Ambulatory end-stage liver disease in Ghana; patient profile and utility of alpha fetoprotein and aspartate aminotransferase: platelet ratio index.
End-stage liver disease (ESLD) is a major burden on public health, particularly in sub-Saharan Africa, where hepatitis B virus (HBV) is an important risk factor. We aimed to describe clinical characteristics of ESLD from cirrhosis or hepatocellular carcinoma (HCC) and the performance of aspartate aminotransferase (AST)-platelet ratio index (APRI) and alpha fetoprotein (AFP) in Ghana. We performed an observational cross-sectional study in outpatient hepatology clinics at three teaching hospitals in Ghana, West Africa. One hundred and forty-one HCC, 216 cirrhosis and 218 chronic HBV patients were recruited by convenience sampling. Sociodemographic, history and examination, laboratory, and disease staging information were shown using descriptive statistics. Performance of the APRI score in diagnosis of cirrhosis and AFP in the diagnosis of HCC was determined using AUROC analysis. Median age at presentation was 44 years for HCC and 46 years for cirrhosis. HBV was found in 69.5% of HCC and 47.2% of cirrhosis cases, and HCV in 6.4% and 3.7% respectively. APRI cut-off of 2 had sensitivity of 45.4% and specificity of 95% in diagnosis of cirrhosis, and cut-off of 1 had sensitivity of 75.9% and specificity of 89%. AUC of AFP was 0.88 (95% CI 0.81-0.94) in diagnosis of HCC. Low monthly income was associated with lower odds of undertaking AFP. Thirty one percent of cirrhotic persons were Child-Pugh C, and 67.9% of HCC patients had advanced or terminal disease at presentation. Our findings emphasize the young age of ESLD patients in Ghana and the advanced nature at presentation. It highlights shortcomings in surveillance and the need for policies to address the burden and improve outcomes in Ghana.


Acute surgical treatment of perforated peptic ulcer in the elderly patients.
Emergency abdominal surgery is associated with high morbidity and mortality rates, especially in the elderly patients, but prompt diagnoses and treatment should not be delayed. We conducted a retrospective review (1) to identify risk factors for morbidity and mortality among elderly patients admitted for emergent surgery of perforated peptic ulcers; and (2) to determine whether there were any differences between those who are 70-79 years old and those 80 years old and older. 94 patients who were older than 70 years old and underwent emergency surgery for perforated peptic ulcer between 2000 and 2004 in our institution were reviewed retrospectively. The following variables were followed: age, sex, comorbidity, previous medications, time from onset of symptoms/signs to surgery, time from arrival in emergent room to surgery, perioperative risks, operative findings, type of operation, morbidity, mortality and length of hospital stay. The age, morbidity, mortality and the length of intensive care unit stay were increased in Group 2 (>80 yrs) than Group 1 (70 to 79 yrs), but they did not achieve significant differences statistically. Time from symptoms/signs to emergency room over 24 hours, American Society of Anesthesiologist grade over IV and limited procedure showed significant contributions to postoperative morbidity on univariate analysis. Comorbidity, time from emergency room to operation room over 12 hours, American Society of Anesthesiologists grade over IV, peri-operative blood transfusion, postoperative morbidity and duration of ICU stays over 5 days were significant factors contributed to mortality on univariate analysis. Further analysis showed comorbidity, peri-operative blood transfusion, and postoperative morbidity were independent and predictive factors of mortality on multivariate model. Although perforated peptic ulcer in the elderly patients is associated with high morbidity and mortality, we should not delay the surgical intervention for patients with advanced age. Timely diagnosis and early surgical management of perforated peptic ulcers was imperative for elderly patients. The abdominal computer tomography was recommended in elderly patients for who had vague and atypical clinical symptoms/signs of perforated peptic ulcer. In addition, more attention should be paid to patients with preoperative comorbidities, peri-operative blood transfusion and post-operative morbidity for which were associated with high post-operative mortality.


Association of the PDYN gene with alcohol dependence and the propensity to drink in negative emotional states.
Synthetic κ-opioid receptor (KOR) agonists induce dysphoric and pro-depressive effects and variations in the KOR (OPRK1) and prodynorphin (PDYN) genes have been shown to be associated with alcohol dependence. We genotyped 23 single nucleotide polymorphisms (SNPs) in the PDYN and OPRK1 genes in 816 alcohol-dependent subjects and investigated their association with: (1) negative craving measured by a subscale of the Inventory of Drug Taking Situations; (2) a self-reported history of depression; (3) the intensity of depressive symptoms measured by the Beck Depression Inventory-II. In addition, 13 of the 23 PDYN and OPRK1 SNPs, which were previously genotyped in a set of 1248 controls, were used to evaluate association with alcohol dependence. SNP and haplotype tests of association were performed. Analysis of a haplotype spanning the PDYN gene (rs6045784, rs910080, rs2235751, rs2281285) revealed significant association with alcohol dependence (p = 0.00079) and with negative craving (p = 0.0499). A candidate haplotype containing the PDYN rs2281285-rs1997794 SNPs that was previously associated with alcohol dependence was also associated with negative craving (p = 0.024) and alcohol dependence (p = 0.0008) in this study. A trend for association between depression severity and PDYN variation was detected. No associations of OPRK1 gene variation with alcohol dependence or other studied phenotypes were found. These findings support the hypothesis that sequence variation in the PDYN gene contributes to both alcohol dependence and the induction of negative craving in alcohol-dependent subjects.


Fetal Congenital Heart Disease Echocardiogram Screening Based on DGACNN: Adversarial One-Class Classification Combined with Video Transfer Learning.
Fetal congenital heart disease (FHD) is a common and serious congenital malformation in children. In Asia, FHD birth defect rates have reached as high as 9.3%. For the early detection of birth defects and mortality, echocardiography remains the most effective method for screening fetal heart malformations. However, standard echocardiograms of the fetal heart, especially four-chamber view images, are difficult to obtain. In addition, the pathophysiological changes in fetal hearts during different pregnancy periods lead to ever-changing two-dimensional fetal heart structures and hemodynamics, and it requires extensive professional knowledge to recognize and judge disease development. Thus, research on the automatic screening for FHD is necessary. In this paper, we proposed a new model named DGACNN that shows the best performance in recognizing FHD, achieving a rate of 85%. The motivation for this network is to deal with the problem that there are insufficient training datasets to train a robust model. There are many unlabeled video slices, but they are tough and time-consuming to annotate. Thus, how to use these un-annotated video slices to improve the DGACNN capability for recognizing FHD, in terms of both recognition accuracy and robustness, is very meaningful for FHD screening. The architecture of DGACNN comprises two parts, that is, DANomaly and GACNN (Wgan-GP and CNN). DANomaly, similar to the ALOCC network, but incorporates cycle adversarial learning to train an end-to-end one-class classification (OCC) network that is more robust and has a higher accuracy than ALOCC in screening video slices. For the GACNN architecture, we use FCH (four chamber heart) video slices at around the end-systole, as screened by DANomaly, to train a WGAN-GP for the purpose of obtaining ideal low-level features that can robustly improve the FHD recognition accuracy. A few annotated video slices, as screened by DANomaly, can also be used for data augmentation so as to improve the FHD recognition further. The experiments show that the DGACNN outperforms other state-of-the-art networks by 1%-20% in recognizing FHD. A comparison experiment shows that the proposed network already outperforms the performance of expert cardiologists in recognizing FHD, reaching 84% in a test. Thus, the proposed architecture has high potential for helping cardiologists complete early FHD screenings.


Randomized phase III intergroup trial of isotretinoin to prevent second primary tumors in stage I non-small-cell lung cancer.
Promising data have suggested that retinoid chemoprevention may help to control second primary tumors (SPTs), recurrence, and mortality of stage I non-small-cell lung cancer (NSCLC) patients. We carried out a National Cancer Institute (NCI) Intergroup phase III trial (NCI #I91-0001) with 1166 patients with pathologic stage I NSCLC (6 weeks to 3 years from definitive resection and no prior radiotherapy or chemotherapy). Patients were randomly assigned to receive a placebo or the retinoid isotretinoin (30 mg/day) for 3 years in a double-blind fashion. Patients were stratified at randomization by tumor stage, histology, and smoking status. The primary endpoint (time to SPT) and the secondary endpoints (times to recurrence and death) were analyzed by log-rank test and the Cox proportional hazards model. All statistical tests were two-sided. After a median follow-up of 3.5 years, there were no statistically significant differences between the placebo and isotretinoin arms with respect to the time to SPTs, recurrences, or mortality. The unadjusted hazard ratio (HR) of isotretinoin versus placebo was 1.08 (95% confidence interval [CI] = 0.78 to 1.49) for SPTs, 0.99 (95% CI = 0.76 to 1.29) for recurrence, and 1.07 (95% CI = 0.84 to 1.35) for mortality. Multivariate analyses showed that the rate of SPTs was not affected by any stratification factor. Rate of recurrence was affected by tumor stage (HR for T(2) versus T(1) = 1.77 [95% CI = 1.35 to 2.31]) and a treatment-by-smoking interaction (HR for treatment-by-current-versus-never-smoking status = 3.11 [95% CI = 1.00 to 9.71]). Mortality was affected by tumor stage (HR for T(2) versus T(1) = 1.39 [95% CI = 1.10 to 1.77]), histology (HR for squamous versus nonsquamous = 1.31 [95% CI = 1.03 to 1.68]), and a treatment-by-smoking interaction (HR for treatment-by-current-versus-never-smoking = 4.39 [95% CI = 1.11 to 17.29]). Mucocutaneous toxicity (P<.001) and noncompliance (40% versus 25% at 3 years) were higher in the isotretinoin arm than in the placebo arm. Isotretinoin treatment did not improve the overall rates of SPTs, recurrences, or mortality in stage I NSCLC. Secondary multivariate and subset analyses suggested that isotretinoin was harmful in current smokers and beneficial in never smokers.


