Acquired haemophilia A diagnosed during pregnancy.
Pregnancy-associated haemophilia A is an uncommon, acquired bleeding disorder which usually presents post-partum; very rarely it may present during pregnancy. No consensus guidelines exist on the management of this condition in pregnancy and very few cases have been reported in the literature. Here we describe the case of a woman presenting with acquired haemophilia A during pregnancy and outline the management of her bleeding disorder. We contrast her case with that of two other women, presenting to the same tertiary referral centre, with acquired haemophilia A presenting post-partum. These cases highlight the heterogeneous management of this condition and how it may be successfully managed in pregnancy.


Multiphase models for simulating hot and slightly miscible DNAPL (dense non-aqueous phase fluids) in a saturated rock fracture under deformation.
In the present study a two dimensional model is first developed to show the behaviour of dense non-aqueous phase liquids (DNAPL) within a rough fracture. To consider the rough fracture, the fracture is imposed with variable apertures along its plane. It is found that DNAPL follows preferential pathways. In next part of the study the above model is further extended for non-isothermal DNAPL flow and DNAPL-water interphase mass transfer phenomenon. These two models are then coupled with joint deformation due to normal stresses. The primary focus of these models is specifically to elucidate the influence of joint alteration due to external stress and fluid pressures on flow driven energy transport and interphase mass transfer. For this, it is assumed that the critical value for joint alteration is associated with external stress and average of water and DNAPL pressures in multiphase system and the temporal and spatial evolution of joint alteration are determined for its further influence on energy transport and miscible phase transfer. The developed model has been studied to show the influence of deformation on DNAPL flow. Further this preliminary study demonstrates the influence of joint deformation on heat transport and phase miscibility via multiphase flow velocities. It is seen that the temperature profile changes and shows higher diffusivity due to deformation and although the interphase miscibility value decreases but the lateral dispersion increases to a considerably higher extent.


MR imaging of periventricular leukomalacia in childhood.
Eight children with clinical and radiologic abnormalities consistent with periventricular leukomalacia were investigated with MR imaging of the brain that employed both inversion-recovery and T2-weighted spin-echo imaging sequences. The more precise delineation of white and gray matter on inversion-recovery images as compared with CT allows a detailed demonstration of the anatomic features of periventricular leukomalacia; specifically, a reduced quantity of white matter in the periventricular region and centrum semiovale and, in more severe cases, cavitated infarcts that replace the immediate periventricular white matter. The T2-weighted spin-echo and short inversion time inversion-recovery images demonstrated abnormally increased signal in white matter that appeared normal on CT scans and only minimally abnormal on conventional inversion-recovery images. These abnormalities most probably represent white matter gliosis that extends beyond the immediate periventricular regions. MR recognition of cerebral white matter abnormalities associated with periventricular leukomalacia may confirm the clinical suspicion of this diagnosis in children with spastic diplegia or quadriplegia.


Impact of lymphedema and arm symptoms on quality of life in breast cancer survivors.
Lymphedema is one of many arm problems reported by breast cancer survivors. Understanding the impact of lymphedema on quality of life requires consideration that arm symptoms may occur with or without lymphedema. It was hypothesized that specific arm symptoms and pain, related or unrelated to lymphedema, would be more associated with quality of life outcomes than arm swelling. The relation of arm swelling and of arm symptoms and associated severity with a range of quality of life outcomes following breast cancer treatment was assessed in a diverse sample of 295 women, 141 of whom had a clinical diagnosis of lymphedema. Arm swelling (as defined by interlimb volume or circumference differences) and lymphedema severity (defined by Common Toxicity Criteria) were less correlated with quality of life than total number of arm symptoms and specific individual symptoms. Pain in the affected arm correlated with poor quality of life outcomes, regardless of arm swelling. When evaluating the impact of lymphedema on quality of life, arm swelling may not be as important as the total number and specific types of arm symptoms present, as these may be more informative about quality of life outcomes in survivors of breast cancer with and without lymphedema.


Nonmedical Use of Prescription Stimulants Among US High School Students to Help Study: Results From a National Survey.
Mixed findings exist regarding extent and efficacy of nonmedical use of prescription stimulants (NMUPS) for study enhancement (SE). This national study of US high school seniors examined NMUPS for SE and addressed risk/benefit questions: To what extent are students reporting NMUPS specifically for SE, and do these individuals demonstrate fewer problem behaviors and superior academic performance? Total of 15 098 US students surveyed (2009-2015) and divided into 4 subgroups: (1) no past-year NMUPS (nonusers), (2) past-year NMUPS to help study (NMUPS-SE only), (3) past-year NMUPS for study/nonstudy motives (NMUPS-SE+ other), and (4) past-year NMUPS for nonstudy motives (NMUPS-nonSE only). Student characteristics (eg, grade point average [GPA]) and substance-related problems (eg, binge drinking) compared between subgroups. Among students who reported past-year NMUPS (n = 781), 7.4% reported NMUPS-SE only, 40.9% NMUPS-SE+ other, and 51.7% NMUPS-nonSE only. Odds of binge drinking, cigarette smoking, marijuana, and opioid nonmedical use significantly higher among all NMUPS subgroups. GPAs significantly lower among subgroups reporting NMUPS nonstudy motives; did not differ between NMUPS-SE only and nonusers. 7% of US high school seniors engaged in NMUPS for SE only (0.4% total population). Findings indicate greater substance-related problems without superior academic performance among NMUPS-SE subgroups.


Cyclebase 3.0: a multi-organism database on cell-cycle regulation and phenotypes.
The eukaryotic cell division cycle is a highly regulated process that consists of a complex series of events and involves thousands of proteins. Researchers have studied the regulation of the cell cycle in several organisms, employing a wide range of high-throughput technologies, such as microarray-based mRNA expression profiling and quantitative proteomics. Due to its complexity, the cell cycle can also fail or otherwise change in many different ways if important genes are knocked out, which has been studied in several microscopy-based knockdown screens. The data from these many large-scale efforts are not easily accessed, analyzed and combined due to their inherent heterogeneity. To address this, we have created Cyclebase--available at http://www.cyclebase.org--an online database that allows users to easily visualize and download results from genome-wide cell-cycle-related experiments. In Cyclebase version 3.0, we have updated the content of the database to reflect changes to genome annotation, added new mRNA and protein expression data, and integrated cell-cycle phenotype information from high-content screens and model-organism databases. The new version of Cyclebase also features a new web interface, designed around an overview figure that summarizes all the cell-cycle-related data for a gene.


Ecology of Freedom: Competitive Tests of the Role of Pathogens, Climate, and Natural Disasters in the Development of Socio-Political Freedom.
Many countries around the world embrace freedom and democracy as part of their political culture. However, culture is at least in part a human response to the ecological challenges that a society faces; hence, it should not be surprising that the degree to which societies regulate the level of individual freedom is related to environmental circumstances. Previous research suggests that levels of societal freedom across countries are systematically related to three types of ecological threats: prevalence of pathogens, climate challenges, and natural disaster threat. Though their incidence overlaps, the literature has not yet provided a competitive test. Drawing upon the ecocultural framework, we tested five rival hypotheses, alternately focused on the above ecological factors and their interactions with economic wealth in explaining country variations in socio-political freedom. Focusing on data from 150 countries, we performed a series of linear mixed-effects regressions predicting freedom in the domains of politics, media, and economy. We found that countries with higher pathogen prevalence were more likely to suppress democracy and media freedom. Economic wealth, however, moderated the effect of pathogen prevalence on economic freedom, with the main effect being only found among wealthy countries, but not among poor countries. In contrast, natural disaster threat predicted political freedom and press freedom only among poor countries, consistent with the idea that disaster threat accompanied by poor resources promote socio-political freedom as a means of increasing collective survival. Throughout our analyses, we found no support for hypotheses based on climatic challenges. In addition, our multilevel approach revealed that country scores for socio-political freedom were highly clustered within world regions, accounting for substantial portions of variance. Overall, the present research offers a nuanced view of the interplay between ecology and wealth in the emergence of socio-political freedom. We discuss new directions in future research considering methodological and theoretical contributions of the present findings.


A subcortical circuit linking the cerebellum to the basal ganglia engaged in vocal learning.
Speech is a complex sensorimotor skill, and vocal learning involves both the basal ganglia and the cerebellum. These subcortical structures interact indirectly through their respective loops with thalamo-cortical and brainstem networks, and directly via subcortical pathways, but the role of their interaction during sensorimotor learning remains undetermined. While songbirds and their song-dedicated basal ganglia-thalamo-cortical circuitry offer a unique opportunity to study subcortical circuits involved in vocal learning, the cerebellar contribution to avian song learning remains unknown. We demonstrate that the cerebellum provides a strong input to the song-related basal ganglia nucleus in zebra finches. Cerebellar signals are transmitted to the basal ganglia via a disynaptic connection through the thalamus and then conveyed to their cortical target and to the premotor nucleus controlling song production. Finally, cerebellar lesions impair juvenile song learning, opening new opportunities to investigate how subcortical interactions between the cerebellum and basal ganglia contribute to sensorimotor learning.


Transcriptional activation of human zeta 2 globin promoter by the alpha globin regulatory element (HS-40): functional role of specific nuclear factor-DNA complexes.
We studied the functional interaction between human embryonic zeta 2 globin promoter and the alpha globin regulatory element (HS-40) located 40 kb upstream of the zeta 2 globin gene. It was shown by transient expression assay that HS-40 behaved as an authentic enhancer for high-level zeta 2 globin promoter activity in K562 cells, an erythroid cell line of embryonic and/or fetal origin. Although sequences located between -559 and -88 of the zeta 2 globin gene were dispensable for its expression on enhancerless plasmids, they were required for the HS-40 enhancer-mediated activity of the zeta 2 globin promoter. Site-directed mutagenesis demonstrated that this HS-40 enhancer-zeta 2 globin promoter interaction is mediated by the two GATA-1 factor binding motifs located at -230 and -104, respectively. The functional domains of HS-40 were also mapped. Bal 31 deletion mapping data suggested that one GATA-1 motif, one GT motif, and two NF-E2/AP1 motifs together formed the functional core of HS-40 in the erythroid-specific activation of the zeta 2 globin promoter. Site-directed mutagenesis further demonstrated that the enhancer function of one of the two NF-E2/AP1 motifs of HS-40 is mediated through its binding to NF-E2 but not AP1 transcription factor. Finally, we did genomic footprinting of the HS-40 enhancer region in K562 cells, adult nucleated erythroblasts, and different nonerythroid cells. All sequence motifs within the functional core of HS-40, as mapped by transient expression analysis, appeared to bind a nuclear factor(s) in living K562 cells but not in nonerythroid cells. On the other hand, only one of the apparently nonfunctional sequence motifs was bound with factors in vivo. In comparison to K562, nucleated erythroblasts from adult human bone marrow exhibited a similar but nonidentical pattern of nuclear factor binding in vivo at the HS-40 region. These data suggest that transcriptional activation of human embryonic zeta 2 globin gene and the fetal/adult alpha globin genes is mediated by erythroid cell-specific and developmental stage-specific nuclear factor-DNA complexes which form at the enhancer (HS-40) and the globin promoters.


