Approximate methods for modeling the scattering properties of nonspherical particles: evaluation of the Wentzel-Kramers-Brillouin method.
Several approximate methods for modeling the electromagnetic (em) scattering properties of nonspherical particles are examined and evaluated. Although some of the approaches are applicable to arbitrary shapes we confine our attention here mainly to spheres and cylinders, for which exact solutions are available for comparisons. Evaluations include comparisons of the computed angular phase function, total extinction efficiency, and backscatter efficiency. Approximate methods investigated include the Rayleigh-Gans (RG) approximation, the Wentzel-Kramers-Brillouin or WKB approximation [and the closely related eikonal approximation (EA)], diffraction theory, and the second-order Shifrin iterative technique. Examples using spheres indicate that for weakly absorbing particles of moderate- to large-size parameters with a real refractive index near unity (i.e., the optically soft case), all models work well in representing the phase function over all scattering angles, with the Shifrin approximation showing the best agreement with the exact solutions. For larger refractive indices, however, the Shifrin approximation breaks down, whereas the WKB method continues to perform relatively well for all scattering angles over a wide range of particle sizes, including those appropriate in both the RG (small particle) and the diffraction (large particle) limits. The relationship between the WKB, eikonal, and anomalous diffraction descriptions of particle extinction is discussed briefly. Backscatter is also discussed in the context of the WKB model, and two modifications to improve the description are included: one to add an internal-reflected internal wave and the other to add a multiplicative scaling factor to preserve the correct backscatter result for strong absorption in the geometric optics limit. A major conclusion of the paper is that the WKB method offers a viable alternative to the more widely used RG and diffraction approximations and is a method that offers significant improvement in accuracy with only a slight increase in mathematical complexity.


Search for meaning, finding meaning and adjustment in women following miscarriage: a longitudinal study.
The purpose of this study was to explore the associations between meaning making and psychological adjustment in 127 women who experienced a miscarriage. A longitudinal and controlled study design was employed. Meaning-making variables and distress outcomes were examined at four, seven and 16 weeks after miscarriage, in two groups of women, one who had medical investigations of the cause of their loss, and a control group receiving standard care. Search for meaning was very common and it declined with time after miscarriage. By seven weeks post-loss, more than half the women reported that they had found meaning/understood why the miscarriage happened. Providing information about the cause of the loss was associated with finding meaning. A decline in the search for meaning and finding meaning at seven weeks post-miscarriage, predicted levels of distress at 16 weeks, whilst controlling for the initial distress and for significant background factors. This study provides support for the notion that search for meaning is very common following negative life events, such as miscarriage, and that finding meaning is important in many peoples' process of adjustment. Providing information about the cause of the loss facilitates finding meaning.


Transcriptome Analysis and Development of SSR Molecular Markers in Glycyrrhiza uralensis Fisch.
Licorice is an important traditional Chinese medicine with clinical and industrial applications. Genetic resources of licorice are insufficient for analysis of molecular biology and genetic functions; as such, transcriptome sequencing must be conducted for functional characterization and development of molecular markers. In this study, transcriptome sequencing on the Illumina HiSeq 2500 sequencing platform generated a total of 5.41 Gb clean data. De novo assembly yielded a total of 46,641 unigenes. Comparison analysis using BLAST showed that the annotations of 29,614 unigenes were conserved. Further study revealed 773 genes related to biosynthesis of secondary metabolites of licorice, 40 genes involved in biosynthesis of the terpenoid backbone, and 16 genes associated with biosynthesis of glycyrrhizic acid. Analysis of unigenes larger than 1 Kb with a length of 11,702 nt presented 7,032 simple sequence repeats (SSR). Sixty-four of 69 randomly designed and synthesized SSR pairs were successfully amplified, 33 pairs of primers were polymorphism in in Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat., Glycyrrhiza glabra L. and Glycyrrhiza pallidiflora Maxim. This study not only presents the molecular biology data of licorice but also provides a basis for genetic diversity research and molecular marker-assisted breeding of licorice.


New Strategies to Enhance Myocardial Regeneration: Expectations and Challenges from Preclinical Evidence.
Nowadays, cardiac regeneration is an emerging topic in the cardiovascular field because of the compelling need for effective therapies for repairing or replacing cardiac tissue damaged by pathological or physiological conditions. Indeed, irreversible myocardial remodeling which follows acute myocardial infarction represents a serious burden of this century. In this context, a great improvement in pharmacological and interventional techniques is accompanied by a big challenge of cardiac regenerative medicine. In the last 20 years, several clinical trials tried to investigate the role of different types of stem cells in promoting cardiac repair. However, the promising results obtained in the preclinical trials have not yet been reproduced in patients. Thus, the development of novel strategies to improve stem cell efficiency became imperative. Here, an overview of the more recent cell types proposed for cardiac regeneration is presented, together with the most interesting approaches to enhance cell regenerative potential as well as cell-free approaches.


Tryptophan residues flanking the second transmembrane helix (TM2) set the signaling state of the Tar chemoreceptor.
The chemoreceptors of Escherichia coli are homodimeric membrane proteins that cluster in patches near the cell poles. They convert environmental stimuli into intracellular signals that control flagellar rotation. The functional domains of a receptor are physically separated by the cell membrane. Chemoeffectors bind to the extracellular (periplasmic) domain, and the cytoplasmic domain mediates signaling and adaptation. These two domains communicate through the second transmembrane helix (TM2) that connects them. In the high-abundance receptors Tar and Tsr, TM2 is flanked by tryptophan residues, which should localize preferentially to the interfacial zone between the polar and hydrophobic layers of the phospholipid bilayer. To investigate the functional significance of the Trp residues that flank TM2 of Tar, we used site-directed mutagenesis to generate the W192A and W209A substitutions. The W192A protein retains full activity in vivo and in vitro, but it increases the K(i) for aspartate in the in vitro assay 3-fold. The W209A replacement eliminates receptor-mediated stimulation of CheA in vitro, and it leads to an increased level of adaptive methylation in vivo. This phenotype in some respects mimics the changes seen upon binding aspartate. Since the W209A substitution may cause the C-terminus of TM2 to protrude farther into the cytoplasm, these results reinforce the hypothesis that aspartate binding causes a similar displacement. Moving Trp to each position from residue 206 to residue 212 generated a wide variety of Tar signaling states that are generally consistent with the predictions of the piston model of transmembrane signaling. None of these receptors was completely locked in one signaling mode, although most showed pronounced signaling biases. Our findings suggest that the Trp residues flanking TM2, especially Trp-209, are important in setting the baseline activity and ligand sensitivity of the Tar receptor. We also conclude that the Tyr-210 residue plays at least an auxiliary role in this control.


Reversible nanostructuration of microfluidic electrode devices by CNT magnetic co-entrapment.
Carbon nanotubes (CNTs) have been extensively used to produce electrodes of enhanced performance but have only been very recently exploited in microfluidic devices. In these cases, CNT electrodes had to be produced prior to device assembly, which might damage the CNT layer. Here, we show a fast and simple method for the reversible nanostructuration of microfluidic electrode devices in situ. The procedure is based on the attachment of single-walled CNTs (SWCNTs) onto the surface of magnetic particles (MPs) and magnetic confinement of the MP/SWCNT composite onto the sensor in a two-step process that provided homogeneous coating. As it is shown, subsequent magnet removal allows MP/SWCNT release and electrode reutilization. Compared to most previously described methods, ours is faster, simpler and also reversible.


Relationships between diurnal blood pressure variation, physical activity, and health-related QOL.
The aim of this study is to clarify the associations between diurnal blood pressure variation, physical activity and health-related quality of life (HRQOL). Ninety-seven volunteers, including 52 hypertensive patients and 45 healthy subjects (average age, 48 years) participated in this study. Twenty-four hour ambulatory blood pressure and heart rate variability were measured using TM2425 (A&D Co). Physical activity was measured using actigraphy, and HRQOL was assessed by a Medical Outcome Study Short-Forum 36-Item Health Survey (SF-36). Awake mean physical activity positively correlated with the nocturnal dip in systolic blood pressure (SBP) (r = 0.242, p < 0.02) and diastolic blood pressure (DBP) (r = 0.219, p < 0.04). The score of physical functioning positively correlated with awake mean physical activity (r = 0.265, p < 0.02). The score of role-emotional also correlated with awake mean physical activity (r = 0.269, p = 0.01). Using multiple regression analysis, the nocturnal dip in SBP was found to be correlated with awake and sleep mean physical activities (p < 0.05, p < 0.05, respectively). In conclusion, physical activity is associated with the nocturnal dip in blood pressure. Moreover, physical activity correlates with some of the factors of HRQOL.


Monitoring anticoagulant control in general practice: comparison of management in areas with and without access to hospital anticoagulant.
Hospital anticoagulant clinics are available only in some areas. There is little information on the contribution of general practitioners to oral anticoagulant monitoring, and whether their management varies with access to hospital clinics. A study was undertaken to compare general practice management of anticoagulant therapy in two health boards with contrasting access to hospital clinics. A postal questionnaire was sent to the senior partners in all 198 practices in Lothian and Fife, Scotland. Lack of access to hospital clinics in Lothian health board resulted in more practices reporting taking sole responsibility for anticoagulant control than in Fife where there was access (P < 0.001). However, there was no significant difference in management policies. Overall, 93% of practices used a protocol for thrombotest target ranges, but 75% had no policy on review frequency and only 2% assessed complication or failure rates. Reduced access to hospital clinics was associated with a decreased likelihood of favouring hospital involvement. Sole responsibility for anticoagulant management was undertaken by 56% of general practices, although only 21% of doctors viewed this as ideal. Most general practitioners felt that they should monitor patients on anticoagulant therapy but should refer to hospital those with problematic control. Access to hospital clinics affected the degree of involvement of general practitioners in oral anticoagulant monitoring, but did not considerably alter their management practice.