Air pollutant emissions from the development, production, and processing of Marcellus Shale natural gas.
The Marcellus Shale is one of the largest natural gas reserves in the United States; it has recently been the focus of intense drilling and leasing activity. This paper describes an air emissions inventory for the development, production, and processing of natural gas in the Marcellus Shale region for 2009 and 2020. It includes estimates of the emissions of oxides of nitrogen (NOx), volatile organic compounds (VOCs), and primary fine particulate matter (< or = 2.5 microm aerodynamic diameter; PM2.5) from major activities such as drilling, hydraulic fracturing, compressor stations, and completion venting. The inventory is constructed using a process-level approach; a Monte Carlo analysis is used to explicitly account for the uncertainty. Emissions were estimated for 2009 and projected to 2020, accounting for the effects of existing and potential additional regulations. In 2020, Marcellus activities are predicted to contribute 6-18% (95% confidence interval) of the NOx emissions in the Marcellus region, with an average contribution of 12% (129 tons/day). In 2020, the predicted contribution of Marcellus activities to the regional anthropogenic VOC emissions ranged between 7% and 28% (95% confidence interval), with an average contribution of 12% (100 tons/day). These estimates account for the implementation of recently promulgated regulations such as the Tier 4 off-road diesel engine regulation and the US. Environmental Protection Agency's (EPA) Oil and Gas Rule. These regulations significantly reduce the Marcellus VOC and NOx emissions, but there are significant opportunities for further reduction in these emissions using existing technologies. The Marcellus Shale is one of the largest natural gas reserves in United States. The development and production of this gas may emit substantial amounts of oxides of nitrogen and volatile organic compounds. These emissions may have special significance because Marcellus development is occurring close to areas that have been designated nonattainment for the ozone standard. Control technologies exist to substantially reduce these impacts. PM2.5 emissions are predicted to be negligible in a regional context, but elemental carbon emissions from diesel powered equipment may be important.


Diurnal variation of von Willebrand factor in plasma: the Bispebjerg study of diurnal variations.
Quantitation of von Willebrand factor (VWF) in plasma is a central element in assessing von Willebrand disease (VWD). VWF activity is known to vary, which has partly been ascribed to biological and preanalytical variation. However, a possible diurnal expression of VWF has not been thoroughly tested. We examined whether VWF antigen and VWF activity in plasma display a diurnal profile in healthy young males, and whether such variation is related to changes in release or elimination. Plasma from 20 healthy young males was collected at 9 time-points over 24 h (15 h of light and 9 h of darkness); the plasma concentration of melatonin was used as an internal control to confirm the normal 24-h rhythms of the individual participants. The data, analyzed by rhythmometric statistics, revealed a significant variation (P = 0.02) and total amplitude of 22.6% in VWF antigen concentrations over the 24-h period. A pronounced variation in VWF activity was also observed, although not significant according to the 24-h statistical model. To examine whether the diurnal pattern was related to changes in elimination or secretion, the ratio between (i) coagulation factor VIII and VWF and (ii) VWF propeptide and VWF was determined. Taken together, the data suggest changes in release and not in clearance. Diurnal variation in von Willebrand antigen and activity in plasma represents an important aspect of the biological variation. Standardized time-of-day plasma sampling for quantitation of VWF in VWD patients seems warranted.


Personal health records in the preclinical medical curriculum: modeling student responses in a simple educational environment utilizing Google Health.
Various problems concerning the introduction of personal health records in everyday healthcare practice are reported to be associated with physicians' unfamiliarity with systematic means of electronically collecting health information about their patients (e.g. electronic health records--EHRs). Such barriers may further prevent the role physicians have in their patient encounters and the influence they can have in accelerating and diffusing personal health records (PHRs) to the patient community. One way to address these problems is through medical education on PHRs in the context of EHR activities within the undergraduate medical curriculum and the medical informatics courses in specific. In this paper, the development of an educational PHR activity based on Google Health is reported. Moreover, student responses on PHR's use and utility are collected and presented. The collected responses are then modelled to relate the satisfaction level of students in such a setting to the estimation about their attitude towards PHRs in the future. The study was conducted by designing an educational scenario about PHRs, which consisted of student instruction on Google Health as a model PHR and followed the guidelines of a protocol that was constructed for this purpose. This scenario was applied to a sample of 338 first-year undergraduate medical students. A questionnaire was distributed to each one of them in order to obtain Likert-like scale data on the sample's response with respect to the PHR that was used; the data were then further analysed descriptively and in terms of a regression analysis to model hypothesised correlations. Students displayed, in general, satisfaction about the core PHR functions they used and they were optimistic about using them in the future, as they evaluated quite high up the level of their utility. The aspect they valued most in the PHR was its main role as a record-keeping tool, while their main concern was related to the negative effect their own opinion might have on the use of PHRs by patients. Finally, the estimate of their future attitudes towards PHR integration was found positively dependent of the level of PHR satisfaction that they gained through their experience (rho=0.524, p<0.001). The results indicate that students support PHRs as medical record keeping helpers and perceive them as beneficial to healthcare. They also underline the importance of achieving good educational experiences in improving PHR perspectives inside such educational activities. Further research is obviously needed to establish the relative long-term effect of education to other methods of exposing future physicians to PHRs.


Cutaneous tissue damage induces long-lasting nociceptive sensitization and regulation of cellular stress- and nerve injury-associated genes in sensory neurons.
Tissue damage is one of the major etiological factors in the emergence of chronic/persistent pain, although mechanisms remain enigmatic. Using incision of the back skin of adult rats as a model for tissue damage, we observed sensitization in a nociceptive reflex enduring to 28days post-incision (DPI). To determine if the enduring behavioral changes corresponded with a long-term impact of tissue damage on sensory neurons, we examined the temporal expression profile of injury-regulated genes and the electrophysiological properties of traced dorsal root ganglion (DRG) sensory neurons. The mRNA for the injury/stress-hub gene Activating Transcription Factor 3 (ATF3) was upregulated and peaked within 4 DPI, after which levels declined but remained significantly elevated out to 28 DPI, a time when the initial incision appears healed and tissue-inflammation largely resolved. Accordingly, stereological image analysis indicated that some neurons expressed ATF3 only transiently (mostly medium-large neurons), while in others it was sustained (mostly small neurons), suggesting cell-type-specific responses. In retrogradely-traced ATF3-expressing neurons, Calcium/calmodulin-dependent protein kinase type IV (CAMK4) protein levels and isolectin-B4 (IB4)-binding were suppressed whereas Growth Associated Protein-43 (GAP-43) and Neuropeptide Y (NPY) protein levels were enhanced. Electrophysiological recordings from DiI-traced sensory neurons 28 DPI showed a significant sensitization limited to ATF3-expressing neurons. Thus, ATF3 expression is revealed as a strong predictor of single cells displaying enduring pain-related electrophysiological properties. The cellular injury/stress response induced in sensory neurons by tissue damage and indicated by ATF3 expression is positioned to contribute to pain which can occur after tissue damage.


TIM-1 Ubiquitination Mediates Dengue Virus Entry.
Dengue virus (DENV) is a major human pathogen causing millions of infections yearly. Despite intensive investigations, a DENV receptor that directly participates in virus internalization has not yet been characterized. Here, we report that the phosphatidylserine receptor TIM-1 is an authentic DENV entry receptor that plays an active role in virus endocytosis. Genetic ablation of TIM-1 strongly impaired DENV infection. Total internal reflection fluorescence microscopy analyses of live infected cells show that TIM-1 is mostly confined in clathrin-coated pits and is co-internalized with DENV during viral entry. TIM-1 is ubiquitinated at two lysine residues of its cytoplasmic domain, and this modification is required for DENV endocytosis. Furthermore, STAM-1, a component of the ESCRT-0 complex involved in intracellular trafficking of ubiquitinated cargos, interacts with TIM-1 and is required for DENV infection. Overall, our results show that TIM-1 is the first bona fide receptor identified for DENV.


MedCo2: Privacy-Preserving Cohort Exploration and Analysis.
Medical studies are usually time consuming, cumbersome and extremely costly to perform, and for exploratory research, their results are also difficult to predict a priori. This is particularly the case for rare diseases, for which finding enough patients is difficult and usually requires an international-scale research. In this case, the process can be even more difficult due to the heterogeneity of data-protection regulations, making the data sharing process particularly hard. In this short paper, we propose MedCo2 (pronounced MedCo square), a distributed system that streamlines the process of a medical study by bridging and enabling both data discovery and data analysis among multiple databases, while protecting data confidentiality and patients' privacy. MedCo2 relies on interactive protocols, homomorphic encryption and differential privacy. It enables the privacy-preserving computations of multiple statistics such as cosine similarity and variance, and the training of machine learning models, on patients that are obliviously selected according to specific criteria among multiple databases.


Genetic screen of a mutant poxvirus library identifies an ankyrin repeat protein involved in blocking induction of avian type I interferon.
Mammalian poxviruses, including vaccinia virus (VACV), have evolved multiple mechanisms to evade the host type I interferon (IFN) responses at different levels, with viral proteins targeting IFN induction, signaling, and antiviral effector functions. Avian poxviruses (avipoxviruses), which have been developed as recombinant vaccine vectors for permissive (i.e., poultry) and nonpermissive (i.e., mammals, including humans) species, encode no obvious equivalents of any of these proteins. We show that fowlpox virus (FWPV) fails to induce chicken beta IFN (ChIFN2) and is able to block its induction by transfected poly(I·C), an analog of cytoplasmic double-stranded RNA (dsRNA). A broad-scale loss-of-function genetic screen was used to find FWPV-encoded modulators of poly(I·C)-mediated ChIFN2 induction. It identified fpv012, a member of a family of poxvirus genes highly expanded in the avipoxviruses (31 in FWPV; 51 in canarypox virus [CNPV], representing 15% of the total gene complement), encoding proteins containing N-terminal ankyrin repeats (ANKs) and C-terminal F-box-like motifs. Under ectopic expression, the first ANK of fpv012 is dispensable for inhibitory activity and the CNPV ortholog is also able to inhibit induction of ChIFN2. FWPV defective in fpv012 replicates well in culture and barely induces ChIFN2 during infection, suggesting that other factors are involved in blocking IFN induction and resisting the antiviral effectors. Nevertheless, unlike parental and revertant viruses, the mutants induce moderate levels of expression of interferon-stimulated genes (ISGs), suggesting either that there is sufficient ChIFN2 expression to partially induce the ISGs or the involvement of alternative, IFN-independent pathways that are also normally blocked by fpv012.