Lack of evidence for the involvement of cytochrome P-450 or other hemoproteins in metabolic activation of glyceryl trinitrate in rabbit aorta.
The present study was conducted to test the hypothesis that glyceryl trinitrate (GTN) metabolic activation in blood vessels is mediated by cytochrome P-450 or some other hemoprotein. The effects of cytochrome P-450 inhibitor [2-dimethylaminoethyl-2,2-diphenyl-n-pentanoate (SKF-525A) and metyrapone] on the response of rabbit aortic rings to GTN was determined by recording cumulative GTN dose-response curves in the presence of either or both inhibitors. The cytochrome P-450 inhibitors did not interfere with the ability of GTN to relax rabbit aortic rings. Treatment of rabbit aortic strips (RAS) with carbon monoxide (CO) was undertaken to elucidate a potential role of ferrous hemoproteins in mediating GTN-induced relaxation. CO did not significantly inhibit GTN-induced vasodilation of RAS. Biotransformation of GTN to glyceryl dinitrates by RAS exposed to CO for 5 min or nitric oxide (NO) for 10 min was determined by incubation of the tissues with 0.5 microM [3H]- or [14C]-GTN in the presence of CO or NO for 10 sec, 30 sec or 2 min. Neither CO nor NO exposure inhibited glyceryl-1,2- or -1,3-dinitrate formation by RAS after the 10-sec or 2-min incubation with RAS. For the 30-sec incubation, NO exposure did inhibit glyceryl dinitrate formation by RAS. This was deemed to be less important than the 10-sec data, the time just before the onset of relaxation. Therefore, emphasis is placed on lack of inhibition by NO at this earlier time point.(ABSTRACT TRUNCATED AT 250 WORDS)


Micromanaging abdominal aortic aneurysms.
The contribution of abdominal aortic aneurysm (AAA) disease to human morbidity and mortality has increased in the aging, industrialized world. In response, extraordinary efforts have been launched to determine the molecular and pathophysiological characteristics of the diseased aorta. This work aims to develop novel diagnostic and therapeutic strategies to limit AAA expansion and, ultimately, rupture. Contributions from multiple research groups have uncovered a complex transcriptional and post-transcriptional regulatory milieu, which is believed to be essential for maintaining aortic vascular homeostasis. Recently, novel small noncoding RNAs, called microRNAs, have been identified as important transcriptional and post-transcriptional inhibitors of gene expression. MicroRNAs are thought to "fine tune" the translational output of their target messenger RNAs (mRNAs) by promoting mRNA degradation or inhibiting translation. With the discovery that microRNAs act as powerful regulators in the context of a wide variety of diseases, it is only logical that microRNAs be thoroughly explored as potential therapeutic entities. This current review summarizes interesting findings regarding the intriguing roles and benefits of microRNA expression modulation during AAA initiation and propagation. These studies utilize disease-relevant murine models, as well as human tissue from patients undergoing surgical aortic aneurysm repair. Furthermore, we critically examine future therapeutic strategies with regard to their clinical and translational feasibility.


Uveitic macular oedema: correlation between optical coherence tomography patterns with visual acuity and fluorescein angiography.
(1) To associate optical coherence tomography (OCT) patterns with particular fluorescein angiography (FA) findings in uveitis patients with macular oedema. (2) To investigate the correlation between tomographic features and visual acuity (VA). (3) To determine the impact of OCT patterns on visual improvement over time. 129 eyes of 90 patients followed for uveitis with macular oedema and adequate media clarity were consecutively included from February to November 2004. We examined the relationship between VA, the fluorescein angiograms grading and central thickness. Mean retinal thickness at the central fovea was 357 (SD 128) microm. The mean VA was 0.5 (logMAR). There was agreement between OCT and FA in 112 eyes (86.8%). In eight eyes (6.2%), serous retinal detachment (SRD) was detected on OCT but not on FA. Nine eyes (7%) had perifoveal leakage on FA without macular thickening on OCT. There were three patterns of macular oedema on OCT: diffuse macular oedema (DMO), cystoid macular oedema (CMO) and SRD. CMO was detected in 61 eyes (50.8%), and in 18 of these eyes (15%), CMO was associated with SRD. DMO was detected in 45 eyes (37.5%), and in eight (6.7%) of these eyes, DMO was associated with SRD. SRD without retinal thickening was present in 14 eyes (11%). Epiretinal membrane was detected by OCT in 27 eyes (21%). Six eyes demonstrated vitreomacular traction (4.7%). There was a correlation between VA and central thickness on OCT (r = 0.407, p = 0.000001). Correlation between VA and central thickness is significant in the group with CMO (r = 0.401, p = 0.001) but not significant in the group with DMO. Furthermore, vision recovery was observed in patients with CMO but was limited in patients with DMO. SRD did not have a significant impact on visual recovery, but its presence reduced the correlation between vision and central thickness. OCT is effective in detection of macular oedema. It allows determination of the distribution of fluid and quantification of retinal thickness particularly in patients with CMO. In these patients, a potential for vision recovery was also identified. DMO was associated with a poor visual prognosis and a poor prognosis for vision recovery. SRD is associated with a high probability of vision recovery when observed alone or underlying CMO eyes. It should be substracted from the central thickness measurement when attempting to correlate central thickness with vision in patients with macular oedema in uveitis.


Review of patient safety incidents reported from critical care units in North-West England in 2009 and 2010.
We categorised and established the rates of patient safety incidents reported during 2009 and 2010 from critical care units in 12 hospital trusts in North-West England. We identified a total of 4219 incidents reported during 127, 467 calendar days of critical care with a median (IQR [range]) of 31 (26-45 [20-57]) incidents per 1000 days per trust. A median (IQR [range]) of 10 (7-13 [3.5-27]) incidents per 1000 days were associated with harm. Pressure sores were the most common cause of harm, with a median (IQR [range]) of 3.9 (1.0-6.6 [0-20.4]) incidents per 1000 days. Only 89 (2.1%) incidents described more than temporary harm, of which 12 were airway related incidents. Five incidents described the use of inappropriate arterial flush solutions. It is possible to compare rates of incident reporting in different trusts over time to determine if different methods of care are associated with different reporting rates. The wide range of reported pressure sore rates suggests that their incidence could be reduced.


Does cigarette use influence adiposity or height in adolescence?
To study the effect of cigarette use on height and adiposity in adolescents. Data on cigarette use were collected every 3 months for 5 years from adolescents initially 12-13 years of age. Height, weight, and triceps skinfold thickness were measured in survey cycles 1, 12, and 19. Multivariate linear regression models were fitted to estimate the association between cigarette use and the anthropometric measures in a dataset that pooled data over two time periods, from survey cycles 1-12 and from survey cycles 12-19. Data were available for 451 boys and 478 girls. Seven percent of boys and 14% of girls smoked > or =30 cigarettes per month on average during the first time period; 9% of boys and 18% of girls smoked > or =30 cigarettes per month on average during the second time period. In boys, a 100-cigarette per month increment in cigarette use over the preceding 2.5 years was independently associated with lower body mass index (-0.4 kg/m(2)) and shorter height (-0.7 cm). In girls, cigarette use was not associated with height or adiposity. While there was no relation in girls, cigarette use appears to decrease body mass index and height in boys. Young girls may be less likely to take up cigarette smoking if tobacco control messages emphasize that cigarette use may not be associated with reduced weight in adolescent females.


Regional Cultures and the Psychological Geography of Switzerland: Person-Environment-Fit in Personality Predicts Subjective Wellbeing.
The present study extended traditional nation-based research on person-culture-fit to the regional level. First, we examined the geographical distribution of Big Five personality traits in Switzerland. Across the 26 Swiss cantons, unique patterns were observed for all traits. For Extraversion and Neuroticism clear language divides emerged between the French- and Italian-speaking South-West vs. the German-speaking North-East. Second, multilevel modeling demonstrated that person-environment-fit in Big Five, composed of elevation (i.e., mean differences between individual profile and cantonal profile), scatter (differences in mean variances) and shape (Pearson correlations between individual and cantonal profiles across all traits; Furr, 2008, 2010), predicted the development of subjective wellbeing (i.e., life satisfaction, satisfaction with personal relationships, positive affect, negative affect) over a period of 4 years. Unexpectedly, while the effects of shape were in line with the person-environment-fit hypothesis (better fit predicted higher subjective wellbeing), the effects of scatter showed the opposite pattern, while null findings were observed for elevation. Across a series of robustness checks, the patterns for shape and elevation were consistently replicated. While that was mostly the case for scatter as well, the effects of scatter appeared to be somewhat less robust and more sensitive to the specific way fit was modeled when predicting certain outcomes (negative affect, positive affect). Distinguishing between supplementary and complementary fit may help to reconcile these findings and future research should explore whether and if so under which conditions these concepts may be applicable to the respective facets of person-culture-fit.


