Extracellular Vesicles Produced by Bifidobacterium longum Export Mucin-Binding Proteins.
Extracellular proteins are important factors in host-microbe interactions; however, the specific factors that enable bifidobacterial adhesion and survival in the gastrointestinal (GI) tract are not fully characterized. Here, we discovered that Bifidobacterium longum NCC2705 cultured in bacterium-free supernatants of human fecal fermentation broth released a myriad of particles into the extracellular environment. The aim of this study was to characterize the physiological properties of these extracellular particles. The particles, approximately 50 to 80 nm in diameter, had high protein and double-stranded DNA contents, suggesting that they were extracellular vesicles (EVs). A proteomic analysis showed that the EVs primarily consisted of cytoplasmic proteins with crucial functions in essential cellular processes. We identified several mucin-binding proteins by performing a biomolecular interaction analysis of phosphoketolase, GroEL, elongation factor Tu (EF-Tu), phosphoglycerate kinase, transaldolase (Tal), and heat shock protein 20 (Hsp20). The recombinant GroEL and Tal proteins showed high binding affinities to mucin. Furthermore, the immobilization of these proteins on microbeads affected the permanence of the microbeads in the murine GI tract. These results suggest that bifidobacterial exposure conditions that mimic the intestine stimulate B. longum EV production. The resulting EVs exported several cytoplasmic proteins that may have promoted B. longum adhesion. This study improved our understanding of the Bifidobacterium colonization strategy in the intestinal microbiome.IMPORTANCEBifidobacterium is a natural inhabitant of the human gastrointestinal (GI) tract. Morphological observations revealed that extracellular appendages of bifidobacteria in complex microbial communities are important for understanding its adaptations to the GI tract environment. We identified dynamic extracellular vesicle (EV) production by Bifidobacterium longum in bacterium-free fecal fermentation broth that was strongly suggestive of differing bifidobacterial extracellular appendages in the GI tract. In addition, export of the adhesive moonlighting proteins mediated by EVs may promote bifidobacterial colonization. This study provides new insight into the roles of EVs in bifidobacterial colonization processes as these bacteria adapt to the GI environment.


Considerations and controversies of AIDS.
The statistical probability of seroconversion is proportional to the number of needlesticks incurred and the likelihood that the needlesticks will be with HIV infected blood. Careful adherence to recommended operating room practices, combined with meticulous attention to handling needles and sharps, should result in few, if any, cases of occupational HIV seroconversion among OR personnel. HIV testing is not feasible in the management of emergency patients; these are often the individuals at highest risk for HIV infection and over whom the surgical team has the least control. Non-operative treatment of HIV-infected patients is not an option; many procedures are performed either to enable the individual to lead a more comfortable, productive life or for diagnostic purposes.


Spike-train communities: finding groups of similar spike trains.
Identifying similar spike-train patterns is a key element in understanding neural coding and computation. For single neurons, similar spike patterns evoked by stimuli are evidence of common coding. Across multiple neurons, similar spike trains indicate potential cell assemblies. As recording technology advances, so does the urgent need for grouping methods to make sense of large-scale datasets of spike trains. Existing methods require specifying the number of groups in advance, limiting their use in exploratory analyses. I derive a new method from network theory that solves this key difficulty: it self-determines the maximum number of groups in any set of spike trains, and groups them to maximize intragroup similarity. This method brings us revealing new insights into the encoding of aversive stimuli by dopaminergic neurons, and the organization of spontaneous neural activity in cortex. I show that the characteristic pause response of a rat's dopaminergic neuron depends on the state of the superior colliculus: when it is inactive, aversive stimuli invoke a single pattern of dopaminergic neuron spiking; when active, multiple patterns occur, yet the spike timing in each is reliable. In spontaneous multineuron activity from the cortex of anesthetized cat, I show the existence of neural ensembles that evolve in membership and characteristic timescale of organization during global slow oscillations. I validate these findings by showing that the method both is remarkably reliable at detecting known groups and can detect large-scale organization of dynamics in a model of the striatum.


Identification and management of poor response to growth-promoting therapy in children with short stature.
Growth hormone (GH) is widely prescribed for children with short stature across a range of growth disorders. Recombinant human (rh) insulin-like growth factor-1 (rhIGF-1) therapy is approved for severe primary IGF-I deficiency - a state of severe GH resistance. Evidence is increasing for an unacceptably high rate of poor or unsatisfactory response to growth-promoting therapy (i.e. not leading to significant catch up growth) in terms of change in height standard deviation score (SDS) and height velocity (HV) in many approved indications. Consequently, there is a need to define poor response and to prevent or correct it by optimizing treatment regimens within accepted guidelines. Recognition of a poor response is an indication for action by the treating physician, either to modify the therapy or to review the primary diagnosis leading either to discontinuation or change of therapy. This review discusses the optimal investigation of the child who is a candidate for GH or IGF-1 therapy so that a diagnosis-based choice of therapy and dosage can be made. The relevant parameters in the evaluation of growth response are described together with the definitions of poor response. Prevention of poor response is addressed by discussion of strategy for first-year management with GH and IGF-1. Adherence to therapy is reviewed as is the recommended action following the identification of the poorly responding patient. The awareness, recognition and management of poor response to growth-promoting therapy will lead to better patient care, greater cost-effectiveness and increased opportunities for clinical benefit.


Mycobacteria and allograft heart valve banking: an international survey.
Since the 1970s many tissue banks have been testing allograft heart valves (HVs) for Mycobacterium tuberculosis (MTB). Donor selection for low risk of tuberculosis (TB) was introduced in the 1980s and appears to have reduced the risk of TB transmission. Regulatory guidance does not specify testing for TB, but does exclude donors with a recent history of TB. This survey of HV international bank practices revealed variations in donor selection, testing and processing of valves. Participant banks (from Europe and the USA) reported that over a period of 15 years, HV tissues from 38,413 donors were banked and 32,289 donors were tested for TB, none being positive. HV-associated tissue from 27,840 donors was stained and underwent microscopy; none of these were positive for acid-fast bacilli (AFB). Non-tuberculosis mycobacteria (NTBM) were detected by culture on 24 HVs. It is recommended that HV banks employ donor selection to exclude donors at risk of TB, to culture material for mycobacteria, and to investigate potential sources when clusters of NTBM are found to facilitate corrective and preventative actions.


ANTIBODY SYNTHESIS AT THE CELLULAR LEVEL. ANTIBODY-INDUCED SUPPRESSION OF 19S AND 7S ANTIBODY RESPONSE.
The suppressing activity of passively transferred antibodies on antibody synthesis against sheep red cells was investigated at the cellular level by the agar-plaque technique developed by Jerne. Humoral antibodies injected prior to the antigen suppressed the appearance of plaque-forming spleen cells producing 19S antibodies completely. Antibodies given during the first 4 days after antigen injection also showed such action, but only after a latency period of 40 hours. The inhibiting efficiency of 7S antibodies was about 100 to 200 times greater than that of 19S antibodies. The results support the conclusion that humoral antibodies inhibit the immune response by removing the stimulus for the proliferation of the antibody producing cells and not by directly depressing antibody synthesis in already committed cells. Passively transferred antibodies inhibited the 7S response if given prior to, or 24 hours after the antigen injection, in analogy with previous results concerning 19S response. In contrast to these previous results on 19S synthesis, antibody transfer had no detectable effect during the early exponential phase of 7S production (5 to 7 days after antigen injection). Only limited inhibition was observed 3 days after the antigen. One possible explanation of this difference is that 7S-producing cells do not divide, or divide at a slow rate. Antigen injection would stimulate the proliferation of 19S-producing cells. Subsequently these would switch to the synthesis of 7S antibodies. These would inhibit the initiation of new 19S-producing cells by combining with the antigen. They would thus suppress the recruitment of their own precursors. A steady state of 7S antibody production by cells with a long lifetime would be the result. This hypothesis ascribes an important regulatory function to 7S antibodies. They would be parts of a feed-back system preventing excessive cell multiplication in response to a single antigen.


CSPP1 stabilizes growing microtubule ends and damaged lattices from the luminal side.
Microtubules are dynamic cytoskeletal polymers, and their organization and stability are tightly regulated by numerous cellular factors. While regulatory proteins controlling the formation of interphase microtubule arrays and mitotic spindles have been extensively studied, the biochemical mechanisms responsible for generating stable microtubule cores of centrioles and cilia are poorly understood. Here, we used in vitro reconstitution assays to investigate microtubule-stabilizing properties of CSPP1, a centrosome and cilia-associated protein mutated in the neurodevelopmental ciliopathy Joubert syndrome. We found that CSPP1 preferentially binds to polymerizing microtubule ends that grow slowly or undergo growth perturbations and, in this way, resembles microtubule-stabilizing compounds such as taxanes. Fluorescence microscopy and cryo-electron tomography showed that CSPP1 is deposited in the microtubule lumen and inhibits microtubule growth and shortening through two separate domains. CSPP1 also specifically recognizes and stabilizes damaged microtubule lattices. These data help to explain how CSPP1 regulates the elongation and stability of ciliary axonemes and other microtubule-based structures.


Hepatic biomarkers of sediment-associated pollution in juvenile turbot, Scophthalmus maximus L.
Hatchery-reared turbot (Scophthalmus maximus L.) were exposed for 3 weeks, under laboratory conditions, to sediment collected from polluted sites in Cork Harbour and a reference site at Ballymacoda, Co. Cork, Ireland. The potential of surficial sediment for inducing hepatic biomarkers was assessed at two levels of biological organisation: expression of cytochrome P450 [Western blotting analysis and 7-ethoxy-resorufin O-dealkylase (EROD), 7-benzoxy resorufin O-dealkylase (BROD), 7-methoxy resorufin O-dealkylase (MROD), 7-pentoxy-resorufin O-dealkylase (PROD) activities] and DNA integrity (Comet assay). Positive controls were generated, either by exposing turbot to cadmium chloride-spiked seawater (Comet assay) or to beta-naphthaflavone by intraperitoneal injection (cytochrome P450 induction). The induction of cytochrome P450 activity (EROD, MROD and PROD) in animals following a 7-day exposure to contaminated sediments was significantly higher than those exposed to reference site sediment and remained elevated thereafter; BROD was not induced. DNA single-strand breaks were also significantly higher following exposure to contaminated sediments throughout the experiment. Although no direct correlation between induction of alkoxyresorufin O-dealkylase activities and a particular chemical class was established, the induction of MROD and PROD activities in fish exposed to sediments containing complex contaminant mixtures, appeared to be more sensitive than conventional EROD activity assays. We conclude from the present laboratory study that S. maximus is a suitable sentinel species for the assessment of moderately contaminated sediments and therefore allows for the further development of this model for future, ecologically relevant, field studies.