A Pilot Study of Aberrant CpG Island Hypermethylation of SPRED1 in Acute Myeloloid Leukemia.
Background: Epigenetic silencing of tumor suppressor genes plays important role in acute myeloid leukemia (AML). Recently, SPRED1, a negative regulator of the RAS MAPK pathway, is identified as a tumour suppressor downregulated in AML. However, little is known regarding its underlying dysregulation in AML. In this study, we investigated methylation status of SPRED1 promoters and their association with mRNA levels in AML. Methods: Methylation level were measured in four regions of SPRED1 (#1: 310 bp ~ 723 bp, #2: 810 bp ~ 1299 bp, #3: 1280 bp ~ 1742 bp and #4: 1715 bp ~ 2059 bp) in a total of 16 patients with de novonon-acute promyelocytic leukemia (non-APL) and three patients who got complete remission after induction treatment using the Sequenom MassARRAY platform. Quantitative real-time polymerase chain reaction (q-RT PCR) was used to analyze SPRED1 mRNA levels. Results: AML patients had a significantly higher average methylation level than controls at regions of #1_CpG_1 (p= 0.04) and #1_CpG_11 (p =0.002). The methylation values for #1_CpG_11 were negatively correlated with mRNA levels (r= -0.558, p=0.013) but there was no significant association between #1_CpG_1 methylation status and mRNA levels (r=-0.103, p=0.675) in AML patients. There was no significant difference in the methylation level when comparing with clinical biochemical parameters and treatment response (p>0.05). Mutations of epigenetic regulation genes such as DNMT3A, TET2 and IDH1/2 were most frequently observed in patients with higher methylation levels. Decreased methylation levels were revealed in three patients who got complete remission. Conclusions: Aberrant methylation statuses of the SPRED1 promoter regions are associated with the downregulation of gene transcription in AML. The methylation level is probably associated with the treatment response of AML. Mutations of epigenetic regulation genes might be involved in the epigenetic aberration of SPRED1.


Association of Urinary Tract Infection in Married Women Presenting with Urinary Incontinence in a Hospital based Population.
Urinary incontinence (UI) is increasingly recognized as a significant health problem, which remains a hygienic as well as social problem. Women have higher risk of developing incontinence in their lifetime compared with men. Urinary tract infection can increase the incidence of incontinence. Present study was undertaken to assess the association of UTI in married women who presented with UI. The present study was aimed to identify the patients (married women) with complaints of UI and determining its association with UTI; and to identify the causative organism for the UTI along with its antimicrobial susceptibility pattern. This is a cross-sectional, non-randomized study of 107 married women with UI, who attended outpatient department in our hospital. Mid-stream urine (MSU) samples were collected from these patients with positive history of incontinence. Screening of urine for significant bacteriuria and culture to identify the etiological agents were performed followed by evaluation of their antimicrobial susceptibility profiles using Kirby Bauer disc diffusion method. Overall 25.2% of patients with incontinence had a positive urine culture. History of UTI was elicited in around 38.3% of patients, among which 15% had positive urine culture and 10.3% of the patients who did not have a history had positive culture. Escherichia coli was the commonest causative organism (66.6) causing UTI, followed by Enterococcus spp. (22.3%), Klebsiella pneumoniae (7.4%) and Proteus mirabilis (3.7%). The antimicrobial susceptibility pattern for Escherichia coli showed high sensitivity to Nitrofurantoin (94.4%) and high resistance to Ampicillin (94.4%). Our study revealed one in every four incontinent patients had UTI and almost half of them suffered from previous episodes of UTI. Thus appropriate correction of the existing UTI can help in the treatment of UI.


[Changes of physiological functions in rats induced by immobilization stress].
A study was conducted on the changes of physiological function in rats due to immobilization stress. Male Fischer rats (SPF) of 32 weeks of age were housed in individual cages for 4 weeks. Then all rats were immobilized by stainless wire mesh for 6 hours daily for 3 days. Blood was collected before the 1st stress, immediately after the 1st stress, immediately after the 3rd stress and the day after the 3rd stress. The results of this experiment were as follows: (1) The total leukocyte counts in the blood of the rats after the 1st trial was significantly higher than that before the 1st trial. (2) The percentage of lymphocytes in the blood after the 1st trial was significantly lower than that before the 1st trial, whereas that of neutrophils was significantly higher. (3) Correlations between phagocytic activity and superoxide production of neutrophils by histochemical NBT reduction assay showed significantly a positive correlation before the 1st trial. However, no significant correlations were observed in immediately after the 1st trial and the 3rd trial. The day after the 3rd trial, a positive correlation was observed again. These correlations showed that an unsuitable state of the neutrophil function was induced by the immobilization stress. (4) Serum biochemical profiles were affected by the immobilization stress. Also, GOT, GPT, LDH, CK and UA were increased after the 1st trial, whereas, TG, TP, ALB and ALP were decreased after the 1st trial. T-CHO was increased only immediately after the 3rd stress. These results suggest that immobilization stress affected blood cells and serum components, and then the host defense and physiological functions were damaged respectively.


Chemopreventive effects of sandalwood oil on skin papillomas in mice.
The essential oil, emulsion or paste of sandalwood (Santalum album L) has been used in India as an ayurvedic medicinal agent for the treatment of inflammatory and eruptive skin diseases. In this investigation, the chemopreventive effects of sandalwood oil (5% in acetone, w/v) on 7,12-dimethylbenz(a)anthracene-(DMBA)-initiated and 12-O-tetradecanoyl phorbol-13-acetate(TPA)-promoted skin papillomas, and TPA-induced ornithine decarboxylase (ODC) activity in CD1 mice were studied. Sandalwood oil treatment significantly decreased papilloma incidence by 67%, multiplicity by 96%, and TPA-induced ODC activity by 70%. This oil could be an effective chemopreventive agent against skin cancer.


Prevalence and Predictors of Driving after Prescription Opioid Use in an Adult ED Sample.
Prescription opioid use and driving is a public health concern given the risks associated with drugged driving, but the issue remains under-studied. We examined the prevalence and correlates of driving after taking prescription opioids (DAPO) among adults seeking emergency department (ED) treatment. Participants (aged 25-60) seeking ED care at a Level I trauma center completed a computerized survey. Validated instruments measured prescription opioid use, driving behaviors, and risky driving. Patients who reported past three-month prescription opioid use and drove at least twice weekly were administered an extended study survey measuring DAPO, depression, pain, and substance use. Among participants completing the screening survey (n = 756; mean age = 42.8 [standard deviation {SD} =10.4]), 37.8% reported past three-month prescription opioid use (30.8% of whom used daily), and 14.7% reported past three-month DAPO. Of screened participants, 22.5% (n = 170) were eligible for the extended study survey. Unadjusted analyses demonstrated that participants reporting DAPO were more likely to use opioids daily (51.1% vs 15.9%) and had higher rates of opioid misuse (mean Current Opioid Misuse Measure score 3.4 [SD = 3.8] vs 1.1 [SD = 2.1]) chronic pain (80.7% vs 42.7%), and driving after marijuana or alcohol use (mean intoxicated driving score 2.1 [SD = 1.3] vs 0.3 [SD = 0.8]) compared to patients not reporting DAPO (all p<0.001). Adjusting for age, gender, employment, and insurance in a logistic regression model, participants reporting DAPO were more likely to report a chronic pain diagnosis (odds ratio [OR] = 3.77, 95% confidence interval [CI], 1.55-9.17), daily opioid use (OR = 3.81, 95% CI, 1.64-8.85), and higher levels of intoxicated driving (OR = 1.62, 95% CI, 1.07-2.45). Alcohol and marijuana use, depression, and opioid misuse were not associated with DAPO in adjusted analyses. Nearly one in six adult patients seeking ED care reported DAPO. The ED may be an important site for interventions addressing opioid-related drugged driving.


An alphaproteobacterium capable of both aerobic and anaerobic anoxygenic photosynthesis but incapable of photoautotrophy: Charonomicrobium ambiphototrophicum, gen. nov., sp. nov.
A facultatively aerobic deep brown coccoid to ovoid bacterium, strain EG17(T), was isolated from a saline effluent stream in the NaCl-dominated brine spring system known as East German Creek in the province of Manitoba, Canada. The strain produced BChl a incorporated into a functional reaction center and two light-harvesting complexes with absorption peaks at 802, 850, and 879 nm. EG17(T) is the first reported anoxygenic phototroph capable of photoheterotrophic growth under both oxic and anoxic conditions. It yielded proportionally the greatest aerobic photosynthetic biomass under oligotrophic conditions. The results of 16S rRNA gene sequence comparisons revealed that EG17(T) was related most closely to the aerobic anoxygenic phototrophs Roseibacterium elongatum (98.3%) and quite distantly to both Dinoroseobacter shibae (95.2%) and Roseicyclus mahoneyensis (94.7%). The DNA G + C content was 65.6 mol%. On the basis of the unique dual aerobic/anaerobic photosynthetic capability, the distinctive spectrophotometric absorption of the photosynthetic apparatus, diagnostic physiological and biochemical traits, and the moderate phylogenetic separation between EG17(T) and its nearest relatives, it is concluded that this microorganism should be classified as a novel genus and species, Charonomicrobium ambiphototrophicum gen. nov., sp. nov., with EG17(T) as the type strain.


Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP.
Elevation of intracellular calcium levels in the presence of normal androgen levels has been implicated in apoptotic prostate cell death. Since the androgen receptor (AR) plays a critical role in the regulation of growth and differentiation of the prostate, it was of interest to determine whether Ca2+ would affect the expression of androgen receptor messenger RNA (mRNA) and protein, thus affecting the ability of androgens to control prostate function. AR-positive human prostate cancer cells, LNCaP, were incubated with either the calcium ionophore A23187 or the intracellular endoplasmic reticulum Ca(2+)-ATPase inhibitor thapsigargin. Subsequently, AR mRNA and protein levels were assessed by Northern and Western blot analysis. Both A23187 and thapsigargin were found to down-regulate steady state AR mRNA levels in a time- and dose-dependent manner. AR mRNA began to decrease after 6-8 h of incubation with 10(-6) M A23187 or 10(-7) M thapsigargin, reaching a nadir at 16 and 10 h of incubation, respectively. In contrast, control mRNA (glyceraldehyde 3-phosphate dehydrogenase) did not change significantly during the treatments with either A23187 or thapsigargin. AR protein levels were found to be decreased after 12 h of incubation with either 10(-6) M A23187 or 10(-7) M thapsigargin. The decrease in AR mRNA and protein seemed to precede apoptosis, since neither A23187 (24 h) nor thapsigargin (30 h) was found to alter cell morphology within the treatment time. Cycloheximide and actinomycin D were unable to change the calcium-mediated decrease in AR mRNA, ruling out the necessity for de novo protein synthesis or a change in mRNA stability. Moreover, the decrease in AR mRNA induced by calcium does not seem to involve protein kinase C- or calmodulin-dependent pathways, since inhibitors of these cellular components had no effect. Nuclear run-on assays demonstrated little or no effects of either A23187 or thapsigargin treatment on AR gene transcription (8 h and 10 h). In conclusion, these studies show that intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.


