The human gut chemical landscape predicts microbe-mediated biotransformation of foods and drugs.
Microbes are nature's chemists, capable of producing and metabolizing a diverse array of compounds. In the human gut, microbial biochemistry can be beneficial, for example vitamin production and complex carbohydrate breakdown; or detrimental, such as the reactivation of an inactive drug metabolite leading to patient toxicity. Identifying clinically relevant microbiome metabolism requires linking microbial biochemistry and ecology with patient outcomes. Here we present MicrobeFDT, a resource which clusters chemically similar drug and food compounds and links these compounds to microbial enzymes and known toxicities. We demonstrate that compound structural similarity can serve as a proxy for toxicity, enzyme sharing, and coarse-grained functional similarity. MicrobeFDT allows users to flexibly interrogate microbial metabolism, compounds of interest, and toxicity profiles to generate novel hypotheses of microbe-diet-drug-phenotype interactions that influence patient outcomes. We validate one such hypothesis experimentally, using MicrobeFDT to reveal unrecognized gut microbiome metabolism of the ovarian cancer drug altretamine.


A Comparison of Laparoscopic Redo Fundoplications for Failed Toupet and Nissen Fundoplications in Children.
We compared laparoscopic redo fundoplications performed for failed laparoscopic Toupet fundoplication (LTF) and failed laparoscopic Nissen fundoplications (LNFs). Redo LTF (R-LTF; n = 4) and redo LNF (R-LNF; n = 6) performed between 2007 and 2014 were assessed retrospectively for severity of intraperitoneal adhesions on a scale of 0-3, identification/preservation of the anterior/posterior/hepatic branches of the vagus nerve (VN), complications, and outcome. Redos were performed after a mean of 34 months in R-LTF and 32 months in R-LNF (P = ns) indicated for sliding hernia (n = 3; 2 with partial wrap dehiscence) and partial wrap dehiscence (n = 1) in R-LTF and sliding hernia (n = 6; 4 with partial wrap dehiscence) in R-LNF. The mean adhesion severity score was 1.5 in R-LTF and 2.5 in R-LNF (P < 0.05). The mean number of VN branches identified/preserved was 2.0 in R-LTF and 0.8 in R-LNF (P < 0.05). Mean operative times and mean blood loss were similar. Intraoperative complications were accidental local trauma (n = 1 in R-LTF and n = 3 in R-LNF, one requiring conversion to open repair) (P = ns). Gastric outlet obstruction developed in two R-LNF cases; both were managed conservatively. There have been no further recurrences to date. Although our series is small, adhesions were less, and identification/preservation of VN was easier during R-LTF.


Combination Therapy with Anti-PD-1, Anti-TIM-3, and Focal Radiation Results in Regression of Murine Gliomas.
Checkpoint molecules like programmed death-1 (PD-1) and T-cell immunoglobulin mucin-3 (TIM-3) are negative immune regulators that may be upregulated in the setting of glioblastoma multiforme. Combined PD-1 blockade and stereotactic radiosurgery (SRS) have been shown to improve antitumor immunity and produce long-term survivors in a murine glioma model. However, tumor-infiltrating lymphocytes (TIL) can express multiple checkpoints, and expression of ≥2 checkpoints corresponds to a more exhausted T-cell phenotype. We investigate TIM-3 expression in a glioma model and the antitumor efficacy of TIM-3 blockade alone and in combination with anti-PD-1 and SRS. C57BL/6 mice were implanted with murine glioma cell line GL261-luc2 and randomized into 8 treatment arms: (i) control, (ii) SRS, (iii) anti-PD-1 antibody, (iv) anti-TIM-3 antibody, (v) anti-PD-1 + SRS, (vi) anti-TIM-3 + SRS, (vii) anti-PD-1 + anti-TIM-3, and (viii) anti-PD-1 + anti-TIM-3 + SRS. Survival and immune activation were assessed. Dual therapy with anti-TIM-3 antibody + SRS or anti-TIM-3 + anti-PD-1 improved survival compared with anti-TIM-3 antibody alone. Triple therapy resulted in 100% overall survival (P < 0.05), a significant improvement compared with other arms. Long-term survivors demonstrated increased immune cell infiltration and activity and immune memory. Finally, positive staining for TIM-3 was detected in 7 of 8 human GBM samples. This is the first preclinical investigation on the effects of dual PD-1 and TIM-3 blockade with radiation. We also demonstrate the presence of TIM-3 in human glioblastoma multiforme and provide preclinical evidence for a novel treatment combination that can potentially result in long-term glioma survival and constitutes a novel immunotherapeutic strategy for the treatment of glioblastoma multiforme. Clin Cancer Res; 23(1); 124-36. ©2016 AACR.


Predictors of endoscopic third ventriculostomy ostomy status in patients who experience failure of endoscopic third ventriculostomy with choroid plexus cauterization.
At failure of endoscopic third ventriculostomy (ETV) with choroid plexus cauterization (CPC), the ETV ostomy may be found to be closed or open. Failure with a closed ostomy may indicate a population that could benefit from evolving techniques to keep the ostomy open and may be candidates for repeat ETV, whereas failure with an open ostomy may be due to persistently abnormal CSF dynamics. This study seeks to identify clinical and radiographic predictors of ostomy status at the time of ETV/CPC failure. The authors conducted a multicenter, retrospective cohort study on all pediatric patients with hydrocephalus who failed initial ETV/CPC treatment between January 2013 and October 2016. Failure was defined as the need for repeat ETV or ventriculoperitoneal (VP) shunt placement. Clinical and radiographic data were collected, and ETV ostomy status was determined endoscopically at the subsequent hydrocephalus procedure. Statistical analysis included the Mann-Whitney U-test, Wilcoxon rank-sum test, t-test, and Pearson chi-square test where appropriate, as well as multivariate logistic regression. Of 72 ETV/CPC failures, 28 patients (39%) had open-ostomy failure and 44 (61%) had closed-ostomy failure. Patients with open-ostomy failure were older (median 5.1 weeks corrected age for gestation [interquartile range (IQR) 0.9-15.9 weeks]) than patients with closed-ostomy failure (median 0.2 weeks [IQR -1.3 to 4.5 weeks]), a significant difference by univariate and multivariate regression. Etiologies of hydrocephalus included intraventricular hemorrhage of prematurity (32%), myelomeningocele (29%), congenital communicating (11%), aqueductal stenosis (11%), cyst/tumor (4%), and other causes (12%). A wider baseline third ventricle was associated with open-ostomy failure (median 15.0 mm [IQR 10.3-18.5 mm]) compared to closed-ostomy failure (median 11.7 mm [IQR 8.9-16.5 mm], p = 0.048). Finally, at the time of failure, patients with closed-ostomy failure had enlargement of their ventricles (frontal and occipital horn ratio [FOHR], failure vs baseline, median 0.06 [IQR 0.00-0.11]), while patients with open-ostomy failure had no change in ventricle size (median 0.01 [IQR -0.04 to 0.05], p = 0.018). Previous CSF temporizing procedures, intraoperative bleeding, and time to failure were not associated with ostomy status at ETV/CPC failure. Older corrected age for gestation, larger baseline third ventricle width, and no change in FOHR were associated with open-ostomy ETV/CPC failure. Future studies are warranted to further define and confirm features that may be predictive of ostomy status at the time of ETV/CPC failure.


Oxidative stress-mediated NFκB phosphorylation upregulates p62/SQSTM1 and promotes retinal pigmented epithelial cell survival through increased autophagy.
p62 is a scaffolding adaptor implicated in the clearance of protein aggregates by autophagy. Reactive oxygen species (ROS) can either stimulate or inhibit NFκB-mediated gene expression influencing cellular fate. We studied the effect of hydrogen peroxide (H2O2)-mediated oxidative stress and NFκB signaling on p62 expression in the retinal pigment epithelium (RPE) and investigated its role in regulation of autophagy and RPE survival against oxidative damage. Cultured human RPE cell line ARPE-19 and primary human adult and fetal RPE cells were exposed to H2O2-induced oxidative stress. The human apolipoprotein E4 targeted-replacement (APOE4) mouse model of AMD was used to study expression of p62 and other autophagy proteins in the retina. p62, NFκB p65 (total, phosphorylated, nuclear and cytoplasmic) and ATG10 expression was assessed by mRNA and protein analyses. Cellular ROS and mitochondrial superoxide were measured by CM-H2DCFDA and MitoSOX staining respectively. Mitochondrial viability was determined using MTT activity. qPCR-array system was used to investigate autophagic genes affected by p62. Nuclear and cytoplasmic levels of NFκB p65 were evaluated after cellular fractionation by Western blotting. We report that p62 is up-regulated in RPE cells under H2O2-induced oxidative stress and promotes autophagic activity. Depletion of endogenous p62 reduces autophagy by downregulation of ATG10 rendering RPE more susceptible to oxidative damage. NFκB p65 phosphorylation at Ser-536 was found to be critical for p62 upregulation in response to oxidative stress. Proteasome inhibition by H2O2 causes p62-NFκB signaling as antioxidant pre-treatment reversed p62 expression and p65 phosphorylation when RPE was challenged by H2O2 but not when by Lactacystin. p62 protein but not RNA levels are elevated in APOE4-HFC AMD mouse model, suggesting reduction of autophagic flux in disease conditions. Our findings suggest that p62 is necessary for RPE cytoprotection under oxidative stress and functions, in part, by modulating ATG10 expression. NFκB p65 activity may be a critical upstream initiator of p62 expression in RPE cells under oxidative stress.


Count: evolutionary analysis of phylogenetic profiles with parsimony and likelihood.
Count is a software package for the analysis of numerical profiles on a phylogeny. It is primarily designed to deal with profiles derived from the phyletic distribution of homologous gene families, but is suited to study any other integer-valued evolutionary characters. Count performs ancestral reconstruction, and infers family- and lineage-specific characteristics along the evolutionary tree. It implements popular methods employed in gene content analysis such as Dollo and Wagner parsimony, propensity for gene loss, as well as probabilistic methods involving a phylogenetic birth-and-death model. Count is available as a stand-alone Java application, as well as an application bundle for MacOS X, at the web site http://www.iro.umontreal.ca/ approximately csuros/gene_content/count.html. It can also be launched using Java Webstart from the same site. The software is distributed under a BSD-style license. Source code is available upon request from the author.