First estimates of the potential cost and cost saving of protecting childhood hearing from damage caused by congenital CMV infection.
Congenital cytomegalovirus (cCMV) is an important cause of childhood deafness, which is modifiable if diagnosed within the first month of life. Targeted screening of infants who do not pass their newborn hearing screening tests in England is a feasible approach to identify and treat cases to improve hearing outcome. To conduct a cost analysis of targeted screening and subsequent treatment for cCMV-related sensorineural hearing loss (SNHL) in an, otherwise, asymptomatic infant, from the perspective of the UK National Health Service (NHS). Using data from the newborn hearing screening programme (NHSP) in England and a recent study of targeted screening for cCMV using salivary swabs within the NHSP, we estimate the cost (in UK pounds (£)) to the NHS. The cost of screening (time, swabs and PCR), assessing, treating and following up cases is calculated. The cost per case of preventing hearing deterioration secondary to cCMV with targeted screening is calculated. The cost of identifying, assessing and treating a case of cCMV-related SNHL through targeted cCMV screening is estimated to be £6683. The cost of improving hearing outcome for an infant with cCMV-related SNHL through targeted screening and treatment is estimated at £14 202. The costs of targeted screening for cCMV using salivary swabs integrated within NHSP resulted in an estimate of cost per case that compares favourably with other screening programmes. This could be used in future studies to estimate the full economic value in terms of incremental costs and incremental health benefits.


Spinal Epidural Hematoma Following Epidural Catheter Removal in a Patient with Postoperative Urgent Coronary Intervention and Intra-Aortic Balloon Pumping (IABP): A Case Report.
BACKGROUND It is still challenging to remove an epidural catheter in a postoperative patient receiving urgent antiplatelet and anticoagulation therapy for acute coronary syndrome. CASE REPORT While under general anesthesia combined with thoracic epidural anesthesia, a 72-year-old male patient underwent right radical nephrectomy for renal cell carcinoma. On postoperative day 1 (POD1), the patient experienced bradycardia and a decrease in blood pressure, and he was diagnosed acute myocardial infarction. Intra-aortic balloon pumping (IABP) was induced for cardiogenic shock, and urgent thrombus aspiration and coronary balloon angioplasty were performed. On POD3, the surgeon removed the epidural catheter under both antiplatelet and anticoagulation therapy. At that time, the platelet count was 45×10⁹/L and the activated partial thromboplastin time (APTT) was 72.2 seconds. Four hours after the epidural catheter was removed, the patient complained of bilateral fatigue in legs and developed a loss of sensation. Six hours after the epidural catheter was removed, he developed motor paralysis and became completely paralyzed in both limbs after 9 hours. At 19 hours after the epidural catheter was removed, emergency magnetic resonance imaging detected a spinal epidural hematoma at the level of Th9-11 with compression of the spinal cord. Emergency laminectomy was performed to decompress and remove the spinal epidural hematoma at 18 hours after the onset of sensorimotor symptoms. After surgery and rehabilitation, these symptoms had only slightly improved. CONCLUSIONS In patients with urgent antithrombotic therapy for urgent percutaneous coronary intervention (PCI) with an IABP for acute coronary syndrome, the epidural catheter should not be removed until the IABP and heparin are discontinued, and platelet counts have recovered.


Cassava traits and end-user preference: Relating traits to consumer liking, sensory perception, and genetics.
Breeding efforts have focused on improving agronomic traits of the cassava plant however little research has been done to enhance the crop palatability. This review investigates the links between cassava traits and end-user preference in relation with sensory characteristics. The main trait is starch and its composition related to the textural properties of the food. Pectin degradation during cooking resulted in increased mealiness. Nutritional components such as carotenoids made the cassava yellow but also altered sweetness and softness; however, yellow cassava was more appreciated by consumers than traditional (white) varieties. Components formed during processing such as organic acids gave fermented cassava products an acidic taste that was appreciated but the fermented smell was not always liked. Anti-nutritional compounds such as cyanogenic glucosides were mostly related to bitter taste. Post-harvest Physiological Deterioration (PPD) affected the overall sensory characteristics and acceptability. Genes responsible for some of these traits were also investigated. Diversity in cassava food products can provide a challenge to identifying acceptance criteria. Socio-economic factors such as gender may also be critical. This review leads to questions in relation to the adaptation of cassava breeding to meet consumer needs and preference in order to maximize income, health and food security.


Output factors of ionization chambers and solid state detectors for mobile intraoperative radiotherapy (IORT) accelerator electron beams.
The electron energy characteristics of mobile intraoperative radiotherapy (IORT) accelerator LIAC® differ from commonly used linear accelerators, thus some of the frequently used detectors can give less accurate results. The aim of this study is to evaluate the output factors (OFs) of several ionization chambers (IC) and solid state detectors (SS) for electron beam energies generated by LIAC® and compare with the output factor of Monte Carlo model (MC) in order to determine the adequate detectors for LIAC® . The OFs were measured for 6, 8, 10, and 12 MeV electron energies with PTW 23343 Markus, PTW 34045 Advanced Markus, PTW 34001 Roos, IBA PPC05, IBA PPC40, IBA NACP-02, PTW 31010 Semiflex, PTW 31021 Semiflex 3D, PTW 31014 Pinpoint, PTW 60017 Diode E, PTW 60018 Diode SRS, SNC Diode EDGE, and PTW 60019 micro Diamond detectors. Ion recombination factors (ksat ) of IC were measured for all applicator sizes and OFs were corrected according to ksat . The measured OFs were compared with Monte Carlo output factors (OFMC ). The measured OFs of IBA PPC05, PTW Advanced Markus, PTW Pinpoint, PTW microDiamond, and PTW Diode E detectors are in good agreement with OFMC . The maximum deviations of IBA PPC05 OFs to OFMC are -1.6%, +1.5%, +1.5%, and +2.0%; for PTW Advanced Markus +1.0%, +1.5%, +2.0%, and +2.0%; for PTW Pinpoint +2.0%, +1.6%, +4.0%, and +2.0%; for PTW microDiamond -1.6%, +2%, +1.1%, and +1.0%; and for PTW Diode E -+1.7%, +1.7%, +1.3%, and +2.5% for 6, 8, 10, and 12 MeV, respectively. PTW Roos, PTW Markus, IBA PPC40, PTW Semiflex, PTW Semiflex 3D, SNC Diode Edge measured OFs with a maximum deviation of +5.6%, +4.5%, +5.6%, +8.1%, +4.8%, and +9.6% with respect to OFMC , while PTW Diode SRS and IBA NACP-02 were the least accurate (with highest deviations -37.1% and -18.0%, respectively). The OFs results of solid state detectors PTW microDiamond and PTW Diode E as well as the ICs with small electrode spacing distance such as IBA PPC05, PTW Advanced Markus and PTW Pinpoint are in excellent agreement with OFMC . The measurements of the other detectors evaluated in this study are less accurate, thus they should be used with caution. Particularly, PTW Diode SRS and IBA NACP-02 are not suitable and their use should be avoided in relative dosimetry measurements under high dose per pulsed (DPP) electron beams.


In silico screening for antibiotic escort molecules to overcome efflux.
Resistance to antibiotics is a growing problem worldwide and occurs in part due to the overexpression of efflux pumps responsible for the removal of antibiotics from bacterial cells. The current study examines complex formation between efflux pump substrates and escort molecules as a criterion for an in silico screening method for molecules that are able to potentiate antibiotic activities. Initially, the SUPERDRUG database was queried to select molecules that were similar to known multidrug resistance (MDR) modulators. Molecular interaction fields generated by GRID and the docking module GLUE were used to calculate the interaction energies between the selected molecules and the antibiotic norfloxacin. Ten compounds forming the most stable complexes with favourable changes to the norfloxacin molecular properties were tested for their potentiation ability by efflux pump modulation assays. Encouragingly, two molecules were proven to act as efflux pump modulators, and hence provide evidence that complex formation between a substrate and a drug can be used for in silico screening for novel escort molecules.


Knowledge and practices of paracetamol administration among caregivers of pediatric age group patients.
Paracetamol is a widely used over the counter drug for pyrexia and mild to moderate pain in all age groups. To assess the knowledge and practices of paracetamol administration among caregivers of the pediatric age group patients attending the university family practice. A clinic-based descriptive cross-sectional survey was carried out among clients attending the family practice of the Faculty of Medicine, University of Kelaniya, Sri Lanka using pretested interviewer administrated questionnaire. Ninety eight caregivers Participated. Majority were females (97%) with a mean age of 32 years. Age of the patients ranged from 1 to 132 (mean: 48 months). The commonest indication for paracetamol was fever (98%) and in 99% of the patients, mother was the administrator. Forty three percent of the children received a supra-therapeutic dose (>15 mg/kg/dose). None exceeded 20 mg/kg/dose. 16% exceeded the recommended dosing frequency. Children above 3 years were at an increased risk of receiving incorrect paracetamol dose (χ(2) =19.55, df=1, P>0.001) A majority (75%) said they followed doctors' advice on paracetamol dose. There was no association between level of education of care giver, deciding dose as directed by doctor and product information leaflet and dosing accuracy. Only one caregiver was able to calculate the paracetamol dose according to weight. A majority (85%) knew about paracetamol poisoning but it was not associated with dosing accuracy. Administration of supratheraputic doses of paracetamol is common and risk increased with child's age. Knowledge on calculating the weight appropriate paracetamol dose is poor. Physicians should educate care givers on judicious use of paracetamol.


CD73 is dispensable for the regulation of inflationary CD8+ T-cells after murine cytomegalovirus infection and adenovirus immunisation.
The ecto-5'-nucleotidase (CD73) is expressed by T-cell subsets, myeloid derived suppressive cells and endothelial cells. It works in conjunction with CD39 to regulate the formation and degradation of adenosine in vivo. Adenosine has previously been shown to suppress the proliferation and cytokine secretion of T-cells and recent evidence suggests that inhibition of CD73 has the potential to enhance T-cell directed therapies. Here we utilised a CD73 knockout mouse model to assess the suppressive ability of CD73 on CD8+ T-cell classical memory and memory "inflation", induced by murine cytomegalovirus (MCMV) infection and adenovirus immunisation. We show that CD73 is dispensable for normal CD8+ T-cell differentiation and function in both models. Thus CD73 as a suppressor of CD8+ T-cells is unlikely to play a deterministic role in the generation and functional characteristics of antiviral memory in these settings.