Trends in diagnosis and control of bovine mastitis: a review.
Mastitis (inflammation of mammary gland) is a most devastating disease condition in terms of economic losses occurring throughout the world. The etiological agents may vary from place to place depending on climate; animal species and animal husbandry and include wide variety of gram positive and gram negative bacteria; and fungi. They may be either contagious viz. Staphylococcus aureus; Streptococcus agalactiae or environmental viz. S. dysgalactiae, S. uberis, Corynebacterium bovis and Coagulase negative Staphylococcus. Conventional diagnostic tests viz. California Mastitis Test (CMT); R-mastitest and Mast-O-test methods are applied under field conditions; whereas somatic cell count and Bulk Tank Somatic Cell Count (BTSCC) are useful for early mastitis detection and detection of sub clinical or chronic mastitis respectively. In vitro culture based diagnosis require further study as they can detect only viable cells. The advent of Polymerase Chain Reaction (PCR) technology along with its various versions like multiplex and real time PCR has improved the rapidity and sensitivity of diagnosis. Circulating micro RNA (miRNA) based diagnosis; immune assay and proteomics based detection along with biochips and biosensors prove to be asset to diagnosticians for advanced diagnosis of this economically important condition. Improvement of milking hygiene; implementation of post-milking teat disinfection; regular control of the milking equipments; implementation of milking order; Improvement of bedding material are the general measures to prevent new cases of mastitis. The use of antibiotics (intramammary infusions; bacteriocins) and herbs (Terminalia spp.) are important for prophylaxis and therapeutics. Vaccines viz. cell based; Recombinant (staphylococcal enterotoxin type C mutant) or chimeric (pauA); live (S. uberis 0140J stain based) and bacterial surface extract based; DNA-based and DNA-protein based have greatly aided in management of bovine mastitis. Quorum sensing and disease resistant breeding using novel biomarkers viz. toll like receptors (TLR) 2 and 4, interleukin (IL) 8; breast cancer type 1 susceptibility protein (BRCA1) and calcium channel voltage-dependent alpha 2/delta sub unit 1 (CACNA2D1) are also indispensable. This mini review gives an overview of all these different aspects that act as trend setters as far as the diagnosis and control of bovine mastitis is concerned to help the diagnosticians; epidemiologists and researchers not to remain ignorant about this grave condition.


A human testis-specific mRNA for phosphoribosylpyrophosphate synthetase that initiates from a non-AUG codon.
Two highly homologous subunits for phosphoribosylpyrophosphate synthetase are encoded by human X-linked genes, PRPS1 and PRPS2 (Taira, M., Kudoh, J., Minoshima, S., Iizasa, T., Shimada, H., Shimizu, Y., Tatibana, M., and Shimizu, N. (1989b) Somat. Cell Mol. Genet. 15, 29-37). These genes are expressed in most tissues, whereas an additional unique mRNA (1.4 kilobases) is present in the testes of rats as well as mice and humans (Taira, M., Iizasa, T., Yamada, K., Shimada, H., and Tatibana, M. (1989a) Biochim. Biophys. Acta 1007, 203-208). In this paper, cDNA cloning revealed that the human testis-specific mRNA was encoded by an autosomal gene, termed PRPS3. RNA blot analysis showed that the expression of this gene began at 4 weeks of age in rats, coinciding with the reported appearance of primary spermatocytes. A cDNA clone of PRPS3 was sequenced and found to encode a predicted product of 317 amino acids which was highly homologous to those of PRPS1 and PRPS2 (94.3% and 91.2% identities, respectively). However, the PRPS3 cDNAs lacked an ATG initiator for translation at the expected position, and instead contained an ACG triplet. In vitro transcription/translation studies, combined with in vitro site-directed mutagenesis experiments, suggested that the ACG codon at this position did serve as a start codon. Analysis of amino-terminal sequence of the radiolabeled PRPS3 product, prepared by in vitro translation, supported the predicted sequence starting with Pro-1, and, in addition, this product was labeled with N-formyl[35S]methionyl-tRNAi. These results suggested that the synthesis of the nascent polypeptide could initiate with methionine at the position corresponding to the ACG codon.


Struck: Structured Output Tracking with Kernels.
Adaptive tracking-by-detection methods are widely used in computer vision for tracking arbitrary objects. Current approaches treat the tracking problem as a classification task and use online learning techniques to update the object model. However, for these updates to happen one needs to convert the estimated object position into a set of labelled training examples, and it is not clear how best to perform this intermediate step. Furthermore, the objective for the classifier (label prediction) is not explicitly coupled to the objective for the tracker (estimation of object position). In this paper, we present a framework for adaptive visual object tracking based on structured output prediction. By explicitly allowing the output space to express the needs of the tracker, we avoid the need for an intermediate classification step. Our method uses a kernelised structured output support vector machine (SVM), which is learned online to provide adaptive tracking. To allow our tracker to run at high frame rates, we (a) introduce a budgeting mechanism that prevents the unbounded growth in the number of support vectors that would otherwise occur during tracking, and (b) show how to implement tracking on the GPU. Experimentally, we show that our algorithm is able to outperform state-of-the-art trackers on various benchmark videos. Additionally, we show that we can easily incorporate additional features and kernels into our framework, which results in increased tracking performance.


Integration of an intensified charge-coupled device (ICCD) camera for accurate spectroscopic measurements.
Intensified charge-coupled devices (ICCD) are used in a great variety of spectroscopic applications, some of them requiring high sensitivity and spectral resolution. The setup, configuration, and featuring of these cameras are fundamental issues in order to acquire high quality spectra. In this work a critical assessment of these detectors is performed and the specific configuration, the optical alignment, featuring, and the dark and shot noise are described and analyzed. Spatial response of the detector usually shows a significant lack of spatial homogeneity and a map of interferences may appear in certain ranges of wavelengths, which damages the quality of the recorded spectra. In this work the spectral resolution and the spatial and spectral sensitivity are also studied. The analysis of the dark current reveals the existence of a smooth but clear spatial dependence. As a final conclusion, the spectra registered with the spectrometer equipped with our ICCD camera allow us to explore and measure accurately spectral line shapes emitted by pulsed plasmas in the visible range and particularly in the ultraviolet (UV) range.


Predictors of accuracy of diagnosis of chronic obstructive pulmonary disease in general practice.
To compare the clinical diagnosis of chronic obstructive pulmonary disease (COPD) with results of post-bronchodilator spirometry in general practice, and examine practitioner, practice and patient characteristics associated with agreement between clinical and spirometric diagnoses. General practitioners from practices in Sydney identified eligible patients aged 40-80 years seen in the past year and prescribed respiratory medications whom they regarded as having COPD. Between November 2006 and April 2008, we collected information on the GPs and their practices, and demographic information, smoking status, comorbidities, respiratory medicines use, vaccination status, quality of life and spirometry results for participating patients. Frequency of COPD diagnosis on spirometry; odds ratios for characteristics associated with agreement between clinical and spirometric diagnoses. 56 GPs from 44 practices participated in the study. Of 1144 eligible patients, 445 were recruited (mean age, 65 years; 49% male). Of these, 257 (57.8%) had post-bronchodilator spirometry consistent with COPD ± asthma, 16 (3.6%) had asthma only, 82 (18.4%) had normal spirometry, and 90 (20.2%) had other spirometric diagnoses. Having a spirometer in the practice was not predictive of agreement between clinical and spirometric diagnoses. Older patient age was significantly associated with correct diagnosis, while higher numbers of comorbidities were associated with misdiagnosis. A substantial proportion of patients clinically identified as having COPD in general practice do not have the condition according to spirometric criteria, with inaccurate diagnosis more common in patients with comorbidities. Policy and practice change is needed to support the use of spirometry in primary care.


Caffeine in plasma and saliva by a radioimmunoassay procedure.
Caffeine was analyzed in human plasma and saliva by a simple, rapid, and sensitive radioimmunoassay procedure. Immunization of rabbits with an antigen prepared by coupling 7-(5-carboxypentyl)-1,3-dimethylxanthine to bovine serum albumin resulted in the formation of antibodies selective for caffeine as opposed to various mono- and dimethylxanthines, mono-, di-, and trimethyluric acids and a variety of common drugs. The radioligand used for competitive binding studies was 7-(2,3-3H2-propyl)-1,3-dimethylxanthine. The procedure permits direct analysis of caffeine in plasma or saliva without extraction. Comparison with a high pressure liquid chromatography method for the analysis of caffeine gave satisfactory results and showed no evidence for interference by metabolites. A caffeine half-life of 4.0 hours determined by the radioimmunoassay was in agreement with previous work. Comparison of human plasma and saliva levels by the radioimmunoassay procedure indicated approximately equal concentrations in the two fluids.


Emergency care research priorities in South Africa.
Emergency care research is rarely undertaken in low- and middle-income countries. A manageable 'road map' for research in South African (SA) emergency care is needed to address research gaps. To identify, collate and prioritise research topics from identified knowledge gaps in emergency care in SA. Seventy-six individuals were invited to participate in a modified Delphi study. Participants were requested to suggest important research topics before rating them. Consensus was achieved when >75% of participants strongly agreed or disagreed. Participants then ranked the agreed statements before selecting the most appropriate methodology relating to study design, funding and collaboration. Three hundred and fifty topics were suggested by 31 participants. Topics were collated into 123 statements before participants rated them. Consensus was achieved for 39 statements. The highest-ranked priority in the prehospital group was to determine which prehospital interventions improve outcomes in critically ill patients. The competence of emergency care providers in performing common lifesaving skills was deemed the most important in clinical emergency care. Implementing and reviewing quality improvement systems scored the highest under general systems and safety management. Only 22 statements achieved consensus regarding study design. The National Department of Health was the preferred funding source, while private organisations and emergency care societies were identified as possible collaborative partners. This study provides expert consensus on priority research areas in emergency care in SA as a guide for emergency care providers to ensure evidence-based care that is relevant to the SA population.


A biaxial rotating bioreactor for the culture of fetal mesenchymal stem cells for bone tissue engineering.
The generation of effective tissue engineered bone grafts requires efficient exchange of nutrients and mechanical stimulus. Bioreactors provide a manner in which this can be achieved. We have recently developed a biaxial rotating bioreactor with efficient fluidics through in-silico modeling. Here we investigated its performance for generation of highly osteogenic bone graft using polycaprolactone-tricalcium phosphate (PCL-TCP) scaffolds seeded with human fetal mesenchymal stem cell (hfMSC). hfMSC scaffolds were cultured in either bioreactor or static cultures, with assessment of cellular viability, proliferation and osteogenic differentiation in vitro and also after transplantation into immunodeficient mice. Compared to static culture, bioreactor-cultured hfMSC scaffolds reached cellular confluence earlier (day 7 vs. day 28), with greater cellularity (2x, p<0.01), and maintained high cellular viability in the core, which was 2000 microm from the surface. In addition, bioreactor culture was associated with greater osteogenic induction, ALP expression (1.5x p<0.01), calcium deposition (5.5x, p<0.001) and bony nodule formation on SEM, and in-vivo ectopic bone formation in immunodeficient mice (3.2x, p<0.001) compared with static-cultured scaffolds. The use of biaxial bioreactor here allowed the maintenance of cellular viability beyond the limits of conventional diffusion, with increased proliferation and osteogenic differentiation both in vitro and in vivo, suggesting its utility for bone tissue engineering applications.