A rare case of oncocytic adrenocortical carcinoma clinically presented as an incidentaloma.
Adrenocortical carcinoma (ACC) is a rare aggressive tumor originating from adrenocortical parenchymal cells and its incidence is approximately 1 per million population per year. An oncocytic ACC is a recently identified entity among the several known histopathological variants of ACC, which is characterized by oncocytic cells, and only a few cases in the available literature have reported this tumor. In contrast to conventional ACCs, oncocytic ACCs usually manifest as solitary lesions presenting in adults without any sex predilection. We report a case of a 70-year-old Japanese man who presented with an incidentally discovered retroperitoneal mass without any evidence of excessive corticosteroid secretion. Laboratory and imaging studies, as well as transgastric endoscopic ultrasound-guided fine needle aspiration failed to establish a definitive diagnosis. Thus, the patient underwent surgical resection of the left-sided peritoneal tumor. Weiss score was positive in 6/9 points and the tumor met two major criteria of the Lin-Weiss-Bisceglia (LWB) system leading to a diagnosis of an oncocytic variant of ACC. Based on our findings in this patient, we conclude that a combination of the Weiss and LWB criteria is required to determine the malignant potential of oncocytic adrenal tumors because ACCs and oncocytomas could be frequently indistinguishable. Careful histopathological examination is pivotal in confirming the oncocytic component in the lesion and hence definitive diagnosis of ACCs.


Dynamic functional reorganization of the motor execution network after stroke.
Numerous studies argue that cortical reorganization may contribute to the restoration of motor function following stroke. However, the evolution of changes during the post-stroke reorganization has been little studied. This study sought to identify dynamic changes in the functional organization, particularly topological characteristics, of the motor execution network during the stroke recovery process. Ten patients (nine male and one female) with subcortical infarctions were assessed by neurological examination and scanned with resting-state functional magnetic resonance imaging across five consecutive time points in a single year. The motor execution network of each subject was constructed using a functional connectivity matrix between 21 brain regions and subsequently analysed using graph theoretical approaches. Dynamic changes in topological configuration of the network during the process of recovery were evaluated by a mixed model. We found that the motor execution network gradually shifted towards a random mode during the recovery process, which suggests that a less optimized reorganization is involved in regaining function in the affected limbs. Significantly increased regional centralities within the network were observed in the ipsilesional primary motor area and contralesional cerebellum, whereas the ipsilesional cerebellum showed decreased regional centrality. Functional connectivity to these brain regions demonstrated consistent alterations over time. Notably, these measures correlated with different clinical variables, which provided support that the findings may reflect the adaptive reorganization of the motor execution network in stroke patients. In conclusion, the study expands our understanding of the spectrum of changes occurring in the brain after stroke and provides a new avenue for investigating lesion-induced network plasticity.


Athletes broad dimensions of dispositional perfectionism: examining changes in life satisfaction and the mediating role of sport-related motivation and coping.
This study examined the associations of dispositional perfectionism, contextual motivation, sport-related coping, goal attainment, and changes in life satisfaction during a sport competition. A sample of 186 athletes completed measures of dispositional perfectionism, contextual motivation, and life satisfaction at Time 1 (before a competition) as well as measures of coping, goal attainment, and life satisfaction at Time 2 (after a competition). Results of structural equation modeling supported a model in which self-determined and non-self-determined motivation partially mediated the relationships between different dimensions of perfectionism and coping. It was also shown that disengagement-oriented coping mediated the negative relationship between evaluative concerns perfectionism and change in life satisfaction. In a similar way, goal attainment mediated the relationships of both task- and disengagement-oriented coping with change in life satisfaction. For the most part, these results are consistent with the motivational properties of evaluative concerns and personal standards perfectionism and with literature regarding coping and self-determination theory.


Prospects of Wannier functions in investigating photonic crystal all-optical devices for signal processing.
Wannier functions derived from Bloch functions have been identified as an efficient means of analyzing the properties of photonic crystals in which localized functions have now opened the door for 2D and 3D structures containing defects to be investigated. In this paper, based on the Maxwell equations in diagonalized form and utilizing Bloch waves we have obtained an equivalent system of algebraic equations in eigenform. By establishing and exploiting several distinct properties of the resulting eigenpairs, we demonstrate an ability to construct Wannier functions associated with the simplest one-dimensional photonic structure. More importantly, the numerical investigation of the inner- and intra-band orthonormality conditions as well as Hilbert space partitioning features shows a capability for multi-resolution analysis that will make all-optical signal processing devices with photonic crystal structures feasible.


Abnormal regurgitation in three cows caused by intrathoracic perioesophageal lesions.
Three Brown Swiss cows with abnormal regurgitation because of a perioesophageal disorder are described. The cows were ill and had poor appetite, salivation and regurgitation of poorly-chewed feed. Collection of rumen juice was successful in one cow, and in another, the tube could be advanced to the level of the 7th intercostal space, and in the third, only saliva could be collected. In one cow, oesophagoscopy revealed a discoloured 10-cm mucosal area with fibrin deposits. Thoracic radiographs were normal. The cows were euthanased and examined postmortem. Cow 1 had a large perioesophageal abscess containing feed material at the level of the thoracic inlet, believed to be the result of a healed oesophageal injury. Cow 2 had an abscess between the oesophagus and trachea 25 cm caudal to the epiglottis with the same presumed aetiology as in cow 1. Cow 3 had a mediastinal carcinoma that enclosed and constricted the oesophagus. Abnormal regurgitation in cattle is usually the result of an oesophageal disorder. Causes of oesophageal disorders vary widely and their identification can be difficult.


A direct test of the differentiation mechanism: REM, BCDMEM, and the strength-based mirror effect in recognition memory.
We explore competing explanations for the reduction in false alarm rate observed when studied items are strengthened. Some models, such as Retrieving Effectively from Memory (REM; Shiffrin & Steyvers, 1997), attribute the false alarm rate reduction to differentiation, a process in which strengthening memory traces at study directly reduces the memory evidence for lure items. Models with no differentiation mechanism, such as the Bind-Cue-Decide Model of Episodic Memory (BCDMEM; Dennis & Humphreys, 2001), explain the false alarm rate reduction in terms of the strength of items expected at retrieval. To contrast these explanations, we separately manipulated item strength at encoding and retrieval. Participants studied mixed lists of weak and strong items. Weak items were always presented once. On separate lists, strong items were either presented twice (Strong-2X) or five times (Strong-5X). Within each strength condition, participants completed separate tests with mixed (strong and weak) targets, pure weak targets, or pure strong targets. They were correctly informed of the type of target on each test. Results showed that false alarm rates decreased from the strong-2X condition to the strong-5X condition for the mixed and pure-strong tests, but not for the pure-weak tests. That is, false alarm rates were determined by the strength of targets appearing on the test, not by the content of the study list. The results support BCDMEM's expectation-based explanation and not REM's differentiation-based explanation.


Protective effect of aqueous extracts from Rhizopus oryzae on liver injury induced by carbon tetrachloride in rats.
Hepatoprotective effects of Rhizopus oryzae/ U-1 aqueous extract (RU) were demonstrated in carbon tetrachloride (CCl4 )-induced liver-injured rats. In order to investigate the RU effects, the rats were administered RU at a dose of 10 or 100 mg/kg of body weight for 10 days before induction of the liver injury by oral administration of CCl4 (125 mg/kg body weight). (i) Pretreatment with RU caused a significant decrease in serum lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities that were increased by the administration of CCl4 . (ii) RU pretreatment (100 mg/kg) increased 5-bromo-2'-deoxyuridine incorporation at 48 h after CCl4 treatment in hepatocytes. (iii) Histological hematoxylin and eosin staining of the liver showed that RU pretreatment reduced the damage induced by CCl4 administration. (iv) Reverse transcriptase PCR analysis showed RU retreatment caused a transient but significant increase in hepatocyte growth factor (HGF) and a sustained and significant increase in insulin-like growth factor-I (IGF-I) gene expression in hepatocytes injured by CCl4 treatment. From these results, we conclude that oral pre-administration of RU was effective to suppress liver injury induced by the subsequent oral CCl4 administration, and RU-induced increase in IGF-I and HGF gene expression may be, even in part, involved in biological actions of RU in rats.


Editorial: Can gluten contribute to irritable bowel syndrome?
Functional gastrointestinal disorders are the most common gastroenterological problem in our society. Changes in gut function, including pain perception, motility, and intestinal permeability, and low-grade inflammation have been described in patients with irritable bowel syndrome (IBS). The triggering factors for the described immunity and gut functional changes in patients with IBS are not completely understood. Similarly to post-infective IBS, some patients with IBS symptoms exhibit immunological evidence of gluten sensitivity but have no overt intestinal mucosal injury. They have symptoms that meet the diagnostic criteria for IBS and respond symptomatically to exclusion of gluten from the diet. Thus, gluten sensitivity may be involved in the pathogenesis of a subgroup of IBS patients. Unfortunately, there remain many unanswered questions regarding the mechanistic link between gluten sensitivity and functional gastrointestinal symptoms.


Glycyrrhizin exerts antioxidative effects in H5N1 influenza A virus-infected cells and inhibits virus replication and pro-inflammatory gene expression.
Glycyrrhizin is known to exert antiviral and anti-inflammatory effects. Here, the effects of an approved parenteral glycyrrhizin preparation (Stronger Neo-Minophafen C) were investigated on highly pathogenic influenza A H5N1 virus replication, H5N1-induced apoptosis, and H5N1-induced pro-inflammatory responses in lung epithelial (A549) cells. Therapeutic glycyrrhizin concentrations substantially inhibited H5N1-induced expression of the pro-inflammatory molecules CXCL10, interleukin 6, CCL2, and CCL5 (effective glycyrrhizin concentrations 25 to 50 µg/ml) but interfered with H5N1 replication and H5N1-induced apoptosis to a lesser extent (effective glycyrrhizin concentrations 100 µg/ml or higher). Glycyrrhizin also diminished monocyte migration towards supernatants of H5N1-infected A549 cells. The mechanism by which glycyrrhizin interferes with H5N1 replication and H5N1-induced pro-inflammatory gene expression includes inhibition of H5N1-induced formation of reactive oxygen species and (in turn) reduced activation of NFκB, JNK, and p38, redox-sensitive signalling events known to be relevant for influenza A virus replication. Therefore, glycyrrhizin may complement the arsenal of potential drugs for the treatment of H5N1 disease.