Identification of a putative glycosyltransferase responsible for the transfer of pseudaminic acid onto the polar flagellin of Aeromonas caviae Sch3N.
Motility in Aeromonas caviae, in a liquid environment (in broth culture), is mediated by a single polar flagellum encoded by the fla genes. The polar flagellum filament of A. caviae is composed of two flagellin subunits, FlaA and FlaB, which undergo O-linked glycosylation with six to eight pseudaminic acid glycans linked to serine and threonine residues in their central region. The flm genetic locus in A. caviae is required for flagellin glycosylation and the addition of pseudaminic acid (Pse) onto the lipopolysaccharide (LPS) O-antigen. However, none of the flm genes appear to encode a candidate glycotransferase that might add the Pse moiety to FlaA/B. The motility-associated factors (Maf proteins) are considered as candidate transferase enzymes, largely due to their conserved proximity to flagellar biosynthesis loci in a number of pathogens. Bioinformatic analysis performed in this study indicated that the genome of A. caviae encodes a single maf gene homologue (maf1). A maf mutant was generated and phenotypic analysis showed it is both nonmotile and lacks polar flagella. In contrast to flm mutants, it had no effect on the LPS O-antigen pattern and has the ability to swarm. Analysis of flaA transcription by reverse transcriptase PCR (RT-PCR) showed that its transcription was unaltered in the maf mutant while a His-tagged version of the FlaA flagellin protein produced from a plasmid was detected in an unglycosylated intracellular form in the maf strain. Complementation of the maf strain in trans partially restored motility, but increased levels of glycosylated flagellin to above wild-type levels. Overexpression of maf inhibited motility, indicating a dominant negative effect, possibly caused by high amounts of glycosylated flagellin inhibiting assembly of the flagellum. These data provide evidence that maf1, a pseudaminyl transferase, is responsible for glycosylation of flagellin and suggest that this event occurs prior to secretion through the flagellar Type III secretion system.


Homogenization of quasi-1d metamaterials and the problem of extended bandwidth.
We derive approximate analytical expressions for the effective permittivity tensor of two-phase metamaterials whose geometry is close to one-dimensional (quasi-one-dimensional metamaterials). Specifically, we consider the metamaterial made of parallel slabs with width given by a linear or parabolic function. Using our approach, the design of epsilon-near-zero, ultra-low and high refractive index metallodielectric metamaterials with extended bandwidth has been demonstrated. In addition, generalizations to the three-dimensional case and some limitations of the presented technique are briefly considered.


The prosociality of married people: Evidence from a large multinational sample.
Single people are more likely to die from COVID-19. Here we study whether this higher death rate could be partly explained by differences in compliance with protective health measures against COVID-19 between single and married people, and the drivers of this marital compliance gap. Data collected from 46,450 respondents in 67 countries reveal that married people are more likely to comply with protective measures than single people. This marital gap in compliance is higher for men (approximately 5%) than for women (approximately 2%). These results are robust across a large range of countries and independent of country level differences with respect to culture, values or infection rates. Prosocial characteristics linked to morality and social belonging explain more than 38% of the marital gap, while individual risk perceptions play a minor role. These findings help explain single people's and particularly single men's greater vulnerability to COVID-19, which in turn can be leveraged to improve the effectiveness of international public policy campaigns aimed at promoting protective health measures.


[Wernicke encephalopathy and dry beriberi; late complications after bariatric surgery performed on a patient with a psychiatric history].
A 38-year-old woman was admitted in a psychiatric hospital because of memory disorders. Dry beriberi and Korsakoff's syndrome, as a late neuropsychiatric manifestation of Wernicke's encephalopathy, were established. The beriberi symptoms decreased slowly, but the patient's memory impairments persisted. These clinical features were caused by a thiamine deficiency. Gastric bypass surgery is associated with an increased risk of thiamine deficiency. Excessive alcohol consumption and poor compliance with vitamin deficiency treatment both contributed to the rapid onset of these disorders. The case we describe demonstrates how important it is that patients who have undergone bariatric surgery are prescribed the correct vitamin supplements and that the treatment is continued for an appropriate period of time; this treatment is particularly important in the case of patients who are also suffering from an comorbid alcohol-related disorder.


The rural mother's experience of caring for a child with a chronic health condition: An integrative review.
To identify and review the literature on rural mothers' experiences in caring for a child with a chronic health condition. Families living with a child who has a chronic health condition experience many challenges; these are often amplified for families living in rural areas, where issues such as the distance from services add further challenges the family must manage. Like many children, rural children with chronic health conditions are primarily cared for by their mothers. The additional strain of geography creates its own unique experiences for mothers who need to access the high-quality care that their child requires. Integrative literature review using the Equator PRISMA guidelines. A search of databases; Cochrane, CINAHL, Ovid, PubMed, ProQuest Health and Medicine, Informit and Scopus for studies published between 2005-2016 using an integrative review approach. A total of 1,484 studies were identified with an additional six studies found through snowballing. The search resulted in seven studies being meeting the inclusion criteria after using the Critical Appraisal Skills Programme. Data from the seven articles were analysed, and the mothers' experiences were synthesised into five themes: "struggling for resources," "barriers in accessing services," "strain of decision-making," "mother's physical and emotional breakdown" and "the daily management of family activities". These five themes formed the basis of this article. The findings indicate that mothers from rural areas face additional barriers related to their rurality, including transportation difficulties, socioeconomic status and social isolation, and are challenged by limited access to specialty medical services, educators and allied health professionals. The literature review outcome will assist in informing nursing practice through identifying and allocating resources to reduce these barriers; rural mother experience will assist in enabling the child to reach their full developmental potential. There is a need for health professionals to understand the challenges and barriers rural mothers face in accessing services. Nurses can assist rural mothers to navigate and access the appropriate services in order to reduce health inequity, increase accessibility to services and reduce rural disadvantage for their child. Nurses and health professionals are in an ideal position to develop future models of care that optimise health outcomes and enable equity and access to services for rural children with chronic conditions similar to those experienced by their urban counterparts.


Ophthalmologic outcome at 30 months' corrected age of a prospective Swedish cohort of children born before 27 weeks of gestation: the extremely preterm infants in sweden study.
Follow-up at 30 months' corrected age reveals eye and visual problems in one-third of children born extremely prematurely (<27 weeks' gestation). To investigate the ophthalmologic outcome of extremely preterm children at 30 months' corrected age. DESIGN, SETTING, AND PARTICIPANTS A prospective, population-based follow-up study (Extremely Preterm Infants in Sweden Study [EXPRESS]) was conducted in Sweden. The population included extremely preterm infants (<27 weeks' gestation) born in Sweden between 2004 and 2007, of whom 491 survived until age 2.5 years. Screening for retinopathy of prematurity (ROP) was performed in the neonatal period. At 30 months' corrected age, an ophthalmologic assessment was performed in 411 of 491 children (83.7%). Visual acuity, manifest strabismus, and refractive errors were evaluated. Visual impairment was identified in 3.1% of the children, and 1.0% were blind. Refractive errors, defined as myopia less than -3 diopters (D), hypermetropia greater than +3 D, astigmatism 2 D or more, and/or anisometropia 2 D or more, were found in 25.6% of the children, and 14.1% had manifest strabismus. There were significant associations between visual impairment and treated ROP (P = .02), cognitive disability (P < .001), and birth weight (P = .02). Multiple regression analyses revealed significant associations between strabismus and treated ROP (P < .001), cognitive disability (P < .01), and cerebral palsy (P = .02). Refractive errors were significantly correlated with severity of ROP (right eye, P < .001; left eye, P < .01). Children who had been treated for ROP had the highest frequency (69.0%) of eye and visual abnormalities. One-third of the extremely prematurely born children in this study had some kind of eye or visual problems, such as visual impairment, strabismus, or major refractive error. Despite being born extremely preterm, the present cohort has a similar prevalence of blindness and visual impairment as in previous Swedish cohorts of children born less prematurely.


Automation to optimise physician treatment of individual patients: examples in psychiatry.
There is widespread agreement by health-care providers, medical associations, industry, and governments that automation using digital technology could improve the delivery and quality of care in psychiatry, and reduce costs. Many benefits from technology have already been realised, along with the identification of many challenges. In this Review, we discuss some of the challenges to developing effective automation for psychiatry to optimise physician treatment of individual patients. Using the perspective of automation experts in other industries, three examples of automation in the delivery of routine care are reviewed: (1) effects of electronic medical records on the patient interview; (2) effects of complex systems integration on e-prescribing; and (3) use of clinical decision support to assist with clinical decision making. An increased understanding of the experience of automation from other sectors might allow for more effective deployment of technology in psychiatry.


Characterization of purified intraembryonic hematopoietic stem cells as a tool to define their site of origin.
Little is known about hematopoietic stem cell (HSC) development from mesoderm. To gain more information on the intraembryonic HSC site of origin, we purified multipotent hematopoietic progenitors from the aorta-gonads-mesonephros (AGM) of mice. This population, expressing c-Kit, AA4.1, CD31, and CD41, but not Flk1, and mainly negative for CD45, proved capable of long-term reconstitution in sublethally irradiated Rag2gammac(-/-) recipients. We assigned the expression of GATA-2, GATA-3, and lmo2 to AGM-HSC, whereas erythromyeloid progenitors express only GATA-2. This unique combination of surface markers and transcription factors could be allocated in the AGM to the intraaortic clusters and the subaortic patches underlying aortic endothelial cells. Taken together, those data indicate that embryonic HSCs (i) differ from their fetal liver and adult counterpart by the low expression of CD45, (ii) do not colocalize with aortic endothelial cells as previously thought, and (iii) are localized, at 10.5 days postcoitum, in the splanchnic mesoderm underlying aortic endothelial cells, within GATA-3(+)CD31(+) cell clusters.


Relative contributions of neutral and non-neutral genetic differentiation to inform conservation of steelhead trout across highly variable landscapes.
Mounting evidence of climatic effects on riverine environments and adaptive responses of fishes have elicited growing conservation concerns. Measures to rectify population declines include assessment of local extinction risk, population ecology, viability, and genetic differentiation. While conservation planning has been largely informed by neutral genetic structure, there has been a dearth of critical information regarding the role of non-neutral or functional genetic variation. We evaluated genetic variation among steelhead trout of the Columbia River Basin, which supports diverse populations distributed among dynamic landscapes. We categorized 188 SNP loci as either putatively neutral or candidates for divergent selection (non-neutral) using a multitest association approach. Neutral variation distinguished lineages and defined broad-scale population structure consistent with previous studies, but fine-scale resolution was also detected at levels not previously observed. Within distinct coastal and inland lineages, we identified nine and 22 candidate loci commonly associated with precipitation or temperature variables and putatively under divergent selection. Observed patterns of non-neutral variation suggest overall climate is likely to shape local adaptation (e.g., potential rapid evolution) of steelhead trout in the Columbia River region. Broad geographic patterns of neutral and non-neutral variation demonstrated here can be used to accommodate priorities for regional management and inform long-term conservation of this species.


