. multicellularis and allowed the microorganisms to maintain their multicellular morphology and cell cycle, confirming for the first time that the entire life cycle of the MMP occurs in a multicellular form. Because Ca. M. multicellularis has a unique multicellular life style, its cultivation is an important achievement for further studies regarding the multicellular evolution in prokaryotes.     

Influence of environmental variables in the efficiency of phage therapy in aquaculture

Aquaculture facilities worldwide continue to experience significant economic losses because of disease caused by pathogenic bacteria, including multidrug-resistant strains. This scenario drives the search for alternative methods to inactivate pathogenic bacteria. Phage therapy is currently considered as a viable alternative to antibiotics for inactivation of bacterial pathogens in aquaculture systems. While phage therapy appears to represent a useful and flexible tool for microbiological decontamination of aquaculture effluents, the effect of physical and chemical properties of culture waters on the efficiency of this technology has never been reported. The present study aimed to evaluate the effect of physical and chemical properties of aquaculture waters (e.g. pH, temperature, salinity and organic matter content) on the efficiency of phage therapy under controlled experimental conditions in order to provide a basis for the selection of the most suitable protocol for subsequent experiments. A bioluminescent genetically transformed Escherichia coli was selected as a model microorganism to monitor real-time phage therapy kinetics through the measurement of bioluminescence, thus avoiding the laborious and time-consuming conventional method of counting colony-forming units (CFU). For all experiments, a bacterial concentration of ≈ 10⁵ CFU ml⁻¹ and a phage concentration of ≈ 10^6–8 plaque forming unit ml⁻¹ were used. Phage survival was not significantly affected by the natural variability of pH (6.5–7.4), temperature (10–25°C), salinity (0–30 g NaCl l⁻¹) and organic matter concentration of aquaculture waters in a temperate climate. Nonetheless, the efficiency of phage therapy was mostly affected by the variation of salinity and organic matter content. As the effectiveness of phage therapy increases with water salt content, this approach appears to be a suitable choice for marine aquaculture systems. The success of phage therapy may also be enhanced in non-marine systems through the addition of salt, whenever this option is feasible and does not affect the survival of aquatic species being cultured.     

Dose‐Dependent Effects and Reversibility of the Injuries Caused by Nandrolone Decanoate in Uterine Tissue and Fertility of Rats

This study is the first to investigate the effects of different doses of nandrolone decanoate (ND) upon uterine tissue and fertility, and if the reproductive alterations can be restored after cessation of the treatment. Wistar female rats were treated with ND at doses of 1.87, 3.75, 7.5, and 15 mg/kg body weight, diluted in vehicle (n = 30/group), or received only mineral oil (control group, n = 45). The animals were divided into three periods of study: ND‐treated receiving a daily subcutaneous injection for 15 consecutive days (1), and treatment with ND followed by 30‐day recovery (2), and 60‐day recovery (3). At the end of each period, five females per group were induced to death to histopathological analysis and the others were allowed to fertility evaluation (at 19th gestational day). Animals that received ND followed by 30‐day recovery exhibited persistent diestrous and marked suppression of reproductive capacity. Conversely, after 60‐day recovery, only lowest doses females (1.87 and 3.75 mg/kg) exhibited restoration of normal estrous cyclicity. Uterine weights were increased after ND treatment similarly to that of the controls after 60‐day recovery. The ND‐treated groups showed histopathological changes in the endometrium, myometrium, and perimetrium, and an increase in the thickness of both muscular and serous layers. Notably, the recovery of uterine tissue after ND treatment was dose‐ and period‐dependent. We reported that administration of ND promoted damage in uterine tissue and fertility of rats, and the recovery periods were insufficient to restore all of the side effects caused by ND under a dose‐dependent response     

Education and WHO Recommendations for Fruit and Vegetable Intake Are Associated with Better Cognitive Function in a Disadvantaged Brazilian Elderly Population: A Population-Based Cross-Sectional Study

Brazil has one of the fastest aging populations in the world and the incidence of cognitive impairment in the elderly is expected to increase exponentially. We examined the association between cognitive impairment and fruit and vegetable intake and associated factors in a low-income elderly population. A cross-sectional population-based study was carried out with 1849 individuals aged 65 or over living in São Paulo, Brazil. Cognitive function was assessed using the Community Screening Instrument for Dementia (CSI-D). Fruit and vegetable intake was assessed with a Food Frequency Questionnaire (FFQ) and categorized into quartiles of intake and into total daily fruit and vegetable intake using the cut-off points for the WHO recommendations (<400grams/day or ≥400 grams/day). The association between cognitive impairment and each quartile of intake, and WHO recommendation levels, was evaluated in two separate multivariate logistic models. The WHO recommendations for daily intakes ≥400 grams/day were significantly associated with 47% decreased prevalence of cognitive impairment. An effect modification was found in both models between cognitive impairment and “years of education and physical activity” and “years of education and blood levels of HDL” So that, having 1 or more years of education and being physically active or having 1 or more years of education and levels higher than 50 mg/dl of HDL-cholesterol strongly decreased the prevalence of cognitive impairment. In this socially deprived population with very low levels of education and physical activity and fruit and vegetable intake, those who attained WHO recommendations, had 1 year or more of education and were physically active had a significantly lower prevalence of cognitive impairment. A more comprehensive understanding of the social determinants of mental health is needed to develop effective public policies in developing countries.     