Independent and combined association of muscle strength and cardiorespiratory fitness in youth with insulin resistance and β-cell function in young adulthood: the European Youth Heart Study.
To examine the independent and combined association of isometric muscle strength of the abdomen and back and cardiorespiratory fitness (CRF) in youth with indices of glucose metabolism in young adulthood among boys and girls from the European Youth Heart Study. We used data from a population-based prospective cohort study among youth followed up for up to 12 years (n = 317). In youth, maximal voluntary contractions during isometric back extension and abdominal flexion were determined using a strain-gauge dynamometer and CRF was obtained from a maximal cycle ergometer test. Insulin resistance (homeostasis model assessment of insulin resistance [HOMA-IR]) and β-cell function (homeostasis model assessment of β-cell function [HOMA-B]) were estimated from fasting serum insulin and glucose that were obtained in youth and at follow-up in young adulthood. For each 1-SD difference in isometric muscle strength (0.16 N/kg) in youth, fasting insulin, HOMA-IR, and HOMA-B in young adulthood changed by -11.3% (95% CI -17.0 to -5.2), -12.2% (-18.2 to -5.7), and -8.9% (-14.4 to -3.0), respectively, in young adulthood after adjustment for CRF and personal lifestyle and demographic factors. Results for CRF were very similar in magnitude, and the magnitude of associations for both exposures was unchanged with additional adjustment for general or abdominal adiposity in youth. Combined associations of muscle strength and CRF with fasting insulin, HOMA-IR, and HOMA-B were additive, and adolescents in the highest sex-specific tertile for both isometric muscle strength and CRF had the lowest levels of these glucose metabolism outcomes. Increasing muscle strength and CRF should be targets in youth primordial prevention strategies of insulin resistance and β-cell dysfunction.


The insecticidal spider toxin SFI1 is a knottin peptide that blocks the pore of insect voltage-gated sodium channels via a large β-hairpin loop.
Spider venoms contain a plethora of insecticidal peptides that act on neuronal ion channels and receptors. Because of their high specificity, potency and stability, these peptides have attracted much attention as potential environmentally friendly insecticides. Although many insecticidal spider venom peptides have been isolated, the molecular target, mode of action and structure of only a small minority have been explored. Sf1a, a 46-residue peptide isolated from the venom of the tube-web spider Segesteria florentina, is insecticidal to a wide range of insects, but nontoxic to vertebrates. In order to investigate its structure and mode of action, we developed an efficient bacterial expression system for the production of Sf1a. We determined a high-resolution solution structure of Sf1a using multidimensional 3D/4D NMR spectroscopy. This revealed that Sf1a is a knottin peptide with an unusually large β-hairpin loop that accounts for a third of the peptide length. This loop is delimited by a fourth disulfide bond that is not commonly found in knottin peptides. We showed, through mutagenesis, that this large loop is functionally critical for insecticidal activity. Sf1a was further shown to be a selective inhibitor of insect voltage-gated sodium channels, consistent with its 'depressant' paralytic phenotype in insects. However, in contrast to the majority of spider-derived sodium channel toxins that function as gating modifiers via interaction with one or more of the voltage-sensor domains, Sf1a appears to act as a pore blocker.


Diabetes as a risk factor for stroke in women compared with men: a systematic review and meta-analysis of 64 cohorts, including 775,385 individuals and 12,539 strokes.
Diabetes mellitus is a major cause of death and disability worldwide and is a strong risk factor for stroke. Whether and to what extent the excess risk of stroke conferred by diabetes differs between the sexes is unknown. We did a systematic review and meta-analysis to estimate the relative effect of diabetes on stroke risk in women compared with men. We systematically searched PubMed for reports of prospective, population-based cohort studies published between Jan 1, 1966, and Dec 16, 2013. Studies were selected if they reported sex-specific estimates of the relative risk (RR) for stroke associated with diabetes, and its associated variability. We pooled the sex-specific RRs and their ratio comparing women with men using random-effects meta-analysis with inverse-variance weighting. Data from 64 cohort studies, representing 775,385 individuals and 12,539 fatal and non-fatal strokes, were included in the analysis. The pooled maximum-adjusted RR of stroke associated with diabetes was 2·28 (95% CI 1·93-2·69) in women and 1·83 (1·60-2·08) in men. Compared with men with diabetes, women with diabetes therefore had a greater risk of stroke--the pooled ratio of RRs was 1·27 (1·10-1·46; I(2)=0%), with no evidence of publication bias. This sex differential was seen consistently across major predefined stroke, participant, and study subtypes. The excess risk of stroke associated with diabetes is significantly higher in women than men, independent of sex differences in other major cardiovascular risk factors. These data add to the existing evidence that men and women experience diabetes-related diseases differently and suggest the need for further work to clarify the biological, behavioural, or social mechanisms involved. None.


Violent video games: The effects of narrative context and reward structure on in-game and postgame aggression.
The potential influence of video game violence on real-world aggression has generated considerable public and scientific interest. Some previous research suggests that playing violent video games can increase postgame aggression. The generalized aggression model (GAM) attributes this to the generalized activation of aggressive schemata. However, it is unclear whether game mechanics that contextualize and encourage or inhibit in-game violence moderate this relationship. Thus, we examined the effects of reward structures and narrative context in a violent video game on in-game and postgame aggression. Contrary to GAM-based predictions, our manipulations differentially affected in-game and postgame aggression. Reward structures selectively affected in-game aggression, whereas narrative context selectively affected postgame aggression. Players who enacted in-game violence through a heroic character exhibited less postgame aggression than players who enacted comparable levels of in-game violence through an antiheroic character. Effects were not attributable to self-activation or character-identification mechanisms, but were consistent with social-cognitive context effects on the interpretation of behavior. These results contradict the GAM's assertion that violent video games affect aggression through a generalized activation mechanism. From an applied perspective, consumer choices may be aided by considering not just game content, but the context in which content is portrayed.


T cell tolerance and activation to a transgene-encoded tumor antigen.
Much has been learned in recent years concerning the nature of tumor antigens recognized by T cells. To apply this knowledge clinically, the nature of the host response to individual and multiple tumor antigens has to be characterized. This will help to define the efficacy of immune surveillance and the immune status of the host following exposure to tumor antigens expressed on pre-neoplastic tissue. To approach these questions, we have developed a transgenic mouse which expresses the tumor-specific antigen P91A. The single amino acid substitution in P91A results in the expression of a new MHC class I (H-2Ld)-binding peptide. In transgenic tissue, the H-2Ld/P91A complex is expressed in isolation from other tumor-associated antigens, allowing definition of the immune response to a single defined tumor antigen, a situation closely analogous to events during tumorigenesis. We show that CD8+ T cell immune surveillance of P91A is ineffective without the introduction of a helper determinant operating through stimulation of CD4+ T cells. Recognition of the isolated P91A tumor antigen on normal tissue by CD8+ T cells is a tolerogenic process. Induction of T cell tolerance suggests tumor antigen-T cell interactions occurring during tumorigenesis may elicit T cell tolerance and hence confound some immunotherapeutic approaches.


The 5S rDNA family evolves through concerted and birth-and-death evolution in fish genomes: an example from freshwater stingrays.
Ribosomal 5S genes are well known for the critical role they play in ribosome folding and functionality. These genes are thought to evolve in a concerted fashion, with high rates of homogenization of gene copies. However, the majority of previous analyses regarding the evolutionary process of rDNA repeats were conducted in invertebrates and plants. Studies have also been conducted on vertebrates, but these analyses were usually restricted to the 18S, 5.8S and 28S rRNA genes. The recent identification of divergent 5S rRNA gene paralogs in the genomes of elasmobranches and teleost fishes indicate that the eukaryotic 5S rRNA gene family has a more complex genomic organization than previously thought. The availability of new sequence data from lower vertebrates such as teleosts and elasmobranches enables an enhanced evolutionary characterization of 5S rDNA among vertebrates. We identified two variant classes of 5S rDNA sequences in the genomes of Potamotrygonidae stingrays, similar to the genomes of other vertebrates. One class of 5S rRNA genes was shared only by elasmobranches. A broad comparative survey among 100 vertebrate species suggests that the 5S rRNA gene variants in fishes originated from rounds of genome duplication. These variants were then maintained or eliminated by birth-and-death mechanisms, under intense purifying selection. Clustered multiple copies of 5S rDNA variants could have arisen due to unequal crossing over mechanisms. Simultaneously, the distinct genome clusters were independently homogenized, resulting in the maintenance of clusters of highly similar repeats through concerted evolution. We believe that 5S rDNA molecular evolution in fish genomes is driven by a mixed mechanism that integrates birth-and-death and concerted evolution.


Inhibition of elastin peptide-mediated angiogenic signaling mechanism(s) in choroidal endothelial cells by the α6(IV)NC1 collagen fragment.
The inhibitory effects and mechanism(s) of type IV collagen α-6 chain-derived noncollagenous domain (α6[IV]NC1 or hexastatin) on elastin-derived peptide (EDP)-activated choroidal endothelial cell migration, kinase signaling, and membrane type 1 metalloproteinase (MT1-MMP) activation are explored. Mouse choroidal endothelial cells (MCECs) were incubated in media with soluble EDPs (kappa elastin, mouse elastin, and Val-Gly-Val-Ala-Pro-Gly [VGVAPG] hexapeptide) for different time intervals with or without α6(IV)NC1. The MCECs proliferation, migration, tube formation, MT1-MMP expression, and angiogenic signaling were analyzed in cells subjected to EDP and α6(IV)NC1 treatments. The MCECs also were subjected to EDPs, and specific inhibitors for evaluation of focal adhesion kinase (FAK) and protein kinase B (Akt) phosphorylation. Kappa elastin, mouse elastin, and VGVAPG enhanced the migration, without affecting the proliferation of MCECs. The α6(IV)NC1 inhibited survival and EDP-activated migration of MCECs. The EDP-activated MCEC tube formation on matrigel also was inhibited by α6(IV)NC1. Further, EDP-activated MT1-MMP expression and FAK/phosphoinositide-3-kinase (PI-3K)/mammalian target of rapamycin (mToR)/Akt phosphorylation in MCECs, were reduced by α6(IV)NC1. The EDP-induced FAK and Akt phosphorylation was blocked by FAK- and Akt-specific inhibitors. The EDPs and α6(IV)NC1 are identified to exhibit opposing effects on MCEC angiogenic behavior and signaling. The α6(IV)NC1 inhibited cell survival, EDP-mediated migration, MT1-MMP expression and, FAK/PI-3K/mToR/Akt phosphorylation in MCECs. This work demonstrates α6(IV)NC1 as a prospective endogenous molecule for the treatment of diseases involving choroidal neovascularization in the eye.