Targeting cancer with peptide aptamers.
A major endeavour in cancer chemotherapy is to develop agents that specifically target a biomolecule of interest. There are two main classes of targeting agents: small molecules and biologics. Among biologics (e.g.: antibodies), DNA, RNA but also peptide aptamers are relatively recent agents. Peptide aptamers are seldom described but represent attractive agents that can inhibit a growing panel of oncotargets including Heat Shock Proteins. Potential pitfalls and coming challenges towards successful clinical trials are presented such as optimizing the delivery of peptide aptamers thanks to Nanotechnology.


Murine coronavirus membrane fusion is blocked by modification of thiols buried within the spike protein.
The envelopes of murine hepatitis virus (MHV) particles are studded with glycoprotein spikes that function both to promote virion binding to its cellular receptor and to mediate virion-cell membrane fusion. In this study, the cysteine-rich spikes were subjected to chemical modification to determine whether such structural alterations impact the virus entry process. Ellman reagent, a membrane-impermeant oxidizing agent which reacts with exposed cysteine residues to effect covalent addition of large thionitrobenzoate moieties, was incubated at 37 degrees C with the JHM strain of MHV. Relative to untreated virus, 1 mM Ellman reagent reduced infectivity by 2 log(10) after 1 h. This level of inhibition was not observed at incubation temperatures below 21 degrees C, suggesting that virion surface proteins undergo thermal transitions that expose cysteine residues to modification by the reagent. Quantitative receptor binding and membrane fusion assays were developed and used to show that Ellman reagent specifically inhibited membrane fusion induced by the MHV JHM spike protein. However, this inhibition was strain specific, because the closely related MHV strain A59 was unaffected. To identify the basis for this strain specificity, spike cDNAs were prepared in which portions encoded either JHM or A59 residues. cDNAs were expressed with vaccinia virus vectors and tested for sensitivity to Ellman reagent in the fusion assays. The results revealed a correlation between the severity of inhibition mediated by Ellman reagent and the presence of a JHM-specific cysteine (Cys-1163). Thus, the presence of this cysteine increases the availability of spikes for a thiol modification that ultimately prevents fusion competence.


Non-coding variability at the APOE locus contributes to the Alzheimer's risk.
Alzheimer's disease (AD) is a leading cause of mortality in the elderly. While the coding change of APOE-ε4 is a key risk factor for late-onset AD and has been believed to be the only risk factor in the APOE locus, it does not fully explain the risk effect conferred by the locus. Here, we report the identification of AD causal variants in PVRL2 and APOC1 regions in proximity to APOE and define common risk haplotypes independent of APOE-ε4 coding change. These risk haplotypes are associated with changes of AD-related endophenotypes including cognitive performance, and altered expression of APOE and its nearby genes in the human brain and blood. High-throughput genome-wide chromosome conformation capture analysis further supports the roles of these risk haplotypes in modulating chromatin states and gene expression in the brain. Our findings provide compelling evidence for additional risk factors in the APOE locus that contribute to AD pathogenesis.


Beta cell mass and growth after syngeneic islet cell transplantation in normal and streptozocin diabetic C57BL/6 mice.
In islet transplantation, nonimmunological factors such as limited growth capacity or increased death rate could reduce the beta cell mass in the graft and lead to failure of the transplant. We studied the evolution of beta cell replication and mass after transplantation of insufficient, minimally sufficient, or excessive islet tissue. Streptozocin diabetic C57BL/6 mice received 150 or 300 syngeneic islets under the kidney capsule and normal mice received 300 islets. In streptozocin diabetic mice 300 islets restored normoglycemia; beta cell replication in transplanted islets was similar to replication in normal pancreas and beta cell mass in the graft remained constant. In contrast, 150 islets were insufficient to achieve normoglycemia; beta cell replication was increased initially but not by 18 or 30 d despite persistent hyperglycemia, and beta cell mass fell progressively. When islets were transplanted into normal recipients, beta cell replication remained normal but beta cells underwent atrophy and mass in the graft was substantially reduced. Therefore, with a successful islet transplant, in diabetic mice beta cell replication and mass remain constant. In contrast, when insufficient islet tissue is transplanted an initial increase in beta cell replication can not compensate for a decline in beta cell mass. When excessive islet tissue is transplanted, beta cell mass is reduced despite normal beta cell replication.


Management of a composite foot defect due to mine explosion using the free fibula osteocutaneous flap.
Free flaps have recently become the first preference for complex foot defects, with many advantages including ability to present suitable and adequate tissue, to enhance blood flow of the extremity, and to decrease risk of osteomyelitis. A 25-year-old male patient was referred to us with a complex injury of the left foot due to mine explosion. We successfully restored the defect with the ipsilateral free vascularized fibula osteocutaneous flap. The flap survived completely. After 8 weeks postoperatively, the patient began to walk. An adequate foot contour was achieved, and both the transverse and longitudinal arches were reestablished. Recovery of the donor site was also good. The free fibula osteocutaneous flap was able to present a composite tissue required in a defective foot, by leaving single donor site morbidity in one session. We believe that this flap may be a preferred option in complex foot defects.


HGF/c-Met pathway has a prominent role in mediating antiapoptotic signals through AKT in epithelial ovarian carcinoma.
The Met receptor tyrosine kinase and its ligand, hepatocyte growth factor (HGF), are overexpressed and/or activated in a variety of human malignancies. However, its role in epithelial ovarian carcinoma (EOC) has not been clearly elucidated. Therefore, we investigated the role of HGF/c-Met signaling pathway in a large series (156) of Saudi EOC patient samples, a panel of cell lines, and xenografts in a NUDE mouse model. Using immunohistochemistry, c-Met overexpression was found in 27.2% Middle Eastern EOC samples and was associated with an advanced tumor stage (P=0.0187). c-Met overexpression was also associated with antiapoptotic markers X-chromosome-linked inhibitors of apoptosis (XIAP) (P=0.0008) and Bcl-XL (P=0.0493) expression. Treatment of EOC cell lines with PHA665752 causes a dose-dependent inhibition of cell viability and induction of apoptosis. Furthermore, PHA665752 treatment causes dephosphorylation of AKT and downregulation of antiapoptotic proteins XIAP and Bcl-XL. In addition, PHA665752-induced apoptosis occurs through activation of Bax-mediated release of cytochrome c and activation of caspases. Finally, co-treatment of EOC with PHA665752 and cisplatin causes augmented effect on apoptosis of EOC cells and resulted in synergistic inhibition of EOC xenograft tumor growth in NUDE mice. These results indicate that c-Met/HGF pathway may be a potential target for therapeutic intervention for treatment of EOC.


Comparative efficacy of fibrinogen and platelet supplementation on the in vitro reversibility of competitive glycoprotein IIb/IIIa receptor-directed platelet inhibition.
Platelet surface glycoprotein IIb/IIIa (alphaIIb/beta3) receptor inhibition, with prevention of fibrinogen binding and platelet aggregation, concomitantly attenuates arterial thrombotic capacity and impairs protective hemostasis, 2 divergent platelet-dependent processes. Because the currently available, Food and Drug Administration-approved small molecule glycoprotein IIb/IIIa receptor antagonists are considered "competitive" inhibitors and there is limited information on the reversibility of platelet inhibition with fibrinogen or platelet supplementation, the following series of in vitro experiments were performed. Washed platelets from 24 healthy volunteers were suspended in tyrodes buffer and incubated with achievable (in vivo) concentrations of either tirofiban or eptifibatide before activation with thrombin receptor agonist peptide (15 micromol/L). Platelet aggregation was inhibited by 40% to 50%, but reversal was achieved with fibrinogen supplementation in a concentration-dependent manner. In a separate series of in vitro experiments, platelet inhibition exceeding 90% was established with tirofiban (average concentration 9.28 microg/L) and eptifibatide (average concentration 95.4 microg/L). Recovery of platelet aggregation to at least 50% was achieved after the addition of fibrinogen (0.76-0.80 g/L), platelets (2.4 x 10(11)/L), or their combination. There was an inverse relationship between plasma baseline fibrinogen and the amount of supplemental fibrinogen needed to restore platelet aggregability (r = -0.60; P <.01). The reversibility of glycoprotein IIb/IIIa-directed platelet inhibition is influenced by cell surface receptor availability and intrinsic pharmacodynamic mechanism of action. Fibrinogen supplementation with fresh frozen plasma or cryoprecipitate either alone or in combination with platelet transfusion represents an important and readily available treatment consideration for restoring hemostatic potential and managing major hemorrhagic complications associated with the administration of small-molecule, competitive glycoprotein IIb/IIIa receptor antagonists.


Natural variation in genes potentially involved in plant architecture and adaptation in switchgrass (Panicum virgatum L.).
Advances in genomic technologies have expanded our ability to accurately and exhaustively detect natural genomic variants that can be applied in crop improvement and to increase our knowledge of plant evolution and adaptation. Switchgrass (Panicum virgatum L.), an allotetraploid (2n = 4× = 36) perennial C4 grass (Poaceae family) native to North America and a feedstock crop for cellulosic biofuel production, has a large potential for genetic improvement due to its high genotypic and phenotypic variation. In this study, we analyzed single nucleotide polymorphism (SNP) variation in 372 switchgrass genotypes belonging to 36 accessions for 12 genes putatively involved in biomass production to investigate signatures of selection that could have led to ecotype differentiation and to population adaptation to geographic zones. A total of 11,682 SNPs were mined from ~ 15 Gb of sequence data, out of which 251 SNPs were retained after filtering. Population structure analysis largely grouped upland accessions into one subpopulation and lowland accessions into two additional subpopulations. The most frequent SNPs were in homozygous state within accessions. Sixty percent of the exonic SNPs were non-synonymous and, of these, 45% led to non-conservative amino acid changes. The non-conservative SNPs were largely in linkage disequilibrium with one haplotype being predominantly present in upland accessions while the other haplotype was commonly present in lowland accessions. Tajima's test of neutrality indicated that PHYB, a gene involved in photoperiod response, was under positive selection in the switchgrass population. PHYB carried a SNP leading to a non-conservative amino acid change in the PAS domain, a region that acts as a sensor for light and oxygen in signal transduction. Several non-conservative SNPs in genes potentially involved in plant architecture and adaptation have been identified and led to population structure and genetic differentiation of ecotypes in switchgrass. We suggest here that PHYB is a key gene involved in switchgrass natural selection. Further analyses are needed to determine whether any of the non-conservative SNPs identified play a role in the differential adaptation of upland and lowland switchgrass.