Photodegradation Pathways of Protein Disulfides: Human Growth Hormone.
Comprehensive product characterization was performed for the photodegradation of protein disulfides, representatively of human growth hormone (somatotropin; hGH), in order to provide a product database, which will be useful for the general analysis of protein stability. HGH was photo-irradiated at λ = 254 and λ > 295 nm and tryptic digests were analyzed by HPLC-MS to investigate light-induced disulfide degradation pathways. A total of 60 products were detected, and structures/tentative structures were assigned to the products by MS2 and MS3 analysis. The main products were reduced Cys residues, dithiohemiacetal, thioether and disulfide scrambling products. In addition, we detected Cys degradation products such as Cys thioaldehyde, dehydroalanine (Dha), Ala, Ser semialdehyde, Ser, S-sulfocysteine, and Gly. Frequently, the tryptic fragments contained more than one modification, i.e. a Cys degradation product in close proximity to a dehydrated amino acid. Several novel cross-links were detected between Cys and Tyr, Cys, Ser and Phe, Cys and Trp, and Trp and Tyr. Photo-induced protein fragmentation was detected specifically at or in close proximity to the disulfide bond between T6 and T16. An in-house packed 75 cm nano-column enabled us to resolve various isomers/diastereomers of the photo-degradation products. A comprehensive analysis of photodegradation products revealed a variety of novel photo-products, including cross-links, originating from disulfide degradation. The mechanisms of product formation are discussed.


Is statin-modified reduction in lipids the most important preventive therapy for cardiovascular disease? A pro/con debate.
The most prescribed medications in the world are statins, lipid modifiers that have been available for over 25 years and amongst the most investigated of all drug classes. With over a million patient years of trial data and publications in the most prestigious medical journals, it is remarkable that quite so much debate remains as to their place in healthcare. They have had a bittersweet passage, with vocal concerns over their possible risks, from suicide to cancer, and allegations that they do not work in women or the elderly, to statements that the whole published dataset, on over 200,000 patients consenting to enter trials, was fatally compromised by being industry-funded by and large. On the other side, there have been billions of dollars spent on generating their evidence base followed by promotion which has returned that investment many times over in profits, and a powerful scientific lobby that argue they are wonder drugs and that continued nihilism on their value risks patient lives. So who is right?


CcpC, a novel regulator of the LysR family required for glucose repression of the citB gene in Bacillus subtilis.
Synergistic carbon catabolite repression of the Bacillus subtilis aconitase (citB) gene by glucose and a source of 2-ketoglutarate is dependent on DNA sequences located upstream of the gene. Mutations in a dyad symmetry element centered at position -66 and in a repeat of the downstream arm of the dyad symmetry at position -27 cause derepressed citB expression. In this work, a protein able to bind to a DNA fragment containing these elements was purified and identified. This protein, named CcpC (Catabolite control protein C), shares sequence similarity with members of the LysR family of transcriptional regulators. In addition to binding to the citB promoter, CcpC bound to the promoter of the citZ gene, which encodes the cell's major citrate synthase and is subject to carbon catabolite repression. In a ccpC null mutant, expression of both citB and citZ was derepressed in glucose-glutamine minimal medium, indicating that CcpC is a negative regulator of citB and citZ gene expression. DNase I footprinting experiments showed that CcpC binds to two sites within the citB promoter region, corresponding to the dyad symmetry and -27 elements. In the presence of citrate, a putative inducer, only the dyad symmetry element was fully protected by CcpC. When the dyad symmetry element was mutated, CcpC was no longer able to bind to either the dyad symmetry or -27 elements. Repression of citB and citZ gene expression during anaerobiosis also proved to be mediated by CcpC.


Improving education and coping of scoliosis patients undergoing surgery, and their families, using e-health.
Healthcare providers have limited time to spend with scoliosis patients who are considering surgery and their families. The purpose of this study was to evaluate an e-health strategy to increase knowledge and coping in patients with scoliosis who are surgical candidates and their families. We enrolled patients with scoliosis who were candidates for surgery and their families. Patients and their families completed the scoliosis knowledge questionnaire, meaning of illness questionnaire, social support and coping questionnaires before and after access to a comprehensive evidence-based scoliosis website ( http://www.aboutkidshealth.ca/scoliosis ). Seventy-four patients and 71 parents completed the evaluation. While both patients and parents improved their knowledge of scoliosis (p = 0.001 and p = 0.003, respectively), the scores of patients were consistently lower than those of the parents both before and after website use (p = 0.0001). Only parents demonstrated a change in the meaning of illness questionnaire, with a small increase in the negative attitude towards illness and a small decrease in the positive attitude towards illness (p = 0002 and p = 0.01, respectively). Of the 12 coping methods examined on the Adolescent Coping Orientation for Problem Experiences (A-COPE) instrument, patients were slightly more likely than parents to use relaxing and solving family problems as tools to cope following website access (p = 0.02 and p = 0.09, respectively). Parents demonstrated no significant changes in the four methods of coping on the Coping Health Inventory for Parents (CHIP) after website exposure. While the majority of patients and parents reported receiving sufficient support, over half of the patients indicated a need for more support in social participation. An evidence-based website increased the knowledge of patients and parents but simply providing access to the website had minimal impact on their coping and perceptions of social support. The website, however, provides users with the opportunity to absorb vital information about scoliosis across several media.


Herpesvirus Infections in KIR2DL2-Positive Multiple Sclerosis Patients: Mechanisms Triggering Autoimmunity.
In multiple sclerosis (MS), there is a possible relationship with viral infection, evidenced by clinical evidence of an implication of infectious events with disease onset and/or relapse. The aim of this research is to study how human herpesvirus (HHVs) infections might dysregulate the innate immune system and impact autoimmune responses in MS. We analyzed 100 MS relapsing remitting patients, in the remission phase, 100 healthy controls and 100 subjects with other inflammatory neurological diseases (OIND) (neuro-lupus) for their immune response to HHV infection. We evaluated NK cell response, levels of HHVs DNA, IgG and pro- and anti-inflammatory cytokines. The results demonstrated that the presence of KIR2DL2 expression on NK cells increased the susceptibility of MS patients to HHV infections. We showed an increased susceptibility mainly to EBV and HHV-6 infections in MS patients carrying the KIR2DL2 receptor and HLA-C1 ligand. The highest HHV-6 viral load was observed in MS patients, with an increased percentage of subjects positive for IgG against HHV-6 in KIR2DL2-positive MS and OIND subjects compared to controls. MS and OIND patients showed the highest levels of IL-8, IL-12p70, IL-10 and TNF-alpha in comparison with control subjects. Interestingly, MS and OIND patients showed similar levels of IL-8, while MS patients presented higher IL-12p70, TNF-alpha and IL-10 levels in comparison with OIND patients. We can hypothesize that HHVs' reactivation, by inducing immune activation via also molecular mimicry, may have the ability to induce autoimmunity and cause tissue damage and consequent MS lesion development.


Point-of-care Hepatitis C virus testing and linkage to treatment in an Australian inner-city emergency department.
In Australia, Hepatitis C Virus (HCV) treatment is declining, despite broad access to direct-acting antiviral medication. People who inject drugs are proportionally over-represented in emergency department presentations. Emergency department assessment of people who have injected drugs for HCV presents an opportunity to engage this marginalised population with treatment. We describe the outcomes of risk-based screening and point-of-care anti-HCV testing for emergency department patients, and linkage to outpatient antiviral treatment. During the three-month study period, consecutive adult patients who presented to the emergency department during the study times were screened for risk factors and offered the OraQuick oral HCV antibody test. Those with reactive results were offered venepuncture in the emergency department for confirmatory testing and direct-acting antiviral treatment in clinic. The main outcome measures were the number and proportion of viremic participants that were linked to the hepatitis clinic, commenced treatment and achieved a sustained viral response. Secondary outcome measures were the proportion (%) of presentations screened that were oral antibody reactive, and the prevalence and type of HCV risk factors. During the study period, 2408 of the 3931 (61%) presentations to the emergency department were eligible for screening. Of these 2408 patients, 1122 (47%) participated, 307 (13%) declined participation and 977 (41%) could not be approached during their time in the emergency department. Among the 1122 participants, 378 (34%) reported at least one risk factor. Subsequently, 368 (97%) of the 378 participants underwent OraQuick anti-HCV test, and 50 (14%) had a reactive result. A risk factor of ever having injected drugs was present in 44 (88%) of participants who were sero-positive. Of the 45 that had blood tested, 30 (67%) were HCV ribonucleic acid (RNA) positive. Three participants died. Of the 27 remaining participants, 10 (37%) commenced treatment and 7 of these 10 (70%) obtained a cure. There was a high rate of homelessness (24%) among anti-HCV positive participants. Among emergency department participants with a risk factor for HCV, positive serology was common using a rapid point-of-care test. A history of injecting drug use was identified as the risk factor with highest yield for positive HCV serology, and is suitable as a single screening question. However, linkage to care post ED presentation was low in this marginalised population. There is a need for new pathways to improve the care cascade for marginalised individuals living with HCV infection.


Lower serum magnesium is associated with vascular calcification in peritoneal dialysis patients: a cross sectional study.
Coronary artery calcification (CAC) is highly prevalent among dialysis patients and is associated with increased cardiovascular and all cause mortality. Magnesium (Mg) inhibits vascular calcification in animal and in-vitro studies but whether the same effect occurs in humans is uncertain. A single centre cross-sectional study of 80 prevalent peritoneal dialysis (PD) patients; on PD only for a minimum of 3 months. A radiologist blinded to patient status calculated their abdominal aortic calcification (AAC) scores on lateral lumbar spine radiographs, a validated surrogate for CAC. Eighty patients provided informed consent and underwent lumbar spine radiography. The mean serum Mg was 0.8 mmol/L (standard deviation 0.2) and mean AAC score 8.9 (minimum 0, maximum 24). A higher serum Mg level was associated with a lower AAC score (R 2 = 0.06, unstandardized coefficient [B] = -7.81, p = 0.03), and remained after adjustment for age, serum phosphate, serum parathyroid hormone, low-density lipoprotein cholesterol, smoking history, and diabetes (model adjusted R 2 = 0.36, serum Mg and AAC score B = -11.44, p = 0.00). This translates to a 0.1 mmol/L increase in serum Mg being independently associated with a 1.1-point decrease in AAC score. Our findings suggest that Mg may inhibit vascular calcification. If this association is replicated across larger studies with serial Mg and vascular calcification measurements, interventions that increase serum Mg and their effect on vascular calcification warrant further investigation in the PD population.