Re-annotation of the CAZy genes of Trichoderma reesei and transcription in the presence of lignocellulosic substrates.
Trichoderma reesei is a soft rot Ascomycota fungus utilised for industrial production of secreted enzymes, especially lignocellulose degrading enzymes. About 30 carbohydrate active enzymes (CAZymes) of T. reesei have been biochemically characterised. Genome sequencing has revealed a large number of novel candidates for CAZymes, thus increasing the potential for identification of enzymes with novel activities and properties. Plenty of data exists on the carbon source dependent regulation of the characterised hydrolytic genes. However, information on the expression of the novel CAZyme genes, especially on complex biomass material, is very limited. In this study, the CAZyme gene content of the T. reesei genome was updated and the annotations of the genes refined using both computational and manual approaches. Phylogenetic analysis was done to assist the annotation and to identify functionally diversified CAZymes. The analyses identified 201 glycoside hydrolase genes, 22 carbohydrate esterase genes and five polysaccharide lyase genes. Updated or novel functional predictions were assigned to 44 genes, and the phylogenetic analysis indicated further functional diversification within enzyme families or groups of enzymes. GH3 β-glucosidases, GH27 α-galactosidases and GH18 chitinases were especially functionally diverse. The expression of the lignocellulose degrading enzyme system of T. reesei was studied by cultivating the fungus in the presence of different inducing substrates and by subjecting the cultures to transcriptional profiling. The substrates included both defined and complex lignocellulose related materials, such as pretreated bagasse, wheat straw, spruce, xylan, Avicel cellulose and sophorose. The analysis revealed co-regulated groups of CAZyme genes, such as genes induced in all the conditions studied and also genes induced preferentially by a certain set of substrates. In this study, the CAZyme content of the T. reesei genome was updated, the discrepancies between the different genome versions and published literature were removed and the annotation of many of the genes was refined. Expression analysis of the genes gave information on the enzyme activities potentially induced by the presence of the different substrates. Comparison of the expression profiles of the CAZyme genes under the different conditions identified co-regulated groups of genes, suggesting common regulatory mechanisms for the gene groups.


Anti-p53 autoantibody in systemic sclerosis: association with limited cutaneous systemic sclerosis.
To determine the prevalence and clinical significance of anti-p53 antibody in patients with systemic sclerosis (SSc). Anti-p53 antibody was examined by ELISA and immunoblotting. Findings were correlated with clinical features of disease and other autoantibodies and compared with other connective tissue diseases as well as normal controls. p53 activity to bind target DNA was evaluated by ELISA using a plate coated with oligonucleotide containing the consensus binding site for p53. IgG anti-p53 antibody levels were elevated in patients with SSc compared to patients with systemic lupus erythematosus (n = 20; p < 0.05), dermatomyositis (n = 21; p < 0.005), atopic dermatitis (n = 17; p < 0.0005), or bullous pemphigoid (n = 10; p < 0.0005) and normal controls (n = 21; p < 0.0005). Remarkably, anti-p53 antibody levels were higher in patients with limited cutaneous SSc (lSSc; n = 30) than those found in patients with diffuse cutaneous SSc (dSSc; n = 40; p < 0.05). IgG or IgM anti-p53 antibody levels did not correlate with the presence or levels of other autoantibodies. IgG anti-p53 antibody was associated with longer disease duration (p < 0.05) and decreased percentage vital capacity (p < 0.05), and correlated negatively with modified Rodnan total skin thickness score (r = -0.352, p < 0.01). Immunoblotting analysis confirmed the presence of IgG anti-p53 antibody in selected patients with SSc. IgG isolated from sera of selected patients with SSc that contained IgG anti-p53 antibody inhibited the p53 activity relative to normal controls. IgG anti-p53 antibody was detected in lSSc and dSSc, and was more prominent in lSSc, indicating that IgG anti-p53 antibody is a novel autoantibody associated with lSSc, a milder form of SSc.


Optimal search strategies for detecting cost and economic studies in EMBASE.
Economic evaluations in the medical literature compare competing diagnosis or treatment methods for their use of resources and their expected outcomes. The best evidence currently available from research regarding both cost and economic comparisons will continue to expand as this type of information becomes more important in today's clinical practice. Researchers and clinicians need quick, reliable ways to access this information. A key source of this type of information is large bibliographic databases such as EMBASE. The objective of this study was to develop search strategies that optimize the retrieval of health costs and economics studies from EMBASE. We conducted an analytic survey, comparing hand searches of journals with retrievals from EMBASE for candidate search terms and combinations. 6 research assistants read all issues of 55 journals indexed by EMBASE for the publishing year 2000. We rated all articles using purpose and quality indicators and categorized them into clinically relevant original studies, review articles, general papers, or case reports. The original and review articles were then categorized for purpose (i.e., cost and economics and other clinical topics) and depending on the purpose as 'pass' or 'fail' for methodologic rigor. Candidate search strategies were developed for economic and cost studies, then run in the 55 EMBASE journals, the retrievals being compared with the hand search data. The sensitivity, specificity, precision, and accuracy of the search strategies were calculated. Combinations of search terms for detecting both cost and economic studies attained levels of 100% sensitivity with specificity levels of 92.9% and 92.3% respectively. When maximizing for both sensitivity and specificity, the combination of terms for detecting cost studies (sensitivity) increased 2.2% over the single term but at a slight decrease in specificity of 0.9%. The maximized combination of terms for economic studies saw no change in sensitivity from the single term and only a 0.1% increase in specificity. Selected terms have excellent performance in the retrieval of studies of health costs and economics from EMBASE.


Feasibility evaluation of an online tool to guide decisions for BRCA1/2 mutation carriers.
Women with BRCA1 or BRCA2 (BRCA1/2) mutations face difficult decisions about managing their high risks of breast and ovarian cancer. We developed an online tool to guide decisions about cancer risk reduction (available at: http://brcatool.stanford.edu ), and recruited patients and clinicians to test its feasibility. We developed questionnaires for women with BRCA1/2 mutations and clinicians involved in their care, incorporating the System Usability Scale (SUS) and the Center for Healthcare Evaluation Provider Satisfaction Questionnaire (CHCE-PSQ). We enrolled BRCA1/2 mutation carriers who were seen by local physicians or participating in a national advocacy organization, and we enrolled clinicians practicing at Stanford University and in the surrounding community. Forty BRCA1/2 mutation carriers and 16 clinicians participated. Both groups found the tool easy to use, with SUS scores of 82.5-85 on a scale of 1-100; we did not observe differences according to patient age or gene mutation. General satisfaction was high, with a mean score of 4.28 (standard deviation (SD) 0.96) for patients, and 4.38 (SD 0.89) for clinicians, on a scale of 1-5. Most patients (77.5 %) were comfortable using the tool at home. Both patients and clinicians agreed that the decision tool could improve patient-doctor encounters (mean scores 4.50 and 4.69, on a 1-5 scale). Patients and health care providers rated the decision tool highly on measures of usability and clinical relevance. These results will guide a larger study of the tool's impact on clinical decisions.


Role of binding site loops in controlling nitric oxide release: structure and kinetics of mutant forms of nitrophorin 4.
Nitrophorins are ferric heme proteins that transport nitric oxide (NO) from blood-sucking insects to victims. NO binding is tighter at lower pH values, as found in the insect salivary gland, and weaker at the pH of the victim's tissue, facilitating NO release and subsequent vasodilation. Previous structural analyses of nitrophorin 4 (NP4) from Rhodnius prolixus revealed a substantial NO-induced conformational change involving the A-B and G-H loops, which rearrange to desolvate the distal pocket and pack nonpolar residues against the heme-ligated NO. Previous kinetic analyses revealed a slow, biphasic, and pH-dependent NO release, which was proposed to be associated with loop movements. In this study, we created NP4 mutants D30A and D30N (A-B loop), D129A/L130A (G-H loop), and T121V (distal pocket). Eight crystal structures were determined, including complexes with NO, NH(3), and imidazole, to resolutions as high as 1.0 A. The NO-induced conformational change is largely abolished in the loop mutants, but retained in T121V. Kinetic analyses using stopped-flow spectroscopy revealed the pH dependence for NO release is eliminated for D129A/L130A, considerably reduced for D30A and D30N, but retained for T121V. NO association rates were increased 2-5-fold for T121V, but were unchanged in the loop mutants. Taken together, our findings demonstrate that the pH dependency for NO release is linked to loop dynamics and that solvent reorganization is apparently rate-limiting for formation of the initial iron-nitrosyl bond. Interestingly, the multiphasic kinetic behavior of rNPs was not affected by mutations, and its cause remains unclear.


A dual role for Ikk alpha in tooth development.
IKK alpha is a component of the I kappa B kinase (IKK) complex that plays a key role in the activation of NF-kappa B. In Ikk alpha mutant mice and mice expressing a transdominant negative mutant of I kappa B alpha (cI kappa B alpha Delta N), molars have abnormal cusps, indicating that Ikk alpha is involved in cusp formation through the NF-kappa B pathway. However, Ikk alpha mutant incisors also have an earlier phenotype where epithelium evaginates outward into the developing oral cavity rather than invaginating into the underlying mesenchyme. A similar evagination of epithelium was also observed in whisker development, suggesting that Ikk alpha contributes to the direction of epithelial growth during the early stages of development in many ectodermal appendages. Since cI kappa B alpha Delta N mice have normal incisor epithelial invagination, Ikk alpha's role appears to be NF-kappa B independent. Changes in Notch1, Notch2, Wnt7b, and Shh expression found in incisor epithelium of Ikk alpha mutants suggest that this NF-kappa B-independent function is mediated by Notch/Wnt/Shh signaling pathways.


Development of a Si-PM-based high-resolution PET system for small animals.
A Geiger-mode avalanche photodiode (Si-PM) is a promising photodetector for PET, especially for use in a magnetic resonance imaging (MRI) system, because it has high gain and is less sensitive to a static magnetic field. We developed a Si-PM-based depth-of-interaction (DOI) PET system for small animals. Hamamatsu 4 × 4 Si-PM arrays (S11065-025P) were used for its detector blocks. Two types of LGSO scintillator of 0.75 mol% Ce (decay time: ∼45 ns; 1.1 mm × 1.2 mm × 5 mm) and 0.025 mol% Ce (decay time: ∼31 ns; 1.1 mm × 1.2 mm × 6 mm) were optically coupled in the DOI direction to form a DOI detector, arranged in a 11 × 9 matrix, and optically coupled to the Si-PM array. Pulse shape analysis was used for the DOI detection of these two types of LGSOs. Sixteen detector blocks were arranged in a 68 mm diameter ring to form the PET system. Spatial resolution was 1.6 mm FWHM and sensitivity was 0.6% at the center of the field of view. High-resolution mouse and rat images were successfully obtained using the PET system. We confirmed that the developed Si-PM-based PET system is promising for molecular imaging research.