Molecular mapping of rice chromosomes.
We report the construction of an RFLP genetic map of rice (Oryza sativa) chromosomes. The map is comprised of 135 loci corresponding to clones selected from a PstI genomic library. This molecular map covers 1,389 cM of the rice genome and exceeds the current classical maps by more than 20%. The map was generated from F2 segregation data (50 individuals) from a cross between an indica and javanica rice cultivar. Primary trisomics were used to assign linkage groups to each of the 12 rice chromosomes. Seventy-eight percent of the clones assayed revealed RFLPs between the two parental cultivars, indicating that rice contains a significant amount of RFLP variation. Strong correlations between size of hybridizing restriction fragments and level of polymorphism indicate that a significant proportion of the RFLPs in rice are generated by insertions/delections. This conclusion is supported by the occurrence of null alleles for some clones (presumably created by insertion or deletion events). One clone, RG229, hybridized to sequences in both the indica and javanica genomes, which have apparently transposed since the divergence of the two cultivars from their last common ancestor, providing evidence for sequence movement in rice. As a by product of this mapping project, we have discovered that rice DNA is less C-methylated than tomato or maize DNA. Our results also suggest the notion that a large fraction of the rice genome (approximately 50%) is single copy.


Potassium transport by turtle colon: active secretion and active absorption.
Measurement of transepithelial potassium fluxes in the absence of transmural electrochemical potential gradients showed that the isolated turtle colon can actively absorb and actively secrete K+. Under short-circuit conditions the active secretory flow was inhibited by mucosal amiloride, whereas the absorptive flow was unaffected by the diuretic. The effects of ouabain and barium on secretory flow were consistent with a simple model involving basolateral uptake by an Na+-K+-ATPase and conductive exist across the apical and basolateral membranes. The active absorptive flux was blocked by mucosal ouabain and by serosal barium. The opposing active flows clearly represented cellular K+ transport, whereas paracellular K+ flows behaved as expected for a free-solution shunt.


The Relationship of Explicit-Implicit Evaluative Discrepancy to Exercise Dropout in Middle-Aged Adults.
Discrepancies between automatically activated associations (i.e., implicit evaluations) and explicit evaluations of motives (measured with a questionnaire) could lead to greater information processing to resolve discrepancies or self-regulatory failures that may affect behavior. This research examined the relationship of health and appearance exercise-related explicit-implicit evaluative discrepancies, the interaction between implicit and explicit evaluations, and the combined value of explicit and implicit evaluations (i.e., the summed scores) to dropout from a yearlong exercise program. Participants (N = 253) completed implicit health and appearance measures and explicit health and appearance motives at baseline, prior to starting the exercise program. The sum of implicit and explicit appearance measures was positively related to weeks in the program, and discrepancy between the implicit and explicit health measures was negatively related to length of time in the program. Implicit exercise evaluations and their relationships to oft-cited motives such as appearance and health may inform exercise dropout.


Microwave-assisted synthesis of novel non-peripherally substituted metallophthalocyanines and their sensing behaviour for a broad range of Lewis bases.
The synthesis of novel, symmetrical, tetrasubstituted metallophthalocyanines (cobalt, zinc, and manganese) bearing four 2-(4-methyl-1,3-thiazol-5-yl)ethoxy units is reported. The new compounds have been characterized using UV-Vis, IR, (1)H NMR, (13)C NMR, and mass spectroscopy data. Photophysical properties of zinc(ii) phthalocyanines were found, including electronic absorption and fluorescence quantum yields. The fluorescence of the complexes was investigated in DMF and it was found that benzoquinone (BQ) was an effective quencher. The response and recovery behaviours of the spin coated films to different analytes, which span a broad range of Lewis bases, have been investigated by means of conductivity measurements. It was observed that the operating temperature had a considerable effect on the gas sensing performance of the sensors investigated. The sensing behaviour of the films for a broad range of Lewis bases and the correlation between the sensor sensitivity and Lewis base enthalpies were investigated. Results show that the sensitivity of the films may be correlated exponentially with the binding enthalpy.


The burden of deliberate self-harm on the critical care unit of a peri-urban referral hospital in the Eastern Cape: a 5-year review of 419 patients.
Buffalo City Municipality (BCM) in the Eastern Cape (EC) has the highest susceptibility to deliberate self-harm (DSH) of any South African city. The EC also has a shortage of critical care beds. OBJECTIVE; This study reviewed DSH admissions over 5 years to the critical care unit (CCU) of Cecilia Makiwane Hospital (CMH), a large peri-urban hospital in the EC. It also examined the financial burden that DSH exerts on public-sector critical care. DSH cases admitted to CMH's CCU between January 2006 and December 2010 were retrospectively reviewed. Patients under 13 years of age were excluded. Age, gender, admission duration, agent used, outcome and toxicology results were recorded. Cost was estimated using the Department of Health 2012 fee schedule. A total of 419 patients, comprising 17% of total CCU admissions, were included in the study. Cholinesterase inhibitors (CIs) were the most common agents ingested (55%). Compared with non-CI groups, CI patients where admitted for twice as long from admission to discharge (p<0.0001), but had a lower mortality rate (p=0.0344). No significant difference was found between gender and survival (p=0.5725) and between the yearly DSH CCU admission means (p=0.052). CI cases cost a minimum of R15 966.29 per admission and DSH CCU cases cost over R1 million per annum. DSH imposes an appreciable burden on the CCU services in the EC. There is a need to better control the unregulated availability of CIs (and related public education) as well as to improve psychiatric and psychological services in the EC rural areas.


Advances in the use of inhalation provocation tests in clinical evaluation.
Recent advances in the use of inhalation provocation tests in the clinical evaluation of asthma have been made with methacholine and histamine tests. The tests can be better standardized and the results more accurately interpreted. The ease of stimulation of bronchoconstriction by methacholine and histamine (bronchial responsiveness) relates closely to the presence and the degree of variable airflow obstruction that occurs spontaneously or is triggered by natural stimuli. Responsiveness can be heightened by bronchial infection and by exposure to allergens or sensitizing chemicals, such as toluene diisocyanate, and can be reduced by treatment. These observations indicate three uses of the tests in clinical practice. First, they can be used to validate the presence of hyperresponsiveness which, when spirometric study is normal, is synonymous with the presence of asthma. Second, they can be used to quantify the severity of hyperresponsiveness which relates closely to the amount of bronchodilator required to treat asthma. Finally, they can be used to measure a change in responsiveness, which is useful to document occupational asthma (defined as hyperresponsiveness caused by materials at work), and to optimize the treatment required to reverse or reduce hyperresponsiveness as much as possible.


US Army Combat Medic Performance With Portable Ultrasound to Detect Sonographic Findings of Pneumothorax in a Cadaveric Model.
Ultrasound, due to recent advances in portability and versatility, has become a valuable clinical adjunct in austere, resource-limited settings and is well demonstrated to be an accurate/efficient means to detect pneumothorax. The purpose of this study was to evaluate the impact of hands-on ultrasound training on ultrasound-naive US Army combat medics' ability to detect sonographic findings of pneumothorax with portable ultrasound in a cadaver model. Ultrasound-naive US Army combat medics assigned to conventional military units were recruited from a single US Army installation and randomized to receive either didactic training only, or "blended" (didactic and hands-on) training on ultrasound detection of pneumothorax. Blinded participants were asked to perform a thoracic ultrasound exam on ventilated human cadaver models. Primary outcome measured was sensitivity and specificity of detecting sonographic findings of pneumothorax between cohorts. Forty-three participants examined a total of 258 hemithoraces. The didactic-only cohort (n = 24) detected sonographic findings of pneumothorax with a sensitivity of 68% and specificity of 57%. The blended cohort (n = 19) detected sonographic findings of pneumothorax with an overall sensitivity of 91% and specificity of 80%. Detection sensitivities were similar between B-mode versus M-mode use. US Army combat medics can use portable U/S to detect sonographic findings of pneumothorax in a human cadaver model with high sensitivity after a brief, blended (didactic and hands-on) training intervention.


2018 Update on Medical Overuse.
Overuse of medical care is a well-recognized problem in health care, associated with patient harm and costs. We sought to identify and highlight original research articles published in 2017 that are most relevant to understanding medical overuse. A structured review of English-language articles published in 2017 was performed, coupled with examination of tables of contents of high-impact journals to identify articles related to medical overuse in adult care. Manuscripts were appraised for their quality, clinical relevance, and impact. A total of 1446 articles were identified, 910 of which addressed medical overuse. Of these, 111 articles were deemed to be the most relevant based on originality, methodologic quality, and scope. The 10 most influential articles were selected by author consensus. Findings included that unnecessary electrocardiograms are common (performed in 22% of patients at low risk) and can lead to a cascade of services, lipid monitoring rarely affects care, patients who were overdiagnosed with cancer experienced anxiety and criticism about not seeking treatment, calcium and vitamin D supplementation does not reduce hip fracture (relative risk, 1.09; 95% CI, 0.85-1.39), and pregabalin does not improve symptoms of sciatica but frequently has adverse effects (40% of patients experienced dizziness). Antipsychotic medications increased the severity of delirium in patients receiving hospice care and were associated with an increased risk of death (hazard ratio, 1.7; P = .003), and robotic-assisted radical nephrectomy was without benefits by being slower and more costly than laparoscopic surgery. High-sensitivity troponin testing often yielded false-positive results, as 16% of patients with positive troponin results in a US hospital had a myocardial infarction. One-third of patients who received a diagnosis of asthma had no evidence of asthma. Restructuring the electronic health record was able to reduce unnecessary testing (from 31.3 to 13.9 low-value tests performed per 100 patient visits). Many current practices were found to represent overuse, with no benefit and potential harms. Other services were used inappropriately. Reviewing these findings and extrapolating to their patients will enable health care professionals to improve the care they provide.