The clinical efficacy of deproteinized bovine bone mineral with 10% collagen in conjunction with localized piezosurgical decortication enhanced orthodontics: A prospective observational study.
Evidence exists on the clinical efficacy and safety of periodontally accelerated osteogenic orthodontics (PAOO) with "Piezocision"-a minimally invasive, flapless alternative to corticotomy for alveolar bone augmentation. Allograft has been extensively studied for alveolar bone augmentation in Piezocision; however, the use of deproteinized bovine bone mineral with 10% collagen (DBBM-C) in Piezocision for PAOO has not been investigated. This study is a prospective, observational, cohort study of 19 patients of Angle Class I malocclusion with a total of 692 teeth assessed for maintenance of health of the periodontal attachment apparatus. Patient-centered pain, sensitivity, and satisfaction outcomes, digital photographs and radiographs, and changes in probing depth, clinical attachment level, width of keratinized tissue, percussion sensitivity, pulp vitality tests, radiographic pathology, and root-crown-ratio were all recorded. Overall treatment was significantly faster (5 to 7 days between clear aligner tray changes), periodontal parameters remained stable, and alveolar bone loss was not observed. Visual analog score for healing, sensitivity/duration, bleeding/duration, swelling/duration, appearance, and inflammation, demonstrated no significant differences between DBBM-C and control (no bone graft) groups. Patient-centered outcomes revealed high levels of satisfaction with Piezocision. Piezocision-treated teeth with DBBM-C tended to exhibit less root resorption, although it was not statistically significant (P = 0.074). Within the limits of the study, our results show that the use of DBBM-C with piezosurgically enhanced orthodontics is effective and safe. This study was not designed to demonstrate equivalence with other materials that might be used in Piezocision. To understand whether there is an advantage to using DBBM-C, additional studies may be required.


Effects of estradiol-drospirenone on ocular and nasal functions in postmenopausal women.
Hormones may have an impact on the incidence and course of dry-eye syndrome, and also nasal obstruction, especially in postmenopausal women. The aim of this study was to investigate the effects of drospirenone 2 mg + estradiol 1 mg (Angeliq®, Bayer AG, Berlin, Germany) on ocular functions and nasal airflow in postmenopausal women. Thirty-four postmenopausal women were recruited for the study. A questionnaire on ocular surface disease index (OSDI) was administered; after routine ophthalmologic examination, tear film break-up time (TBUT) and Schirmer tests were performed. Intraocular pressure was measured with a Goldmann applanation tonometer. Nasal airflow resistance values were measured using rhinomanometry. Participants were submitted to oral daily treatment with drospirenone 2 mg + estradiol 1 mg. The duration of the study was 6 months and the participants were studied in the basal condition and after 6 months of therapy. The OSDI score and intraocular pressure were significantly higher before treatment than after treatment (p<0.001); however, the results of the TBUT and Schimer test were significantly lower (p<0.05). Rhinomanometric values were better during drospirenone 2 mg + estradiol 1 mg therapy than those observed at baseline (p<0.001). Oral daily treatment with drospirenone 2 mg + estradiol 1 mg reduces the risks for ocular diseases and nasal obstruction in postmenopausal women.


Public health interventions in midwifery: a systematic review of systematic reviews.
Maternity care providers, particularly midwives, have a window of opportunity to influence pregnant women about positive health choices. This aim of this paper is to identify evidence of effective public health interventions from good quality systematic reviews that could be conducted by midwives. Relevant databases including MEDLINE, Pubmed, EBSCO, CRD, MIDIRS, Web of Science, The Cochrane Library and Econlit were searched to identify systematic reviews in October 2010. Quality assessment of all reviews was conducted. Thirty-six good quality systematic reviews were identified which reported on effective interventions. The reviews were conducted on a diverse range of interventions across the reproductive continuum and were categorised under: screening; supplementation; support; education; mental health; birthing environment; clinical care in labour and breast feeding. The scope and strength of the review findings are discussed in relation to current practice. A logic model was developed to provide an overarching framework of midwifery public health roles to inform research policy and practice. This review provides a broad scope of high quality systematic review evidence and definitively highlights the challenge of knowledge transfer from research into practice. The review also identified gaps in knowledge around the impact of core midwifery practice on public health outcomes and the value of this contribution. This review provides evidence for researchers and funders as to the gaps in current knowledge and should be used to inform the strategic direction of the role of midwifery in public health in policy and practice.


The influence of transformational leadership on employee well-being: results from a survey of companies in the information and communication technology sector in Germany.
The aim of this study was to investigate the impact of a transformational leadership style on employee well-being. Data for the study were taken from an on-line survey of all employees from six German companies in the information and communication technology sector. The data obtained were analyzed using multiple logistic regressions. Analysis of data from a total of 318 employees (response rate 58.4%) revealed a significant relationship between transformational leadership and employee well-being (P = 0.00; OR = 1.13; 95% CI = 1.05 to 1.22). A higher degree of perceived transformational leadership is associated with greater well-being. The results of this study suggest that a transformational leadership style, which both conveys a sense of trust and meaningfulness and individually challenges and develops employees, also has a positive effect on employee well-being.


Prevalence and reporting of recruitment, randomisation and treatment errors in clinical trials: A systematic review.
Background/aims In clinical trials, it is not unusual for errors to occur during the process of recruiting, randomising and providing treatment to participants. For example, an ineligible participant may inadvertently be randomised, a participant may be randomised in the incorrect stratum, a participant may be randomised multiple times when only a single randomisation is permitted or the incorrect treatment may inadvertently be issued to a participant at randomisation. Such errors have the potential to introduce bias into treatment effect estimates and affect the validity of the trial, yet there is little motivation for researchers to report these errors and it is unclear how often they occur. The aim of this study is to assess the prevalence of recruitment, randomisation and treatment errors and review current approaches for reporting these errors in trials published in leading medical journals. Methods We conducted a systematic review of individually randomised, phase III, randomised controlled trials published in New England Journal of Medicine, Lancet, Journal of the American Medical Association, Annals of Internal Medicine and British Medical Journal from January to March 2015. The number and type of recruitment, randomisation and treatment errors that were reported and how they were handled were recorded. The corresponding authors were contacted for a random sample of trials included in the review and asked to provide details on unreported errors that occurred during their trial. Results We identified 241 potentially eligible articles, of which 82 met the inclusion criteria and were included in the review. These trials involved a median of 24 centres and 650 participants, and 87% involved two treatment arms. Recruitment, randomisation or treatment errors were reported in 32 in 82 trials (39%) that had a median of eight errors. The most commonly reported error was ineligible participants inadvertently being randomised. No mention of recruitment, randomisation or treatment errors was found in the remaining 50 of 82 trials (61%). Based on responses from 9 of the 15 corresponding authors who were contacted regarding recruitment, randomisation and treatment errors, between 1% and 100% of the errors that occurred in their trials were reported in the trial publications. Conclusion Recruitment, randomisation and treatment errors are common in individually randomised, phase III trials published in leading medical journals, but reporting practices are inadequate and reporting standards are needed. We recommend researchers report all such errors that occurred during the trial and describe how they were handled in trial publications to improve transparency in reporting of clinical trials.


Fe limitation decreases transcriptional regulation over the diel cycle in the model diatom Thalassiosira pseudonana.
Iron (Fe) is an important growth factor for diatoms and its availability is further restricted by changes in the carbonate chemistry of seawater. We investigated the physiological attributes and transcriptional profiles of the diatom Thalassiosira pseudonana grown on a day: night cycle under different CO2/pH and iron concentrations, that in combination generated available iron (Fe') concentrations of 1160, 233, 58 and 12 pM. We found the light-dark conditions to be the main driver of transcriptional patterns, followed by Fe' concentration and CO2 availability, respectively. At the highest Fe' (1160 pM), 55% of the transcribed genes were differentially expressed between day and night, whereas at the lowest Fe' (12 pM), only 28% of the transcribed genes displayed comparable patterns. While Fe limitation disrupts the diel expression patterns for genes in most central metabolism pathways, the diel expression of light- signaling molecules and glycolytic genes was relatively robust in response to reduced Fe'. Moreover, we identified a non-canonical splicing of transcripts encoding triose-phosphate isomerase, a key-enzyme of glycolysis, generating transcript isoforms that would encode proteins with and without an active site. Transcripts that encoded an active enzyme maintained a diel expression at low Fe', while transcripts that encoded the non-active enzyme lost the diel expression. This work illustrates the interplay between nutrient limitation and transcriptional regulation over the diel cycle. Considering that future ocean conditions will reduce the availability of Fe in many parts of the oceans, our work identifies some of the regulatory mechanisms that may shape future ecological communities.


Prehospital Delay, Precipitants of Admission, and Length of Stay in Patients With Exacerbation of Heart Failure.
Factors that precipitate hospitalization for exacerbation of heart failure provide targets for intervention to prevent hospitalizations. To describe demographic, clinical, behavioral, and psychosocial factors that precipitate admission for exacerbation of heart failure and assess the relationships between precipitating factors and delay before hospitalization, and between delay time and length of hospital stay. All admissions in 12 full months to a tertiary medical center were reviewed if the patient had a discharge code related to heart failure. Data on confirmed admissions for exacerbation of heart failure were included in the study. Electronic and paper medical records were reviewed to identify how long it took patients to seek care after they became aware of signs and symptoms, factors that precipitated exacerbation, and discharge details. Exacerbation of heart failure was confirmed in 482 patients. Dyspnea was the most common symptom (92.5% of patients), and 20.3% of patients waited until they were severely dyspneic before seeking treatment. The most common precipitating factor was poor medication adherence. Delay times from symptom awareness to seeking treatment were shorter in patients who had a recent change in medicine for heart failure, renal failure, or poor medication adherence and longer in patients with depressive symptoms and hypertension. Depressive symptoms, recent change in heart failure medicine, renal failure, poor medication adherence, and hypertension are risk factors for hospitalizations for exacerbation of heart failure.


