Evaluation of a Rapid Molecular Drug-Susceptibility Test for Tuberculosis.
Fluoroquinolones and second-line injectable drugs are the backbone of treatment regimens for multidrug-resistant tuberculosis, and resistance to these drugs defines extensively drug-resistant tuberculosis. We assessed the accuracy of an automated, cartridge-based molecular assay for the detection, directly from sputum specimens, of Mycobacterium tuberculosis with resistance to fluoroquinolones, aminoglycosides, and isoniazid. We conducted a prospective diagnostic accuracy study to compare the investigational assay against phenotypic drug-susceptibility testing and DNA sequencing among adults in China and South Korea who had symptoms of tuberculosis. The Xpert MTB/RIF assay and sputum culture were performed. M. tuberculosis isolates underwent phenotypic drug-susceptibility testing and DNA sequencing of the genes katG, gyrA, gyrB, and rrs and of the eis and inhA promoter regions. Among the 308 participants who were culture-positive for M. tuberculosis, when phenotypic drug-susceptibility testing was used as the reference standard, the sensitivities of the investigational assay for detecting resistance were 83.3% for isoniazid (95% confidence interval [CI], 77.1 to 88.5), 88.4% for ofloxacin (95% CI, 80.2 to 94.1), 87.6% for moxifloxacin at a critical concentration of 0.5 μg per milliliter (95% CI, 79.0 to 93.7), 96.2% for moxifloxacin at a critical concentration of 2.0 μg per milliliter (95% CI, 87.0 to 99.5), 71.4% for kanamycin (95% CI, 56.7 to 83.4), and 70.7% for amikacin (95% CI, 54.5 to 83.9). The specificity of the assay for the detection of phenotypic resistance was 94.3% or greater for all drugs except moxifloxacin at a critical concentration of 2.0 μg per milliliter (specificity, 84.0% [95% CI, 78.9 to 88.3]). When DNA sequencing was used as the reference standard, the sensitivities of the investigational assay for detecting mutations associated with resistance were 98.1% for isoniazid (95% CI, 94.4 to 99.6), 95.8% for fluoroquinolones (95% CI, 89.6 to 98.8), 92.7% for kanamycin (95% CI, 80.1 to 98.5), and 96.8% for amikacin (95% CI, 83.3 to 99.9), and the specificity for all drugs was 99.6% (95% CI, 97.9 to 100) or greater. This investigational assay accurately detected M. tuberculosis mutations associated with resistance to isoniazid, fluoroquinolones, and aminoglycosides and holds promise as a rapid point-of-care test to guide therapeutic decisions for patients with tuberculosis. (Funded by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, and the Ministry of Science and Technology of China; ClinicalTrials.gov number, NCT02251327 .).


Inequality and changes in women's use of maternal health-care services in Tajikistan.
Using recently available survey data for Tajikistan, this study explores changes in the pattern of maternal health care during the last decade and the extent to which inequalities in access to that care have emerged. In particular, the links between poverty and women's educational status and the use of maternal health-care services are investigated. The survey findings demonstrate a significant decline in the use of maternal health-care services in Tajikistan since the country gained independence from the Soviet Union in 1991. They show changes in the location of delivery and the person providing assistance, with a clear shift away from giving birth in a medical facility toward giving birth at home. More than two-fifths of all women who gave birth in the year prior to the survey delivered their baby at home. Women from the poorest quintile are three times more likely than women from the richest quintile to undergo a home delivery without a trained assistant.


Zingiber officinale Mitigates Brain Damage and Improves Memory Impairment in Focal Cerebral Ischemic Rat.
Cerebral ischemia is known to produce brain damage and related behavioral deficits including memory. Recently, accumulating lines of evidence showed that dietary enrichment with nutritional antioxidants could reduce brain damage and improve cognitive function. In this study, possible protective effect of Zingiber officinale, a medicinal plant reputed for neuroprotective effect against oxidative stress-related brain damage, on brain damage and memory deficit induced by focal cerebral ischemia was elucidated. Male adult Wistar rats were administrated an alcoholic extract of ginger rhizome orally 14 days before and 21 days after the permanent occlusion of right middle cerebral artery (MCAO). Cognitive function assessment was performed at 7, 14, and 21 days after MCAO using the Morris water maze test. The brain infarct volume and density of neurons in hippocampus were also determined. Furthermore, the level of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in cerebral cortex, striatum, and hippocampus was also quantified at the end of experiment. The results showed that cognitive function and neurons density in hippocampus of rats receiving ginger rhizome extract were improved while the brain infarct volume was decreased. The cognitive enhancing effect and neuroprotective effect occurred partly via the antioxidant activity of the extract. In conclusion, our study demonstrated the beneficial effect of ginger rhizome to protect against focal cerebral ischemia.