Warfarin use and long-term outcomes in patients with acute myocardial infarction and atrial fibrillation.
Warfarin use in patients with acute myocardial infarction (AMI) and atrial fibrillation (AF) remains challenging. We describe use of warfarin up to 1 year after hospitalization among patients with AMI and AF according to stroke and bleeding risk, and identify factors associated with long-term mortality in this population. Patients with AMI and AF who underwent cardiac catheterization during their AMI hospitalization in 1995-2007 were identified from the Duke Databank for Cardiovascular Disease. Warfarin use at discharge, 6 months, and 1 year as well as long-term vital status were assessed by surveys. Rates of warfarin use were presented according to CHADS2 and CHA2DS2VASc stroke and ATRIA bleeding risk scores. Cox proportional hazards modeling was used to determine whether warfarin use at discharge was independently associated with 1-year mortality. A total of 879 patients hospitalized with AMI with AF were identified. Median age was 72 (25th, 75th percentiles: 64, 79), and median follow-up was 4.1 years (1.3, 7.4). The rate of warfarin use at discharge was 24 % and did not differ by CHADS2, CHA2DS2VASc, or ATRIA risk scores. Warfarin use remained similar at 6 months (26 %) and 1 year (27 %). Long-term mortality was high and did not differ by whether warfarin was or was not prescribed at discharge (72 and 71 %, respectively). Factors associated with 1-year mortality were history of heart failure (HR 1.58, 95 % CI 1.32-1.90), higher Charlson comorbidity index (HR 1.19, 95 % CI 1.11-1.28), and older age (HR 1.03 per 1-year increase, 95 % CI 1.02-1.05). Warfarin use at discharge among patients hospitalized for AMI who had comorbid AF was low and remained low at 1 year. Warfarin use at hospital discharge was not associated with either 1-year mortality or long-term mortality.


Accumulation parameters and seasonal trends for PCBs in temperate and boreal forest plant species.
The concentration of polychlorinated biphenyls (PCBs) in the air and vegetation was measured periodically in two alpine forests, during the growing season. Foliage samples from nine plant species typical of the temperate and boreal environment were collected and analyzed. Leaf concentrations of tri- and tetra-CBs showed fast response times with changing temperature and gas-phase concentrations, suggesting that a partitioning equilibrium is approached relatively rapidly (few days) in the field. Heavier compounds showed kinetically limited accumulation trends, not reaching equilibrium during the growing season. Results were used to estimate the bioconcentration factors or equilibrium plant/air partition coefficient (KPA) for each species. Values of log KPA (calculated on a mass/volume basis) ranged between 0.78 and 1.96 and were correlated to the log KOA. Uptake trends of the higher chlorinated compounds showed intraspecific differences which were partially explained by the specific leaf area (SLA).


A novel human WD protein, h-beta TrCp, that interacts with HIV-1 Vpu connects CD4 to the ER degradation pathway through an F-box motif.
HIV-1 Vpu interacts with CD4 in the endoplasmic reticulum and triggers CD4 degradation, presumably by proteasomes. Human beta TrCP identified by interaction with Vpu connects CD4 to this proteolytic machinery, and CD4-Vpu-beta TrCP ternary complexes have been detected by coimmunoprecipitation. beta TrCP binding to Vpu and its recruitment to membranes require two phosphoserine residues in Vpu essential for CD4 degradation. In beta TrCP, WD repeats at the C terminus mediate binding to Vpu, and an F box near the N terminus is involved in interaction with Skp1p, a targeting factor for ubiquitin-mediated proteolysis. An F-box deletion mutant of beta TrCP had a dominant-negative effect on Vpu-mediated CD4 degradation. These data suggest that beta TrCP and Skp1p represent components of a novel ER-associated protein degradation pathway that mediates CD4 proteolysis.


ABO blood group and COVID-19: a review on behalf of the ISBT COVID-19 Working Group.
Growing evidence suggests that ABO blood group may play a role in the immunopathogenesis of SARS-CoV-2 infection, with group O individuals less likely to test positive and group A conferring a higher susceptibility to infection and propensity to severe disease. The level of evidence supporting an association between ABO type and SARS-CoV-2/COVID-19 ranges from small observational studies, to genome-wide-association-analyses and country-level meta-regression analyses. ABO blood group antigens are oligosaccharides expressed on red cells and other tissues (notably endothelium). There are several hypotheses to explain the differences in SARS-CoV-2 infection by ABO type. For example, anti-A and/or anti-B antibodies (e.g. present in group O individuals) could bind to corresponding antigens on the viral envelope and contribute to viral neutralization, thereby preventing target cell infection. The SARS-CoV-2 virus and SARS-CoV spike (S) proteins may be bound by anti-A isoagglutinins (e.g. present in group O and group B individuals), which may block interactions between virus and angiotensin-converting-enzyme-2-receptor, thereby preventing entry into lung epithelial cells. ABO type-associated variations in angiotensin-converting enzyme-1 activity and levels of von Willebrand factor (VWF) and factor VIII could also influence adverse outcomes, notably in group A individuals who express high VWF levels. In conclusion, group O may be associated with a lower risk of SARS-CoV-2 infection and group A may be associated with a higher risk of SARS-CoV-2 infection along with severe disease. However, prospective and mechanistic studies are needed to verify several of the proposed associations. Based on the strength of available studies, there are insufficient data for guiding policy in this regard.


Tropomyosin-related kinase B in the mesolimbic dopamine system: region-specific effects on cocaine reward.
Previous studies found that brain-derived neurotrophic factor (BDNF) derived from nucleus accumbens (NAc) neurons can mediate persistent behavioral changes that contribute to cocaine addiction. To further investigate BDNF signaling in the mesolimbic dopamine system, we analyzed tropomyosin-related kinase B (TrkB) messenger RNA (mRNA) and protein changes in the NAc and ventral tegmental area (VTA) in rats following 3 weeks of cocaine self-administration. To study the role of BDNF-TrkB activity in the VTA and NAc in cocaine reward, we used localized viral-mediated Cre recombinase expression in floxed BDNF and floxed TrkB mice to knockdown BDNF or TrkB in the VTA and NAc in cocaine place conditioning tests and TrkB in the NAc in cocaine self-administration tests. We found that 3 weeks of active cocaine self-administration significantly increased TrkB protein levels in the NAc shell, while yoked (passive) cocaine exposure produced a similar increase in the VTA. Localized BDNF knockdown in either region reduced cocaine reward in place conditioning, whereas only TrkB knockdown in the NAc reduced cocaine reward. In mice self-administering cocaine, TrkB knockdown in the NAc produced a downward shift in the cocaine self-administration dose-response curve but had no effect on the acquisition of cocaine or sucrose self-administration. Together, these data suggest that BDNF synthesized in either VTA or NAc neurons is important for maintaining sensitivity to cocaine reward but only BDNF activation of TrkB receptors in the NAc mediates this effect. In addition, up-regulation of NAc TrkB with chronic cocaine use could promote the transition to more addicted biological states.


Effect of Sahaja yoga practice on seizure control & EEG changes in patients of epilepsy.
The effect of Sahaja yoga meditation on seizure control and electroencephalographic alterations was assessed in 32 patients of idiopathic epilepsy. The subjects were randomly divided into 3 groups. Group I (n = 10) practised Sahaja yoga for 6 months, Group II (n = 10) practised exercises mimicking Sahaja yoga for 6 months and Group III (n = 12) served as the epileptic control group. Group I subjects reported a 62 per cent decrease in seizure frequency at 3 months and a further decrease of 86 per cent at 6 months of intervention. Power spectral analysis of EEG showed a shift in frequency from 0-8 Hz towards 8-20 Hz. The ratios of EEG powers in delta (D), theta (T), alpha (A) and beta (B) bands i.e., A/D, A/D + T, A/T and A + B/D + T were increased. Per cent D power decreased and per cent A increased. No significant changes in any of the parameters were found in Groups II and III, indicating that Sahaja yoga practice brings about seizure reduction and EEG changes. Sahaja yoga could prove to be beneficial in the management of patients of epilepsy.


Prospective State and Trait Predictors of Daily Medication Adherence Behavior in HIV.
Many persons living with HIV (PLWH) are nonadherent to medication. Trait level measures that ask about predictors of adherence in the abstract may not adequately capture state level daily variability that more directly impacts adherence. This preliminary study was designed to test six predictors of electronically monitored adherence at both the state and trait levels and to compare their relative effects. Using a smartphone, 87 PLWH completed randomly cued daily surveys on thoughts, mood, stress, coping, social support, and treatment motivation. All participants also completed baseline surveys on each construct. These state and trait variables were tested as prospective predictors of next-day adherence in multilevel models, and their relative importance was quantified. The analysis sample consisted of 53 PLWH who stored their most frequent antiretroviral medication in a bottle that time-stamped openings to measure adherence. Higher state level motivation, OR = 1.55, 95% CI [1.07, 2.24], and negative mood, OR = 1.33, 95% CI [1.07, 1.63], predicted greater adherence the following day. Importantly, these effects were only found at the state level. Trait level control beliefs predicted greater adherence, OR = 1.65, 95% CI [1.17, 2.35], but contrary to prediction, validated trait level measures of mood, stress, coping, social support, and motivation did not. Trait and state level measures predicted adherence, but there were differences between them. Motivation for treatment and negative mood predicted adherence when measured the preceding day, but not as aggregate measures. At the trait level, only control beliefs predicted adherence. Researchers should consider state level variations in mood and motivation as possible explanations for nonadherence. Interventions could be developed to target state level variables.