Purification and characterization of poly(A) polymerase from Saccharomyces cerevisiae.
Poly(A) polymerase was purified 22,000-fold to homogeneity from a whole cell extract of Saccharomyces cerevisiae with a yield of 22%. The enzyme is a monomeric polypeptide with a denatured molecular weight of 63,000. Incorporation of labeled ATP into acid-precipitable material by the purified enzyme proceeds faster with manganese than with magnesium ions. Various RNA homopolymers as well as Escherichia coli tRNA or rRNA can serve as primers. An RNA that terminates at the natural poly(A) site of the CYC1 gene is not more efficiently elongated than several nonspecific substrates, indicating the requirement for additional factors to provide specificity. Elongation of the primer is distributive. Covering of a poly(A) primer with poly(A)-binding protein reduces the enzyme's activity more than 10-fold.


Provider experiences with the large-scale 'Helping Babies Breathe' training programme in Tanzania.
Worldwide, there has been renewed emphasis on reducing neonatal mortality in low-resource countries. The Helping Babies Breathe (HBB) programme has been shown to reduce newborn deaths. The aim of this study is to present provider-level perceptions and experiences of the HBB programme implemented at-scale in Tanzania and identify key lessons learned for scalability in similar and other settings. Focus group discussions with HBB-trained providers were conducted using a prospective longitudinal study design between October 2013 and May 2015. A semi-structured discussion guide was used to facilitate the focus groups which were held 4-6 weeks and 4-6 months post-HBB training. Data were managed using NVivo software and analysed thematically. A total of 222 focus group discussions were conducted in 252 trained facilities and involved 599 providers across 15 regions of Tanzania. Birth attendants reported that the training programme helped increase knowledge, skills and confidence, and that the provided equipment simplified resuscitation. Supportive supervision and regular follow-up visits were considered critical for skills retention. On the other hand, the brief 1-day training in Tanzania, small financal incentives, intra-facility rotations of trained attendants, staff shortages, limited rescucitation spaces and mastery of the bag-and-mask were considered challenges to the HBB programme in Tanzania. The HBB programme was largely very well received during its first at-scale implementation in Tanzania. Addressing the main challenges cited by participants, particularly the training duration, may increase provider satisfaction with the HBB training programme.


Real-time reception of multi-gigabit coherent optical OFDM signals.
Coherent Optical OFDM (CO-OFDM) has been demonstrated for delivering superior performance in spectral efficiency, receiver sensitivity, and polarization-dispersion resilience. Fueled by the rapid advancement in semiconductor technology, high-speed field-programmable gate arrays (FPGA) and analogue-to-digital-converters/digital-to-analogue converters (ADC/DACs) have been increasingly adopted for digital signal processing in optical communications. In this paper, we report the first FPGA-based real-time implementation of coherent optical OFDM (CO-OFDM) receiver with a transmission rate up to 3.1 Gb/s. Several basic aspects of CO-OFDM signal processing are described in detail, and the BER sensitivity performance are evaluated in real-time. To the best of our knowledge, we have achieved the record real-time reception date rate for a single-input single-output (SISO) coherent OFDM signal, in either RF domain or optical domain.


Lung cancer: interobserver agreement on interpretation of pulmonary findings at low-dose CT screening.
To evaluate agreement among radiologists on the interpretation of pulmonary findings at low-dose computed tomographic (CT) screening examinations for lung cancer. Institutional review board approval and informed consent were obtained. HIPAA guidelines were followed. Sixteen radiologists from the 10 National Lung Screening Trial screening centers of the National Cancer Institute's Lung Screening Study network reviewed image subsets from 135 baseline low-dose screening CT examinations in 135 trial participants (89 men, 46 women; mean age, 62.7 years +/- 5.4 [standard deviation]). Interpretations were classified into one of four of the following categories: noncalcified nodule 4 mm or larger in greatest transverse dimension (positive screening result); noncalcified nodule smaller than 4 mm in greatest transverse dimension (negative screening result); calcified, benign nodule (negative screening result); or no nodule (negative screening result). A recommendation for follow-up evaluation was obtained for each case. Interobserver agreement was evaluated by using the multirater kappa statistic and by using response frequencies and descriptive statistics. Multirater kappa values ranged from 0.58 (for agreement among all four classifications; 95% confidence interval: 0.55, 0.61) to 0.64 (for agreement on classification as a positive or negative screening result; 95% confidence interval: 0.62, 0.65). The average percentage of reader pairs in agreement on the screening result per case (percentage agreement) was 82%. There was wide variation in the total number of abnormalities detected and classified as pulmonary nodules, with differences of up to more than twofold among radiologists. For cases classified as positive, multirater kappa for follow-up recommendations was 0.35. Interobserver agreement was moderate to substantial; potential for considerable improvement exists. Clinical trial registration no. NCT00047385.


Intimin from enteropathogenic Escherichia coli restores murine virulence to a Citrobacter rodentium eaeA mutant: induction of an immunoglobulin A response to intimin and EspB.
The formation of attaching and effacing (A/E) lesions is central to the pathogenesis of enteropathogenic Escherichia coli (EPEC)-mediated disease in humans and Citrobacter rodentium (formerly C. freundii biotype 4280)-mediated transmissible colonic hyperplasia in mice. Closely related outer membrane proteins, known as intimins, are required for formation of the A/E lesion by both EPEC (Int(EPEC)) and C. rodentium (Int(CR)). A secreted protein, EspB (formally EaeB), is also necessary for A/E-lesion formation. Here we report that expression of a cloned Int(EPEC), encoded by plasmid pCVD438, restores murine virulence to an intimin-deficient mutant of C. rodentium DBS255. Replacement of Cys937 with Ala abolished the ability of the cloned EPEC intimin to complement the deletion mutation in DBS255. Ultrastructural examination of tissues from wild-type C. rodentium and DBS255(pCVD438)-infected mice revealed multiple A/E lesion on infected cells and loss of contact between enterocytes and basement membrane. Histological investigation showed that although both wild-type C. rodentium and DBS255(pCVD438) colonized the descending colon and induced colonic hyperplasia in orally infected 21-day-old mice, the latter strain adhered to epithelial cells located deeper within crypts. Nonetheless, infection with the wild-type strain was consistently more virulent, as indicated by a higher mortality rate. All the surviving mice, challenged with either wild-type C. rodentium or DBS255(pCVD438), developed a mucosal immunoglobulin A response to intimin and EspB. These results show that C. rodentium infection provides a relevant, simple, and economic model to investigate the role of EPEC proteins in the formation of A/E lesions in vivo and in intestinal disease.


The relationship between female genital mutilation and HIV transmission in sub-Saharan Africa.
Female genital mutilation (FGM) is an age-old practice that has since been linked with many health problems. This review aims to highlight some of the controversies trailing the relationship between FGM and HIV transmission in sub-Saharan Africa. A literature search was conducted on the subject matter. This was done using articles published in English while limiting the geographical coverage to sub-Saharan Africa. Three themes were noted. These themes include: Direct causal link between FGM and HIV transmission; indirect causal link between FGM and HIV transmission and a negative or no association between FGM and HIV transmission. While many of the arguments are within scientific reasoning, the researches supporting the views seem to lack the necessary objectivity. This study underscored the need for a more objective lens in viewing and conducting research on the relationship between FGM and HIV transmission in sub-Saharan Africa.


Exploring ComQXPA quorum-sensing diversity and biocontrol potential of Bacillus spp. isolates from tomato rhizoplane.
Bacillus subtilis is a widespread and diverse bacterium t exhibits a remarkable intraspecific diversity of the ComQXPA quorum-sensing (QS) system. This manifests in the existence of distinct communication groups (pherotypes) that can efficiently communicate within a group, but not between groups. Similar QS diversity was also found in other bacterial species, and its ecological and evolutionary meaning is still being explored. Here we further address the ComQXPA QS diversity among isolates from the tomato rhizoplane, a natural habitat of B. subtilis, where these bacteria likely exist in their vegetative form. Because this QS system regulates production of anti-pathogenic and biofilm-inducing substances such as surfactins, knowledge on cell-cell communication of this bacterium within rhizoplane is also important from the biocontrol perspective. We confirm the presence of pherotype diversity within B. subtilis strains isolated from a rhizoplane of a single plant. We also show that B. subtilis rhizoplane isolates show a remarkable diversity of surfactin production and potential plant growth promoting traits. Finally, we discover that effects of surfactin deletion on biofilm formation can be strain specific and unexpected in the light of current knowledge on its role it this process.


Inhibition of HIV Env binding to cellular receptors by monoclonal antibody 2G12 as probed by Fc-tagged gp120.
During natural HIV infection, an array of host receptors are thought to influence virus attachment and the kinetics of infection. In this study, to probe the interactions of HIV envelope (Env) with various receptors, we assessed the inhibitory properties of various anti-Env monoclonal antibodies (mAbs) in binding assays. To assist in detecting Env in attachment assays, we generated Fc fusions of full-length wild-type gp120 and several variable loop-deleted gp120s. Through investigation of the inhibition of Env binding to cell lines expressing CD4, CCR5, DC-SIGN, syndecans or combinations thereof, we found that the broadly neutralizing mAb, 2G12, directed to a unique carbohydrate epitope of gp120, inhibited Env-CCR5 binding, partially inhibited Env-DC-SIGN binding, but had no effect on Env-syndecan association. Furthermore, 2G12 inhibited Env attachment to primary monocyte-derived dendritic cells, that expressed CD4 and CCR5 primary HIV receptors, as well as DC-SIGN, and suggested that the dual activities of 2G12 could be valuable in vivo for inhibiting initial virus dissemination and propagation.


Expansion of alpha-galactosylceramide-stimulated Valpha24+ NKT cells cultured in the absence of animal materials.
Valpha24+ NKT is an innate lymphocyte with potential antitumor activity. Clinical applications of Valpha24+ natural killer (NK) T cells, which are innate lymphocytes with potential antitumor activity, require their in vitro expansion. To avoid the potential dangers posed to patients by fetal bovine serum (FBS), the authors evaluated non-FBS culture conditions for the selective and efficient expansion of human Valpha24+ NKT cells. Mononuclear cells (MNCs) and plasma from the peripheral blood of normal healthy donors were used before and after G-CSF mobilization. MNCs and plasma separated from apheresis products were also used. MNCs were cultured for 12 days in AIM-V medium containing alpha-galactosylceramide (alpha-GalCer) (100 ng/mL) and IL-2 (100 U/mL) supplemented with FBS, autologous plasma, or autologous serum. The cultured cells were collected and their surface markers, intracellular cytokines, and cytotoxicity were evaluated. The highest expansion ratio for Valpha24+ NKT cells was obtained from G-CSF-mobilized MNCs cultured in medium containing 5% autologous plasma. Cultures containing MNCs and autologous plasma obtained before and after G-CSF mobilization had approximately 350-fold and 2,000-fold expansion ratios, respectively. These results suggest that G-CSF mobilization conferred a proliferative advantage to Valpha24+ NKT cells by modifying the biology of cells and plasma factors. Expanded Valpha24+ NKT cells retained their surface antigen expression and production of IFN-gamma and exhibited CD1d-independent cytotoxicity against tumor cells. Valpha24+ NKT cells can be efficiently expanded from G-CSF-mobilized peripheral blood MNCs in non-FBS culture conditions with alpha-GalCer and IL-2.