Fibrinogen is a ligand for the Staphylococcus aureus microbial surface components recognizing adhesive matrix molecules (MSCRAMM) bone sialoprotein-binding protein (Bbp).
Microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) are bacterial surface proteins mediating adherence of the microbes to components of the extracellular matrix of the host. On Staphylococci, the MSCRAMMs often have multiple ligands. Consequently, we hypothesized that the Staphylococcus aureus MSCRAMM bone sialoprotein-binding protein (Bbp) might recognize host molecules other than the identified bone protein. A ligand screen revealed that Bbp binds human fibrinogen (Fg) but not Fg from other mammals. We have characterized the interaction between Bbp and Fg. The binding site for Bbp was mapped to residues 561-575 in the Fg Aα chain using recombinant Fg chains and truncation mutants in Far Western blots and solid-phase binding assays. Surface plasmon resonance was used to determine the affinity of Bbp for Fg. The interaction of Bbp with Fg peptides corresponding to the mapped residues was further characterized using isothermal titration calorimetry. In addition, Bbp expressed on the surface of bacteria mediated adherence to immobilized Fg Aα. Also, Bbp interferes with thrombin-induced Fg coagulation. Together these data demonstrate that human Fg is a ligand for Bbp and that Bbp can manipulate the biology of the Fg ligand in the host.


Evaluation of a geriatrics primary care model using prospective matching to guide enrollment.
Few definitive guidelines exist for rigorous large-scale prospective evaluation of nonrandomized programs and policies that require longitudinal primary data collection. In Veterans Affairs (VA) we identified a need to understand the impact of a geriatrics primary care model (referred to as GeriPACT); however, randomization of patients to GeriPACT vs. a traditional PACT was not feasible because GeriPACT has been rolled out nationally, and the decision to transition from PACT to GeriPACT is made jointly by a patient and provider. We describe our study design used to evaluate the comparative effectiveness of GeriPACT compared to a traditional primary care model (referred to as PACT) on patient experience and quality of care metrics. We used prospective matching to guide enrollment of GeriPACT-PACT patient dyads across 57 VA Medical Centers. First, we identified matches based an array of administratively derived characteristics using a combination of coarsened exact and distance function matching on 11 identified key variables that may function as confounders. Once a GeriPACT patient was enrolled, matched PACT patients were then contacted for recruitment using pre-assigned priority categories based on the distance function; if eligible and consented, patients were enrolled and followed with telephone surveys for 18 months. We successfully enrolled 275 matched dyads in near real-time, with a median time of 7 days between enrolling a GeriPACT patient and a closely matched PACT patient. Standardized mean differences of < 0.2 among nearly all baseline variables indicates excellent baseline covariate balance. Exceptional balance on survey-collected baseline covariates not available at the time of matching suggests our procedure successfully controlled many known, but administratively unobserved, drivers of entrance to GeriPACT. We present an important process to prospectively evaluate the effects of different treatments when randomization is infeasible and provide guidance to researchers who may be interested in implementing a similar approach. Rich matching variables from the pre-treatment period that reflect treatment assignment mechanisms create a high quality comparison group from which to recruit. This design harnesses the power of national administrative data coupled with collection of patient reported outcomes, enabling rigorous evaluation of non-randomized programs or policies.


Predictors of persistent methicillin-resistant Staphylococcus aureus bacteraemia in patients treated with vancomycin.
The high prevalence of methicillin-resistant Staphylococcus aureus (MRSA) coupled with an increase in vancomycin use have induced vancomycin tolerance in MRSA, adversely affecting the outcome of MRSA bacteraemia. This study aimed to identify predictors of persistent MRSA bacteraemia (PMRSAB) in patients treated with vancomycin. A retrospective, case-control study was performed at a university hospital in Korea from January 2006 to February 2009. Subjects included 96 patients who had MRSA bacteraemia and received vancomycin under therapeutic drug monitoring. We compared the clinical characteristics, management and outcomes of cases with PMRSAB (>or=7 days, n = 31) with controls with non-PMRSAB (<or=3 days, n = 32). Vancomycin MICs were determined by the Vitek 2 system. Of 96 patients with MRSA bacteraemia, MRSA isolates from 21 patients (21.9%) showed a vancomycin MIC of 2 mg/L. Independent predictors of PMRSAB were: retention of implicated medical devices [odds ratio (OR), 10.35; 95% confidence interval (CI), 1.03-104.55]; MRSA infection of at least two sites (OR, 10.24; 95% CI, 1.72-61.01); and vancomycin MIC of 2 mg/L (OR, 6.34; 95% CI, 1.21-33.09). The frequency of side effects and mean trough serum vancomycin concentrations were not significantly different between the two groups. Sixteen patients with PMRSAB subsequently received teicoplanin +/- arbekacin, linezolid or quinupristin/dalfopristin, due to vancomycin failure or intolerance. To minimize the risk of PMRSAB, early removal of implicated devices and evaluation for metastatic infections should be encouraged. Alternative antibiotic therapy is warranted for infections due to isolates with elevated vancomycin MICs, as well as for the high rates of side effects.


Nurses' encounters with children in child and school health care: negotiated guidance within a given frame.
Throughout childhood, children take part in health visits according to a health-monitoring programme. The visits are aimed to promote the children's development and health and to strengthen them to take own responsibility for their health. Nurses' actions when encountering children at these visits are not explored to any great extent. Exploring nurses' actions can facilitate their reflections on their actions towards children and thereby promote children's involvement in such visits. The aim of this study was to explore nurses' actions when encountering children at health visits. A qualitative explorative design, based on 30 video recordings of health visits in child and school health care, was used in this study. These visits were ordinary real-life health visits. The data were subjected to qualitative content analysis. The right to conduct video recordings during health visits was approved by appropriate research ethics committees. The findings show that nurses, in order to carry out the health visits, encounter children through negotiated guidance. This guidance is understood as the process through which the nurses reach agreement with the children, and is comprised of directed and pliable strategies. At one moment, the nurse can use a directed strategy to inform the child and at the next moment a pliable strategy to provide the child space within the given frame, the health-monitoring programme. By using these strategies intertwined, the nurse can provide the child space within the given frame and, at the same time, fulfil his/her responsibility to promote children's health and development. The results highlight nurses' challenging and complex assignment of guiding children to promote their engagement in the health visits, thereby enabling the nurses to promote the children's health and development according to the national health-monitoring programme.


SECM for imaging and detection of latent fingerprints.
Fingerprint analysis is one of the most important methods used for personal identification of evidence found at a crime scene for forensic purposes. Using scanning electrochemical microscopy (SECM) imaging, researchers can visualize the ultrastructure of human fingerprints on wet porous and non-porous surfaces by combining with silver-staining or multi-metal-deposition (MMD) technology. SECM allows investigators to image chemical activities of fingerprint surfaces with an impressively high resolution, such as the third level valuable information for confirming an identification. This methodology takes a significant advantage of the high sensitivity of SECM towards the small variation of electrochemical reaction rates at the substrate surface. In this review, we highlight the recent breakthroughs in ultrasensitive imaging and detection of latent fingerprints with a special focus on a novel application of SECM. We will also discuss our perspectives on future research directions.


Performance and Validation of a Novel Biomarker-Based Stroke Risk Score for Atrial Fibrillation.
Atrial fibrillation is associated with increased but variable risk of stroke. Our aim was to validate the recently developed biomarker-based ABC (age, biomarkers [high-sensitivity troponin and N-terminal fragment B-type natriuretic peptide], and clinical history of prior stroke/transient ischemic attack)-stroke risk score and compare its performance with the CHA2DS2VASc and ATRIA (Anticoagulation and Risk Factors in Atrial Fibrillation) risk scores. The ABC-stroke score includes age, biomarkers (N-terminal fragment B-type natriuretic peptide and high-sensitivity cardiac troponin), and clinical history (prior stroke). This validation was based on 8356 patients, 16 137 person-years of follow-up, and 219 adjudicated stroke or systemic embolic events in anticoagulated patients with atrial fibrillation in the RE-LY study (Randomized Evaluation of Long-Term Anticoagulation Therapy). Levels of N-terminal fragment B-type natriuretic peptide, high-sensitivity cardiac troponin T (hs-cTnT), and high-sensitivity cardiac troponin I (hs-cTnI) were determined in plasma samples obtained at study entry. The ABC-stroke score was well calibrated with 0.76 stroke/systemic embolic events per 100 person-years in the predefined low (<1%/y) risk group, 1.48 in the medium (1%-2%/y) risk group, and 2.60 in the high (>2%/y) risk group for the ABC-stroke score with hs-cTnT. Hazard ratios for stroke/systemic embolic events were 1.95 for medium- versus low-risk groups, and 3.44 for high- versus low-risk groups. ABC-stroke score achieved C indices of 0.65 with both hs-cTnT and hs-cTnI, in comparison with 0.60 for CHA2DS2VASc (P=0.004 for hs-cTnT and P=0.022 hs-cTnI) and 0.61 for ATRIA scores (P=0.005 hs-cTnT and P=0.034 for hs-cTnI). The biomarker-based ABC-stroke score was well calibrated and consistently performed better than both the CHA2DS2VASc and ATRIA stroke scores. The ABC score should be considered an improved decision support tool in the care of patients with atrial fibrillation. URL: http://www.clinicaltrials.gov. Unique identifiers: ARISTOTLE, NCT00412984; RE-LY, NCT00262600.


Antibiotic-Induced Changes to the Host Metabolic Environment Inhibit Drug Efficacy and Alter Immune Function.
Bactericidal antibiotics alter microbial metabolism as part of their lethality and can damage mitochondria in mammalian cells. In addition, antibiotic susceptibility is sensitive to extracellular metabolites, but it remains unknown whether metabolites present at an infection site can affect either treatment efficacy or immune function. Here, we quantify local metabolic changes in the host microenvironment following antibiotic treatment for a peritoneal Escherichia coli infection. Antibiotic treatment elicits microbiome-independent changes in local metabolites, but not those distal to the infection site, by acting directly on host cells. The metabolites induced during treatment, such as AMP, reduce antibiotic efficacy and enhance phagocytic killing. Moreover, antibiotic treatment impairs immune function by inhibiting respiratory activity in immune cells. Collectively, these results highlight the immunomodulatory potential of antibiotics and reveal the local metabolic microenvironment to be an important determinant of infection resolution.