Motuporamine Derivatives as Antimicrobial Agents and Antibiotic Enhancers against Resistant Gram-Negative Bacteria.
Dihydromotuporamine C and its derivatives were evaluated for their in vitro antimicrobial activities and antibiotic enhancement properties against Gram-negative bacteria and clinical isolates. The mechanism of action of one of these derivatives, MOTU-N44, was investigated against Enterobacter aerogenes by using fluorescent dyes to evaluate outer-membrane depolarization and permeabilization. Its efficiency correlated with inhibition of dye transport, thus suggesting that these molecules inhibit drug transporters by de-energization of the efflux pump rather than by direct interaction of the molecule with the pump. This suggests that depowering the efflux pump provides another strategy to address antibiotic resistance.


Helicobacter pylori in apparently healthy children aged 0-12 years in urban Kampala, Uganda: a community-based cross sectional survey.
Helicobacter pylori is one of the most common causes of bacterial infection in human beings. Studies have showed a high prevalence of Helicobacter pylori among people in low-income countries and colonization early in life. A monoclonal antigen test, performed on faeces, HpSA ImmunoCardSTAT, has a high sensitivity, specificity and accuracy and the faecal test can be performed in all ages, also in resource-limited settings. The main objective of this study was to determine the prevalence and factors associated with Helicobacter pylori colonization in apparently healthy children aged 0-12 years in urban Kampala, Uganda. We tested 427 apparently healthy children, age 0-12 years (211 males, 216 females), in a cross sectional survey for Helicobacter pylori colonization using HpSA ImmunoCardSTAT. A short standardized interview with socio-demographic information and medical history was used to assess risk factors. The overall prevalence of Helicobacter pylori in the 427 children was 44.3% (189 out of 427). Early colonization was common, 28.7%, in children younger than 1 year of age. The age specific rates were 46.0% in children age 1- < 3 years, 51.7% in children age 3- < 6 years, 54.8% in children age 6- < 9 years and 40.0% in children age 9- < 12 years. There was a significant difference in prevalence by gender; female 38.5% versus male 49.8% and by type of housing; permanent house 38.5% versus semi-permanent house 48.6%. Congestive living and education level of the female caretaker showed a clear trend for a difference in prevalence. Factors independently associated with Helicobacter pylori colonization included: drugs taken last three months, using a pit latrine, sources of drinking water and wealth index. The prevalence of Helicobacter pylori colonization among urban Ugandan children is high at an early age and increases with age. The impact of Helicobacter pylori colonization on children's health in Uganda needs to be further clarified.


Reproducibility and sensitivity of 36 methods to quantify the SARS-CoV-2 genetic signal in raw wastewater: findings from an interlaboratory methods evaluation in the U.S.
In response to COVID-19, the international water community rapidly developed methods to quantify the SARS-CoV-2 genetic signal in untreated wastewater. Wastewater surveillance using such methods has the potential to complement clinical testing in assessing community health. This interlaboratory assessment evaluated the reproducibility and sensitivity of 36 standard operating procedures (SOPs), divided into eight method groups based on sample concentration approach and whether solids were removed. Two raw wastewater samples were collected in August 2020, amended with a matrix spike (betacoronavirus OC43), and distributed to 32 laboratories across the U.S. Replicate samples analyzed in accordance with the project's quality assurance plan showed high reproducibility across the 36 SOPs: 80% of the recovery-corrected results fell within a band of ±1.15 log10 genome copies per L with higher reproducibility observed within a single SOP (standard deviation of 0.13 log10). The inclusion of a solids removal step and the selection of a concentration method did not show a clear, systematic impact on the recovery-corrected results. Other methodological variations (e.g., pasteurization, primer set selection, and use of RT-qPCR or RT-dPCR platforms) generally resulted in small differences compared to other sources of variability. These findings suggest that a variety of methods are capable of producing reproducible results, though the same SOP or laboratory should be selected to track SARS-CoV-2 trends at a given facility. The methods showed a 7 log10 range of recovery efficiency and limit of detection highlighting the importance of recovery correction and the need to consider method sensitivity when selecting methods for wastewater surveillance.


Role of nucleoside/nucleotide analogues and low-dose hepatitis B immune globülin in prophylaxis of hepatitis B recurrence among cadaveric liver transplant recipients.
Hepatitis B core antibody (HBcAb) positivity of the donor or the recipient may pose a risk of hepatitis B virus (HBV) reactivation following liver transplantation (LT). We retrospectively investigated patient survival and reactivation among recipients who were given low-dose Hepatitis B Immune Globulin (HBIG) plus antiviral agent (AV) versus AV only. Records of cadaveric LT recipients, between 2013 and 2016, with positive Hepatitis B surface Antigen (HBsAg) and/or HBcAb and recipients who had received LT from HBcAb-positive donors were reviewed. Patient characteristics and clinical data were extracted. Donor variables were retrieved from the United Network of Organ Sharing (UNOS) database. HBIG (1560 IU/mL) Intravenous (IV) was intraoperatively administered with three daily doses. Entecavir 1 mg daily was also given. STATA was used for statistical analysis. There were 53 recipients; 39 (73.6%) were male with a median age of 59 y. HCV was the major indication in 30 (55.6%) patients. There were 28 recipients (52.8%) who received HBIG plus AV and 25 (47.2%) received AV only. The Model of End Stage Liver Disease (MELD) score between the groups were similar. Survival rates at 6, 12, and 24 months were 100% (n=53), 93.2% (n=44), and 100.0% (n=26), respectively. There was no reactivation; two recipients in the AV group and one in the HBIG plus AV group died within 12 months. This study supports the use of low-dose HBIG and AV for post-LT prophylaxis to be as effective as conventionally used high-dose HBIG (9600 IU) plus AV. Future prospective larger studies are warranted to examine the potential benefits of using AV alone without HBIG.


Transmission-blocking activity of tafenoquine (WR-238605) and artelinic acid against naturally circulating strains of Plasmodium vivax in Thailand.
The sporontocidal activity of tafenoquine (WR-238605) and artelinic acid was determined against naturally circulating isolates of Plasmodium vivax in western Thailand. Primaquine was used as a negative control and a dihydroacridine-dione (WR-250547) was used as a positive control. Laboratory-reared Anopheles dirus mosquitoes were infected with P. vivax by allowing mosquitoes to feed on blood (placed in an artificial-membrane feeding apparatus) collected from gametocytemic volunteers reporting to local malaria clinics in Tak province, Thailand. Four days post-infection, mosquitoes were refed on uninfected mice treated 90 minutes earlier with a given drug. Drug activity was determined by assessing oocyst and sporozoite development. Neither primaquine nor artelinic acid affected oocyst or sporozoite development at a dose of 100 mg of base drug/kg of mouse body weight. In contrast, tafenoquine and WR-250547 affected sporogonic development at doses as low as 25.0 and 0.39 mg/kg, respectively. The potential role of these compounds in the prevention of malaria transmission is discussed, as are alternative strategies for the use of transmission-blocking antimalarial drugs.


Clinical causes and treatment of the thrombotic storm.
Thrombotic storm represents an extreme prothrombotic phenotype, characterized by multiple thrombotic events affecting diverse vascular beds occurring over a brief period of time. Thrombotic events involve venous and arterial circulation, including unusual locations, such as cerebral sinus venous thrombosis, intra-abdominal thromboembolic occlusions and microvascular events. Some patients will have antiphospholipid antibodies, but a significant number have no identifiable hypercoagulable state. The mainstay of treatment consists of anticoagulant therapy, although some patients appear to benefit from the addition of immunomodulatory therapies. Other disorders that share this thrombotic storm phenotype include catastrophic antiphospholipid syndrome, spontaneous heparin-induced thrombocytopenia and similar aggressive clinical disorders. Ongoing studies are focused on identifying underlying genetic factors that may predispose patients to develop this extreme clinical phenotype.


PU.1 opposes IL-7-dependent proliferation of developing B cells with involvement of the direct target gene bruton tyrosine kinase.
Deletion of genes encoding the E26 transformation-specific transcription factors PU.1 and Spi-B in B cells (CD19-CreΔPB mice) leads to impaired B cell development, followed by B cell acute lymphoblastic leukemia at 100% incidence and with a median survival of 21 wk. However, little is known about the target genes that explain leukemogenesis in these mice. In this study we found that immature B cells were altered in frequency in the bone marrow of preleukemic CD19-CreΔPB mice. Enriched pro-B cells from CD19-CreΔPB mice induced disease upon transplantation, suggesting that these were leukemia-initiating cells. Bone marrow cells from preleukemic CD19-CreΔPB mice had increased responsiveness to IL-7 and could proliferate indefinitely in response to this cytokine. Bruton tyrosine kinase (BTK), a negative regulator of IL-7 signaling, was reduced in preleukemic and leukemic CD19-CreΔPB cells compared with controls. Induction of PU.1 expression in cultured CD19-CreΔPB pro-B cell lines induced Btk expression, followed by reduced STAT5 phosphorylation and early apoptosis. PU.1 and Spi-B regulated Btk directly as shown by chromatin immunoprecipitation analysis. Ectopic expression of BTK was sufficient to induce apoptosis in cultured pro-B cells. In summary, these results suggest that PU.1 and Spi-B activate Btk to oppose IL-7 responsiveness in developing B cells.


Investigation of estrogen activity in the raw and treated waters of riverbank infiltration using a yeast estrogen screen and chemical analysis.
Exposure to various endocrine disrupting chemicals (EDCs) can lead to adverse effects on reproductive physiology and behavior in both animals and humans. An adequate strategy for the prevention of environmental contamination and eliminating the effects of them must be established. Chemicals with estrogenic activity were selected, and the effectiveness of their removal during the purification processes in two drinking water treatment plants (DWTPs) using riverbank infiltrated water was determined. Thirty-five water samples in two sampling campaigns throughout different seasons were collected and screened with a yeast estrogen test; furthermore, bisphenol A (BPA), 17ß-estradiol (E2) and ethinyl-estradiol (EE2) content were measured using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Our results confirm that estrogenic compounds are present in sewage effluents and raw surface river water of DWTPs. Very low estrogen activity and pg/L concentrations of BPA and E2 were detected during drinking water processing and occasionally in drinking water. Based on this study, applied riverbank filtration and water treatment procedures do not seem to be suitable for the total removal of estrogenic chemicals. Local contamination could play an important role in increasing the BPA content of the drinking water at the consumer endpoint.