IMproving PArticipation of patients in Clinical Trials--rationale and design of IMPACT.
One of the most commonly reported problems of randomised trials is that recruitment is usually slower than expected. Trials will cost more and take longer, thus delaying the use of the results in clinical practice, and incomplete samples imply decreased statistical power and usefulness of its results. We aim to identify barriers and facilitators for successful patient recruitment at the level of the patient, the doctor and the hospital organization as well as the organization and design of trials over a broad range of studies. We will perform two cohort studies and a case-control study in The Netherlands. The first cohort study will report on a series of multicenter trials performed in a nationwide network of clinical trials in obstetrics and gynaecology. A questionnaire will be sent to all clinicians recruiting for these trials to identify determinants--aggregated at centre level--for the recruitment rate. In a case control-study nested in this cohort we will interview patients who refused or consented participation to identify factors associated with patients' consent or refusal. In a second cohort study, we will study trials that were prospectively registered in the Netherlands Trial Register. Using a questionnaire survey we will assess whether issues on hospital organization, trial organization, planning and trial design were associated with successful recruitment, i.e. 80% of the predefined number of patients recruited within the planned time. This study will provide insight in barriers and facilitators for successful patient recruitment in trials. The results will be used to provide recommendations and a checklist for individual trialists to identify potential pitfalls for recruitment and judge the feasibility prior to the start of the study. Identified barriers and motivators coupled to evidence-based interventions can improve recruitment of patients in clinical trials.


[Epidemiological profile of traffic-related disability in Peru, 2012].
To describe the epidemiological profile of people living with disabilities due to traffic accidents (TA) in Peru. Secondary analysis of the National Survey Specialized on Disability (ENEDIS) of 2012 and an ecological analysis of TA records of the Ministry of Transportation and Communications was done. Disability by traffic accidents (DAT) was reported by 49,036 persons; 81.3% of whom live in urban areas. The most frequent disability was limited locomotion and skill (77.4%), followed by visual impairment (22.9%). Dependence for activities was reported in 44.7% of persons with disabilities. The regions with the highest prevalence of TA have a higher prevalence of disability by traffic accidents (Spearman coefficient: 0.426, p=0.034). Most of disability due to TA is found in urban areas, correspond to males and consist of persons in economically productive age. The most common form of disability is in locomotion. Most of individuals do not receive any form of rehabilitation, which accentuates health inequity related to traffic accidents.


Cervical cancer beliefs and pap test screening practices among Chinese American immigrants.
To examine beliefs and Pap test utilization among Chinese American women, the largest Asian female population in the United States. Cross-sectional descriptive, correlational study. Metropolitan areas of Portland, OR. 100 foreign-born Chinese women aged 40 years and older. Three questionnaires were modified, translated, combined, and pretested. Participants completed the self-administered questionnaire in a group setting. Utilization of Pap test screening, health beliefs, and cultural and sociodemographic variables. Sixty-eight percent reported having a Pap test within the prior three years (adherence), and 84% reported ever having a Pap test. The odds of Pap test use and adherence decreased with increasing age. Women with insurance or a regular healthcare provider had better odds of Pap test use and adherence. Older age, older age when a participant moved to the United States, and increased modesty were negatively associated with ever having had a Pap test. Age and cultural beliefs influence Pap test use and adherence. The strength of provider recommendation and healthcare access as predictors suggest areas for interventions designed to increase screening for cervical cancer. Nurses play a vital role in preventive health care, especially with the growing number of advanced practice nurses delivering primary care. Primary healthcare providers should be reminded of their influential role in increasing adherence to cancer screening. Further health policy action is necessary to extend screening coverage to those who do not have adequate health insurance.


Redefining electrosensitivity: A new literature-supported model.
In critically examining literature on electrohypersensitivity and the reported somatic responses to anthropogenic modulated radiofrequency radiation (RFR) exposure, it becomes apparent that electrohypersensitivity is one part of a range of consequences. Current evidence on the necessity of considering patients' overall health status leads us to propose a new model in which electrohypersensitivity is but part of the electrosensitive status inherent in being human. We propose the likelihood and type of response to environmental RFR include i) a linear somatic awareness continuum, ii) a non-linear somatic response continuum, and iii) the extent of each individual's capacity to repair damage (linked to homeostatic response). We anticipate this last, dynamic, aspect is inextricably linked to the others through the autonomic nervous system. The whole is dependent upon the status of the interconnected immune and inflammatory systems. This holistic approach leads us to propose various outcomes. For most, their body maintains homeostasis by routine repair. However, some develop electrohypersensitivity either due to RFR exposure or as an ANS-mediated, unconscious response (aka nocebo effect), or both. We suggest RFR exposure may be one factor in the others developing an auto-immune disease or allergy. A few develop delayed catastrophic disease such as glioma. This model gives the blanket term ElectroMagnetic Illness (EMI) to all RFR-related conditions. Thus, EHS appears to be one part of a range of responses to a novel and rapidly changing evolutionary situation.


Effect of multiple patient reminders in improving diabetic retinopathy screening. A randomized trial.
To determine whether multiple mailed patient reminders can produce an increase in the rate of diabetic retinal examinations (DRE) over that seen with a single reminder. All diabetic members > or = 18 years who were enrolled in a large network-based health maintenance organization (HMO) in California from August 1996 to July 1997 were identified using claims and pharmacy databases. Members who had no record of DRE in the HMO's claims database were then randomized into two groups. Both groups received mailed educational materials and a reminder to obtain the examination. Their physician groups also received a letter explaining the program, current guidelines for DRE, and a list of their diabetes patients with their DRE status. The single intervention group received no additional reminders. The multiple intervention group received additional reminders at 3, 6, and 9 months after baseline if they continued with no record of service, as determined from the claims database. The study cohort comprised 19,523 diabetic members, which were randomized into single (n = 9,614) and multiple (n = 9,909) intervention groups. There was an increase in monthly DRE rates after the intervention in August 1996 for both intervention groups. After the second reminder was sent to the multiple intervention group, the percentage of diabetic members receiving DRE was higher than the single intervention group. Rates before and after the third intervention were not significantly different, nor were monthly differences found. There was a significant difference in overall annual DRE rates between the groups (P = 0.023). Multiple patient reminders are more effective than single reminders in improving DRE rates in a managed care setting. However, the improvement noted was clinically small and appeared only after the second reminder; no incremental improvement was seen with additional reminders. Resources used for multiple reminders aimed at diabetic retinopathy might better be spent on other approaches to reducing complications of diabetes.


Photographic and luminometric detection of luciferase reporter phages for drug susceptibility testing of clinical Mycobacterium tuberculosis isolates.
Luciferase reporter phages (LRPs) have proven to be efficient tools for drug susceptibility testing of Mycobacterium tuberculosis. Luminometric detection of LRP activity offers higher sensitivity and quantitative results, while a Polaroid film detection method offers a "low-tech" inexpensive alternative that is called the Bronx box. In this work we evaluated, improved, and compared the performance of the luminometer and the Bronx box formats for drug susceptibility testing with LRPs by using 51 clinical isolates of M. tuberculosis, with the agar proportion method (PM) serving as reference. The sensitivity in detecting resistance to isoniazid and rifampin, antibiotics that define multidrug resistance (MDR), was 100% for both methods. The turnaround time for results was reduced from 3 weeks for PM to 54 or 94 h for luminometry or the Bronx box, respectively. These results support the utility of LRPs as a screening test for the surveillance of MDR tuberculosis.


Homeopathy for treatment of irritable bowel syndrome.
Irritable bowel syndrome (IBS) is a common, chronic disorder that leads to decreased health-related quality of life and work productivity. Evidence-based treatment guidelines have not been able to give guidance on the effects of homeopathic treatment for IBS because no systematic reviews have been carried out to assess the effectiveness of homeopathic treatment for IBS. Two types of homeopathic treatment were evaluated in this systematic review. In clinical homeopathy a specific remedy is prescribed for a specific condition. This differs from individualised homeopathic treatment, where a homeopathic remedy based on a person's individual symptoms is prescribed after a detailed consultation. To assess the effectiveness and safety of homeopathic treatment for treating IBS. We searched MEDLINE, the Cochrane Central Register of Controlled Trials, EMBASE, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), the Allied and Complementary Medicine Database (AMED), Cochrane IBD/FBD Group Specialised Register, Cochrane Complementary Medicine Field Specialised Register and the database of the Homeopathic Library (Hom-inform) from inception to February 2013. Randomised controlled trials (RCTs), cohort and case-control studies that compared homeopathic treatment with placebo, other control treatments, or usual care, in adults with IBS were considered for inclusion. Two authors independently assessed the risk of bias and extracted data. The primary outcome was global improvement in IBS. The overall quality of the evidence supporting this outcome was assessed using the GRADE criteria. We calculated the mean difference (MD) and 95% confidence interval (CI) for continuous outcomes and the risk ratio (RR) and 95% CI for dichotomous outcomes. Three RCTs (213 participants) were included. No cohort or case-control studies were identified. Two studies published in 1976 and 1979 compared clinical homeopathy (homeopathic remedy) to placebo for constipation-predominant IBS. One study published in 1990 compared individualised homeopathic treatment (consultation plus remedy) to usual care (defined as high doses of dicyclomine hydrochloride, faecal bulking agents and diet sheets asking the patient to take a high fibre diet) for the treatment of IBS in female patients. Due to the low quality of reporting in the included studies the risk of bias in all three studies was unclear on most criteria and high for some criteria. A meta-analysis of two small studies (129 participants with constipation-predominant IBS) found a statistically significant difference in global improvement between the homeopathic remedy asafoetida and placebo at a short-term follow-up of two weeks. Seventy-three per cent of patients in the homeopathy group improved compared to 45% of placebo patients (RR 1.61, 95% CI 1.18 to 2.18). There was no statistically significant difference in global improvement between the homeopathic remedies asafoetida plus nux vomica and placebo. Sixty-eight per cent of patients in the homeopathy group improved compared to 52% of placebo patients (1 study, N = 42, RR 1.31, 95% CI 0.80 to 2.15). GRADE analyses rated the overall quality of the evidence for the outcome global improvement as very low due to high or unknown risk of bias, short-term follow-up and sparse data. There was no statistically significant difference found between individualised homeopathic treatment and usual care (1 RCT, N = 20) for the outcome "feeling unwell", where the participant scored how "unwell" they felt before, and after treatment (MD 0.03; 95% CI -3.16 to 3.22). None of the included studies reported on adverse events. A pooled analysis of two small studies suggests a possible benefit for clinical homeopathy, using the remedy asafoetida, over placebo for people with constipation-predominant IBS. These results should be interpreted with caution due to the low quality of reporting in these trials, high or unknown risk of bias, short-term follow-up, and sparse data. One small study found no statistically difference between individualised homeopathy and usual care (defined as high doses of dicyclomine hydrochloride, faecal bulking agents and diet sheets advising a high fibre diet). No conclusions can be drawn from this study due to the low number of participants and the high risk of bias in this trial. In addition, it is likely that usual care has changed since this trial was conducted. Further high quality, adequately powered RCTs are required to assess the efficacy and safety of clinical and individualised homeopathy compared to placebo or usual care.