Self-assembly of natural and synthetic drug amphiphiles into discrete supramolecular nanostructures.
Molecular assembly provides an effective approach to construct discrete supramolecular nanostructures of various sizes and shapes in a simple manner. One important technological application of the resulting nanostructures is their potential use as anticancer drug carriers to facilitate targeted delivery to tumour sites and consequently to improve clinical outcomes. In this carrier-assisted delivery strategy, anticancer drugs have been almost exclusively considered as the cargo to be carried and delivered, and their potential as molecular building blocks has been largely ignored. In this discussion, we report the use of anticancer drugs as molecular building units to create discrete supramolecular nanostructures that contain a high and quantitative drug loading and also have the potential for self-delivery. We first show the direct assembly of two amphiphilic drug molecules (methotrexate and folic acid) into discrete nanostructures. Our results reveal that folic acid exhibits rich self-assembly behaviour via Hoogsteen hydrogen bonding under various solvent conditions, whereas methotrexate is unable to assemble into any well-defined nanostructures under the same conditions, despite its similar chemical structure. Considering the low water solubility of most anticancer drugs, hydrophilic segments must be conjugated to the drug in order to bestow the necessary amphiphilicity. We have demonstrated this for camptothecin through the attachment of beta-sheet-forming peptides with overall hydrophilicity. We found that the intermolecular interactions among camptothecin segments and those among beta-sheet peptides act together to define the formation of stable one-dimensional nanostructures in dilute solutions, giving rise to nanotubes or nanofibers depending upon the processing conditions used. These results lead us to believe that self-assembly of drugs into discrete nanostructures not only offers an innovative way to craft self-delivering anticancer drugs, but also extends the paradigm of using molecular assembly as a toolbox to achieve functional nanostructures, to a new area which is specifically focused on the direct assembly of functional molecules (e.g. drugs, or imaging agents) into nanostructures of their own.


The velocity of climate change.
The ranges of plants and animals are moving in response to recent changes in climate. As temperatures rise, ecosystems with 'nowhere to go', such as mountains, are considered to be more threatened. However, species survival may depend as much on keeping pace with moving climates as the climate's ultimate persistence. Here we present a new index of the velocity of temperature change (km yr(-1)), derived from spatial gradients ( degrees C km(-1)) and multimodel ensemble forecasts of rates of temperature increase ( degrees C yr(-1)) in the twenty-first century. This index represents the instantaneous local velocity along Earth's surface needed to maintain constant temperatures, and has a global mean of 0.42 km yr(-1) (A1B emission scenario). Owing to topographic effects, the velocity of temperature change is lowest in mountainous biomes such as tropical and subtropical coniferous forests (0.08 km yr(-1)), temperate coniferous forest, and montane grasslands. Velocities are highest in flooded grasslands (1.26 km yr(-1)), mangroves and deserts. High velocities suggest that the climates of only 8% of global protected areas have residence times exceeding 100 years. Small protected areas exacerbate the problem in Mediterranean-type and temperate coniferous forest biomes. Large protected areas may mitigate the problem in desert biomes. These results indicate management strategies for minimizing biodiversity loss from climate change. Montane landscapes may effectively shelter many species into the next century. Elsewhere, reduced emissions, a much expanded network of protected areas, or efforts to increase species movement may be necessary.


Does the Solvent in a Dispersant Impact the Efficiency of Crude-Oil Dispersion?
Dispersants, used in the mitigation of oil spills, are mixtures of amphiphilic molecules (surfactants) dissolved in a solvent. The recent large-scale use of dispersants has raised environmental concerns regarding the safety of these materials. In response to these concerns, our lab has developed a class of eco-friendly dispersants based on blends of the food-grade surfactants, soy lecithin (L) and Tween 80 (T), in a solvent. We have shown that these "L/T dispersants" are very efficient at dispersing crude oil into seawater. The solvent for dispersants is usually selected based on factors like toxicity, volatility, or viscosity of the overall mixture. However, with regard to the dispersion efficiency of crude oil, the solvent is considered to play a negligible role. In this paper, we re-examine the role of solvent in the L/T system and show that it can actually have a significant impact on the dispersion efficiency. That is, the dispersion efficiency can be altered from poor to excellent simply by varying the solvent while keeping the same blend of surfactants. We devise a systematic procedure for selecting the optimal solvents by utilizing Hansen solubility parameters. The optimal solvents are shown to have a high affinity for crude oil and limited hydrophilicity. Our analysis further enables us to identify solvents that combine high dispersion efficiency, good solubility of the L/T surfactants, a low toxicity profile, and a high flash point.


OX40 ligation of CD4+ T cells enhances virus-specific CD8+ T cell memory responses independently of IL-2 and CD4+ T regulatory cell inhibition.
We have previously shown that CD4(+) T cells are required to optimally expand viral-specific memory CD8(+) CTL responses using a human dendritic cell-T cell-based coculture system. OX40 (CD134), a 50-kDa transmembrane protein of the TNFR family, is expressed primarily on activated CD4(+) T cells. In murine models, the OX40/OX40L pathway has been shown to play a critical costimulatory role in dendritic cell/T cell interactions that may be important in promoting long-lived CD4(+) T cells, which subsequently can help CD8(+) T cell responses. The current study examined whether OX40 ligation on ex vivo CD4(+) T cells can enhance their ability to "help" virus-specific CTL responses in HIV-1-infected and -uninfected individuals. OX40 ligation of CD4(+) T cells by human OX40L-IgG1 enhanced the ex vivo expansion of HIV-1-specific and EBV-specific CTL from HIV-1-infected and -uninfected individuals, respectively. The mechanism whereby OX40 ligation enhanced help of CTL was independent of the induction of cytokines such as IL-2 or any inhibitory effect on CD4(+) T regulatory cells, but was associated with a direct effect on proliferation of CD4(+) T cells. Thus, OX40 ligation on CD4(+) T cells represents a potentially novel immunotherapeutic strategy that should be investigated to treat and prevent persistent virus infections, such as HIV-1 infection.


Compound heterozygous mutations in the SUR1 (ABCC 8) subunit of pancreatic K(ATP) channels cause neonatal diabetes by perturbing the coupling between Kir6.2 and SUR1 subunits.
KATP channels regulate insulin secretion by coupling β-cell metabolism to membrane excitability. These channels are comprised of a pore-forming Kir6.2 tetramer which is enveloped by four regulatory SUR1 subunits. ATP acts on Kir6.2 to stabilize the channel closed state while ADP (coordinated with Mg(2+)) activates channels via the SUR1 domains. Aberrations in nucleotide-binding or in coupling binding to gating can lead to hyperinsulinism or diabetes. Here, we report a case of diabetes in a 7-mo old child with compound heterozygous mutations in ABCC8 (SUR1[A30V] and SUR1[G296R]). In unison, these mutations lead to a gain of KATP channel function, which will attenuate the β-cell response to increased metabolism and will thereby decrease insulin secretion. (86)Rb(+) flux assays on COSm6 cells coexpressing the mutant subunits (to recapitulate the compound heterozygous state) show a 2-fold increase in basal rate of (86)Rb(+) efflux relative to WT channels. Experiments on excised inside-out patches also reveal a slight increase in activity, manifested as an enhancement in stimulation by MgADP in channels expressing the compound heterozygous mutations or homozygous G296R mutation. In addition, the IC 50 for ATP inhibition of homomeric A30V channels was increased ~6-fold, and was increased ~3-fold for both heteromeric A30V+WT channels or compound heterozygous (A30V +G296R) channels. Thus, each mutation makes a mechanistically distinct contribution to the channel gain-of-function that results in neonatal diabetes, and which we predict may contribute to diabetes in related carrier individuals.


Evaluation of a Home-Printable Vision Screening Test for Telemedicine.
Many ophthalmology appointments have been converted to telemedicine assessments. The use of a printed vision chart for ophthalmology telemedicine appointments that can be used by people who are excluded from digital testing has yet to be validated. To evaluate the repeatability of visual acuity measured using the Home Acuity Test (HAT) and the agreement between the HAT and the last in-clinic visual acuity. This diagnostic study was conducted from May 11 to 22, 2020, among 50 control participants and 100 adult ophthalmology outpatients who reported subjectively stable vision and were attending routine telemedicine clinics. Bland-Altman analysis of corrected visual acuity measured with the HAT was compared with the last measured in-clinic visual acuity on a conventional Early Treatment Diabetic Retinopathy Study logMAR chart. For control participants, repeatability of the HAT and agreement with standard logMAR visual acuity measurement. For ophthalmology outpatients, agreement with the last recorded in-clinic visual acuity and with the International Classification of Diseases and Related Health Problems, 11th Revision visual impairment category. A total of 50 control participants (33 [66%] women; mean [SD] age, 36.0 [10.8] years) and 100 ophthalmology patients with a wide range of diseases (65 [65%] women; mean [SD] age, 55.3 [22.2] years) were recruited. For control participants, mean (SD) test-retest difference in the HAT line score was -0.012 (0.06) logMAR, with limits of agreement (LOA) between -0.13 and 0.10 logMAR. The mean (SD) difference in visual acuity compared with conventional vision charts was -0.14 (0.14) logMAR (range, -0.4 to 0.18 log MAR) (-7 letters) in controls, with LOA of -0.41 to 0.12 logMAR (-20 to 6 letters). For ophthalmology outpatients, the mean (SD) difference in visual acuity was -0.10 (0.17) logMAR (range, -0.5 to 0.3 logMAR) (1 line on a conventional logMAR sight chart), with the HAT indicating poorer visual acuity than the previous in-clinic test, and LOA of -0.44 to 0.23 logMAR (-22 to 12 letters). There was good agreement in the visual impairment category for ophthalmology outpatients (Cohen κ = 0.77 [95% CI, 0.74-0.81]) and control participants (Cohen κ = 0.88 [95% CI, 0.88-0.88]). This study suggests that the HAT can be used to measure visual acuity by telephone for a wide range of ophthalmology outpatients with diverse conditions. Test-retest repeatability is relatively high, and agreement in the visual impairment category is good for this sample, supporting the use of printed charts in this context.