Direct and buffering effects of social support among gynecologic cancer survivors.
There are few studies of QoL among long-term gynecologic cancer survivors; available data suggest significant sequelae of disease and treatment. Research clarifying circumstances that improve difficult survivorship trajectories is lacking. The present study examines whether social support moderates the relationship between physical functioning and psychological outcomes by testing the stress-buffering hypothesis. Participants (N = 260) were gynecologic cancer survivors (cervical, n = 47; endometrial, n = 133; ovarian, n = 69; vulvar, n = 11). Compromised physical health was conceptualized as multidimensional. Social support (SNI, PSS-Fa, PSS-Fr, ISEL) was tested as a buffer of adverse psychological outcomes (IES-R, CES-D). Results for traumatic stress provided evidence for buffering; whereas social support was of general benefit for depressive symptoms. Effects varied by source and type of support. These results suggest that circumstances for gynecologic cancer survivors burdened with physical symptoms may be worse for those with fewer support resources, providing needed insight into a common target of psychosocial interventions for cancer survivors.


Clinical relevance of MMP-9, MMP-2, TIMP-1 and TIMP-2 in colorectal cancer.
The main aim of this study was to evaluate the clinical relevance of Gelatinases in colorectal cancer (CRC). Ninety-five CRCs and their paired normal tissues were investigated to detect total levels of MMP-9, MMP-2, and the tissue inhibitors TIMP-1 and TIMP-2. Also, pro-MMP and MMP activity, and potential associations with clinical parameters were estimated. MMP-9, MMP-2 and TIMP-1 levels were greater in CRCs than in normal tissues, differences being significant for MMP-9 and TIMP-1. However, TIMP-2 showed significantly lower levels in tumour samples. Moreover, significant differences in the state of activation between gelatinases were found. TIMP-1 low levels were significantly associated with poor clinical outcome of patients. According to these data, different roles have to be attributed to MMP-2 and MMP-9 in CRC progression. Moreover, TIMP-1 level evaluation emerges as the main prognostic factor in relation to Gelatinases A and B activity in CRC.


Replication and segregation of plasmids containing cis-acting regulatory sites of silent mating-type genes in Saccharomyces cerevisiae are controlled by the SIR genes.
In Saccharomyces cerevisiae, two cis-acting regulatory sites called E and I flank the silent mating-type gene, HMRa, and mediate SIR-dependent transcriptional repression of the a1-a2 promoters. It has been shown previously that the E and I sites have plasmid replicator (ARS) activity. We show in this report that the ARS activity of the E and I sites is governed by the SIR genotype of the cell. In wild-type cells, a plasmid carrying the E site from HMRa (HMR E) in the vector YIp5 exhibited very high mitotic stability at a copy number of approximately 25 per cell. However, in sir2, sir3, or sir4 mutants, plasmids with HMR E had the low mitotic stability characteristic of plasmids containing ARS1, a SIR-independent replicator. Elevated mitotic stability of plasmids that carry HMR E is due to a segregation mechanism provided by SIR and HMR E. In sir2 and sir4 mutants, the plasmid copy number was significantly lowered, suggesting that these gene products also participate in the replication of plasmids carrying HMR E. The phenotype of point mutations introduced at an 11-base-pair ARS consensus sequence present at HMR E indicated that this sequence is functional but not absolutely required for autonomous replication of the plasmid and that it is not required for SIR-dependent mitotic stabilization. A plasmid carrying both a centromere and HMR E exhibited reduced mitotic stability in wild-type cells. This destabilization appeared to be due to antagonism between the segregation functions provided by the centromere and by HMR E.


The CPLANE protein Intu protects kidneys from ischemia-reperfusion injury by targeting STAT1 for degradation.
Intu is known as a ciliogenesis and planar polarity effector (CPLANE) protein. Although roles for Intu have been reported during embryonic development and in the context of developmental disorders, its function and regulation in adult tissues remain poorly understood. Here we show that ablation of Intu specifically in kidney proximal tubules aggravates renal ischemia-reperfusion injury, and leads to defective post-injury ciliogenesis. We identify signal transducer and activator of transcription 1 (STAT1) as a novel interacting partner of Intu. In vitro, Intu and STAT1 colocalize at the centriole/basal body area, and Intu promotes proteasomal degradation of STAT1. During cell stress, Intu expression preserves cilia length and cell viability, and these actions are antagonized by STAT1 expression. Thus, we propose a role for Intu in protecting cells and tissues after injury by targeting STAT1 for degradation and maintaining primary cilia.


Incidence of and risk factors for hepatocellular carcinoma in primary biliary cirrhosis: national data from Japan.
Primary biliary cirrhosis (PBC) primarily affects females and is rarely complicated by hepatocellular carcinoma (HCC). Although the HCC incidence in PBC patients is low, several characteristics and risk factors associated with its development have been reported. In this study, national data concerning the current status of carcinogenesis in PBC patients in Japan are reviewed. Using data from two national questionnaire surveys, we investigated the clinicopathological findings associated with HCC in PBC patients. According to the data of all reviewed PBC patients, the HCC incidence was 2.4% (71/2946). The HCC incidence by gender was 5.1% (19/370) in males and 2.0% (52/2576) in females, and the proportion of males was 26.7%. Prognosis was significantly poorer in the PBC patients with HCC than in those without. Multivariate analysis of risk factors associated with HCC by gender revealed histological stage at the time of PBC diagnosis as an independent risk factor associated with the development of HCC in females, but not in males. Furthermore, data from another national survey of 178 PBC patients with HCC (male/female = 49/129; proportion of males 27.5%) revealed that the duration between the diagnosis of PBC and that of HCC was significantly shorter in males than in females. In addition, histological stage at the time of HCC diagnosis was an independent risk factor for HCC in females, whereas no risk factors were identified in males. these data indicate that males are at risk of developing HCC at any histological stage of PBC. Therefore, male PBC patients in particular should be carefully screened for HCC from the early stages of PBC.


Analysis of performance and age of the fastest 100-mile ultra-marathoners worldwide.
The performance and age of peak ultra-endurance performance have been investigated in single races and single race series but not using worldwide participation data. The purpose of this study was to examine the changes in running performance and the age of peak running performance of the best 100-mile ultra-marathoners worldwide. The race times and ages of the annual ten fastest women and men were analyzed among a total of 35,956 finishes (6,862 for women and 29,094 for men) competing between 1998 and 2011 in 100-mile ultra-marathons. The annual top ten performances improved by 13.7% from 1,132±61.8 min in 1998 to 977.6±77.1 min in 2011 for women and by 14.5% from 959.2±36.4 min in 1998 to 820.6±25.7 min in 2011 for men. The mean ages of the annual top ten fastest runners were 39.2±6.2 years for women and 37.2±6.1 years for men. The age of peak running performance was not different between women and men (p>0.05) and showed no changes across the years. These findings indicated that the fastest female and male 100-mile ultra-marathoners improved their race time by ∼14% across the 1998-2011 period at an age when they had to be classified as master athletes. Future studies should analyze longer running distances (>200 km) to investigate whether the age of peak performance increases with increased distance in ultra-marathon running.


Detection of iron-depositing Pedomicrobium species in native biofilms from the Odertal National Park by a new, specific FISH probe.
Iron-depositing bacteria play an important role in technical water systems (water wells, distribution systems) due to their intense deposition of iron oxides and resulting clogging effects. Pedomicrobium is known as iron- and manganese-oxidizing and accumulating bacterium. The ability to detect and quantify members of this species in biofilm communities is therefore desirable. In this study the fluorescence in situ hybridization (FISH) method was used to detect Pedomicrobium in iron and manganese incrusted biofilms. Based on comparative sequence analysis, we designed and evaluated a specific oligonucleotide probe (Pedo 1250) complementary to the hypervariable region 8 of the 16S rRNA gene for Pedomicrobium. Probe specificities were tested against 3 different strains of Pedomicrobium and Sphingobium yanoikuyae as non-target organism. Using optimized conditions the probe hybridized with all tested strains of Pedomicrobium with an efficiency of 80%. The non-target organism showed no hybridization signals. The new FISH probe was applied successfully for the in situ detection of Pedomicrobium in different native, iron-depositing biofilms. The hybridization results of native bioflims using probe Pedo_1250 agreed with the results of the morphological structure of Pedomicrobium bioflims based on scanning electron microscopy.


An adeno-associated virus vector efficiently and specifically transduces mouse skeletal muscle.
Expression of a therapeutic gene in the skeletal muscle is a practical strategy to compensate a patients' insufficient circulating factor. Its clinical application requires a muscle-targeting vector capable of inducing a continuous high-level transgene expression. We modified an adeno-associated virus serotype 2 (AAV2) vector expressing luciferase from the mouse muscle creatine kinase gene promoter-enhancer (Ckm). First, AAVS1 insulator was inserted into the vector genome for transcriptional enhancement. This increased transduction of mouse quadriceps muscle by 11-fold at 4 weeks after intramuscular injection. Second, two capsid modifications were combined (21F capsid): incorporation of a segment of AAV1 capsid to produce a hybrid capsid and substitution of a tyrosine with a phenylalanine. Use of 21F capsid increased muscle transduction further by 18-fold, resulting in 200-fold higher efficacy than that of the unmodified vector. Compared with a vector having human elongation factor 1α promoter which showed similar efficacy in the muscle, this vector having Ckm transduced non-muscle organs less efficiently after intravenous administration. The AAV2 vector composed of the modified genome and capsid provides a backbone to develop a clinical vector expressing a therapeutic gene in the muscle.