Minimum set of mutations needed to optimize cyan fluorescent proteins for live cell imaging.
Cyan fluorescent proteins (CFPs) are widely used as FRET donors in genetically encoded biosensors for live cell imaging. Recently, cyan variants with greatly improved fluorescence quantum yields have been developed by large scale random mutagenesis. We show that the introduction of only two mutations, T65S and H148G, is able to confer equivalent performances on the popular form ECFP, leading to Aquamarine (QY = 0.89, τ(f) = 4.12 ns). Besides an impressive pH stability (pK(1/2) = 3.3), Aquamarine shows a very low general sensitivity to its environment, and undetectable photoswitching reactions. Aquamarine gives efficient and bright expression in different mammalian cell systems, with a long and single exponential intracellular fluorescence lifetime mostly insensitive to the fusion or the subcellular location of the protein. Aquamarine was also able to advantageously replace the CFP donor in the FRET biosensor AKAR for ratiometric measurements of protein kinase A activity. The performances of Aquamarine show that only two rounds of straightforward single point mutagenesis can be a quick and efficient way to optimize the donor properties in FRET-based biosensors.


Variable radiological lung nodule evaluation leads to divergent management recommendations.
Radiological evaluation of incidentally detected lung nodules on computed tomography (CT) influences management. We assessed international radiological variation in 1) pulmonary nodule characterisation; 2) hypothetical guideline-derived management; and 3) radiologists' management recommendations.107 radiologists from 25 countries evaluated 69 CT-detected nodules, recording: 1) first-choice composition (solid, part-solid or ground-glass, with percentage confidence); 2) morphological features; 3) dimensions; 4) recommended management; and 5) decision-influencing factors. We modelled hypothetical management decisions on the 2005 and updated 2017 Fleischner Society, and both liberal and parsimonious interpretations of the British Thoracic Society 2015 guidelines.Overall agreement for first-choice nodule composition was good (Fleiss' κ=0.65), but poorest for part-solid nodules (weighted κ 0.62, interquartile range 0.50-0.71). Morphological variables, including spiculation (κ=0.35), showed poor-to-moderate agreement (κ=0.23-0.53). Variation in diameter was greatest at key thresholds (5 mm and 6 mm). Agreement for radiologists' recommendations was poor (κ=0.30); 21% disagreed with the majority. Although agreement within the four guideline-modelled management strategies was good (κ=0.63-0.73), 5-10% of radiologists would disagree with majority decisions if they applied guidelines strictly.Agreement was lowest for part-solid nodules, while significant measurement variation exists at important size thresholds. These variations resulted in generally good agreement for guideline-modelled management, but poor agreement for radiologists' actual recommendations.


JAK-STAT1/3-induced expression of signal sequence-encoding proopiomelanocortin mRNA in lymphocytes reduces inflammatory pain in rats.
Proopiomelanocortin (POMC)-derived beta-endorphin1-31 from immune cells can inhibit inflammatory pain. Here we investigated cytokine signaling pathways regulating POMC gene expression and beta-endorphin production in lymphocytes to augment such analgesic effects. Interleukin-4 dose-dependently elevated POMC mRNA expression in naïve lymph node-derived cells in vitro, as determined by real-time PCR. This effect was neutralized by janus kinase (JAK) inhibitors. Transfection of Signal Transducer and Activator of Transcription (STAT) 1/3 but not of STAT6 decoy oligonucleotides abolished interleukin-4 induced POMC gene expression. STAT3 was phosphorylated in in vitro interleukin-4 stimulated lymphocytes and in lymph nodes draining inflamed paws in vivo. Cellular beta-endorphin increased after combined stimulation with interleukin-4 and concanavalin A. Consistently, in vivo reduction of inflammatory pain by passively transferred T cells improved significantly when donor cells were pretreated with interleukin-4 plus concanavalin A. This effect was blocked by naloxone-methiodide. Interleukin-4 can amplify endogenous opioid peptide expression mediated by JAK-STAT1/3 activation in mitogen-activated lymphocytes. Transfer of these cells leads to inhibition of inflammatory pain via activation of peripheral opioid receptors.


The spectrum of acridine resistant mutants of bacteriophage T4 reveals cryptic effects of the tsL141 DNA polymerase allele on spontaneous mutagenesis.
Mutations in the ac gene of bacteriophage T4 confer resistance to acridine-inhibition of phage development. Previous studies had localized the ac gene region; we show that inactivation of T4 Open Reading Frame 52.2 confers the Acr phenotype. Thus, 52.2 is ac. The resistance mechanism is unknown. The ac gene provides a convenient forward mutagenesis assay. Its compact size (156 bp) simplifies mutant sequencing and diverse mutant types are found: base substitutions leading to missense or nonsense codons, in-frame deletions or duplications within the coding sequence, deletion or duplication frameshifts, insertions, complex mutations, and large deletions extending into neighboring sequences. Comparisons of spontaneous mutagenesis between phages bearing the wild-type or tsL141 alleles of DNA polymerase demonstrate that the impact of the mutant polymerase is cryptic when total spontaneous mutant frequencies are compared, but the DNA sequences of the ac mutants reveal a substantial alteration of fidelity by the mutant polymerase. The patterns of base substitution mutagenesis suggest that some site-specific mutation rate effects may reflect hotspots for mutagenesis arising by different mechanisms. A new class of spontaneous duplication mutations, having sequences inconsistent with misaligned pairing models, but consistent with nick-processing errors, has been identified at a hotspot in ac.


Tunable liquid-filled lens integrated with aspherical surface for spherical aberration compensation.
A novel liquid-filled lens design is presented. During fabrication, high precision single point diamond turning (SPDT) is introduced into standard soft lithography process to fabricate an aspherical surface constituting one end of lens. This enables the spherical aberration associated with the operation of the conventional liquid-filled lenses to be compensated for. Through flexibly optimizing this surface contour, it can be designed to work within particular working regions with improved optical quality. At the same time, the deformable elastic membrane is still adopted at the other end of the lens, thus preserving the high focal length tunability. This proof of concept and the performance of the proposed lens have been demonstrated using the lateral shearing interferometry experiment..


An epidemiological description of a folk illness: a study of empacho in Guatemala.
Although anthropologists have provided descriptions of many folk illnesses, few have systematically evaluated their prevalence and determined who is at greatest risk for acquiring them. This report attempts to provide a systematic description of the folk illness empacho including the symptoms that define it. Illness prevalence was estimated and subpopulations at greatest risk were identified from illness histories collected from a random sample of households in rural Guatemala. Empacho was found to constitute a distinct cluster of symptoms: diarrhea, vomiting, headache, and lack of appetite. It differed from other gastrointestinal illnesses in that headaches were more likely and stomachaches were less likely to be reported. Empacho was highly prevalent and occurred in adults and children. Further, results showed that although empacho was frequently diagnosed by residents, folk healers were rarely consulted for any illness. Nevertheless, a strong association exists between a household diagnosis of empacho and the use of folk healers by those households (p less than .001).


Entamoeba shows reversible variation in ploidy under different growth conditions and between life cycle phases.
Under axenic growth conditions, trophozoites of Entamoeba histolytica contain heterogenous amounts of DNA due to the presence of both multiple nuclei and different amounts of DNA in individual nuclei. In order to establish if the DNA content and the observed heterogeneity is maintained during different growth conditions, we have compared E. histolytica cells growing in xenic and axenic cultures. Our results show that the nuclear DNA content of E. histolytica trophozoites growing in axenic cultures is at least 10 fold higher than in xenic cultures. Re-association of axenic cultures with their bacterial flora led to a reduction of DNA content to the original xenic values. Thus switching between xenic and axenic growth conditions was accompanied by significant changes in the nuclear DNA content of this parasite. Changes in DNA content during encystation-excystation were studied in the related reptilian parasite E. invadens. During excystation of E. invadens cysts, it was observed that the nuclear DNA content increased approximately 40 fold following emergence of trophozoites in axenic cultures. Based on the observed large changes in nuclear size and DNA content, and the minor differences in relative abundance of representative protein coding sequences, rDNA and tRNA sequences, it appears that gain or loss of whole genome copies may be occurring during changes in the growth conditions. Our studies demonstrate the inherent plasticity and dynamic nature of the Entamoeba genome in at least two species.