Tea consumption and bone health in Chinese adults: a population-based study.
Tea is a worldwide drink with controversial effect on bone health. The sex-specific associations are unrevealed among general population. This study showed that prolonged moderate tea consumption benefited bone health in women, while no additional benefit with stronger tea. However, tea consumption was not associated with bone health in men. Tea consumption has been shown a potentially beneficial effect on bone health in postmenopausal women. However, little is known about such association in men, and whether stronger tea instead harms bone health due to elevated urinary excretion of calcium associated with caffeine in the tea. The aim of this study was to examine the association between various metrics of tea consumption and bone health. The present study included 20,643 participants from the China Kadoorie Biobank (CKB), who have finished both baseline survey (2004-2008) and a re-survey (2013-2014). They were aged 38-86 years at re-survey. Tea consumption was self-reported at both baseline and re-survey. Bone mineral density (BMD) was measured using calcaneal quantitative ultrasound once at re-survey. Compared with non-consumers, prolonged weekly tea consumers in women was associated with higher calcaneus BMD measures, with β (95% CI) of 0.98 (0.22, 1.74) for BUA, 4.68 (1.74, 7.61) for SOS, and 1.95 (0.81, 3.10) for SI. Among prolonged weekly tea consumers, no linear increase in BMD measures with the amount of tea leaves added was observed. The SOS and SI were higher in consumers with tea leaves 3.0-5.9 g/day than in those with < 3.0 g/day, but were reduced to non-significant for those with ≥ 6.0 g/day. Tea consumption was not associated with calcaneus BMD measures in men. Prolonged moderate tea consumption benefited bone health in women but not in men. For stronger tea consumption with more tea leaves added, neither benefit nor harm to bone health was observed.


Origins of regulated cell-to-cell variability.
Single-cell measurements and lineage-tracing experiments are revealing that phenotypic cell-to-cell variability is often the result of deterministic processes, despite the existence of intrinsic noise in molecular networks. In most cases, this determinism represents largely uncharacterized molecular regulatory mechanisms, which places the study of cell-to-cell variability in the realm of molecular cell biology. Further research in the field will be important to advance quantitative cell biology because it will provide new insights into the mechanisms by which cells coordinate their intracellular activities in the spatiotemporal context of the multicellular environment.


Communication of Information about Genetic Risks: Putting Families at the Center.
Genetic information is a family affair. With the expansion of genomic technologies, many new causal genes and variants have been established and the potential for molecular diagnoses increased, with implications not only for patients but also their relatives. The need for genetic counseling and intrafamilial circulation of information on genetic risks grew accordingly. Also, the amount and, particularly, the complexity of the information to convey multiplied. Sharing information about genetic risks with family members, however, has never been an easy matter and often becomes a source of personal and familial conflicts and distress. Ethical requisites generally prevent healthcare professionals from directly contacting their consultands' relatives (affected or still at risk), who often feel unsupported throughout that process. We discuss here the communication of genetic risks to family members. We first consider genomic testing as a basis for family-centered health care, as opposed to a predominant focus on the individual. We reviewed the literature on sharing genetic risk information with family members, and the associated ethical issues for professionals. Some clinical cases are presented and discussed, and key issues for meeting the needs of individuals and families are addressed. We argue that genetic information is inextricably linked to the family and that communicating about genetic risks is a process grounded within the broader milieu of family relationships and functioning. We conclude for the need for a more family-centered approach and interventions that can promote sensitive attitudes to the provision of genetic information to and within the family, as well as its inclusion in educational and training programmes for genetic healthcare professionals.


Wine glasses and hourglasses: Non-adaptive complexity of vesicle traffic in microbial eukaryotes.
Microbial eukaryotes present a stunning diversity of endomembrane organization. From specialized secretory organelles such as the rhoptries and micronemes of apicomplexans, to peroxisome-derived metabolic compartments such as the glycosomes of kinetoplastids, different microbial taxa have explored different solutions to the compartmentalization and processing of cargo. The basic secretory and endocytic system, comprising the ER, Golgi, endosomes, and plasma membrane, as well as diverse taxon-specific specialized endomembrane organelles, are coupled by a complex network of cargo transport via vesicle traffic. It is tempting to connect form to function, ascribing biochemical roles to each compartment and vesicle of such a system. Here we argue that traffic systems of high complexity could arise through non-adaptive mechanisms via purely physical constraints, and subsequently be exapted for various taxon-specific functions. Our argument is based on a Boolean mathematical model of vesicle traffic: we specify rules of how compartments exchange vesicles; these rules then generate hypothetical cells with different types of endomembrane organization. Though one could imagine a large number of hypothetical vesicle traffic systems, very few of these are consistent with molecular interactions. Such molecular constraints are the bottleneck of a metaphorical hourglass, and the rules that make it through the bottleneck are expected to generate cells with many special properties. Sampling at random from among such rules represents an evolutionary null hypothesis: any properties of the resulting cells must be non-adaptive. We show by example that vesicle traffic systems generated in this random manner are reminiscent of the complex trafficking apparatus of real cells.


Outcome Prediction in Patients with Severe COVID-19 Requiring Extracorporeal Membrane Oxygenation-A Retrospective International Multicenter Study.
The role of veno-venous extracorporeal membrane oxygenation therapy (V-V ECMO) in severe COVID-19 acute respiratory distress syndrome (ARDS) is still under debate and conclusive data from large cohorts are scarce. Furthermore, criteria for the selection of patients that benefit most from this highly invasive and resource-demanding therapy are yet to be defined. In this study, we assess survival in an international multicenter cohort of COVID-19 patients treated with V-V ECMO and evaluate the performance of several clinical scores to predict 30-day survival. This is an investigator-initiated retrospective non-interventional international multicenter registry study (NCT04405973, first registered 28 May 2020). In 127 patients treated with V-V ECMO at 15 centers in Germany, Switzerland, Italy, Belgium, and the United States, we calculated the Sequential Organ Failure Assessment (SOFA) Score, Simplified Acute Physiology Score II (SAPS II), Acute Physiology And Chronic Health Evaluation II (APACHE II) Score, Respiratory Extracorporeal Membrane Oxygenation Survival Prediction (RESP) Score, Predicting Death for Severe ARDS on V‑V ECMO (PRESERVE) Score, and 30-day survival. In our study cohort which enrolled 127 patients, overall 30-day survival was 54%. Median SOFA, SAPS II, APACHE II, RESP, and PRESERVE were 9, 36, 17, 1, and 4, respectively. The prognostic accuracy for all these scores (area under the receiver operating characteristic-AUROC) ranged between 0.548 and 0.605. The use of scores for the prediction of mortality cannot be recommended for treatment decisions in severe COVID-19 ARDS undergoing V-V ECMO; nevertheless, scoring results below or above a specific cut-off value may be considered as an additional tool in the evaluation of prognosis. Survival rates in this cohort of COVID-19 patients treated with V‑V ECMO were slightly lower than those reported in non-COVID-19 ARDS patients treated with V-V ECMO.


Quantifying transcription factor kinetics: at work or at play?
Transcription factors (TFs) interact dynamically in vivo with chromatin binding sites. Here we summarize and compare the four different techniques that are currently used to measure these kinetics in live cells, namely fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), single molecule tracking (SMT) and competition ChIP (CC). We highlight the principles underlying each of these approaches as well as their advantages and disadvantages. A comparison of data from each of these techniques raises an important question: do measured transcription kinetics reflect biologically functional interactions at specific sites (i.e. working TFs) or do they reflect non-specific interactions (i.e. playing TFs)? To help resolve this dilemma we discuss five key unresolved biological questions related to the functionality of transient and prolonged binding events at both specific promoter response elements as well as non-specific sites. In support of functionality, we review data suggesting that TF residence times are tightly regulated, and that this regulation modulates transcriptional output at single genes. We argue that in addition to this site-specific regulatory role, TF residence times also determine the fraction of promoter targets occupied within a cell thereby impacting the functional status of cellular gene networks. Thus, TF residence times are key parameters that could influence transcription in multiple ways.


Genome-wide association study to identify genetic determinants of severe asthma.
The genetic basis for developing asthma has been extensively studied. However, association studies to date have mostly focused on mild to moderate disease and genetic risk factors for severe asthma remain unclear. To identify common genetic variants affecting susceptibility to severe asthma. A genome-wide association study was undertaken in 933 European ancestry individuals with severe asthma based on Global Initiative for Asthma (GINA) criteria 3 or above and 3346 clean controls. After standard quality control measures, the association of 480 889 genotyped single nucleotide polymorphisms (SNPs) was tested. To improve the resolution of the association signals identified, non-genotyped SNPs were imputed in these regions using a dense reference panel of SNP genotypes from the 1000 Genomes Project. Then replication of SNPs of interest was undertaken in a further 231 cases and 1345 controls and a meta-analysis was performed to combine the results across studies. An association was confirmed in subjects with severe asthma of loci previously identified for association with mild to moderate asthma. The strongest evidence was seen for the ORMDL3/GSDMB locus on chromosome 17q12-21 (rs4794820, p=1.03×10((-8)) following meta-analysis) meeting genome-wide significance. Strong evidence was also found for the IL1RL1/IL18R1 locus on 2q12 (rs9807989, p=5.59×10((-8)) following meta-analysis) just below this threshold. No novel loci for susceptibility to severe asthma met strict criteria for genome-wide significance. The largest genome-wide association study of severe asthma to date was carried out and strong evidence found for the association of two previously identified asthma susceptibility loci in patients with severe disease. A number of novel regions with suggestive evidence were also identified warranting further study.


High-Throughput HIV-Cell Fusion Assay for Discovery of Virus Entry Inhibitors.
HIV-1 initiates infection by merging its envelope membrane with the target cell membrane, a process that is mediated by the viral Env glycoprotein following its sequential binding to CD4 and coreceptors, CXCR4 or CCR5. Although HIV-1 fusion has been a target for antiviral therapy, the virus has developed resistance to drugs blocking the CCR5 binding or Env refolding steps of this process. This highlights the need for novel inhibitors. Here, we adapted and optimized an enzymatic HIV-cell fusion assay, which reports the transfer of virus-encapsulated β-lactamase into the cytoplasm, to high-throughput screening (HTS) with a 384-well format. The assay was robustly performed in HTS format and was validated by the pilot screen of a small library of pharmacologically active compounds. Several hits identified by screening included a prominent cluster of purinergic receptor antagonists. Functional studies demonstrated that P2X1 receptor antagonists selectively inhibited HIV-1 fusion without affecting the fusion activity of an unrelated virus that enters cells through an endocytic route. The inhibition of HIV-cell fusion by P2X1 antagonists was not through downmodulation of the cell surface expression of CD4 or coreceptors, thus implicating P2X1 receptor in the HIV-1 fusion step. The ability of these antagonists to inhibit viruses regardless of their coreceptor (CXCR4 or CCR5) preference indicates that fusion is blocked at a late step downstream of coreceptor binding. A future large-scale screening campaign for HIV-1 fusion inhibitors, using the above functional readout, will likely reveal novel classes of inhibitors and suggest potential targets for antiviral therapy.


