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Background: There is a paucity of literature regarding the microbial etiology of community-acquired pneumonia (CAP) in India. The current study was aimed to study the microbial etiology of hospitalized adults with CAP. Methods: The study was conducted in a 700-bedded North Indian hospital. Consecutive adults admitted with CAP over a period of 2 years from 2013 to 2015 were recruited for the study, and apart from clinical evaluation underwent various microbiological studies in the form of blood culture, sputum culture, urinary antigen for pneumococcus and Legionella, serology for Mycoplasma and Chlamydia and real-time reverse transcriptase polymerase chain reaction for influenza viruses. Radiographic studies were performed in all patients and repeated as required. The patients were treated with standard antibiotic/antiviral therapy and outcomes were recorded. Results: A total of 225 patients (median age: 59 years) were enrolled. Streptococcus pneumoniae was the most common organism found (30.5%), followed by Legionella pneumophila (17.5%), influenza viruses (15.4%), Mycoplasma pneumoniae (7.2%), Chlamydia pneumonia (5.5%), Mycobacterium tuberculosis (4.8%), Klebsiella pneumoniae (4.8%), methicillin-resistant Staphylococcus aureus (3.5%), Pseudomonas aeruginosa (3.1%), methicillin-sensitive S. aureus (1.7%), and Acinetobacter sp. (0.8%) with 4% of patients having multiple pathogens etiologies. High Pneumonia Severity Index score correlated with the severity and outcome of the CAP but was not predictive of any definite etiological pathogen. In-hospital mortality was 8%. Conclusion: Streptococcus pneumoniae, Legionella, and influenza constitute the most common etiological agents for north Indian adults with CAP requiring hospitalization. Appropriate antibiotic therapy and preventive strategies such as influenza and pneumococcal vaccination need to be considered in appropriate groups.

Platelet thrombosis under arterial conditions remains a large clinical problem. Previous in vitro experiments have concentrated on early adherence without thrombotic occlusion. We have developed a controllable hemodynamic system that creates intravascular thrombosis to occlusion. Lightly heparinized (3.5 USP units/mL), whole, porcine blood is perfused through a 1.5 mm inner diameter, tubular, collagen-coated stenosis. The microscopic growth of thrombus is optically recorded using a high resolution CCD camera. Occlusive thrombus is examined using microcomputed tomography and histology. Thrombus consistently formed in the throat of the stenosis where wall shear rates were greatest. Rapid platelet accumulation (RPA) reached rates as high as 13.7 μm 3  μm −2  min −1 . Total occlusion of flow occurred after 17 ± 2.6 min ( n  = 6). The average thrombus volume accumulation of 7.8 ± 3.5 μm 3  μm 2  min −1 occurred under very high wall shear rates exceeding 100,000 s −1 . Significant volumes of thrombus did not form until 7.6 ± 3.6 min after the onset of flow, a delay consistent with activation of adherent mural platelets. Platelets did not accumulate with large volume for normal wall shear rates <2000 s −1 . Very high wall shear rates stimulate the capture of millions of circulating platelets with exposure times <2 ms in an arterial stenosis.

تدور الرواية حول حكاية غريق أمضى عشرة أيام عائما على متن زورق دون طعام أو شراب. ونصب بطلا قوميا، ثم تهاوت عليه قبلات ملكات الجمال ؛ فأصبح ثريا بفضل ما قام بتصويره من إعلانات. وعقب نشر تفاصيل الحكاية تفجرت الفضيحة ؛ حيث كان الفوز والتكريم والثروة من نصيب الغريق، أما الصحفي الذي أخذ على عاتقه جمع أطراف الحكاية، فقد كان مصيره النفي والتشريد.

Root-knot nematodes (RKNs) are obligate endoparasites that maintain a biotrophic relationship with their hosts over a period of several weeks and induce the differentiation of root cells into specialized feeding cells. Nematode effectors synthesized in the oesophageal glands and injected into the plant tissue through the syringe-like stylet certainly play a central role in these processes. In a search for nematode effectors, we used comparative genomics on expressed sequence tag (EST) datasets to identify Meloidogyne incognita genes encoding proteins potentially secreted upon the early steps of infection. We identified three genes specifically expressed in the oesophageal glands of parasitic juveniles that encode predicted secreted proteins. One of these genes, Mi-EFF1 is a pioneer gene that has no similarity in databases and a predicted nuclear localization signal. We demonstrate that RKNs secrete Mi-EFF1 within the feeding site and show Mi-EFF1 targeting to the nuclei of the feeding cells. RKNs were previously shown to secrete proteins in the apoplasm of infected tissues. Our results show that nematodes sedentarily established at the feeding site also deliver proteins within plant cells through their stylet. The protein Mi-EFF1 injected within the feeding cells is targeted at the nuclei where it may manipulate nuclear functions of the host cell.