Early surgical intervention or watchful waiting for the management of asymptomatic mitral regurgitation: a systematic review and meta-analysis.
Discordance between studies drives continued debate regarding the best management of asymptomatic severe mitral regurgitation (MR). The aim of the present study was to conduct a systematic review and meta-analysis of management plans for asymptomatic severe MR, and compare the effectiveness of a strategy of early surgery to watchful waiting. A systematic review was performed using the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Studies were excluded if they: (I) lacked a watchful waiting cohort; (II) included symptomatic patients; or (III) included etiologies other than degenerative mitral valve disease. The primary outcome of the study was all-cause mortality at 10 years. Secondary outcomes included operative mortality, repair rate, repeat mitral valve surgery, and development of new atrial fibrillation. Five observational studies were eligible for review and three were included in the pooled analysis. In asymptomatic patients without class I triggers (symptoms or ventricular dysfunction), pooled analysis revealed a significant reduction in long-term mortality with an early surgery approach [hazard ratio (HR) =0.38; 95% confidence interval (CI): 0.21-0.71]. This survival benefit persisted in a sub-group analysis limited to patients without class II triggers (atrial fibrillation or pulmonary hypertension) [relative risk (RR) =0.85; 95% CI: 0.75-0.98]. Aggregate rates of operative mortality did not differ between treatment arms (0.7% vs. 0.7% for early surgery vs. watchful waiting). However, significantly higher repair rates were achieved in the early surgery cohorts (RR =1.10; 95% CI: 1.02-1.18). Despite disagreement between individual studies, the present meta-analysis demonstrates that a strategy of early surgery may improve survival and increase the likelihood of mitral valve repair compared with watchful waiting. Early surgery may also benefit patients when instituted prior to the development of class II triggers.


Structural and Functional Analysis of E. coli Cyclopropane Fatty Acid Synthase.
Cell membranes must adapt to different environments. In Gram-negative bacteria, the inner membrane can be remodeled directly by modification of lipids embedded in the bilayer. For example, when Escherichia coli enters stationary phase, cyclopropane fatty acid (CFA) synthase converts most double bonds in unsaturated inner-membrane lipids into cyclopropyl groups. Here we report the crystal structure of E. coli CFA synthase. The enzyme is a dimer in the crystal and in solution, with each subunit containing a smaller N-domain that associates tightly with a larger catalytic C-domain, even following cleavage of the inter-domain linker or co-expression of each individual domain. Efficient catalysis requires dimerization and proper linkage of the two domains. These findings support an avidity-based model in which one subunit of the dimer stabilizes membrane binding, while the other subunit carries out catalysis.


Taking it to another level: do personality-based human capital resources matter to firm performance?
Drawing on the attraction-selection-attrition perspective, strategic human resource management (SHRM) scholarship, and recent human capital research, this study explores organization-level emergence of personality (i.e., personality-based human capital resources) and its direct, interactive, and (conditional) indirect effects on organization-level outcomes based on data from 6,709 managers across 71 firms. Results indicate that organization-level mean emotional stability, extraversion, and conscientiousness are positively related to organization-level managerial job satisfaction and labor productivity but not to financial performance. Furthermore, organization-level mean and variance in emotional stability interact to predict all three organization-level outcomes, and organization-level mean and variance in extraversion interact to predict firm financial performance. Specifically, the positive effects of organization-level mean emotional stability and extraversion are stronger when organization-level variance in these traits is lower. Finally, organization-level mean emotional stability, extraversion, and conscientiousness are all positively related to firm financial performance indirectly via labor productivity, and the indirect effects are more positive when organization-level variance in those personality traits is lower. Overall, the findings suggest that personality-based human capital resources demonstrate tangible effects on organization-level outcomes. Theoretical and practical implications of these findings are discussed along with study limitations and future research directions.


Viral etiologies of lower respiratory tract infections among Egyptian children under five years of age.
Lower respiratory tract infections (LRTI) are responsible for a considerable number of deaths among children, particularly in developing countries. In Egypt and the Middle East region, there is a lack of data regarding the viral causes of LRTI. In this study, we aimed to identify the relative prevalence of various respiratory viruses that contribute to LRTIs in young children. Although, nucleic acid-based methods have gained importance as a sensitive tool to determine the viral infections, their use is limited because of their prohibitive cost in low-income countries. Therefore, we applied three different laboratory methods, and presented the different virus prevalence patterns detected by each method. We collected nasopharyngeal aspirate samples, demographic data and, clinical data from 450 children under five years of age who presented with LRTI at Abou El Reesh hospital in Cairo during a one-year period. To identify the viral causes of the LRTI we used direct fluorescence assay, real-time reverse-transcriptase polymerase chain reaction (rt-RT-PCR), and shell vial culture. We tested for eight major respiratory viruses. Two hundred sixty-nine patients (59.9%) had a viral infection, among which 10.8% had a co-infection with two or more viruses. By all three methods, respiratory syncytial virus (RSV) was the most predominant, and parainfluenza virus type 2 (HPIV-2), influenza B virus (FLUBV) were the least predominant. Other viral prevalence patterns differed according to the detection method used. The distribution of various viruses among different age groups and seasonal distribution of the viruses were also determined. RSV and human adenovirus were the most common respiratory viruses detected by rt-RT-PCR. Co-infections were found to be frequent among children and the vast majority of co-infections were detected by nucleic acid-based detection assays.


Systematic observation of physical activity in afterschool programs: preliminary findings from Movin' Afterschool intervention.
National and state organizations have called upon afterschool programs (3-6 PM, ASP) to promote physical activity (PA). Few strategies exist that ASPs can use to increase the PA of children enrolled. This study evaluated a policy-level intervention (Movin' Afterschool, MAS) designed to increase PA through staff implemented policy-level changes and ongoing technical support. Twelve preexisting community-based ASPs serving 580 children (5-12 yrs, 57% girls) were invited to take part in MAS. Evaluation of children's PA, staff behaviors (engaged or promote PA, other ASP tasks, general supervising), and environmental features (equipment, organized PA) at baseline (Fall 2010) and postassessment (Spring 2011) were collected using SOPLAY (System for Observing Play and Leisure Activity in Youth) for boys and girls, separately. Random effects models evaluated changes in PA categories (sedentary, walking, vigorous). The percentage of boys and girls sedentary decreased by 11.8% and 11.4%, respectively. Girls' walking increased by 6.9% while boys' vigorous PA increased by 6.5%. Greater increases in vigorous activity were observed as postassessment in organized activities for boys and during indoor activities for girls. Findings indicate a policy-level approach targeting staff training and ongoing technical support can produce notable increases in PA within the ASP setting.


Symptoms of anxiety and depression in a volunteer twin population. The etiologic role of genetic and environmental factors.
We examined the etiologic role of genetic and environmental factors in 14 symptoms of anxiety and depression reported by 3,798 pairs of adult twins from the Australian National Health and Medical Research Council Twin Register. Multifactorial multiple-threshold models fit the individual symptom scores well. For a substantial majority of the symptoms, the variance in liability was best explained by only genetic factors and environmental influences specific to the individual, where 33% to 46% of the variance was due to genetic factors. For four symptoms, it was not possible to choose definitively between models that, in addition to specific environment, included genetic vs familial environmental effects. These results provide strong evidence for the role of genetic factors in the etiology of symptoms of anxiety and depression as reported in a general population. Evidence for an etiologic role of familial environmental factors was much weaker. If familial environmental factors play any role in the production of these symptoms, they are more important in symptoms of depression than of anxiety, and the factors that predispose to these symptoms are only modestly correlated in males and females.


Do racial and ethnic differences in contraceptive attitudes and knowledge explain disparities in method use?
Sustained efforts have not attenuated racial and ethnic disparities in unintended pregnancy and effective contraceptive use in the United States. The roles of attitudes toward contraception, pregnancy and fertility remain relatively unexplored. Knowledge of contraceptive methods and attitudes about contraception, pregnancy, childbearing and fertility were assessed among 602 unmarried women aged 18-29 at risk for unintended pregnancy who participated in the 2009 National Survey of Reproductive and Contraceptive Knowledge. The contribution of attitudes to racial and ethnic disparities in effective method use was assessed via mediation analysis, using a series of regression models. Blacks and Latinas were more likely than whites to believe that the government encourages contraceptive use to limit minority populations (odds ratio, 2.5 for each). Compared with white women, Latinas held more favorable attitudes toward pregnancy (2.5) and childbearing (coefficient, 0.3) and were more fatalistic about the timing of pregnancy (odds ratio, 2.3); blacks were more fatalistic about life in general (2.0). Only one attitude, skepticism that the government ensures contraceptive safety, was associated with contraceptive use (0.7), but this belief did not differ by race or ethnicity. Although blacks and Latinas used less effective methods than whites (0.3 and 0.4, respectively), attitudes did not explain disparities. Lower contraceptive knowledge partially explained Latinas' use of less effective methods. Providing basic information about effective methods might help to decrease ethnic disparities in use. Research should examine other variables that might account for these disparities, including health system characteristics and provider behavior.


Uninterrupted rivaroxaban vs. uninterrupted vitamin K antagonists for catheter ablation in non-valvular atrial fibrillation.
VENTURE-AF is the first prospective randomized trial of uninterrupted rivaroxaban and vitamin K antagonists (VKAs) in patients with non-valvular atrial fibrillation (NVAF) undergoing catheter ablation (CA). Trial size was administratively set at 250, the protocol-specified target. Events were independently and blindly adjudicated. We randomly assigned 248 NVAF patients to uninterrupted rivaroxaban (20 mg once-daily) or to an uninterrupted VKA prior to CA and for 4 weeks afterwards. The primary endpoint was major bleeding events after CA. Secondary endpoints included thromboembolic events (composite of stroke, systemic embolism, myocardial infarction, and vascular death) and other bleeding or procedure-attributable events. Patients were 59.5 ± 10 years of age, 71% male, 74% paroxysmal AF, and had a CHA2DS2-VASc score of 1.6. The average total heparin dose used to manage activated clotting time (ACT) was slightly higher (13 871 vs. 10 964 units; P < 0.001) and the mean ACT level attained slightly lower (302 vs. 332 s; P < 0.001) in rivaroxaban and VKA arms, respectively. The incidence of major bleeding was low (0.4%; 1 major bleeding event). Similarly, thromboembolic events were low (0.8%; 1 ischemic stroke and 1 vascular death). All events occurred in the VKA arm and all after CA. The number of any adjudicated events (26 vs. 25), any bleeding events (21 vs. 18), and any other procedure-attributable events (5 vs. 5) were similar. In patients undergoing CA for AF, the use of uninterrupted oral rivaroxaban was feasible and event rates were similar to those for uninterrupted VKA therapy. Clinicaltrials.gov trial registration number is NCT01729871.