High Stability of the Epigenome in Drosophila Interspecific Hybrids.
Interspecific hybridization is often seen as a genomic stress that may lead to new gene expression patterns and deregulation of transposable elements (TEs). The understanding of expression changes in hybrids compared with parental species is essential to disentangle their putative role in speciation processes. However, to date we ignore the detailed mechanisms involved in genomic deregulation in hybrids. We studied the ovarian transcriptome and epigenome of the Drosophila buzzatii and Drosophila koepferae species together with their F1 hybrid females. We found a trend toward underexpression of genes and TE families in hybrids. The epigenome in hybrids was highly similar to the parental epigenomes and showed intermediate histone enrichments between parental species in most cases. Differential gene expression in hybrids was often associated only with changes in H3K4me3 enrichments, whereas differential TE family expression in hybrids may be associated with changes in H3K4me3, H3K9me3, or H3K27me3 enrichments. We identified specific genes and TE families, which their differential expression in comparison with the parental species was explained by their differential chromatin mark combination enrichment. Finally, cis-trans compensatory regulation could also contribute in some way to the hybrid deregulation. This work provides the first study of histone content in Drosophila interspecific hybrids and their effect on gene and TE expression deregulation.


Combined in vivo metabolic effects of quetiapine and methadone in brain and blood of rats.
Changes in pharmacokinetics and endogenous metabolites may underlie additive biological effects of concomitant use of antipsychotics and opioids. In this study, we employed untargeted metabolomics analysis and targeted analysis to examine the changes in drug metabolites and endogenous metabolites in the prefrontal cortex (PFC), midbrain, and blood of rats following acute co-administration of quetiapine and methadone. Rats were divided into four groups and received cumulative increasing doses of quetiapine (QTP), methadone (MTD), quetiapine + methadone (QTP + MTD), or vehicle (control). All samples were analyzed using liquid chromatography-mass spectrometry (LC-MS). Our findings revealed increased levels of the quetiapine metabolites: Norquetiapine, O-dealkylquetiapine, 7-hydroxyquetiapine, and quetiapine sulfoxide, in the blood and brain when methadone was present. Our study also demonstrated a decrease in methadone and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in the rat brain when quetiapine was present. Despite these findings, there were only small differences in the levels of 225-296 measured endogenous metabolites due to co-administration compared to single administrations. For example, N-methylglutamic acid, glutaric acid, p-hydroxyphenyllactic acid, and corticosterone levels were significantly decreased in the brain of rats treated with both compounds. Accumulation of serotonin in the midbrain was additionally observed in the MTD group, but not in the QTP + MTD group. In conclusion, this study in rats suggests a few but important additive metabolic effects when quetiapine and methadone are co-administered.


Encouraging responses in sexual and relationship violence prevention: what program effects remain 1 year later?
Colleges and universities are high-risk settings for sexual and relationship violence. To address these problems, institutions of higher education have implemented prevention programs, many of which train students as potential bystanders who can step in to help diffuse risky situations, identify and challenge perpetrators, and assist victims. The impact of bystander sexual and relationship violence prevention programs on long-term behavior of bystanders has remained a key unanswered question for those who seek to offer the most effective programs as well as for policy makers. In this study, the researchers experimentally evaluated the effectiveness of the Bringing in the Bystander® in-person program. Participants were 948 1st-year college students of whom 47.8% were women and 85.2% identified as White (15% also identified as Hispanic in a separate question) between the ages of 18 and 24 at two universities (one a rural, primarily residential campus and the other an urban, highly commuter campus) in the northeastern United States. To date, this is the first study to have found positive behavior changes as long-lasting as 1 year following an educational workshop focusing on engaging bystanders in preventing sexual and relationship violence. Even so, many questions remain to be answered about prevention and intervention of this type. More prospective research is needed on bystander-focused prevention of these forms of violence to help understand and better predict the complicated relationships both between and among the attitudes and behaviors related to preventing sexual and relationship violence. In this regard, we make specific recommendations for designing and evaluating programs based on our findings relating to the importance of moderators, especially two key understudied ones, readiness to help and opportunity to intervene.


Cell trafficking of endothelial progenitor cells in tumor progression.
Blood vessel formation plays an essential role in many physiologic and pathologic processes, including normal tissue growth and healing, as well as tumor progression. Endothelial progenitor cells (EPC) are a subtype of stem cells with high proliferative potential that are capable of differentiating into mature endothelial cells, thus contributing to neovascularization in tumors. In response to tumor-secreted cytokines, EPCs mobilize from the bone marrow to the peripheral blood, home to the tumor site, and differentiate to mature endothelial cells and secrete proangiogenic factors to facilitate vascularization of tumors. In this review, we summarize the expression of surface markers, cytokines, receptors, adhesion molecules, proteases, and cell signaling mechanisms involved in the different steps (mobilization, homing, and differentiation) of EPC trafficking from the bone marrow to the tumor site. Understanding the biologic mechanisms of EPC cell trafficking opens a window for new therapeutic targets in cancer.


Family history of type 2 diabetes is associated with increased carotid artery intimal-medial thickness in Mexican Americans.
To evaluate whether the joint risk of diabetes and atherosclerosis tracked within families, we assessed the correlation between a family history of diabetes and intimal-medial thickness (IMT) of the common carotid artery (CCA). Study subjects included 620 nondiabetic individuals from 24 families enrolled in the San Antonio Family Heart Study. The thickness of the far walls of the CCA was measured by B-mode ultrasonography. Statistical analyses included familial correlations to account for the nonindependence of family data. After adjusting for sex, age, and IMT reader effects, the heritability of IMT in this population was 16% (P = 0.009). Using a more comprehensive family history score that accounted for diabetes status of the individual's parents and older siblings, we observed a significant correlation between family history score and increased CCA IMT (0.006 mm increase in CCA IMT for every point increase of diabetes family history score, P = 0.016). This association remained even after further adjustment for BMI, smoking, and fasting insulin and glucose levels. After adjusting for several cardiovascular risk factors, the mean CCA IMT in those with high family history scores for diabetes was still 0.037 mm thicker than those with low family history scores for diabetes (P = 0.040). These results suggest that the genetic contribution to CCA IMT variability is modest. Also, the small increase in subclinical atherosclerosis observed even among nondiabetic Mexican Americans with a positive family history of diabetes is probably transmitted along with the risk of diabetes through shared etiologic risk factors between diabetes and cardiovascular disease.


Adaptive fingerprint image enhancement with emphasis on preprocessing of data.
This article proposes several improvements to an adaptive fingerprint enhancement method that is based on contextual filtering. The term adaptive implies that parameters of the method are automatically adjusted based on the input fingerprint image. Five processing blocks comprise the adaptive fingerprint enhancement method, where four of these blocks are updated in our proposed system. Hence, the proposed overall system is novel. The four updated processing blocks are: 1) preprocessing; 2) global analysis; 3) local analysis; and 4) matched filtering. In the preprocessing and local analysis blocks, a nonlinear dynamic range adjustment method is used. In the global analysis and matched filtering blocks, different forms of order statistical filters are applied. These processing blocks yield an improved and new adaptive fingerprint image processing method. The performance of the updated processing blocks is presented in the evaluation part of this paper. The algorithm is evaluated toward the NIST developed NBIS software for fingerprint recognition on FVC databases.


Climatic shocks associate with innovation in science and technology.
Human history is shaped by landmark discoveries in science and technology. However, across both time and space the rate of innovation is erratic: Periods of relative inertia alternate with bursts of creative science and rapid cascades of technological innovations. While the origins of the rise and fall in rates of discovery and innovation remain poorly understood, they may reflect adaptive responses to exogenously emerging threats and pressures. Here we examined this possibility by fitting annual rates of scientific discovery and technological innovation to climatic variability and its associated economic pressures and resource scarcity. In time-series data from Europe (1500-1900CE), we indeed found that rates of innovation are higher during prolonged periods of cold (versus warm) surface temperature and during the presence (versus absence) of volcanic dust veils. This negative temperature-innovation link was confirmed in annual time-series for France, Germany, and the United Kingdom (1901-1965CE). Combined, across almost 500 years and over 5,000 documented innovations and discoveries, a 0.5°C increase in temperature associates with a sizable 0.30-0.60 standard deviation decrease in innovation. Results were robust to controlling for fluctuations in population size. Furthermore, and consistent with economic theory and micro-level data on group innovation, path analyses revealed that the relation between harsher climatic conditions between 1500-1900CE and more innovation is mediated by climate-induced economic pressures and resource scarcity.


Predictors and outcomes of scleroderma renal crisis: the high-dose versus low-dose D-penicillamine in early diffuse systemic sclerosis trial.
The reported frequency of scleroderma M01-R renal crisis (SRC) in diffuse systemic sclerosis (SSc; scleroderma) is 15-20%. Early use of angiotensin-converting enzyme (ACE) inhibitors has markedly improved outcome. The present analysis reexamines the prognostic factors for and outcome of SRC in a prospective cohort of patients with early diffuse SSc. We retrospectively evaluated the cohort of SSc patients who participated in the High-Dose Versus Low-Dose D-Penicillamine in Early Diffuse SSc trial. Patients with diffuse cutaneous scleroderma were enrolled if their disease duration was <18 months. Because the trial failed to show a difference between treatment groups, the data were pooled. One hundred thirty-four SSc patients entered the observation period a mean +/- SD of 0.8 +/- 0.3 years after onset of SSc. SRC occurred in 18 patients a mean +/- SD of 0.9 +/- 1.1 years after entry. During a mean +/- SD 4.0 +/- 1.1 years of followup after entry, 9 of the 18 patients died (mean +/- SD 0.6 +/- 0.9 years after SRC onset). Baseline characteristics that predicted SRC included a modified Rodnan skin thickness score of >or=20 (P < 0.01), enlarged cardiac silhouette on radiograph (P = 0.04), large joint contractures (wrist, elbow, knee) (P = 0.008), and prednisone use at entry (P = 0.01). Baseline characteristics that did not predict SRC included age, sex, race, Health Assessment Questionnaire score, fist closure, handspread, lung involvement, muscle weakness, erythrocyte sedimentation rate, and platelet count. In 5 of 10 subjects for whom at least 2 sequential skin scores were available, skin scores increased significantly (P = 0.012) in the 6 months before onset of SRC. SRC occurred in 13% of patients soon (mean 11 months) after entry into the cohort. Predictors of SRC identified in this study included higher than average skin score, prednisone use at study entry, large joint contractures, and heart enlargement. Our data suggest, however, that low-dose prednisone alone was not associated with the onset of SRC, except in the appropriate clinical setting. Although ACE inhibitors and dialysis are now readily available, SRC continues to be associated with poor survival (in this study, 50% of patients with SRC died).