Fluctuation pressure of biomembranes in planar confinement.
The fluctuation pressure of a lipid-bilayer membrane is important for the stability of lamellar phases and the adhesion of membranes to surfaces. In contrast to many theoretical studies, which predict a decrease of the pressure with the cubed inverse distance between the membranes, Freund suggested very recently a linear inverse distance dependence [Proc. Natl. Acad. Sci. USA 110, 2047 (2013)]. We address this discrepancy by performing Monte Carlo simulations for a membrane model discretized on a square lattice and employ the wall theorem to evaluate the pressure for a single membrane between parallel walls. For distances that are small compared with the lattice constant, the pressure indeed depends on the inverse distance as predicted by Freund. For intermediate distances, the pressure depends on the cubed inverse distance as predicted by Helfrich [Z. Naturforsch. A 33, 305 (1978)]. Here, the crossover length between the two regimes is a molecular length scale. Finally, for distances large compared with the mean squared fluctuations of the membrane, the entire membrane acts as a soft particle and the pressure on the walls again depends linearly on the inverse distance.


Radiological study of pleural changes in relation to mesothelioma in Turkey.
In some villages in central Turkey pleural changes occur as a result of environmental exposure to mineral fibres. In most cases the fibre is asbestos but in some cases the non-asbestos fibre erionite, a zeolite, is responsible. The incidence of malignant mesothelioma is much higher in "erionite villages" than in "asbestos villages" despite similar frequencies of pleural changes. In this study chest radiographs from 466 people from asbestos villages, 549 from erionite villages, and 382 controls were compared. The frequency of pleural calcification was about the same in the two groups of villages studied, but the minor fissures were visible to a greater degree in erionite cases. In people from erionite villages "atypical" pleural calcification, due to calcification of the visceral rather than the parietal pleura, was more common. These differences may indicate that the fibres have different lengths and diameters.


The tissue distribution of the B7-2 costimulator in mice: abundant expression on dendritic cells in situ and during maturation in vitro.
B7-2 is a recently discovered, second ligand for the CTLA-4/CD28, T cell signaling system. Using the GL-1 rat monoclonal antibody (mAb), we monitored expression of B7-2 on mouse leukocytes with an emphasis on dendritic cells. By cytofluorography, little or no B7-2 was detected on most cell types isolated from spleen, thymus, peritoneal cavity, skin, marrow, and blood. However, expression of B7-2 could be upregulated in culture. In the case of epidermal and spleen dendritic cells, which become highly immunostimulatory for T cells during a short period of culture, the upregulation of B7-2 was dramatic and did not require added stimuli. Lipopolysaccharide did not upregulate B7-2 levels on dendritic cells, in contrast to macrophages and B cells. By indirect immunolabeling, the level of staining with GL-1 mAb exceeded that seen with rat mAbs to several other surface molecules including intercellular adhesion molecule 1, B7-1, CD44, and CD45, as well as new hamster mAbs to CD40, CD48, and B7-1/CD80. Of these accessory molecules, B7-2 was a major species that increased in culture, implying a key role for B7-2 in the functional maturation of dendritic cells. B7-2 was the main (> 90%) CTLA-4 ligand on mouse dendritic cells. When we applied GL-1 to tissue sections of a dozen different organs, clear-cut staining with B7-2 antigen was found in many. B7-2 staining was noted on liver Kupffer cells, interstitial cells of heart and lung, and profiles in the submucosa of the esophagus. B7-2 staining was minimal in the kidney and in the nonlymphoid regions of the gut, and was not observed at all in the brain. In the tongue, only rare dendritic cells in the oral epithelium were B7-2+, but reactive cells were scattered about the interstitial spaces of the muscle. In all lymphoid tissues, Gl-1 strongly stained certain distinct regions that are occupied by dendritic cells and by macrophages. For dendritic cells, these include the thymic medulla, splenic periarterial sheaths, and lymph node deep cortex; for macrophages, the B7-2-rich regions included the splenic marginal zone and lymph node subcapsular cortex. Splenic B7-2+ cells were accessible to labeling with GL-1 mAb given intravenously. Dendritic cell stimulation of T cells (DNA synthesis) during the mixed leukocyte reaction was significantly (35-65%) blocked by GL-1.(ABSTRACT TRUNCATED AT 400 WORDS)


Bullous dermatosis on legs of elderly: A new clinical entity?
A lot of diseases occur on the skin of elderly persons. We report four elderly cases of bullous dermatosis that did not meet various differential diagnoses. Japanese, heart failure, atrophic skin and leg edema probably due to aging, as well as flaccid or tense bullae localized in legs were the common factors to our patients. Such conditions may be increased in coming aging society. Accordingly, it is worth regarding such symptom as the new clinical entity, which may comfort patients with similar condition and attract further attention.


Cardiac catheterisation guided by MRI in children and adults with congenital heart disease.
Fluoroscopically guided cardiac catheterisation is an essential tool for diagnosis and treatment of congenital heart disease. Drawbacks include poor soft tissue visualisation and exposure to radiation. We describe the first 16 cases of a novel method of cardiac catheterisation guided by MRI with radiographic support. In our cardiac catheterisation laboratory, we combine magnetic resonance and radiographic imaging facilities. We used MRI to measure flow and morphology, and real-time MRI sequences to visualise balloon angiographic catheters. 12 patients underwent diagnostic cardiac catheterisation, two had interventional cardiac catheterisations, and for two patients, MRI was used to plan radiofrequency ablation for treatment of tachyarrhythmias. In 14 patients, some or all of the cardiac catheterisation was guided by MRI. In two patients undergoing radiofrequency ablation, catheters were manipulated with use of fluoroscopic guidance and outcome was assessed with MRI. All patients received lower amounts of radiation than controls. There was some discrepancy between pulmonary vascular resistance calculated by flow derived from MRI and the traditional Fick method. We were able to superimpose fluoroscopic images of electro physiology electrode catheters on the three dimensional MRI of the cardiac anatomy. We have shown that cardiac catheterisation guided by MRI is safe and practical in a clinical setting, allows better soft tissue visualisation, provides more pertinent physiological information, and results in lower radiation exposure than do fluoroscopically guided procedures. MRI guidance could become the method of choice for diagnostic cardiac catheterisation in patients with congenital heart disease, and an important tool in interventional cardiac catheterisation and radiofrequency ablation.


Exploring determinants for quality of life among older people in pain and in need of help for daily living.
The aim of the study was to investigate the quality of life and related factors among older people who are in pain and in need of help to manage daily living. To intervene against the low quality of life in nursing care knowledge about factors affecting it is needed and this is especially important for vulnerable people such as those who suffer from pain and who are in need of help to manage daily living. Five hundred and twenty-six people, aged 75-102 years participated in this study. Those in pain reported a significantly higher degree of all complaints and lower quality of life in all measures compared with those not in pain. Overall quality of life was associated with mobility problems, sleeping problems and depressed mood, while health-related quality of life was associated with living in special accommodations, walking problems, mobility problems and fatigue. Those in need of help to manage daily living and in pain seem to be at higher risk of lowered quality of life than those not in pain and the lower quality of life among those in pain is probably caused by the complex of complaints rather than pain per se. Daily nursing care should identify and treat the complex of complaints related to pain as well as pain itself, to improve everyday life and quality of life for older people in pain.


Lifelong low-phylloquinone intake is associated with cognitive impairments in old rats.
In a previous report, we showed vitamin K to preferentially accumulate in brain regions rich in white matter and to positively correlate with certain sphingolipids. In rodents, pharmacological vitamin K deficiency has resulted in behavioral perturbations. To gain insight on the role of vitamin K status on brain function, we investigated learning abilities (Morris water maze), motor activity (open field), and anxiety (elevated plus maze) in distinct groups of 6-, 12-, and 20-mo-old female Sprague-Dawley rats that had been fed diets containing low (L; ~80 μg/kg diet), adequate (A; ~500 μg/kg diet), or high (H; ~2000 μg/kg diet) levels of phylloquinone (μg/kg diet; n = 9-12/diet) since weaning. In 20-mo-old rats, sphingolipids (cerebroside, sulfatide, sphingomyelin, ceramide, and gangliosides), phylloquinone, and menaquinone-4 were also assessed in cerebellum, midbrain, pons medulla, striatum, and hippocampus. Lifetime consumption of a low-vitamin K diet resulted in cognitive deficits in the 20-mo-old rats, with those in the L group having longer latencies than those in the H group (P < 0.05); this was associated with higher concentrations of ceramides in the hippocampus (P < 0.05) and lower gangliosides in the pons medulla and midbrain (P < 0.05). The low-vitamin K diet did not affect cognition at 6 and 12 mo of age, nor did it affect motor activity or anxiety at any age. Although much remains to be elucidated about the mechanism of action of vitamin K in cognition, this report points to vitamin K as an important nutritional factor contributing to cognitive health during aging.


POD-1/Tcf21 overexpression reduces endogenous SF-1 and StAR expression in rat adrenal cells.
During gonad and adrenal development, the POD-1/capsulin/TCF21transcription factor negatively regulates SF-1/NR5A1expression, with higher SF-1 levels being associated with increased adrenal cell proliferation and tumorigenesis. In adrenocortical tumor cells, POD-1 binds to the SF-1 E-box promoter region, decreasing SF-1 expression. However, the modulation of SF-1 expression by POD-1 has not previously been described in normal adrenal cells. Here, we analyzed the basal expression of Pod-1 and Sf-1 in primary cultures of glomerulosa (G) and fasciculata/reticularis (F/R) cells isolated from male Sprague-Dawley rats, and investigated whether POD-1 overexpression modulates the expression of endogenous Sf-1 and its target genes in these cells. POD-1 overexpression, following the transfection of pCMVMycPod-1, significantly decreased the endogenous levels of Sf-1 mRNA and protein in F/R cells, but not in G cells, and also decreased the expression of the SF-1 target StAR in F/R cells. In G cells overexpressing POD-1, no modulation of the expression of SF-1 targets, StAR and CYP11B2, was observed. Our data showing that G and F/R cells respond differently to ectopic POD-1 expression emphasize the functional differences between the outer and inner zones of the adrenal cortex, and support the hypothesis that SF-1 is regulated by POD-1/Tcf21 in normal adrenocortical cells lacking the alterations in cellular physiology found in tumor cells.