Regulatory domain phosphorylation to distinguish the mechanistic basis underlying acute CFTR modulators.
Modulator compounds intended to overcome disease-causing mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) show significant promise in clinical testing for cystic fibrosis. However, the mechanism(s) of action underlying these compounds are not fully understood. Activation of CFTR ion transport requires PKA-regulated phosphorylation of the regulatory domain (R-D) and dimerization of the nucleotide binding domains. Using a newly developed assay, we evaluated nine compounds including both CFTR potentatiators and activators discovered via various high-throughput screening strategies to acutely augment CFTR activity. We found considerable differences in the effects on R-D phosphorylation. Some (including UC(CF)-152) stimulated robust phosphorylation, and others had little effect (e.g., VRT-532 and VX-770). We then compared CFTR activation by UC(CF)-152 and VRT-532 in Ussing chamber studies using two epithelial models, CFBE41o(-) and Fischer rat thyroid cells, expressing various CFTR forms. UC(CF)-152 activated wild-type-, G551D-, and rescued F508del-CFTR currents but did not potentiate cAMP-mediated CFTR activation. In contrast, VRT-532 moderately activated CFTR short-circuit current and strongly potentiated forskolin-mediated current. Combined with the result that UC(CF)-152, but not VRT-532 or VX-770, acts by increasing CFTR R-D phosphorylation, these findings indicate that potentiation of endogenous cAMP-mediated activation of mutant CFTR is not due to a pathway involving augmented R-D phosphorylation. This study presents an assay useful to distinguish preclinical compounds by a crucial mechanism underlying CFTR activation, delineates two types of compound able to acutely augment CFTR activity (e.g., activators and potentiators), and demonstrates that a number of different mechanisms can be successfully employed to activate mutant CFTR.


SETDB1 is required for intestinal epithelial differentiation and the prevention of intestinal inflammation.
The intestinal epithelium is a rapidly renewing tissue which plays central roles in nutrient uptake, barrier function and the prevention of intestinal inflammation. Control of epithelial differentiation is essential to these processes and is dependent on cell type-specific activity of transcription factors which bind to accessible chromatin. Here, we studied the role of SET Domain Bifurcated Histone Lysine Methyltransferase 1, also known as ESET (SETDB1), a histone H3K9 methyltransferase, in intestinal epithelial homeostasis and IBD. We investigated mice with constitutive and inducible intestinal epithelial deletion of Setdb1, studied the expression of SETDB1 in patients with IBD and mouse models of IBD, and investigated the abundance of SETDB1 variants in healthy individuals and patients with IBD. Deletion of intestinal epithelial Setdb1 in mice was associated with defects in intestinal epithelial differentiation, barrier disruption, inflammation and mortality. Mechanistic studies showed that loss of SETDB1 leads to de-silencing of endogenous retroviruses, DNA damage and intestinal epithelial cell death. Predicted loss-of-function variants in human SETDB1 were considerably less frequently observed than expected, consistent with a critical role of SETDB1 in human biology. While the vast majority of patients with IBD showed unimpaired mucosal SETDB1 expression, comparison of IBD and non-IBD exomes revealed over-representation of individual rare missense variants in SETDB1 in IBD, some of which are predicted to be associated with loss of function and may contribute to the pathogenesis of intestinal inflammation. SETDB1 plays an essential role in intestinal epithelial homeostasis. Future work is required to investigate whether rare variants in SETDB1 contribute to the pathogenesis of IBD.


Spin and energy analyzed photoemission: a feasibility analysis.
New scientific opportunities, particularly for investigation of surface magnetism, will be provided by spin and energy analyzed photoemission. Electron-optical conservation laws and phase space concepts are summarized and applied to determine the feasiblity of an experiment consisting of a photoemitter in a magnetic field, a photoelectron energy analyzer and an electron spin analyzer. For the example of photoemission from a Ni crystal using He I resonance radiation and typical parameters for the energy and spin analyzers, a final signal count rate of approximately 220 counts/s is calculated. Ways to increase the count rate by orders of magnitude are described. In particular, a new experimental configuration is suggested which may avoid the large reduction in count rate caused by the magnetic field.


Size and sociodemographic characteristics of the Afghan refugee population in Pakistan.
Some recent data are presented on the size and selected sociodemographic characteristics of the Afghan refugee population in Pakistan. Although the official figures show that there were 3.27 million registered Afghan refugees in Pakistan, it is estimated that the actual number may be as high as 3.6 million. There is an excess of females over males, mainly due to war-related activities and excessive casualties particularly among males. While infant and childhood mortality rates are declining and are lower than the levels prevalent in Pakistan, as well as in Afghanistan during the pre-war period, the fertility levels among Afghan refugees seem very high indeed.


Proteasome activity imaging and profiling characterizes bacterial effector syringolin A.
Syringolin A (SylA) is a nonribosomal cyclic peptide produced by the bacterial pathogen Pseudomonas syringae pv syringae that can inhibit the eukaryotic proteasome. The proteasome is a multisubunit proteolytic complex that resides in the nucleus and cytoplasm and contains three subunits with different catalytic activities: β1, β2, and β5. Here, we studied how SylA targets the plant proteasome in living cells using activity-based profiling and imaging. We further developed this technology by introducing new, more selective probes and establishing procedures of noninvasive imaging in living Arabidopsis (Arabidopsis thaliana) cells. These studies showed that SylA preferentially targets β2 and β5 of the plant proteasome in vitro and in vivo. Structure-activity analysis revealed that the dipeptide tail of SylA contributes to β2 specificity and identified a nonreactive SylA derivative that proved essential for imaging experiments. Interestingly, subcellular imaging with probes based on epoxomicin and SylA showed that SylA accumulates in the nucleus of the plant cell and suggests that SylA targets the nuclear proteasome. Furthermore, subcellular fractionation studies showed that SylA labels nuclear and cytoplasmic proteasomes. The selectivity of SylA for the catalytic subunits and subcellular compartments is discussed, and the subunit selectivity is explained by crystallographic data.


Fucoidan Extracted from Hijiki Protects Brain Microvessel Endothelial Cells Against Diesel Exhaust Particle Exposure-Induced Disruption.
This study was performed to evaluate the protective effects of fucoidan against the decreased function of primary cultured bovine brain microvessel endothelial cells (BBMECs) after exposure to diesel exhaust particles (DEPs). BBMECs were extracted from bovine brains and cultured until confluent. To evaluate the function of BBMECs, we performed a permeability test using cell-by-cell equipment and by Western blot analysis for zonular occludens-1 (ZO-1), which is a tight junction protein of BMECs, and evaluated oxidative stress in BBMECs using the DCFH-DA assay and the CUPRAC-BCS assay. The increased oxidative stress in BBMECs following DEP exposure was suppressed by fucoidan. In addition, permeability of BBMECs induced by DEP exposure was decreased by fucoidan treatment. Our results showed that fucoidan protects against BBMEC disruption induced by DEP exposure. This study provides evidence that fucoidan might protect the central nervous system (CNS) against DEP exposure.


Mycobacterium tuberculosis DNA gyrase as a target for drug discovery.
Bacterial DNA gyrase is an important target of antibacterial agents, including fluoroquinolones. In most bacterial species, fluoroquinolones inhibit DNA gyrase and topoisomerase IV and cause bacterial cell-death. Other naturally occurring bacterial DNA gyrase inhibitors, such as novobiocin, are also known to be effective as antibacterial agents. DNA gyrase is an ATP-dependent enzyme that acts by creating a transient double-stranded DNA break. It is unique in catalyzing the negative supercoiling of DNA and is essential for efficient DNA replication, transcription, and recombination. DNA gyrase is a tetrameric A2B2 protein. The A subunit carries the breakage-reunion active site, whereas the B subunit promotes ATP hydrolysis. The M. tuberculosis genome analysis has identified a gyrB-gyrA contig in which gyrA and gyrB encode the A and B subunits, respectively. There is no evidence that M. tuberculosis has homologs of the topoisomerase IV, parC and parE genes, which are present in most other bacteria. Newer fluoroquinolones, including moxifloxacin and gatifloxacin, exhibit potent activity against M. tuberculosis, and show potential to shorten the duration for TB treatment. Resistance to fluoroquinolones remains uncommon in clinical isolates of M. tuberculosis. M. tuberculosis DNA gyrase is thus a validated target for anti-tubercular drug discovery. Inhibitors of this enzyme are also active against non-replicating mycobacteria, which might be important for the eradication of persistent organisms. A novel inhibitor of M. tuberculosis DNA gyrase would be effective against multi-drug resistant (MDR)-TB, and it could also be effective against fluoroquinolone-resistant M. tuberculosis.


Family History of Cancer and Risk of Biliary Tract Cancers: Results from the Biliary Tract Cancers Pooling Project.
Background: Although some familial cancer syndromes include biliary tract cancers (BTCs; cancers of the gallbladder, intrahepatic and extrahepatic bile ducts, and ampulla of Vater), the few studies that have examined the relationships between family history of cancer (FHC) and BTCs have reported inconclusive findings. The objective of this study was to investigate the associations of FHC with risk of BTC in the Biliary Tract Cancers Pooling Project (BiTCaPP).Methods: We used Cox proportional hazards regressions models to estimate HRs and 95% confidence intervals for associations between FHC (any, first-degree, in female relative, in male relative, relative with gastrointestinal cancer, and relative with hormonally related cancer) and BTC risk by anatomic site within the biliary tract, adjusting for sex and race/ethnicity. Sensitivity analyses were conducted that restricted to studies reporting cholecystectomy data and to people without a history of cholecystectomy.Results: Data on FHC were available from 12 prospective studies within BiTCaPP, which collectively contributed 2,246 cases (729 gallbladder, 345 intrahepatic and 615 extrahepatic bile duct, and 385 ampulla of Vater cancers) with 21,706,107 person-years of follow-up. A marginal, inverse association between FHC and gallbladder cancer was driven to the null when analysis was restricted to studies reporting cholecystectomy data and to people without a history of cholecystectomy. FHC was not associated with risk of BTC at the other anatomic sites.Conclusions: These findings do not support an association between FHC and BTCs.Impact: In a study of 1.5 million people, FHC is not a risk factor for BTCs. Cancer Epidemiol Biomarkers Prev; 27(3); 348-51. ©2018 AACR.