The experience of seeking, gaining and maintaining employment after traumatic spinal cord injury and the vocational pathways involved.
Vocational potential in people with spinal cord injury (SCI) are unrealised with rates of employment substantially lower than in the labour force participation of the general population and the pre-injury employment rates. To understand the experience and pathway of people achieving employment outcome after traumatic spinal cord injury by; classifying participants into employment outcome groups of stable, unstable and without employment; identifying pre and post-injury pathways for participants in each group and, exploring the experiences of people of seeking, gaining and maintaining employment. Thirty-one participants were interviewed. Mixed methods approach including interpretive phenomenological analysis and vocational pathway mapping of quantitative data. The most common pathway identified was from study and work pre-injury to stable employment post-injury. Four super-ordinate themes were identified from the interpretive phenomenological analysis; expectations of work, system impacts, worker identity and social supports. Implications for clinical practice include fostering cultural change, strategies for system navigation, promotion of worker identity and optimal use of social supports. The findings increase insight and understanding of the complex experience of employment after spinal cord injury. There is opportunity to guide experimental research, policy development and education concerning the complexity of the return to work experience and factors that influence pathways.


Idiopathic Brainstem Neuronal Chromatolysis (IBNC): a novel prion protein related disorder of cattle?
The epidemic form of Bovine Spongiform Encephalopathy (BSE) is generally considered to have been caused by a single prion strain but at least two strain variants of cattle prion disorders have recently been recognized. An additional neurodegenerative condition, idiopathic brainstem neuronal chromatolysis and hippocampal sclerosis (IBNC), a rare neurological disease of adult cattle, was also recognised in a sub-set of cattle submitted under the BSE Orders in which lesions of BSE were absent. Between the years of 1988 and 1991 IBNC occurred in Scotland with an incidence of 7 cases per 100,000 beef suckler cows over the age of 6 years. When the brains of 15 IBNC cases were each tested by immunohistochemistry, all showed abnormal labelling for prion protein (PrP). Immunohistological labelling for PrP was also present in the retina of a single case available for examination. The pattern of PrP labelling in brain is distinct from that seen in other ruminant prion diseases and is absent from brains with other inflammatory conditions and from normal control brains. Brains of IBNC cattle do not reveal abnormal PrP isoforms when tested by the commercial BioRad or Idexx test kits and do not reveal PrPres when tested by Western blotting using stringent proteinase digestion methods. However, some weakly protease resistant isoforms of PrP may be detected when tissues are examined using mild proteinase digestion techniques. The study shows that a distinctive neurological disorder of cattle, which has some clinical similarities to BSE, is associated with abnormal PrP labelling in brain but the pathology and biochemistry of IBNC are distinct from BSE. The study is important either because it raises the possibility of a significant increase in the scope of prion disease or because it demonstrates that widespread and consistent PrP alterations may not be confined to prion diseases. Further studies, including transmission experiments, are needed to establish whether IBNC is a condition in which prion protein is abnormally regulated or it is yet a further example of an infectious cattle prion disease.


Engineering of β-mannanase from Aspergillus niger to increase product selectivity towards medium chain length mannooligosaccharides.
Mannooligosaccharides (MOSs) are one of the most commonly used biomass-derived feed additives. The effectiveness of MOS varies with the length of oligosaccharides, medium length MOSs such as mannotetraose and mannopentaose being the most efficient. This study aims at improving specificity of β-mannanase from Aspergillus niger toward the desirable product size through rational-based enzyme engineering. Tyr 42 and Tyr 132 were mutated to Gly to extend the substrate binding site, allowing higher molecular weight MOS to non-catalytically bind to the enzyme. Hydrolysis product content was analyzed by high-performance anion-exchange chromatography with pulsed amperometric detection. Instead of mannobiose, the enzyme variants yielded mannotriose and mannotetraose as the major products, followed by mannobiose and mannopentaose. Overall, 42% improvement in production yield of highly active mannotetraose and mannopentaose was achieved. This validates the use of engineered β-mannanase to selectively produce larger MOS, making them promising candidates for large-scale MOS enzymatic production process.


Repeatability of a questionnaire to assess respiratory symptoms in smokers.
To evaluate the repeatability of a questionnaire designed to assess change in respiratory symptoms 90 smokers were interviewed on two occasions. The questionnaire included questions from the Medical Research Council questionnaire on respiratory symptoms, questions on acute chest illness and cough and phlegm production in the preceding two weeks, a modification of Field's card system for estimating frequency of cough, and an objective assessment of the presence of phlegm--the loose cough sign. The study was carried out in two parts. During the first part 30 male smokers were interviewed by one observer and then re-interviewed 1 to 2 hours later by a different observer. During the second part 60 subjects were interviewed and then after a period of 1 to 10 days re-interviewed by the same observer. The results showed that the within-subject variation representing the measurement error for Field's card system was 15.1% of the between-subject variation and was adequately Normal to justify the use of standard analytical techniques. Similar results were obtained from questions on cough and phlegm scored between 1 and 5, although the variation in this case was rather less Normal. In general, the between-observer, within-observer, and within-subject repeatability were satisfactory for all parts of the questionnaire with the exception of the loose cough sign which had a relatively low prevalence. There was no evidence of an observer order effect and there were no important systematic differences due to lapses in time or different observers.The findings indicate that the techniques such as the cough scoring system may be used to permit studies of respiratory symptoms via questionnaire methods to be much smaller than those required to detect equivalent differences in prevalences.


Photon upconversion sensitized nanoprobes for sensing and imaging of pH.
Acidic pH inside cells indicates cellular dysfunctions such as cancer. Therefore, the development of optical pH sensors for measuring and imaging intracellular pH is a demanding challenge. The available pH-sensitive probes are vulnerable to e.g. photobleaching or autofluorescence background in biological materials. Our approach circumvents these problems due to near infrared excitation and upconversion photoluminescence. We introduce a nanosensor based on upconversion resonance energy transfer (UC-RET) between an upconverting nanoparticle (UCNP) and a fluorogenic pH-dependent dye pHrodo™ Red that was covalently bound to the aminosilane surface of the nanoparticles. The sensitized fluorescence of the pHrodo™ Red dye increases strongly with decreasing pH. By referencing the pH-dependent emission of pHrodo™ Red with the pH-insensitive upconversion photoluminescence of the UCNP, we developed a pH-sensor which exhibits a dynamic range from pH 7.2 to 2.5. The applicability of the introduced pH nanosensor for pH imaging was demonstrated by imaging the two emission wavelengths of the nanoprobe in living HeLa cells with a confocal fluorescence microscope upon 980 nm excitation. This demonstrates that the presented pH-nanoprobe can be used as an intracellular pH-sensor due to the unique features of UCNPs: excitation with deeply penetrating near-infrared light, high photostability, lack of autofluorescence and biocompatibility due to an aminosilane coating.


Computer-Based Cognitive Rehabilitation Interventions for Traumatic Brain Injury: A Critical Review of the Literature.
Computer-based interventions have been developed to improve cognitive performance after mild traumatic brain injury; however, a thorough evaluation of this body of research has not been addressed in the literature. The aim of this study was to provide a synthesis and critical review of current research studies that have tested the efficacy of computer-based interventions on cognitive performance after mild traumatic brain injury. A critical review was conducted by identifying relevant studies in the electronic databases PubMed/MEDLINE, PsycInfo, and CINAHL from 2011 to the present. Because of the limited number of publications focused exclusively on mild traumatic brain injury, research studies that assessed the impact of computer-based interventions on cognitive outcomes in populations with acquired brain injury were included. Of the 58 studies identified, only 10 publications included participants with mild traumatic brain injury. Overall, the identified studies did not use a standard method for assessing the severity of traumatic brain injury, and many studies included participants with a wide variety of etiologies for acquired brain injury and used multiple measures of cognitive performance, which made comparisons difficult across studies. In addition to small sample sizes, the study samples were heterogeneous in regard to the number of previous traumatic brain injuries, time elapsed since injury, and age and gender distributions. Preinjury comorbidities that may affect cognitive performance, such as depression, anxiety, or learning disabilities, were often not assessed. There is weak evidence that computer-based interventions can improve working memory and cognitive function in individuals after mild traumatic brain injury. Because of the low-quality evidence, seminal questions remain regarding the optimal format, dosage, timing, and duration of computer-based intervention for improving cognitive performance. Future studies should focus on using a strong research design, such as a prospective, longitudinal, repeated-measures study, with an adequate number of participants who meet mild traumatic brain injury criteria. Preinjury comorbidities, cognitive reserve, time since injury, age, and gender should be addressed in the design because there may be differences in recovery time and mechanisms of cognitive plasticity among populations. Overall, computer-based interventions seem promising as an approach to improve working memory in individuals with acquired brain injury. There is no evidence that currently available interventions are specific to mild traumatic brain injury. Well-designed research studies with adequate sample sizes are needed to assess the effect of computer-based interventions on cognitive performance after mild traumatic brain injury.


The feasibility of electronic tracking devices in dementia: a telephone survey and case series.
Patients with dementia who go out unaccompanied are at risk of accidents or getting lost. It is not known whether they could benefit from electronic tracking devices or whether such devices are practically feasible. The likely demand for an electronic tracking device was assessed by means of a telephone survey of a convenience sample of 99 carers. The practical feasibility of a tracking system was assessed in 24 patients with dementia. The telephone survey suggested that 20% of patients were at continuing risk of traffic accidents and 45% were at continuing risk of getting lost. About 7% could have benefited from using the device at the time of survey and a further 11% could have benefited at an earlier point in their illness. In the feasibility study, only nine patients consistently used the device. In two patients, it was successfully used in a search. One patient was injured by a passing vehicle when he had got lost out of range of the device. A major barrier to using the device was recognizing the risk of getting lost before it happened. Significant numbers of patients are at risk. Electronic tracking devices may occasionally be useful in carefully selected cases.