Epidemiology and patterns of musculoskeletal motorcycle injuries in the USA.
Motorcycles have become an increasingly popular mode of transportation despite their association with a greater risk for injury compared with automobiles. Whereas the recent incidence of annual passenger vehicle fatalities in the United States of America (USA) has progressively declined, motorcycle fatalities have steadily increased in the past 11 years. Although motorcycle injuries (MIs) have been studied, to the author's knowledge there are no published reports on MIs in the USA during this 11-year period. Methods : Study data were derived from a prospectively collected Level I trauma center database. Data sampling included motorcycle crash injury evaluations for the 10-year period ending on 31 August 2008. This retrospective analysis included patient demographic and medical data, helmet use, Glasgow coma scale (GCS) score, injury severity score (ISS), length of hospital stay (LOS), specific injury diagnosis, and death. Data statistics were analyzed using the Spearman correlation coefficient, Kruskal-Wallis tests, and logistic regression. The study identified 1252 motorcycle crash injuries. Helmets were worn by 40.7% of patients for which helmet data were available. The rates of the most common orthopedic injuries were tibia/fibula (19.01%), spine (16.21%), and forearm (10.14%) fractures. The most common non-orthopedic motorcycle crash injuries were concussions (21.09%), skull fractures (8.23%), face fractures (13.66%), and hemo- and pneumothorax (8.79%). There was a significant correlation between greater age and higher ISS (r=0.21, P<0.0001) and longer LOS (r=0.22, P<0.0001). Older patients were also less likely to wear a helmet (OR=0.99, 95% CI: 0.98, 0.997), associated with a significantly higher risk for death (after adjustment for helmet use OR=1.03, 95% CI: 1.00, 1.05). All patients without helmets had a significantly lower GCS score (P=0.0001) and a higher mortality rate (after adjustment for patient demographic data OR=2.28, 95% CI: 1.13, 4.58). Conclusion : Compared with historical reports, the prevalence of skull, face, spine, and pelvis fractures have increased in American motorcycle crashes. Compared to recent European studies, the incidence of USA skull and face fractures is much higher, while the incidence of USA spine and pelvis fractures is more comparable; however, this is not associated with increased in-hospital mortality.


Effect modification by sex and baseline CD4+ cell count among adults receiving combination antiretroviral therapy in Botswana: results from a clinical trial.
The Tshepo study was the first clinical trial to evaluate outcomes of adults receiving nevirapine (NVP)-based versus efavirenz (EFV)-based combination antiretroviral therapy (cART) in Botswana. This was a 3 year study (n=650) comparing the efficacy and tolerability of various first-line cART regimens, stratified by baseline CD4(+): <200 (low) vs. 201-350 (high). Using targeted maximum likelihood estimation (TMLE), we retrospectively evaluated the causal effect of assigned NNRTI on time to virologic failure or death [intent-to-treat (ITT)] and time to minimum of virologic failure, death, or treatment modifying toxicity [time to loss of virological response (TLOVR)] by sex and baseline CD4(+). Sex did significantly modify the effect of EFV versus NVP for both the ITT and TLOVR outcomes with risk differences in the probability of survival of males versus the females of approximately 6% (p=0.015) and 12% (p=0.001), respectively. Baseline CD4(+) also modified the effect of EFV versus NVP for the TLOVR outcome, with a mean difference in survival probability of approximately 12% (p=0.023) in the high versus low CD4(+) cell count group. TMLE appears to be an efficient technique that allows for the clinically meaningful delineation and interpretation of the causal effect of NNRTI treatment and effect modification by sex and baseline CD4(+) cell count strata in this study. EFV-treated women and NVP-treated men had more favorable cART outcomes. In addition, adults initiating EFV-based cART at higher baseline CD4(+) cell count values had more favorable outcomes compared to those initiating NVP-based cART.


Associations Between Khat Use and HIV Risk and Status Among Voluntary Counseling and Testing Center Clients in Addis Ababa, Ethiopia.
Ethiopia is 1 of 15 countries that account for nearly 75% of all people living with HIV. Khat use, a common practice in Ethiopia and sub-Saharan Africa, has gained attention as a potential risk factor for HIV. Our objective was to evaluate associations between khat use and demographic and risk-taking characteristics (alcohol use, sexual behavior), and also associations between HIV status and these characteristics among voluntary counseling and testing clients in Addis Ababa, Ethiopia. This cross-sectional study was conducted among Pro Pride voluntary counseling and testing center clients, aged 18 to 49 years, from November 2009 to March 2010. All clients were approached, and 98.8% of these (684) completed a self-administered questionnaire. Associations between khat use, alcohol use, multiple sexual partners, and HIV status were assessed using logistic regression models. Using khat in the past 90 days (current khat use) was significantly associated with being Muslim, being male, alcohol use, and having a greater number of sex partners in one's lifetime. The adjusted odds of current khat use was almost 7-fold higher in those having 4 or more sexual partners in their lifetime (adjusted odds ratio 6.89, 95% confidence interval 3.87, 12.25) as compared with those with 1 or none. HIV-positive status was significantly associated with age, employment, marital status, number of sex partners in one's lifetime, and khat use. Having used khat in one's lifetime, but not currently, past khat use, was associated with over a 2-fold increased adjusted odds (adjusted odds ratio 2.64, 95% confidence interval 1.13, 6.19) of being HIV-infected. Our findings highlight associations between current khat use and the modifiable factors of number of lifetime sexual partners and alcohol use. In addition, we observed the association between HIV status and past khat use after adjusting for age, marital status, and number of sex partners in one's lifetime.


Known structures and unknown mechanisms of TMEM16 scramblases and channels.
The TMEM16 family of membrane proteins is composed of both Ca2+-gated Cl- channels and Ca2+-dependent phospholipid scramblases. The functional diversity of TMEM16s underlies their involvement in numerous signal transduction pathways that connect changes in cytosolic Ca2+ levels to cellular signaling networks. Indeed, defects in the function of several TMEM16s cause a variety of genetic disorders, highlighting their fundamental pathophysiological importance. Here, we review how our mechanistic understanding of TMEM16 function has been shaped by recent functional and structural work. Remarkably, the recent determination of near-atomic-resolution structures of TMEM16 proteins of both functional persuasions has revealed how relatively minimal rearrangements in the substrate translocation pathway are sufficient to precipitate the dramatic functional differences that characterize the family. These structures, when interpreted in the light of extensive functional analysis, point to an unusual mechanism for Ca2+-dependent activation of TMEM16 proteins in which substrate permeation is regulated by a combination of conformational rearrangements and electrostatics. These breakthroughs pave the way to elucidate the mechanistic bases of ion and lipid transport by the TMEM16 proteins and unravel the molecular links between these transport activities and their function in human pathophysiology.


Optimization of sequence alignment for simple sequence repeat regions.
Microsatellites, or simple sequence repeats (SSRs), are tandemly repeated DNA sequences, including tandem copies of specific sequences no longer than six bases, that are distributed in the genome. SSR has been used as a molecular marker because it is easy to detect and is used in a range of applications, including genetic diversity, genome mapping, and marker assisted selection. It is also very mutable because of slipping in the DNA polymerase during DNA replication. This unique mutation increases the insertion/deletion (INDELs) mutation frequency to a high ratio - more than other types of molecular markers such as single nucleotide polymorphism (SNPs).SNPs are more frequent than INDELs. Therefore, all designed algorithms for sequence alignment fit the vast majority of the genomic sequence without considering microsatellite regions, as unique sequences that require special consideration. The old algorithm is limited in its application because there are many overlaps between different repeat units which result in false evolutionary relationships. To overcome the limitation of the aligning algorithm when dealing with SSR loci, a new algorithm was developed using PERL script with a Tk graphical interface. This program is based on aligning sequences after determining the repeated units first, and the last SSR nucleotides positions. This results in a shifting process according to the inserted repeated unit type.When studying the phylogenic relations before and after applying the new algorithm, many differences in the trees were obtained by increasing the SSR length and complexity. However, less distance between different linage had been observed after applying the new algorithm. The new algorithm produces better estimates for aligning SSR loci because it reflects more reliable evolutionary relations between different linages. It reduces overlapping during SSR alignment, which results in a more realistic phylogenic relationship.


Management of pediatric obesity: a lifestyle modification approach.
Over the last decades, pediatric obesity has become a global epidemic with worldwide estimates as high as 43 million children and adolescents affected, and this number is rising at an exponential rate. With pediatric obesity comes a host of co-morbidities including impaired glucose tolerance, dyslipidemia, hypertension, and impaired liver function. Treatment of this population has proven to be challenging for many reasons. For patients, a new baseline exists consisting of an increasingly sedentary lifestyle as well as a lack of availability of affordable healthy alternatives. In addition, there is an overwhelming presence of energy-dense foods. For physicians, there are many issues including lack of time, training, and reimbursement. The most efficacious and reliable way to treat this population and its co-morbidities is with a healthy, balanced lifestyle consisting of a realistic diet plan and exercise regimen. The is the cornerstone of therapy in the Center for Obesity And its Complications in Health (COACH) clinic which is Children's Medical Center's (Dallas, TX) strategy to combat and treat pediatric overweight and obesity. Lifestyle changes of diet and exercise plans are tailored to each individual's interests and metabolic needs in COACH which is a multi-disciplinary clinic. Additionally, co-morbidities are screened for and treated aggressively to help prevent long-term complications of overweight and obesity. If others do similar interventions in their communities, this global epidemic has the possibility of more positive outcomes than those currently projected.


Single cell RNA sequencing identifies unique inflammatory airspace macrophage subsets.
Macrophages are well recognized for their dual roles in orchestrating inflammatory responses and regulating tissue repair. In almost all acutely inflamed tissues, 2 main subclasses of macrophages coexist. These include embryonically derived resident tissue macrophages and BM-derived recruited macrophages. While it is clear that macrophage subsets categorized in this fashion display distinct transcriptional and functional profiles, whether all cells within these categories and in the same inflammatory microenvironment share similar functions or whether further specialization exists has not been determined. To investigate inflammatory macrophage heterogeneity on a more granular level, we induced acute lung inflammation in mice and performed single cell RNA sequencing of macrophages isolated from the airspaces during health, peak inflammation, and resolution of inflammation. In doing so, we confirm that cell origin is the major determinant of alveolar macrophage (AM) programing, and, to our knowledge, we describe 2 previously uncharacterized, transcriptionally distinct subdivisions of AMs based on proliferative capacity and inflammatory programing.