Excellent visualization of nuclei was obtained here using a whole-mount procedure adapted to provide high-resolution images of large, irregularly shaped nuclei. The procedure is based on tissue clearing, and fluorescent staining of nuclear DNA with the dye propidium iodide. The method developed for standard confocal imaging was applied to large multicellular root swellings, named galls, induced in plant hosts by the root-knot nematode Meloidogyne incognita. Here, we performed a functional analysis, and examined the nuclear structure in giant feeding cells overexpressing the cell cycle inhibitor Kip-related protein 4 (KRP4). Ectopic KRP4 expression in galls led to aberrant nuclear structure, disturbing giant cell expansion and nematode reproduction. In vivo live-cell imaging of GFP-KRP4 demonstrated that this protein co-localizes to chromosomes from prophase to late anaphase during cell cycle progression. The data presented here suggest the involvement of KRP4 during mitotic progression in plant cells. The detailed results obtained using confocal analysis also demonstrate the potential utility of a rapid, easy-to-use clearing method for the analysis of the nuclei of certain Arabidopsis mutants and other complex plant nuclei.

NADPH-cytochrome P450 reductase (CPR) is the unique redox partner of microsomal cytochrome P450s (CYPs). CPR exists in a conformational equilibrium between open and closed conformations throughout its electron transfer (ET) function. Previously, we have shown that electrostatic and flexibility properties of the hinge segment of CPR are critical for ET. Three mutants of human CPR were studied (S243P, I245P and R246A) and combined with representative human drug-metabolizing CYPs (isoforms 1A2, 2A6 and 3A4). To probe the effect of these hinge mutations different experimental approaches were employed: CYP bioactivation capacity of pre-carcinogens, enzyme kinetic analysis, and effect of the ionic strength and cytochrome b(5) (CYB5) on CYP activity. The hinge mutations influenced the bioactivation of pre-carcinogens, which seemed CYP isoform and substrate dependent. The deviations of Michaelis-Menten kinetic parameters uncovered tend to confirm this discrepancy, which was confirmed by CYP and hinge mutant specific salt/activity profiles. CPR/CYB5 competition experiments indicated a less important role of affinity in CPR/CYP interaction. Overall, our data suggest that the highly flexible hinge of CPR is responsible for the existence of a conformational aggregate of different open CPR conformers enabling ET-interaction with structural varied redox partners.

Large and systematic mRNA library design disentangles the complex sequence determinants of translation efficiency in bacteria. Comparative analyses of natural and mutated sequences have been used to probe mechanisms of gene expression, but small sample sizes may produce biased outcomes. We applied an unbiased design-of-experiments approach to disentangle factors suspected to affect translation efficiency in E. coli. We precisely designed 244,000 DNA sequences implementing 56 replicates of a full factorial design to evaluate nucleotide, secondary structure, codon and amino acid properties in combination. For each sequence, we measured reporter transcript abundance and decay, polysome profiles, protein production and growth rates. Associations between designed sequences properties and these consequent phenotypes were dominated by secondary structures and their interactions within transcripts. We confirmed that transcript structure generally limits translation initiation and demonstrated its physiological cost using an epigenetic assay. Codon composition has a sizable impact on translatability, but only in comparatively rare elongation-limited transcripts. We propose a set of design principles to improve translation efficiency that would benefit from more accurate prediction of secondary structures in vivo .

Wildfire activity is expected to increase across the Mediterranean Basin because of climate change. However, the effects of future climate change on the combinations of atmospheric conditions that promote wildfire activity remain largely unknown. Using a fire-weather based classification of wildfires, we show that future climate scenarios point to an increase in the frequency of two heat-induced fire-weather types that have been related to the largest wildfires in recent years. Heat-induced fire-weather types are characterized by compound dry and warm conditions occurring during summer heatwaves, either under moderate (heatwave type) or intense (hot drought type) drought. The frequency of heat-induced fire-weather is projected to increase by 14% by the end of the century (2071-2100) under the RCP4.5 scenario, and by 30% under the RCP8.5, suggesting that the frequency and extent of large wildfires will increase throughout the Mediterranean Basin. Wildfire is a complex phenomenon that occurs when three conditions are met: available fuel, an ignition source (due to lightning or human activities) and weather conditions conducive to fires (fire weather) 1. Climate and weather are important drivers of wildfire activity across a range of timescales 2-5 , and, consequently, current and potential future climate-induced changes in wildfire activity might threaten ecosystems and human well-being 6. In most Euro-Mediterranean countries, wildfire activity has been declining owing to management and suppression measures undertaken since the 1980s 7. However, some recent extreme wildfire events, including those that occurred in 2016 in France 8 , 2017 in Spain and Portugal 9 and 2018 in Greece 10 have highlighted the limits of wildfire suppression capabilities under exceptional fire-weather conditions. Furthermore, studies show that wildfire activity is expected to increase across the Mediterranean Basin due to climate change 11,12. However, how the combinations of climate and weather conditions that promote the largest wildfires will respond to climate change remain largely unknown. Climate and weather are both drivers of wildfire activity. Soil moisture deficit over days to months increases fuel aridity and flammability 6,13 while a number of synoptic weather conditions associated to different combinations of short-term and instantaneous meteorological fields (precipitation, temperature, relative humidity and wind speed) influence wildfire behaviour 5,14. Most of the largest wildfires occur when these conditions are met 8. For instance, the combination of extreme drought with extreme wind or heatwaves have both been identified as crucial factors in the occurrence of crown wildfires in Mediterranean forests and shrublands 15-17. In this study, we focus on the frequency of current and future weather and climate conditions associated with wildfires in four countries (France, Portugal, Greece, Tunisia) of the Mediterranean basin covering most of its diverse biogeographic and climatic conditions (Supplementary Figs. S1-S3). Building on the insights gained open