The Finnish Diabetes Risk Score is associated with insulin resistance and progression towards type 2 diabetes.
The Finnish Diabetes Risk Score (FINDRISC) questionnaire is a practical screening tool to estimate the diabetes risk and the probability of asymptomatic type 2 diabetes. In this study we evaluated the usefulness of the FINDRISC to predict insulin resistance in a population at increased diabetes risk. Data of 771 and 526 participants in a cross-sectional survey (1996) and a cohort study (1997-2000), respectively, were used for the analysis. Data on the FINDRISC and oral glucose tolerance test parameters were available from each participant. The predictive value of the FINDRISC was cross-sectionally evaluated using the area under the curve-receiver operating characteristics method and by correlation analyses. A validation of the cross-sectional results was performed on the prospective data from the cohort study. The FINDRISC was significantly correlated with markers of insulin resistance. The receiver operating characteristics-area under the curve for the prediction of a homeostasis model assessment insulin resistance index of more than five was 0.78 in the cross-sectional survey and 0.74 at baseline of the cohort study. Moreover, the FINDRISC at baseline was significantly associated with disease evolution (P < 0.01), which was defined as the change of glucose tolerance during the 3 yr follow-up. The results indicate that the FINDRISC can be applied to detect insulin resistance in a population at high risk for type 2 diabetes and predict future impairment of glucose tolerance.


Child care as an untapped setting for obesity prevention: state child care licensing regulations related to nutrition, physical activity, and media use for preschool-aged children in the United States.
Child care is a potential setting for obesity prevention; 8.6 million preschool-aged children participated in child care in 2001. Each US state creates and enforces its own child care licensing regulations. We analyzed obesity-related child care licensing regulations of US states. We downloaded state licensing regulations for children in child care centers (CCCs), small family child care homes (SFHs), and large family or group child care homes (LFGHs) in each state and the District of Columbia (collectively referred to as "states") in 2006 from national and state Web sites. We conducted a quantitative content analysis to identify 13 coding dimensions related to nutrition, physical activity, and media use. We found variability among and within states. CCCs were the most heavily regulated and had the most specific regulations, followed by LFGHs. SFHs had the fewest and most general regulations. Just 2 states, Michigan and West Virginia, specified that CCC menus should be consistent with the Dietary Guidelines for Americans. Only 12 states had regulations that limited foods of low nutritional value in CCCs. Thirty-six states required that children have daily outdoor activity time in CCCs; only 9 states set specific minimum lengths of time that children should be outdoors each day. Eight states set quantified time limits on screen time per day or per week in SFHs. Opportunities exist for strengthening state licensing regulations to prevent childhood obesity. The increasing prevalence of childhood obesity underscores the urgency for state policy efforts to create child care environments that foster healthful eating and participation in physical activity.


Brain structural maturation and the foundations of cognitive behavioral development.
Thorough knowledge of normal neural foundations for cognitive behavioral development is fundamental to understand the mechanisms of both neurodevelopmental disorders and normal adaptation. This review aims at identifying the trends in MRI studies published within the last 18 months illuminating maturational structural brain foundations for normal cognitive behavioral development. Development is coordinated within neurocognitive systems, with predictable functional correlates. There is great individual variability within the normal range. Relationships between brain and cognitive variance at any given age are moderate, and appear to be of a complex and dynamic nature. Importantly, current studies point to a dimensional component to cognitive and behavioral psychopathology in which differences among healthy and clinical developmental groups exist along a continuum. Finally, factors influencing and detectable in early development are likely to have lifespan consequences. Brain development is highly coordinated, but the normal individual variation at any given age is substantial. Relationships between brain and cognitive measures are typically moderate and may fluctuate with age. A dimensional component to neural foundations for multiple developmental disorders makes the study of normal individual brain differences in development even more important to understand both normal and clinical cognitive behavioral outcomes throughout life.


Non-ruminative processing reduces overgeneral autobiographical memory retrieval in students.
It has been suggested that overgeneral memory (OGM) represents a vulnerability marker for depression [Williams, J. M. G., Barnhofer, T., Crane, C., Hermans, D., Raes, F., Watkins, E., et al. (2007). Autobiographical memory specificity and emotional disorder. Psychological Bulletin, 133, 122-148]. One important underlying mechanism involved is rumination [e.g., Watkins, E., & Teasdale, J. D. (2001). Rumination and overgeneral memory in depression: Effects of self-focus and analytic thinking. Journal of Abnormal Psychology, 110, 353-357; Watkins, E., & Teasdale, J. D. (2004). Adaptive and maladaptive self-focus in depression. Journal of Affective Disorders, 82, 1-8]. It is as yet unclear to what extent the relationship between rumination and OGM also applies to nonclinical groups. The present study investigated this relationship in a nonclinical student sample, using an innovative sentence completion procedure to assess OGM. As hypothesized, the experimental induction of a concrete, process-focused (or non-ruminative) thinking style (n=102) led to less OGMs as compared to the experimental induction of an abstract, evaluative (or ruminative) thinking style (n=93). The present results add to the accumulating body of evidence that abstract, evaluative (or ruminative) thinking is a crucial underlying process of OGM, and expand prior literature by extending this idea to nonclinical individuals and by using a new procedure to assess OGM.


Whole-Genome Sequence of Mycoplasma bovis Strain Ningxia-1.
A genome sequence of the Mycoplasma bovis Ningxia-1 strain was tested by Pacific Biosciences (PacBio) single-molecule real-time (SMRT) sequencing technology. The strain was isolated from a lesioned calf lung in 2013 in Pengyang, Ningxia, China. The single circular chromosome of 1,033,629 bp shows differences between complete Mycoplasma bovis genome in insertion-like sequences (ISs), integrative conjugative elements (ICEs), lipoproteins (LPs), variable surface lipoproteins (VSPs), pathogenicity islands (PAIs), etc.


Distinct subcellular localisations of the putative inositol 1,3,4,5-tetrakisphosphate receptors GAP1IP4BP and GAP1m result from the GAP1IP4BP PH domain directing plasma membrane targeting.
Inositol 1,3,4,5-tetrakisphosphate (IP4), is a ubiquitous inositol phosphate that has been suggested to function as a second messenger. Recently, we purified and cloned a putative IP4 receptor, termed GAP1(IP4BP)[1], which is also a member of the GAP1 family of GTPase-activating proteins for the Ras family of GTPases. A homologue of GAP1(IP4BP), called GAP1(m), has been identified [2] and here we describe the cloning of a GAP1(m) cDNA from a human circulating-blood cDNA library. We found that a deletion mutant of GAP1(m), in which the putative phospholipid-binding domains (C2A and C2B) have been removed, binds to IP4 with a similar affinity and specificity to that of the corresponding GAP1(IP4BP) mutant. Expression studies of the proteins in either COS-7 or HeLa cells showed that, whereas GAP1(IP4BP) is located solely at the plasma membrane, GAP1(m) seems to have a distinct perinuclear localisation. By mutational analysis, we have shown that the contrast in subcellular distribution of these two closely related proteins may be a function of their respective pleckstrin homology (PH) domains. This difference in localisation has fundamental significance for our understanding of the second messenger functions of IP4.


Increasing n-butanol production with Saccharomyces cerevisiae by optimizing acetyl-CoA synthesis, NADH levels and trans-2-enoyl-CoA reductase expression.
n-Butanol can serve as an excellent gasoline substitute. Naturally, it is produced by some Clostridia species which, however, exhibit only limited suitability for industrial n-butanol production. The yeast Saccharomyces cerevisiae would be an ideal host due to its high robustness in fermentation processes. Nevertheless, n-butanol yields and titers obtained so far with genetically engineered yeast strains are only low. In our recent work, we showed that n-butanol production via a clostridial acetoacetyl-CoA-derived pathway in engineered yeast was limited by the availability of coenzyme A (CoA) and cytosolic acetyl-CoA. Increasing their levels resulted in a strain producing up to 130 mg/L n-butanol under anaerobic conditions. Here, we show that under aerobic conditions. this strain can even produce up to 235 mg/L n-butanol probably due to a more efficient NADH re-oxidation. Nevertheless, expression of a bacterial water-forming NADH oxidase (nox) significantly reduced n-butanol production although it showed a positive effect on growth and glucose consumption. Screening for an improved version of an acetyl-CoA forming NAD+-dependent acetylating acetaldehyde dehydrogenase, adhEA267T/E568K/R577S, and its integration into n-butanol-producing strain further improved n-butanol production. Moreover, deletion of the competing NADP+-dependent acetaldehyde dehydrogenase Ald6 had a superior effect on n-butanol formation. To increase the endogenous supply of CoA, amine oxidase Fms1 was overexpressed together with pantothenate kinase coaA from Escherichia coli, and could completely compensate the beneficial effect on n-butanol synthesis of addition of pantothenate to the medium. By overexpression of each of the enzymes of n-butanol pathway in the n-butanol-producing yeast strain, it turned out that trans-2-enoyl-CoA reductase (ter) was limiting n-butanol production. Additional overexpression of ter finally resulted in a yeast strain producing n-butanol up to a titer of 0.86 g/L and a yield of 0.071 g/g glucose. By further optimizing substrate supply and redox power in the form of coenzyme A, acetyl-CoA and NADH, n-butanol production with engineered yeast cells could be improved to levels never reached before with S. cerevisiae via an acetoacetyl-CoA-derived pathway in synthetic medium. Moreover, our results indicate that the NAD+/NADH redox balance and the trans-2-enoyl-CoA reductase reaction seem to be bottlenecks for n-butanol production with yeast.