Patient-derived glioblastoma cells show significant heterogeneity in treatment responses to the inhibitor-of-apoptosis-protein antagonist birinapant.
Resistance to temozolomide (TMZ) greatly limits chemotherapeutic effectiveness in glioblastoma (GBM). Here we analysed the ability of the Inhibitor-of-apoptosis-protein (IAP) antagonist birinapant to enhance treatment responses to TMZ in both commercially available and patient-derived GBM cells. Responses to TMZ and birinapant were analysed in a panel of commercial and patient-derived GBM cell lines using colorimetric viability assays, flow cytometry, morphological analysis and protein expression profiling of pro- and antiapoptotic proteins. Responses in vivo were analysed in an orthotopic xenograft GBM model. Single-agent treatment experiments categorised GBM cells into TMZ-sensitive cells, birinapant-sensitive cells, and cells that were insensitive to either treatment. Combination treatment allowed sensitisation to therapy in only a subset of resistant GBM cells. Cell death analysis identified three principal response patterns: Type A cells that readily activated caspase-8 and cell death in response to TMZ while addition of birinapant further sensitised the cells to TMZ-induced cell death; Type B cells that readily activated caspase-8 and cell death in response to birinapant but did not show further sensitisation with TMZ; and Type C cells that showed no significant cell death or moderately enhanced cell death in the combined treatment paradigm. Furthermore, in vivo, a Type C patient-derived cell line that was TMZ-insensitive in vitro and showed a strong sensitivity to TMZ and TMZ plus birinapant treatments. Our results demonstrate remarkable differences in responses of patient-derived GBM cells to birinapant single and combination treatments, and suggest that therapeutic responses in vivo may be greatly affected by the tumour microenvironment.


Cisplatin-tethered gold nanospheres for multimodal chemo-radiotherapy of glioblastoma.
Glioblastoma multiforme (GBM) remains the most aggressive and challenging brain tumour to treat. We report the first successful chemo-radiotherapy on patient derived treatment resistant GBM cells using a cisplatin-tethered gold nanosphere. After intracellular uptake, the nanosphere effects DNA damage which initiates caspase-mediated apoptosis in those cells. In the presence of radiation, both gold and platinum of cisplatin, serve as high atomic number radiosensitizers leading to the emission of ionizing photoelectrons and Auger electrons. This resulted in enhanced synergy between cisplatin and radiotherapy mediated cytotoxicity, and photo/Auger electron mediated radiosensitisation leading to complete ablation of the tumour cells in an in vitro model system. This study demonstrates the potential of designed nanoparticles to target aggressive cancers in the patient derived cell lines providing a platform to move towards treatment strategies.


IGHV-unmutated and IGHV-mutated chronic lymphocytic leukemia cells produce activation-induced deaminase protein with a full range of biologic functions.
Clonal evolution occurs during the course of chronic lymphocytic leukemia (CLL) and activation-induced deaminase (AID) could influence this process. However, this possibility has been questioned in CLL because the number of circulating AID mRNA(+) cells is exceedingly low; synthesis of AID protein by blood CLL cells has not been demonstrated; the full range of AID functions is lacking in unmutated CLL (U-CLL), and no prospective analysis linking AID expression and disease severity has been reported. The results of the present study show that circulating CLL cells and those within secondary lymphoid tissues can make AID mRNA and protein. This production is related to cell division because more AID mRNA was detected in recently divided cells and AID protein was limited to the dividing fraction and was up-regulated on induction of cell division. AID protein was functional because AID(+) dividing cells exhibited more double-stranded DNA breaks, IGH class switching, and new IGHV-D-J mutations. Each of these actions was documented in U-CLL and mutated CLL (M-CLL). Furthermore, AID protein was associated with worse patient outcome and adverse cytogenetics. We conclude that the production of fully functional AID protein by U-CLL and M-CLL cells could be involved in clonal evolution of the disease.


Comparison of two PCR-based human papillomavirus genotyping methods.
We compared two consensus primer PCR human papillomavirus (HPV) genotyping methods for the detection of individual HPV genotypes and carcinogenic HPV genotypes as a group, using a stratified sample of enrollment cervical specimens from sexually active women participating in the NCI/Costa Rica HPV16/18 Vaccine Efficacy Trial. For the SPF(10) method, DNA was extracted from 0.1% of the cervical specimen by using a MagNA Pure LC instrument, a 65-bp region of the HPV L1 gene was targeted for PCR amplification by using SPF(10) primers, and 25 genotypes were detected by reverse-line blot hybridization of the amplicons. For the Linear Array (LA) method, DNA was extracted from 0.5% of the cervical specimen by using an MDx robot, a 450-bp region of the HPV L1 gene was targeted for PCR amplification by using PGMY09/11 L1 primers, and 37 genotypes were detected by reverse-line blot hybridization of the amplicons. Specimens (n = 1,427) for testing by the LA method were randomly selected from strata defined on the basis of enrollment test results from the SPF(10) method, cytology, and Hybrid Capture 2. LA results were extrapolated to the trial cohort (n = 5,659). The LA and SPF(10) methods detected 21 genotypes in common; HPV16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66, -68, and -73 were considered the carcinogenic HPV genotypes. There was no difference in the overall results for grouped detection of carcinogenic HPV by the SPF(10) and LA methods (35.3% versus 35.9%, respectively; P = 0.5), with a 91.8% overall agreement and a kappa value of 0.82. In comparisons of individual HPV genotypes, the LA method detected significantly more HPV16, HPV18, HPV39, HPV58, HPV59, HPV66, and HPV68/73 and less HPV31 and HPV52 than the SPF(10) method; inclusion of genotype-specific testing for HPV16 and HPV18 for those specimens testing positive for HPV by the SPF(10) method but for which no individual HPV genotype was detected abrogated any differences between the LA and SPF(10) methods. The LA method detected more carcinogenic-HPV-genotype infections per specimen than the SPF(10) method (P < 0.001). In conclusion, the LA method and the SPF(10) method with HPV16 and HPV18 genotype-specific detection among ungenotyped HPV-positive specimens were comparable for detection of HPV16 and HPV18, the two HPV genotypes targeted by current prophylactic HPV vaccines. Both approaches are suitable for monitoring the impact of HPV16/18 vaccines in clinical trials.


SAMHD1: at the crossroads of cell proliferation, immune responses, and virus restriction.
SAMHD1 is a triphosphohydrolase enzyme that controls the intracellular level of deoxyribonucleoside triphosphates (dNTPs) and plays a role in innate immune sensing and autoimmune disease. SAMHD1 has also been identified as an intrinsic virus restriction factor, inactivated through degradation by HIV-2 Vpx or through a post-transcriptional regulatory mechanism. Phosphorylation of SAMHD1 by cyclin-dependent kinases has been strongly associated with inactivation of the virus restriction mechanism, providing an association between virus replication and cell proliferation. Tight regulation of cell proliferation suggests that viruses, particularly HIV-1 replication, latency, and reactivation, may be similarly controlled by multiple checkpoint mechanisms that, in turn, regulate dNTP levels. In this review, we discuss how SAMHD1 is a viral restriction factor, the mechanism associated with viral restriction, the pathway leading to its inactivation in proliferating cells, and how strategies aimed at controlling virus restriction could lead to a functional cure for HIV.


Integrating virtual patients into courses: follow-up seminars and perceived benefit.
The use of virtual patients (VPs) suggests promising effects on student learning. However, currently empirical data on how best to use VPs in practice are scarce. More knowledge is needed regarding aspects of integrating VPs into a course, of which student acceptance is one key issue. Several authors call for looking beyond technology to see VPs in relation to the course context. The follow-up seminar is proposed as an important aspect of integration that warrants investigation. A cross-sectional explanatory study was performed in a clinical clerkship introduction course at four teaching hospitals affiliated to the same medical faculty. The VP-related activities were planned collaboratively by teachers from all four settings. However, each setting employed a different strategy to follow up the activity in the course. Sixteen questionnaire items were grouped into three scales pertaining to: perceived benefit of VPs; wish for more guidance on using VPs, and wish for assessment and feedback on VPs. Scale scores were compared across the four settings, which were ranked according to the level of intensity of students' processing of cases during VP follow-up activities. The perceived benefit of VPs and their usage were higher in the two intense-use settings compared with the moderate- and low-intensity settings. The wish for more guidance was high in the low- and one of the high-intensity settings. Students in all settings displayed little interest in more assessment and feedback regarding VPs. High case processing intensity was related to positive perceptions of the benefit of VPs. However, the low interest in more assessment and feedback on the use of VPs indicates the need to clearly communicate the added value of the follow-up seminar. The findings suggest that a more intense follow-up pays off in terms of the benefit perceived by students. This study illustrates the need to consider VPs from the perspective of a holistic course design and not as isolated add-ons.


Kinetic and biochemical analyses on the reaction mechanism of a bacterial ATP-citrate lyase.
The prokaryotic ATP-citrate lyase is considered to be a key enzyme of the carbon dioxide-fixing reductive tricarboxylic acid (RTCA) cycle. Kinetic examination of the ATP-citrate lyase from the green sulfur bacterium Chlorobium limicola (Cl-ACL), an alpha(4)beta(4) heteromeric enzyme, revealed that the enzyme displayed typical Michaelis-Menten kinetics toward ATP with an apparent K(m) value of 0.21 +/- 0.04 mm. However, strong negative cooperativity was observed with respect to citrate binding, with a Hill coefficient (n(H)) of 0.45. Although the dissociation constant of the first citrate molecule was 0.057 +/- 0.008 mm, binding of the first citrate molecule to the enzyme drastically decreased the affinity of the enzyme for the second molecule by a factor of 23. ADP was a competitive inhibitor of ATP with a K(i) value of 0.037 +/- 0.006 mm. Together with previous findings that the enzyme catalyzed the reaction only in the direction of citrate cleavage, these kinetic features indicated that Cl-ACL can regulate both the direction and carbon flux of the RTCA cycle in C. limicola. Furthermore, in order to gain insight on the reaction mechanism, we performed biochemical analyses of Cl-ACL. His273 of the alpha subunit was indicated to be the phosphorylated residue in the catalytic center, as both catalytic activity and phosphorylation of the enzyme by ATP were abolished in an H273A mutant enzyme. We found that phosphorylation of the subunit was reversible. Nucleotide preference for activity was in good accordance with the preference for phosphorylation of the enzyme. Although residues interacting with nucleotides in the succinyl-CoA synthetase from Escherichia coli were conserved in AclB, AclA alone could be phoshorylated with the same nucleotide specificity observed in the holoenzyme. However, AclB was necessary for enzyme activity and contributed to enhance phosphorylation and stabilization of AclA.