Climate Change and Health on the U.S. Gulf Coast: Public Health Adaptation is Needed to Address Future Risks.
The impacts of climate change on human health have been documented globally and in the United States. Numerous studies project greater morbidity and mortality as a result of extreme weather events and other climate-sensitive hazards. Public health impacts on the U.S. Gulf Coast may be severe as the region is expected to experience increases in extreme temperatures, sea level rise, and possibly fewer but more intense hurricanes. Through myriad pathways, climate change is likely to make the Gulf Coast less hospitable and more dangerous for its residents, and may prompt substantial migration from and into the region. Public health impacts may be further exacerbated by the concentration of people and infrastructure, as well as the region's coastal geography. Vulnerable populations, including the very young, elderly, and socioeconomically disadvantaged may face particularly high threats to their health and well-being. This paper provides an overview of potential public health impacts of climate variability and change on the Gulf Coast, with a focus on the region's unique vulnerabilities, and outlines recommendations for improving the region's ability to minimize the impacts of climate-sensitive hazards. Public health adaptation aimed at improving individual, public health system, and infrastructure resilience is urgently needed to meet the challenges climate change may pose to the Gulf Coast in the coming decades.


Collective electric and magnetic plasmonic resonances in spherical nanoclusters.
We report an investigation on the optical properties of three-dimensional nanoclusters (NCs) made by spherical constellations of metallic nanospheres arranged around a central dielectric sphere, which can be realized and assembled by current state-of-the-art nanochemistry techniques. This type of NCs supports collective plasmon modes among which the most relevant are those associated with the induced electric and magnetic resonances. Combining a single dipole approximation for each nanoparticle and the multipole spherical-wave expansion of the scattered field, we achieve an effective characterization of the optical response of individual NCs in terms of their scattering, absorption, and extinction efficiencies. By this approximate model we analyze a few sample NCs identifying the electric and magnetic resonance frequencies and their dependence on the size and number of the constituent nanoparticles. Furthermore, we discuss the effective electric and magnetic polarizabilities of the NCs, and their isotropic properties. A homogenization method based on an extension of the Maxwell Garnett model to account for interaction effects due to higher order multipoles in dense packed arrays is applied to a distribution of NCs showing the possibility of obtaining metamaterials with very large, small, and negative values of permittivity and permeability, and even negative index.


ESX-1-induced apoptosis is involved in cell-to-cell spread of Mycobacterium tuberculosis.
Apoptosis modulation is a procedure amply utilized by intracellular pathogens to favour the outcome of the infection. Nevertheless, the role of apoptosis during infection with Mycobacterium tuberculosis, the causative agent of human tuberculosis, is subject of an intense debate and still remains unclear. In this work, we describe that apoptosis induction in host cells is clearly restricted to virulent M. tuberculosis strains, and is associated with the capacity of the mycobacteria to secrete the 6 kDa early secreted antigenic target ESAT-6 bothunder in vitro and in vivo conditions. Remarkably, only apoptosis-inducing strains are able to propagate infection into new cells, suggesting that apoptosis is used by M. tuberculosis as a colonization mechanism. Finally, we demonstrate that in vitro modulation of apoptosis affects mycobacterial cell-to-cell spread capacity, establishing an unambiguous relationship between apoptosis and propagation of M. tuberculosis. Our data further indicate that BCG and MTBVAC vaccines are inefficient in inducing apoptosis and colonizing new cells, correlating with the strong attenuation profile of these strains previously observed in vitro and in vivo.


Dietary zinc-deficiency and its recovery responses in the thermogenesis of rats.
The aim of this study was to elucidate whether dietary zinc-deficiency and its recovery play a role in controlling autonomic thermoregulation. We investigated the effects of dietary zinc-deficiency and its recovery on autonomic thermoregulation by measuring the rectal temperature, an index of deep body temperature. The weaned male Sprague Dawley rats were randomly divided into the dietary zinc-deficient diet (0.6 mg zinc/kg diet) group and the control diet (35.2 mg zinc/kg diet) group, and were fed for 4 weeks. In the recovery period, the rats of two groups were fed with the control diet for 3 weeks. The rectal temperature was significantly decreased throughout the period of zinc-deficiency and the hypothermic responses during the experimental period were recovered to the control group levels at least within 1 week in the recovery process from dietary zinc-deficiency. These results suggest that the deep temperature in rats was clearly associated with the dietary zinc intake levels.


HPMV: human protein mutation viewer - relating sequence mutations to protein sequence architecture and function changes.
Next-generation sequencing advances are rapidly expanding the number of human mutations to be analyzed for causative roles in genetic disorders. Our Human Protein Mutation Viewer (HPMV) is intended to explore the biomolecular mechanistic significance of non-synonymous human mutations in protein-coding genomic regions. The tool helps to assess whether protein mutations affect the occurrence of sequence-architectural features (globular domains, targeting signals, post-translational modification sites, etc.). As input, HPMV accepts protein mutations - as UniProt accessions with mutations (e.g. HGVS nomenclature), genome coordinates, or FASTA sequences. As output, HPMV provides an interactive cartoon showing the mutations in relation to elements of the sequence architecture. A large variety of protein sequence architectural features were selected for their particular relevance to mutation interpretation. Clicking a sequence feature in the cartoon expands a tree view of additional information including multiple sequence alignments of conserved domains and a simple 3D viewer mapping the mutation to known PDB structures, if available. The cartoon is also correlated with a multiple sequence alignment of similar sequences from other organisms. In cases where a mutation is likely to have a straightforward interpretation (e.g. a point mutation disrupting a well-understood targeting signal), this interpretation is suggested. The interactive cartoon can be downloaded as standalone viewer in Java jar format to be saved and viewed later with only a standard Java runtime environment. The HPMV website is: http://hpmv.bii.a-star.edu.sg/ .


Allozyme diversity in Macbridea alba (Lamiaceae), an endemic florida mint.
Macbridea alba is a herbaceous perennial mint endemic to the panhandle region of Florida. We used starch gel electrophoresis to describe allozyme diversity and genetic structure in this federally threatened plant. Ten populations were analyzed, with an average sample size of 47 plants (range 41-48 plants) per population. Of the 22 loci analyzed, 11 (50%) were polymorphic, with an average of 36.2% of the loci polymorphic within populations. Gene diversity measures for the species (Hes = 0.121) and for the populations (Hep = 0.099) were slightly higher than means found for other surveyed endemic plants. Compared to nine previously analyzed woody mints, however, M. alba is genetically depauperate. Little genetic structure was evident within the species, with 92% of the total genetic variation found within populations. Genetic identities between population pairs were high (mean I = 0.98). The perennial life habit of M. alba and long-distance pollination by bumblebees may contribute to the maintenance of genetic diversity within this threatened species.


Pasteurized and nanofiltered, plasma-derived C1 esterase inhibitor concentrate for the treatment of hereditary angioedema.
Hereditary angioedema (HAE) is a relatively rare autosomal dominant disorder that is typically characterized by recurrent episodes of edema in various body locations. It is most commonly caused by an inherited deficiency of functionally active C1 esterase inhibitor (C1-INH). Replacement therapy with a human plasma-derived C1-INH concentrate is recommended for the treatment and prophylaxis of acute attacks of HAE due to C1-INH deficiency (HAE-C1-INH). This article will discuss the current therapies available for the treatment of HAE-C1-INH, latest treatment guidelines, results of several studies demonstrating the efficacy and safety of the plasma-derived, pasteurized and nanofiltered C1-INH concentrate Berinert(®) (CSL Behring GmBH, Marburg, Germany), and future perspectives for the treatment and management of HAE-C1-INH.


Magnetic resonance imaging in the evaluation of patients for percutaneous vertebroplasty.
Osteoporosis and osteoporotic compression fractures of the vertebral bodies are major health problems facing women and older people of both sexes. In the last several years, percutaneous vertebroplasty has been developed as a treatment for pain caused by vertebral body compression fractures and primary or metastatic neoplasms. A large part of the success of this procedure depends on correct patient selection. As such, magnetic resonance imaging (MRI) plays a vital role in this process. In this review, the clinical evaluation of patients considered for vertebroplasty, the role of MRI in the pretreatment process, the postvertebroplasty appearance of the spine on MRI, and the future applications, such as real-time guidance with MR imaging, will be discussed.


A new human acute monocytic leukemia cell line SHI-1 with t(6;11)(q27;q23), p53 gene alterations and high tumorigenicity in nude mice.
Human leukemia cell lines are of great value in leukemia research. Thus far 36 leukemia cell lines carrying the 11q23 translocation and MLL rearrangements, including two cell lines with t(6;11)(q27;q23) and an MLL-AF6 fusion gene have been described. We have established a new monocytic cell line with t(6;11), designated SHI-1, and herein describe its biological characteristics. Mononuclear cells isolated from the bone marrow of a patient with acute monocytic leukemia (AML-M5b) at relapse were inoculated and passaged by liquid culture. The biological features of the cell line were characterized by morphological assays, flow cytometry, cytogenetic analysis, reverse transcription polymerase chain reaction (RT-PCR), direct sequencing, fluorescence in situ hybridization (FISH), clonogenic culture, quantitative fluorescent PCR, zymography, short tandem repeating sequences-PCR (STR-PCR), multiplex-FISH (M-FISH), and tumorigenic capacity in nude mice. The SHI-1 cell line has been maintained in continuous culture without any external cytokines for three years. The morphology and immunoprofile of the cells show typical features of monocytic lineage. Karyotypic analysis demonstrated a t(6;11)(q27;q23) translocation accompanied by a deletion of 17p, which are the same abnormalities as were seen in the leukemia cells of this patient in relapse. The MLL-AF6 fusion transcript and the loss of one p53 allele were proven by chromosome painting, FISH and RT-PCR analysis in both SHI-1 cells and the primary leukemia cells. A point mutation of ATC-->ACC at codon 195 of exon 6 in another p53 allele was found by direct sequencing of DNA in SHI-1 cells as well as in the primary leukemia cells. Neither Epstein-Barr virus nor mycoplasma was detected in SHI-1 cells. Tumor masses were found in all sixteen mice 9-19 days after subcutaneous injection of SHI-1 cells. DNA fingerprinting confirmed the authenticity of the cell line. SHI-1 is a new monocytic leukemia cell line with the t(6;11) translocation, p53 gene alterations, and high tumorigenicity in nude mice. It could be a valuable tool in the study of leukemogenesis.