First record of Stegomyia albopicta in Turkey determined by active ovitrap surveillance and DNA barcoding.
Despite its confirmed establishment in neighboring Greece and Bulgaria, the presence of the Oriental invasive species Stegomyia albopicta (Skuse) (=Aedes albopictus) has never been confirmed in Turkey. Active surveillance for this container-breeding species was carried out using oviposition traps at 15 discrete sites in the towns of Ipsala (n=8 sites), Kesan (n=5) (Edirne District), and Malkara (n=2) (Tekirdag District) in the Thrace region of northwestern Turkey, from May 23 through November 10, 2011. Eggs collected were reared to the fourth larval instar and adult stages where possible to facilitate integrated morphological and molecular species identification. DNA barcodes (658 bp of the mitochondrial cytochrome c oxidase I [COI] gene) were compared with all four potentially invasive Stegomyia species: St. aegypti, St. albopicta, St. cretina, and St. japonica. Sequences generated for samples collected in Thrace Region were herein confirmed as St. albopicta, the first record of this vector species in Turkey. Eggs of St. albopicta were detected in two discrete localities: (1) In the grounds of a restaurant in Kesan (in week 36), and (2) in the customs area of the Turkish-Greek border at Ipsala (in weeks 32 and 38). Multiple detection of St. albopicta eggs indicates the possible establishment of the species in northwestern Turkey. Finding this important disease vector has implications for public health and requires the implementation of active vector monitoring programs and targeted vector suppression strategies to limit the spread of this invasive vector species in Turkey.


Low bit rate video coding using robust motion vector regeneration in the decoder.
In this paper, we present a novel coding technique that makes use of the nonstationary characteristics of an image sequence displacement field to estimate and encode motion information. We utilize an MPEG style codec in which the anchor frames in a sequence are encoded with a hybrid approach using quadtree, DCT, and wavelet-based coding techniques. A quadtree structured approach is also utilized for the interframe information. The main objective of the overall design is to demonstrate the coding potential of a newly developed motion estimator called the coupled linearized MAP (CLMAP) estimator. This estimator can be used as a means for producing motion vectors that may be regenerated at the decoder with a coarsely quantized error term created in the encoder. The motion estimator generates highly accurate motion estimates from this coarsely quantized data. This permits the elimination of a separately coded displaced frame difference (DFD) and coded motion vectors. For low bit rate applications, this is especially important because the overhead associated with the transmission of motion vectors may become prohibitive. We exploit both the advantages of the nonstationary motion estimator and the effective compression of the anchor frame coder to improve the visual quality of reconstructed QCIF format color image sequences at low bit rates. Comparisons are made with other video coding methods, including the H.261 and MPEG standards and a pel-recursive-based codec.


Effects of Cabergoline on Thyroid Hormones and Semen Quality of Dog.
In this study, the concentration of prolactin, FT4, and semen quality were investigated in 5 clinically healthy fertile mixed-breed dogs (1-3 years old) treated with cabergoline (5μg/kg) during 9 weeks. Semen analysis was performed for volume, live or dead, concentration, and motility factors every week. The results indicated that cabergoline administration caused a minor but significant reduction of the mean prolactin concentration (P < .05) and did not affect the secretion of FT4 (P > .05). Further, there was not a significant effect of cabergoline on semen quality, statistically. We could not find any relationship between prolactin and FT4 concentration and changes in semen characteristics.


No evidence for increased transmissibility from recurrent mutations in SARS-CoV-2.
COVID-19 is caused by the coronavirus SARS-CoV-2, which jumped into the human population in late 2019 from a currently uncharacterised animal reservoir. Due to this recent association with humans, SARS-CoV-2 may not yet be fully adapted to its human host. This has led to speculations that SARS-CoV-2 may be evolving towards higher transmissibility. The most plausible mutations under putative natural selection are those which have emerged repeatedly and independently (homoplasies). Here, we formally test whether any homoplasies observed in SARS-CoV-2 to date are significantly associated with increased viral transmission. To do so, we develop a phylogenetic index to quantify the relative number of descendants in sister clades with and without a specific allele. We apply this index to a curated set of recurrent mutations identified within a dataset of 46,723 SARS-CoV-2 genomes isolated from patients worldwide. We do not identify a single recurrent mutation in this set convincingly associated with increased viral transmission. Instead, recurrent mutations currently in circulation appear to be evolutionary neutral and primarily induced by the human immune system via RNA editing, rather than being signatures of adaptation. At this stage we find no evidence for significantly more transmissible lineages of SARS-CoV-2 due to recurrent mutations.


Association of copper levels in the hair with gray matter volume, mean diffusivity, and cognitive functions.
Although copper plays a critical role in normal brain functions and development, it is known that excess copper causes toxicity. Here we investigated the associations of copper levels in the hair with regional gray matter volume (rGMV), mean diffusivity (MD), and cognitive differences in a study cohort of 924 healthy young adults. Our findings showed that high copper levels were associated mostly with low cognitive abilities (low scores on the intelligence test consisting of complex speed tasks, involving reasoning task, a complex arithmetic task, and a reading comprehension task) as well as lower reverse Stroop interference, high rGMV over widespread areas of the brain [mainly including the bilateral lateral and medial parietal cortices, medial temporal structures (amygdala, hippocampus, and parahippocampal gyrus), middle cingulate cortex, orbitofrontal cortex, insula, perisylvian areas, inferior temporal lobe, temporal pole, occipital lobes, and supplementary motor area], as well as high MD of the right substantia nigra and bilateral hippocampus, which are indicative of low density in brain tissues. These results suggest that copper levels are associated with mostly aberrant cognitive functions, greater rGMV in extensive areas, greater MD (which are indicative of low density in brain tissues) in subcortical structures in the healthy young adults, possibly reflecting copper's complex roles in neural mechanisms.


Changing interpretations of emotional expressions in working memory with aging.
[Correction Notice: An Erratum for this article was reported online in Emotion on Jun 24 2019 (see record 2019-34942-001). In the article, the plots for Figure 3a shifted incorrectly to the right. The error bars should be centered on 10, 30, 50, 70, and 90. The corrected figure is present in the erratum.] Working memory (WM) shows significant decline with age. It is interesting to note that some research has suggested age-related impairments can be reduced in tasks that involve emotion-laden stimuli. However, only a few studies have explored how WM for emotional material changes in aging. Here we developed a novel experimental task to compare and contrast how emotional material is represented in older versus younger adults. The task enabled us to separate overall WM accuracy from emotional biases in the content of affective representations in WM. We found that, in addition to overall decline in WM performance, older adults showed a systematic positivity bias in representing information in WM relative to younger adults (positivity effect). They remembered fearful faces as being less fearful than younger adults and interpreted ambiguous facial expressions more positively. The findings show that aging brings a type of positivity bias when picking up affective information for guiding future behavior. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Ligand control of interaction in vivo between ecdysteroid receptor and ultraspiracle ligand-binding domain.
Ecdysteroids (Ecs) enhance the formation of Ec receptor-ultraspiracle protein (EcR-USP) heterodimers which regulate gene transcription. To study EcR-USP heterodimerization, fusion proteins were constructed from the LBDs (ligand-binding domains) of Drosophila EcR or USP and the activation or DNA-binding region of GAL4 respectively. Reporter gene ( lacZ ) activation was fully dependent on the co-expression of the two fusion proteins and thus constitutes a reliable measure for the interaction in vivo between the two LBDs in the yeast cell. To identify structures involved in heterodimerization, a total of 27 point mutations were generated in the EcR and USP LBDs at selected sites. Heterodimerization and its inducibility by ligand were mainly affected by mutations in the dimerization interface and in the ligand-binding pocket of EcR respectively. However, also mutations not located in these structures or even in the LBD of USP influenced ligand-dependent heterodimerization. Together with previously reported ligand-binding studies, the existence of such local, intra- and inter-molecular mutation effects suggest that ligand-induced dimerization results from a synergistic interaction between ligand-binding and heterodimerization functions in EcR LBD, and that it depends on global features of the LBDs of EcR and USP and on their mutual surface compatibility.


Dietary Diversity and Child Development in the Far West of Nepal: A Cohort Study.
Poverty adversely affects child development through multiple pathways in low- and middle-income countries. Relationships between diet and child development are poorly understood. In this study, we aimed to explore these associations in a longitudinal cohort of 305 children in rural Nepal (baseline mean age 14 months), evaluating dietary diversity and the consumption of specific food groups at three timepoints over 1.5 years. Child development was assessed using the Ages and Stages questionnaire-version 3 (ASQ-3). Associations between the number of days that children consumed minimum dietary diversity (MDD) (≥4/8 items) and specific food groups over time (range 0-3) and total and subscale ASQ scores at age 23-38 months were estimated using multiple linear and logistic regression, dichotomizing scores at the lowest quartile. After adjusting for confounders, each additional day of consuming MDD was associated with a 35% reduction in the odds of low total ASQ score [OR 0.65, 95% CI (0.46, 0.92)]. The consumption of animal source foods [OR 0.64, (0.46, 0.89)], and vegetables/fruits [OR 0.60, (0.41, 0.90), but not processed foods [OR 0.99, (0.62, 1.59)] was associated with lower odds of low total development. Vegetables, fruits and animal source foods may be important for child development in this setting.


Cardiorespiratory fitness and long-term risk of sleep apnea: A national cohort study.
Sleep apnea is increasing in prevalence, and is an important cause of cardiometabolic diseases and mortality worldwide. Its only established modifiable risk factor is obesity; however, up to half of all sleep apnea cases may occur in non-obese persons, and hence there is a pressing need to identify other modifiable risk factors to facilitate more effective prevention. We sought to examine, for the first time, cardiorespiratory fitness in relation to the risk of sleep apnea, independent of obesity. A national cohort study was conducted to examine cardiorespiratory fitness in all 1,547,478 Swedish military conscripts during 1969-1997 (97%-98% of all 18-year-old men) in relation to risk of sleep apnea through 2012 (maximum age 62 years). Cardiorespiratory fitness was measured as maximal aerobic workload in Watts, and sleep apnea was identified from nationwide outpatient and inpatient diagnoses. A total of 44,612 (2.9%) men were diagnosed with sleep apnea in 43.7 million person-years of follow-up. Adjusting for age, height, weight, socioeconomic factors and family history of sleep apnea, low cardiorespiratory fitness at age 18 years was associated with a significantly increased risk of sleep apnea in adulthood (lowest versus highest cardiorespiratory fitness tertile: incidence rate ratio, 1.44; 95% confidence interval, 1.40-1.49; p < 0.001; continuous cardiorespiratory fitness per 100 Watts: incidence rate ratio, 0.71; 95% confidence interval, 0.70-0.73; p < 0.001). An increased risk was observed even among men with normal body mass index (lowest versus highest cardiorespiratory fitness tertile: incidence rate ratio, 1.30; 95% confidence interval, 1.26-1.35; p < 0.001). These findings identify low cardiorespiratory fitness early in life as a new modifiable risk factor for development of sleep apnea in adulthood.