N-of-1 Randomized Trials of Ultra-Micronized Palmitoylethanolamide in Older Patients with Chronic Pain.
Ultra-micronized palmitoylethanolamide (um-PEA) represents an attractive option for chronic pain control in complex older patients at higher risk of adverse effects with traditional analgesics. The aim of this study was to determine the effectiveness of um-PEA versus placebo on chronic pain intensity and function in individual geriatric patients. We performed randomized, blinded N-of-1 trials with two 3-week um-PEA versus placebo comparisons, separated by 2-week washout periods. The study included outpatients aged ≥ 65 years with chronic, non-cancer, non-ischemic pain in the back, joints, or limbs. Patients were randomized to Um-PEA 600 mg or placebo twice daily. Pain intensity was measured using an 11-point visual numeric scale. Functional impairment was measured using a Back Pain Functional Scale. Impact of each N-of-1 trial was measured on the clinician's intention to treat and confidence. Ten of 11 eligible patients consented over 7 months [all female, mean age 83.2 years (SD 4.6)]. Three patients interrupted the trial: one had diarrhea (under placebo), one for low adherence, and one for intercurrent pneumonia. A small statistically significant effect in favor of um-PEA was seen at the mixed method analyses in two patients (effect size equal to 8% of the baseline pain). A statistically significant impact on function was found in one patient. After the trial, um-PEA was prescribed to four patients; in two patients the clinician changed their pre-trial intention to treat; the clinician confidence in the treatment plan either increased (5) or remained the same (2). Our experience confirmed that N-of-1 trials may help make personalized evidence-based decisions in complex older patients, with special feasibility considerations. CLINICALTRIALS.GOV: NCT02699281.


Microbial Communities' Characterization in Urban Recreational Surface Waters Using Next Generation Sequencing.
Microbial communities in surface waters used for recreational purposes are indicators of contamination and risk of contact with human pathogens. Hence, monitoring microbial communities in recreational waters is important for potential public health threats to humans. Such monitoring is rare in Colombia, even in its capital, Bogotá, the most populous city in the country. This city encompasses metropolitan and linear parks with recreational water bodies that are used frequently by the public, and the presence of pathogens can compromise the health of the citizens. Therefore, we examined the bacterial, and eukaryotic communities in urban recreational lakes (URL) in four metropolitan parks in Bogotá, Colombia. Samples from four metropolitan parks (Los Novios, Simon Bolivar, El Tunal, and Timiza) and one stream contaminated with sewage from a linear park (El Virrey) were collected. We used amplicon next-generation sequencing of the 16S-rRNA gene and 18S-rRNA gene to characterize microbial communities followed by bioinformatics analyses. In addition, general water quality parameters-pH, hardness, acidity, alkalinity, dissolved oxygen, and nitrites-were recorded using a commercial kit. Genera of pathogens, including Legionella, Pseudomonas, Mycobacterium, Candida, and Naegleria, were found in lake waters. The stream El Virrey was, however, the only surface water that showed an abundance of fecal bacteria, often associated with low oxygen concentrations. All water bodies showed a predominance of fungal phyla, except for the lake at Timiza. This lake showed the highest pH, and its ecological dynamics are likely different from other water bodies. Likewise, some URLs displayed a greater abundance of cyanobacteria, including toxin-producing species. Algal genera associated with eutrophication were predominant among primary producing microorganisms. This study shows for the first time the description of the bacterial and eukaryotic communities of some URLs and a stream in Bogotá. The URLs and the stream harbored various pathogens that might pose a risk to the citizen's health.


Content-balancing strategy in bifactor computerized adaptive patient-reported outcome measurement.
Most multidimensional patient-reported outcomes (PRO) measures are lengthy to complete. Computerized adaptive testing (CAT) that selects the most informative items can potentially reduce respondent burden without sacrificing measurement accuracy. The commonly used maximum Fisher information item selection method has been reported to lead to highly unbalanced item bank usage and potentially imprecise trait estimation. This study employs the content-balancing strategy in a bifactor-modeled CAT item selection and examines its impact on measurement accuracy and item bank usage. Item responses from a population-based SF-36 survey were first calibrated using the bifactor graded response model. Four post hoc CATs using items and responses from the SF-36 data set were then created. The content-balancing strategy was adopted in the item selection procedure of the bifactor-modeled CAT. The measurement accuracy and usage of items of the CAT were compared between the tests with and without the content-balancing strategy. The results indicate that the CAT implemented with the content-balancing strategy offers a better overall measurement accuracy of both the general health status and the two health domains (physical and mental) of the SF-36. The content-balancing strategy helps the CAT-PRO to balance the selection of items and achieve improved measurement accuracy. Its implementation in real-time CAT administration to measure multidimensional PRO traits merits further studies.


Membrane potential of Plasmodium-infected erythrocytes.
The membrane potential (Em) of normal and Plasmodium chabaudi-infected rat erythrocytes was determined from the transmembrane distributions of the lipophilic anion, thiocyanate (SCN), and cation, triphenylmethylphosphonium (TPMP). The SCN- and TPMP-measured Em of normal erythrocytes are -6.5 +/- 3 mV and -10 +/- 4 mV, respectively. The TPMP-measured Em of infected cells depended on parasite developmental stage; "late" stages (schizonts and gametocytes) were characterized by a Em = -35 mV "early stages (ring and copurifying noninfected) by a low Em (-16 mV). The SCN-determined Em of infected cells was -7 mV regardless of parasite stage. Studies with different metabolic inhibitors including antimycin A, a proton ionophore (carbonylcyanide m-chlorophenylhydrazone [CCCP] ), and a H+ -ATPase inhibitor (N,N'-dicyclohexylcarbodiimide, [DCCD] ) indicate that SCN monitors the Em across the erythrocyte membrane of infected and normal cells whereas TPMP accumulation reflects the Em across the plasma membranes of both erythrocyte and parasite. These inhibitor studies also implicated proton fluxes in Em-generation of parasitized cells. Experiments with weak acids and bases to measure intracellular pH further support this proposal. Methylamine distribution and direct pH measurement after saponin lysis of erythrocyte membranes demonstrated an acidic pH for the erythrocyte matrix of infected cells. The transmembrane distributions of weak acids (acetate and 5,5-dimethyloxazolidine-2,4-dione) indicated a DCCD-sensitive alkaline compartment. The combined results suggest that the intraerythrocyte parasite Em and delta pH are in part the consequence of an electrogenic proton pump localized to the parasite plasma membrane.


Predicting radiographic hip osteoarthritis from range of movement.
The primary objective was to test the hypothesis that new attenders in primary care with hip pain and radiographic osteoarthritis (OA) have a decreased range of movement compared with those without OA. The secondary objective was to define the planes of movement and thresholds that were the most discriminatory for OA. Men and women aged 40 yr and over presenting with a new episode of hip pain were recruited from 36 general practices across the UK. A standardized radiographic and clinical examination was performed. The discriminating ability of the range of movement for each plane to identify those with radiographic OA was assessed using receiver operating characteristic curves. New hip pain attenders with radiographic OA had restricted movement at the hip compared with those without radiographic change. Restriction in internal rotation was the most predictive and flexion the least predictive of radiographic OA. At this cut-off, restriction in any single plane had a sensitivity of 86% for moderate and 100% for severe OA (specificity was 54 and 42% respectively). Restriction in all three planes had greater discrimination (sensitivity was 33% for mild to moderate OA and 54% for severe OA; specificity was 93 and 88% respectively). Restriction in range of movement was predictive of the presence of OA in these new presenters to primary care with hip pain, and the results of this examination could be used to inform decisions regarding radiography.


Work-related traumatic dental injuries: Prevalence, characteristics and risk factors.
The prevalence of work-related oral trauma is underestimated because minor dental injuries are often not reported in patients with several injuries in different parts of the body. In addition, little data are available regarding their characteristics. The aim of this epidemiological study was to determine the prevalence, types, and characteristics of occupational traumatic dental injuries (TDIs) in a large working community. Work-related TDIs that occurred during the period between 2011 and 2013 in the District of Genoa (Northwest of Italy, 0.86 million inhabitants) were analyzed. Patients' data were obtained from the National Institute for Insurance against Accidents at Work database. During the 2 year period, 112 TDIs (345 traumatized teeth) were recorded. The prevalence was 5.6‰ of the total amount of occupational trauma. The highest prevalence was found in the fourth and fifth decades of life (OR=3.6, P < .001), and males were injured more often than females (70.5% vs 29.5%, OR=2.8, P < .001). Service and office workers represented 52% of the sample, and construction/farm/factory workers and craftsmen were 48%. TDIs involved only teeth and surrounding tissue in 66% of cases, or in combination with another maxillofacial injury in 34%. They were statistically associated with construction/farm/factory workers group (Chi squared P < .01). Crown fracture was recorded in 34.5% of cases, subluxation/luxation in 10.7%, avulsion in 9%, root fracture in 3.8%, and concussion in 3.5%. Thirty-two subjects (28.6%, 133 teeth, OR=4.3, P < .001) presented at least 1 traumatized tooth with previous dental treatment. Among 212 (61.4%) traumatized teeth, 67.5% were upper incisors, 17.5% were lower incisors, 3.3% were upper canines, 1.9% were lower canines, and 9.9% were bicuspids and molars. Work-related TDIs had a low overall prevalence, and fractures were the most frequent dental injury. Age, gender, and preexisting dental treatments represented risk factors for work-related TDIs.


Outcomes of tamoxifen chemoprevention for breast cancer in very high-risk women: a cost-effectiveness analysis.
To estimate the effects on survival, quality-adjusted survival, and health care costs of using tamoxifen for primary prevention in subgroups of women at very high risk for breast cancer. A decision analysis was performed using a hypothetical cohort of women that included subgroups with atypical hyperplasia, Gail risk greater than 5, lobular carcinoma-in-situ, or two or more first-degree relatives with breast cancer. Data sources were the Breast Cancer Prevention Trial, the Surveillance, Epidemiology, and End-Results program, time trade-off preference ratings, the Group Health Cooperative of Puget Sound, and the United States Health Care Financing Administration. Our model predicted that tamoxifen would prolong the average survival of cohort members initiating use at ages 35, 50, and 60 years by 70, 42, and 27 days, respectively. It would prolong survival even more for those in the higher-risk groups, especially those with atypical hyperplasia (202, 89, and 45 days). Tamoxifen use was also projected to extend quality-adjusted survival by 158, 80, and 50 days in the atypical hyperplasia group. For younger women in the highest risk groups, chemoprevention with tamoxifen was estimated to have cost savings or be cost-effective, both with and without quality adjustments. Chemoprevention with tamoxifen may be particularly beneficial to women with atypical hyperplasia, 5-year Gail model risk greater than 5%, lobular carcinoma-in-situ, or two or more first-degree relatives with breast cancer. The benefits may be greater if tamoxifen is initiated before age 50 years rather than after and if the breast cancer risk reduction conferred by tamoxifen lasts longer than 5 years. For women with a very high risk of invasive breast cancer, chemoprevention with tamoxifen seems to be cost-effective.