Plant virus particles carrying tumour antigen activate TLR7 and Induce high levels of protective antibody.
Induction of potent antibody is the goal of many vaccines targeted against infections or cancer. Modern vaccine designs that use virus-like particles (VLP) have shown efficacy for prophylactic vaccination against virus-associated cancer in the clinic. Here we used plant viral particles (PVP), which are structurally analogous to VLP, coupled to a weak idiotypic (Id) tumour antigen, as a conjugate vaccine to induce antibody against a murine B-cell malignancy. The Id-PVP vaccine incorporates a natural adjuvant, the viral ssRNA, which acts via TLR7. It induced potent protective anti-Id antibody responses in an in vivo mouse model, superior to the "gold standard" Id vaccine, with prevalence of the IgG2a isotype. Combination with alum further increased antibody levels and maintained the IgG2a bias. Engagement of TLR7 in vivo was followed by secretion of IFN-α by plasmacytoid dendritic cells and by activation of splenic CD11chi conventional dendritic cells. The latter was apparent from up-regulation of co-stimulatory molecules and from secretion of a wide range of inflammatory cytokines and chemokines including the Th1-governing cytokine IL-12, in keeping with the IgG2a antibody isotype distribution. PVP conjugates are a novel cancer vaccine design, offering an attractive molecular form, similar to VLP, and providing T-cell help. In contrast to VLP, they also incorporate a safe "in-built" ssRNA adjuvant.


Structure and dynamics of aquaporin-1.
Structural response of a AQP1 is examined by a coarse-grained model with a phenomenological interaction potential with a knowledge-based residue-residue interaction (derived from an ensemble of protein structures in PDB). The thermal response of the protein chain exhibits an unexpected characteristics in its native phase where the radius of gyration of the protein decreases on raising the temperature. The radius of gyration of AQP1 increases on increasing the temperature before saturating to a random-coil morphology in denatured phase at high temperatures. Three regions of persistent globularization are identified, toward the end segments 1M-25V and 250V-269K and a narrow region in the middle 155A-163D along the backbone. Varying the temperature leads to a systematic redistribution of self-organizing residues with globular and fibrous morphologies with an effective dimension D~2 (random coil) at high temperature and D~3 (globular conformation) in native phase. A preliminary analysis is also presented on the effect of a crowded membrane environment on the protein structure by incorporating effective solute constituents. Conformation of the protein is found to be pinned by selective binding of solute to specific targets; the matrix directed structure differs considerably from that of a protein in a generic solvent. The structure of AQP1 can be controlled by temperature and constitutive elements of the underlying matrix.


Prevalence of subdural collections in children with macrocrania.
The relationship between enlarged subarachnoid spaces and subdural collections is poorly understood and creates challenges for clinicians investigating the etiology of subdural collections. The purpose of this study was to determine the prevalence of subdural collections on cross sectional imaging in children with macrocephaly correlating with subarachnoid space enlargement. The radiology information system of a large pediatric medical center was reviewed for "macrocrania" and "macrocephaly" on reports of cranial MRI/CT examinations in children <24 months of age, over a 24-month period. Head circumference was obtained from the clinical record. Studies were reviewed blindly for subdural collection presence and subarachnoid space size. Children with prior cranial surgery, parenchymal abnormalities, hydrocephalus, or conditions predisposing to parenchymal volume loss were excluded. Chart review was performed on those with subdural collections. Imaging from 177 children with enlarged head circumference was reviewed. Nine were excluded, for a final cohort of 168 subjects (108 with enlarged subarachnoid space). Subdural collections were identified in 6 (3.6%), all with enlarged subarachnoid space (6/108, 5.6%). In 4, subdural collections were small, homogeneous, and nonhemorrhagic. In 2, the collections were complex (septations or hemorrhage). Two children were reported as victims of child abuse (both with complex collections). No definitive etiology was established in the other cases. The prevalence of subdural collections in imaged children with macrocrania was 3.6%, all occurring in children with enlarged subarachnoid space. Our results suggest that enlarged subarachnoid space can be associated with some subdural collections in this cohort. Despite this, we believe that unexpected subdural collections in children should receive close clinical evaluation for underlying causes, including abusive head trauma.


Complete Reduction for Pilon Fracture Can Make Complete Failure.
The surgical management of distal intra-articular comminuted fracture of the tibia (pilon fracture) is difficult because complications frequently develop. The minimally invasive plate osteosynthesis technique is generally accepted for this type of fracture. In this study, complications developed after open reduction and internal fixation using multiple miniplates for accurate reduction of small fracture fragments. Therefore, when we use this technique, we need to pay attention to the development of complications such as nonunion, avascular necrosis, and osteomyelitis by the disruption of both endosteal blood supply by fracture and periosteal blood supply during approach or reduction.


[Study of the Mechanisms of Maintaining the Transparency of the Lens and Treatment of Its Related Diseases for Making Anti-cataract and/or Anti-presbyopia Drugs].
The lens of the eye is an avascular and anuclear tissue that serves to focus objects on the retina. Cataract is opacity within the clear lens that changes the transparency and refractive index of the lens causing significant visual impairments. These impairments can severely restrict the ability to carry out daily activities. Cataracts is common among elderly person occurring in more than 80% of patients aged 80 or older. Notably, we have recently identified key compounds that are effective against cataract formation. Presbyopia is also an ocular disease that typically develops in people over the age of 45 while affecting almost 100% of people over the age of 65. Recent research suggests that age-related changes in hydrostatic pressure of the lens controlled by Na/K ATPase contribute to the development of presbyopia. In the lens, Na/K ATPase has been shown to be regulated by transient receptor potential cation channels, vanilloid 1 (TRPV1) and 4, thus suggesting the potential role of TRPV1 and TRPV4 in the development of presbyopia. This review article summarizes data obtained from our laboratory with my colleagues highlighting the critical role of aquaporin 0 (AQP0) in maintaining a healthy lens redox environment, key molecules that delay the onset of cataract in vivo, as well as potential mechanisms of lens hydrostatic pressure control that may be associated with presbyopia.


Assembly and channel opening of outer membrane protein in tripartite drug efflux pumps of Gram-negative bacteria.
Gram-negative bacteria are capable of expelling diverse xenobiotic substances from within the cell by use of three-component efflux pumps in which the energy-activated inner membrane transporter is connected to the outer membrane channel protein via the membrane fusion protein. In this work, we describe the crystal structure of the membrane fusion protein MexA from the Pseudomonas aeruginosa MexAB-OprM pump in the hexameric ring arrangement. Electron microscopy study on the chimeric complex of MexA and the outer membrane protein OprM reveals that MexA makes a tip-to-tip interaction with OprM, which suggests a docking model for MexA and OprM. This docking model agrees well with genetic results and depicts detailed interactions. Opening of the OprM channel is accompanied by the simultaneous exposure of a protein structure resembling a six-bladed cogwheel, which intermeshes with the complementary cogwheel structure in the MexA hexamer. Taken together, we suggest an assembly and channel opening model for the MexAB-OprM pump. This study provides a better understanding of multidrug resistance in Gram-negative bacteria.


Sequence-specific intramembrane proteolysis: identification of a recognition motif in rhomboid substrates.
Members of the widespread rhomboid family of intramembrane proteases cleave transmembrane domain (TMD) proteins to regulate processes as diverse as EGF receptor signaling, mitochondrial dynamics, and invasion by apicomplexan parasites. However, lack of information about their substrates means that the biological role of most rhomboids remains obscure. Knowledge of how rhomboids recognize their substrates would illuminate their mechanism and might also allow substrate prediction. Previous work has suggested that rhomboid substrates are specified by helical instability in their TMD. Here we demonstrate that rhomboids instead primarily recognize a specific sequence surrounding the cleavage site. This recognition motif is necessary for substrate cleavage, it determines the cleavage site, and it is more strictly required than TM helix-destabilizing residues. Our work demonstrates that intramembrane proteases can be sequence specific and that genome-wide substrate prediction based on their recognition motifs is feasible.


Impossibility of coexistence of three pure and simple competitors in configurations of three interconnected chemostats.
It is well established that pure and simple microbial competitors cannot coexist at a steady state if their environment is homogeneous. For the case of two microbial populations competing purely and simply in two interconnected chemostats having time-invariant input(s), it is known from the literature that a stable steady state of coexistence arises in domains of the operating parameters space and is attributed to the spatial heterogeneities of the system, which allow a different species to have the competitive advantage in each one of the two sub-environments. This article investigates whether the aforementioned result can be extended to the case of three species competing in three interconnected vessels. By studying all possible alternate configurations of the three-chemostat system, it is shown that coexistence of the three species is impossible, except possibly for some discrete values of the operating parameters. Some potential explanations for the results are discussed.


Protective effect of neonatal BCG vaccines against tuberculous meningitis.
Neonatal BCG vaccination reduces the risk of tuberculosis and provides protection higher than 80% against the development of meningeal and miliary tuberculosis in newborns. Tuberculosis meningitis remains a major problem and also an important cause of death in some countries. In countries with high and moderate incidence of tuberculosis, prevention from the most severe complications of tuberculosis can be achieved only with a high coverage of the universal BCG neonatal immunization, being higher than 98% in the cohort of newborns. The decrease in BCG immunization coverage within immunization program during the year 2003 in Bosnia and Herzegovina influenced the increase in tuberculous meningitis. During 2002, when coverage with BCG vaccination in cohort of newborns was 90%, the incidence rate of tuberculous meningitis was 19. 04%oo. With the 68% decrease in BCG immunization coverage in the cohort of newborns in Bosnia and Herzegovina during the year 2003, the incidence of tuberculous meningitis raised to 33 33%oo. It has been proven that the 22% decrease of the neonatal BCG immunization coverage in the cohort of newborns /vaccination program of children/ caused 175 times higher number of the tuberculous meningitis cases. Newborns affected by the tuberculous meningitis were not BCG vaccinated. BCG vaccine provided effective protection against tuberculous meningitis, as well against the death of newborns caused by tuberculosis.


Updated European Association of Urology Guidelines on Renal Cell Carcinoma: Immune Checkpoint Inhibition Is the New Backbone in First-line Treatment of Metastatic Clear-cell Renal Cell Carcinoma.
Recent randomised trials have demonstrated a survival benefit for a front-line ipilimumab and nivolumab combination therapy, and pembrolizumab and axitinib combination therapy in metastatic clear-cell renal cell carcinoma. The European Association of Urology Guidelines Panel has updated its recommendations based on these studies. PATIENT SUMMARY: Pembrolizumab plus axitinib is a new standard of care for patients diagnosed with kidney cancer spread outside the kidney and who did not receive any prior treatment for their cancer (treatment naïve). This applies to all risk groups as determined by the International Metastatic Renal Cell Carcinoma Database Consortium criteria.