Ovarian masses during adolescence: clinical, ultrasonographic and pathologic findings, serum tumor markers and endocrinological profile.
This retrospective study was undertaken to assess the epidemiological and clinical features, laboratory findings, preoperative work-up, surgical treatment and pathologic findings in adolescents with ovarian cysts. All adolescents who were referred to our institution and had been operated on because of a diagnosed ovarian mass, from January 1997 to June 2003, were included in this study. Forty-four cases of women with an ovarian mass were retrospectively analyzed. These patients, aged between 12 and 21 years, had 47 ovarian masses (three patients had bilateral lesions), of which 49% were non-neoplastic and 51% were neoplastic. Of the neoplastic lesions, 62.5% were germ cell tumors, 20.8% were epithelial and 16.7% were sex cord-stromal tumors. Of the neoplastic tumors, 95.8% were benign while 4.2% were malignant. Procedures included 30 operative laparoscopies (68.20%) and 14 exploratory laparotomies (31.8%). Simple excision of the ovarian cyst was performed in 39 cases (88.6%). According to our study, most of the adolescents with an ovarian cyst underwent an operation because of a neoplastic lesion. The majority of ovarian tumors occurring in adolescents are non-epithelial in origin, and germ cell tumors are the most common histological type. Ultrasound examination is one of the most important diagnostic tools. Preoperative diagnostic approach of these patients should always include careful history taking, physical examination, imaging and evaluation of serum assays. Gynecologists who care for young girls must be familiar with the differential diagnosis of ovarian masses whose surgical treatment should be conservative when appropriate, so that hormonal status and future fertility are not compromised in this group of patients.


Cross immunity and vaccination against multiple microparasite strains.
We explore the equilibrium properties of a series of compartmental, ODE models describing the interaction between different strains of pathogen. The interaction is conceptualized as acting through shared antigens: infection and recovery from one strain leaves the host with a primed immune response against subsequent strains. The models consider the effect of this priming on susceptibility (the ability to be infected) and transmission (the ability to infect) in an SIR model. In these models, the specific past history of infection is encapsulated in different susceptible compartments within the model. In a third, SIS, model, specific past history is not included, but strains have differential abilities to infect previously infected hosts. Equilibrium results include criteria for the coexistence of strains. For the SIR models, the region of coexistence defined by parameters shrinks as the effect of strains on each other (increased antigenic similarity) increases. For the SIS model, coexistence depends critically on the rate at which complete susceptibility is recovered following infection, and coexisting strains must have differential abilities to infect completely and partially susceptible hosts. Interestingly, this model provides analogies to commensalism (the first species gains from the presence of the second; the second neither gains nor loses from the interaction) and symbiosis (the presence of both species benefits the other). Additionally, we show that the maximum number of coexisting strains is two in this model. The effect of vaccination depends on the initial strain structure, the ability of vaccination to mount protection to both strains and the coverage. Vaccination may allow a previously excluded strain to coexist or exist alone, and may allow a previously rarer strain to become more common with the possibility of increasing incidence of disease. We discuss the dynamics of these models, compare model results to observed patterns and consider additional model structures. The importance of these results to specific multi-strain pathogens, in particular rotavirus, is considered.


Planning forest restoration within private land holdings with conservation co-benefits at the landscape scale.
Forest loss is mainly due to the conversion of forest to agriculture, mostly in private lands. Forest restoration is a global priority, yet restoration targets are ambitious and budget-limited. Therefore, assessing the outcome of alternative decisions on land-use within private lands is paramount to perform cost-effective restoration. We present a novel framework that incorporates spatial planning for forest restoration within private lands with conservation co-benefits at the landscape scale. As a case study, we used three real landscapes of 10.000 ha with differing amounts of forest cover in the Atlantic Forest region of Brazil, and three hypothetical animal species with different dispersal abilities. We estimated the total amount of forest that landholders must restore to comply with the Native Vegetation Protection Law, which requires landholders to reforest 20% of their land within a 20-year time frame. We compared the cost-effectiveness of five restoration strategies based on the improvement in habitat availability and restoration costs. The most cost-effective strategy depends on a landscape's initial amount of forest cover and the species of concern. We revealed that spatial planning for restoration in private lands increased habitat availability up to 12 times more than random restoration, which was always the least cost-effective strategy. Cost-effective large-scale restoration in Brazil depends on public policies that assist landholders to comply with the law and on prioritizing areas for restoration within private lands. We show that by adding habitat availability as target in spatial prioritization, benefits for biodiversity can be hastened at low additional cost, even in real world scenarios with severe spatial constraints. Despite constraints, spatially planned restoration for law compliance in Brazil increased landscape permeability by creating corridors and stepping stones. Our framework should be used to plan restoration in Brazilian private lands and can be customized for other regions worldwide.


End-to-End Differentiable Learning of Protein Structure.
Predicting protein structure from sequence is a central challenge of biochemistry. Co-evolution methods show promise, but an explicit sequence-to-structure map remains elusive. Advances in deep learning that replace complex, human-designed pipelines with differentiable models optimized end to end suggest the potential benefits of similarly reformulating structure prediction. Here, we introduce an end-to-end differentiable model for protein structure learning. The model couples local and global protein structure via geometric units that optimize global geometry without violating local covalent chemistry. We test our model using two challenging tasks: predicting novel folds without co-evolutionary data and predicting known folds without structural templates. In the first task, the model achieves state-of-the-art accuracy, and in the second, it comes within 1-2 Å; competing methods using co-evolution and experimental templates have been refined over many years, and it is likely that the differentiable approach has substantial room for further improvement, with applications ranging from drug discovery to protein design.


Novel pathogenic variants in SLCO2A1 causing autosomal dominant primary hypertrophic osteoarthropathy.
Primary hypertrophic osteoarthropathy (PHO), or pachydermoperiostosis, is characterized by a clinical association including digital clubbing, periostosis and pachydermia. SLCO2A1 and HPGD genes are both responsible for PHO. The pathology is classically defined as an autosomal recessive disorder with clinical variability ranging from a mild to more severe phenotype. However, the hypothesis for an autosomal dominant form suggested for a long time was only demonstrated for the first time in 2021 for SLCO2A1. We aimed to detect a second pathogenic variant by a deep sequencing of the entire SLCO2A1 and HPGD genes, associated with functional transcription analysis in PHO patients harboring only one heterozygous variant. Among 10 PHO patients, 4 presented a single pathogenic or probably pathogenic novel variant in SLCO2A1 in heterozygous status (NM_005630.3: c.234+1G > A, c.1523_1524delCT, c.1625G > A and c.31delC), and the others carried homozygous pathogenic variants. For heterozygous forms, we found no additional pathogenic variant in HPGD or SLCO2A1. PHO can be a dominant form with age at disease onset later than that for the recessive form. This dominant form is not exceptional in young adults. In conclusion, both modes of inheritance of PHO explain the clinical variability and the difference in age at disease onset. Molecular analysis is especially required in the incomplete form to distinguish it from secondary hypertrophic osteoarthropathy.


Use of a versatile high efficiency multiparallel denuder for the sampling of PAHs in ambient air: gas and particle phase concentrations, particle size distribution and artifact formation.
The design and performance of a multiparallel plate denuder able to operate at low and high-flow (3-30 L/min) for the collection of polycyclic aromatic hydrocarbon (PAH) vapor is described. The denuder, in combination with a micro orifice uniform deposit impactor (MOUDI) was used to assess processes of artifact formation in MOUDIs used with and without an upstream denuder. Duplicate sampling trains with an upstream denuder showed good repeatability of the measured gas and particle-phase concentrations and low breakthrough in the denuder (3.5-15%). The PAH size distributions within undenuded and denuded MOUDIs were studied. Use of the denuder altered the measured size distribution of PAHs toward smaller sizes, but both denuded and undenuded systems are subject to sampling artifacts.


Effects of slow-release urea and rumen-protected methionine and histidine on performance of dairy cows.
This experiment was conducted with the objective to investigate the effects of slow-release urea and rumen-protected (RP) Met and His supplementation of a metabolizable protein (MP)-deficient diet (according to NRC, 2001) on lactation performance of dairy cows. Sixty lactating Holstein cows were used in a 10-wk randomized complete block-design trial. Cows were fed a covariate diet for 2 wk and then assigned to one of the following treatments for an 8-wk experimental period: (1) MP-adequate diet [AMP; 107% of MP requirements, based on the National Research Council (NRC, 2001)]; (2) MP-deficient diet (DMP; 95% of MP requirements); (3) DMP supplemented with slow-release urea (DMPU); (4) DMPU supplemented with RPMet (DMPUM); and (5) DMPUM supplemented with RPHis (DMPUMH). Total-tract apparent digestibility of dry matter, organic matter, neutral detergent fiber, and crude protein, and urinary N and urea-N excretions were decreased by DMP, compared with AMP. Addition of slow-release urea to the DMP diet increased urinary urea-N excretion. Dry matter intake (DMI) and milk yield (on average 44.0±0.9kg/d) were not affected by treatments, except DMPUMH increased DMI and numerically increased milk yield, compared with DMPUM. Milk true protein concentration and yield were increased and milk fat concentration tended to be decreased by DMPUMH, compared with DMPUM. Cows gained less body weight on the DMP diet, compared with AMP. Plasma concentrations of His and Lys were not affected by treatments, whereas supplementation of RPMet increased plasma Met concentration. Plasma concentration of 3-methylhistidine was or tended to be higher for DMP compared with AMP and DMPU, respectively. Addition of RPHis to the DMPUM diet tended to increase plasma glucose and creatinine. In conclusion, feeding a 5% MP-deficient diet (according to NRC, 2001) did not decrease DMI and yields of milk and milk components, despite a reduction in nutrient digestibility. Supplementation of RPHis increased DMI and milk protein concentration and yield. These results are in line with our previous data and suggest that His may have a positive effect on voluntary feed intake and milk production and composition in high-yielding dairy cows fed MP-deficient diets.