Improving the Manchester Triage System for Pediatric Emergency Care: An International Multicenter Study

Objectives

This multicenter study examines the performance of the Manchester Triage System (MTS) after changing discriminators, and with the addition use of abnormal vital sign in patients presenting to pediatric emergency departments (EDs).

Design

International multicenter study

Settings

EDs of two hospitals in The Netherlands (2006–2009), one in Portugal (November–December 2010), and one in UK (June–November 2010).

Patients

Children (<16years) triaged with the MTS who presented at the ED.

Methods

Changes to discriminators (MTS 1) and the value of including abnormal vital signs (MTS 2) were studied to test if this would decrease the number of incorrect assignment. Admission to hospital using the new MTS was compared with those in the original MTS. Likelihood ratios, diagnostic odds ratios (DORs), and c-statistics were calculated as measures for performance and compared with the original MTS. To calculate likelihood ratios and DORs, the MTS had to be dichotomized in low urgent and high urgent.

Results

60,375 patients were included, of whom 13% were admitted. When MTS 1 was used, admission to hospital increased from 25% to 29% for MTS ‘very urgent’ patients and remained similar in lower MTS urgency levels. The diagnostic odds ratio improved from 4.8 (95%CI 4.5–5.1) to 6.2 (95%CI 5.9–6.6) and the c-statistic remained 0.74. MTS 2 did not improve the performance of the MTS.

Conclusions

MTS 1 performed slightly better than the original MTS. The use of vital signs (MTS 2) did not improve the MTS performance.     

Classical and epigenetic approaches to metabolite diversification in filamentous fungi

Studies on fungal metabolites have produced an overwhelming expectation concerning the production of novel bioactive compounds for pharmaceutical applications. The adding of various biosynthetic precursors and the changing of nutritional components in the fermentation medium can change biosynthesis pathways, also leading to the production of novel metabolites. In addition, several growing conditions can be classically manipulated to modify fungal metabolite profiles. Recently, modern genome sequence tools have shown that not all gene clusters are regularly expressed in conventional growing conditions, thus expanding the possibilities of modulating the chemical metabolite profiles produced by filamentous fungi. This review discusses and exemplifies classical and epigenetic tools successfully applied to diversify metabolite production and to produce fungal metabolites from silent metabolic pathways.     

Anti-inflammatory effects of inosine in allergic lung inflammation in mice: evidence for the participation of adenosine A2A and A3 receptors

Inosine, a naturally occurring purine formed from the breakdown of adenosine, is associated with immunoregulatory effects. Evidence shows that inosine modulates lung inflammation and regulates cytokine generation. However, its role in controlling allergen-induced lung inflammation has yet to be identified. In this study, we aimed to investigate the role of inosine and adenosine receptors in a murine model of lung allergy induced by ovalbumin (OVA). Intraperitoneal administration of inosine (0.001–10 mg/kg, 30 min before OVA challenge) significantly reduced the number of leukocytes, macrophages, lymphocytes and eosinophils recovered in the bronchoalveolar lavage fluid of sensitized mice compared with controls. Interestingly, our results showed that pre-treatment with the selective A_2A receptor antagonist (ZM241385), but not with the selective A_2B receptor antagonist (alloxazine), reduced the inhibitory effects of inosine against macrophage count, suggesting that A_2A receptors mediate monocyte recruitment into the lungs. In addition, the pre-treatment of mice with selective A₃ antagonist (MRS3777) also prevented inosine effects against macrophages, lymphocytes and eosinophils. Histological analysis confirmed the effects of inosine and A_2A adenosine receptors on cell recruitment and demonstrated that the treatment with ZM241385 and alloxazine reverted inosine effects against mast cell migration into the lungs. Accordingly, the treatment with inosine reduced lung elastance, an effect related to A₂ receptors. Moreover, inosine reduced the levels of Th₂-cytokines, interleukin-4 and interleukin-5, an effect that was not reversed by A_2A or A_2B selective antagonists. Our data show that inosine acting on A_2A or A₃ adenosine receptors can regulate OVA-induced allergic lung inflammation and also implicate inosine as an endogenous modulator of inflammatory processes observed in the lungs of asthmatic patients.     

Microdeletion 11q13.1.q13.2 in a patient presenting with developmental delay,facial dysmorphism,and esophageal atresia: Possible role of the GSTP1 gene in esophagus malformation

BACKGROUND: Esophageal atresia is a major congenital malformation characterized by a complete interruption of the esophageal continuity. It is frequently observed in associations and syndromes. As an isolated finding, it has a multifactorial etiology whose genetic factors are poorly known. Recently, the GST family, especially the GSTM1 null genotype (but not the GSTP1 polymorphism I105V), has been associated with esophageal atresia. These enzymes play a role in phase II detoxification of xenobiotics. Here we present the clinical and molecular findings observed in a patient suggesting that the loss of the GSTP1 allele might predispose to this malformation. CASE: We describe a patient presenting with esophageal atresia associated with developmental delay and facial dysmorphism, whose mother used tobacco and alcohol during the first 2 months of her pregnancy. Microdeletion/microduplication analysis was performed using comparative genomic hybridization and a 180K Agilent array. It detected a de novo 2 Mb chromosome 11q13.1.q13.2 deletion. CONCLUSION: The deleted chromosomal segment includes the GSTP1 gene. We hypothesize that the deletion of one GSTP1 allele (an isoform highly expressed in embryonic tissues), associated with specific environmental factors, such as tobacco and alcohol, could cause the esophageal atresia observed in our patient. Birth Defects Research (Part A) 97:463–466, 2013. © 2013 Wiley Periodicals, Inc.