Phenotypic variation in molecular mimicry between Helicobacter pylori lipopolysaccharides and human gastric epithelial cell surface glycoforms. Acid-induced phase variation in Lewis(x) and Lewis(y) expression by H. Pylori lipopolysaccharides.
Helicobacter pylori is an important gastroduodenal pathogen of humans whose survival in the gastric environment below pH 4 is dependent on bacterial production of urease, whereas above pH 4 urease-independent mechanisms are involved in survival, but that remain to be elucidated fully. Previous structural investigations on the lipopolysaccharides (LPSs) of H. pylori have shown that the majority of these surface glycolipids express partially fucosylated, glucosylated, or galactosylated N-acetyllactosamine (LacNAc) O-polysaccharide chains containing Lewis(x) (Le(x)) and/or Lewis(y) (Le(y)), although some strains also express type 1 determinants, Lewis(a), Lewis(b), and H-1 antigen. In this study, we investigated acid-induced changes in the structure and composition of LPS and cellular lipids of the genome-sequenced strain, H. pylori 26695. When grown in liquid medium at pH 7, the O-chain consisted of a type 2 LacNAc polysaccharide, which was glycosylated with alpha-1-fucose at O-3 of the majority of N-acetylglucosamine residues forming Le(x) units, including chain termination by a Le(x) unit. However, growth in liquid medium at pH 5 resulted in production of a more complex O-chain whose backbone of type 2 LacNAc units was partially glycosylated with alpha L-fucose, thus forming Le(x), whereas the majority of the nonfucosylated N-acetylglucosamine residues were substituted at O-6 by alpha-D-galactose residues, and the chain was terminated by a Le(y) unit. In contrast, detailed chemical analysis of the core and lipid A components of LPS and analysis of cellular lipids did not show significant differences between H. pylori 26695 grown at pH 5 and 7. Although putative molecular mechanisms affecting Le(x) and Le(y) expression have been investigated previously, this is the first report identifying an environmental trigger inducing phase variation of Le(x) and Le(y) in H. pylori that can aid adaptation of the bacterium to its ecological niche.


Unraveling Anthocyanin Bioavailability for Human Health.
This review considers the bioavailability of health-protective anthocyanin pigments from foods, in light of the multiple molecular structures and complicated traffic patterns taken by anthocyanins both as flavonoid metabolites and as phenolic acid metabolites within the body. Anthocyanins have generally been considered to have notoriously poor bioavailability, based on the very low levels typically detected in routine human blood draws after ingestion. Although some investigations have assessed anthocyanin bioavailability solely based on the measurement of parent anthocyanins or phenolic acid breakdown products, more recent research has increasingly revealed the presence, qualitative diversity, relatively high concentrations, and tenacity of molecular intermediates of anthocyanins that retain the unique flavonoid C6-C3-C6 backbone structure. We argue that the persistence of anthocyanin metabolites suggests enterohepatic recycling, leading to prolonged residence time, and supports the notion that anthocyanins are far more bioavailable than previously suggested.


Complementary and non-coincident increases in heart rate variability and irregularity during fetal development.
1. Two distinct notions of variability have been defined to assess heart rate: deviation from a constant output (SD) and irregularity. One statistical measure of irregularity is approximate entropy, with greater irregularity corresponding to larger approximate entropy values. The specific aims of this investigation were to determine the manner in which SD and irregularity evolve during fetal development and whether this evolution is coincident or distinct. 2. Fetal heart rate was computed in 14 males and 17 females for 15 min of undisturbed recording using a fetal actocardiograph at 4 week intervals from 20 to 36 weeks gestation. 3. Mean heart rate decreased significantly with gestational ages (P < 0.05). SD increased significantly from 4.4 +/- 0.3 ms (SEM) at 20 weeks to 7.7 +/- 0.4 ms at 36 weeks (P < 0.05) and was similar between male and female fetuses (P = 0.57). Fixed approximate entropy increased significantly from 0.47 +/- 0.04 to 0.78 +/- 0.03, paralleling the change in SD (P < 0.01). Notably, normalized approximate entropy, which decorrelates SD from regularity, increased significantly with gestational age (P < 0.01) for males, while it remained relatively constant for females (P = 0.68). Approximate entropy was significantly lower at 20 weeks in males than females (P < 0.05); however, the values were similar by 28 weeks gestation. 4. Our results demonstrate that variability increases, and that irregularity increases, independently in male fetuses but not female fetuses, consistent with an increase in the coupling of emerging networks with gestational age. The present study demonstrates the complementary information obtained by analyses of both measures of variability and regularity. This reinforces a difference in gender-based development as noted in the separate context of fetal lung maturation.


Adherence to Treatment Guidelines in Heart Failure Patients in the Top End Region of Northern Territory.
Heart failure (HF) is associated with significant morbidity and mortality and recurrent hospitalisations, particularly in the Indigenous Australians of the Northern Territory. In remote Northern Australia, the epidemiology is less clear but anecdotal evidence suggests it may be worse. In addition, some anecdotal evidence suggests that prognostic pharmacological therapy could also be underutilised. Minimal HF data exists in the remote and Indigenous settings, making this study unique. A retrospective cohort review of pharmacological management of 99 patients from 1 January 2014 to 31 December 2014 was performed. Ninety-nine (99) patients were identified. 59.6% were non-Indigenous vs 40.4% Indigenous. The majority was male (69.7%). Indigenous patients were younger; median age was 51.4 (43.4-60.6) vs 70.5 (62.2-77.0), p<0.001. Major causes of HF were coronary artery disease (61%) and dilated cardiomyopathy (27%). Associated comorbidities included hypertension (52%), dyslipidaemia (38%), diabetes mellitus (40%) and atrial fibrillation (25%). The use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers (ACEI/ARB) and β-blocker was 68% and 87%, respectively. Forty-one (41) patients not on an ACEI/ARB and/or β-blocker were identified. Seventeen (17) of those patients (42%) did not receive an ACEI/ARB because of renal failure. Four (4) patients (10%) did not take a β-blocker due to hypotension. Fourteen (14) patients (34%) were not prescribed an ACEI/ARB and/or β-blocker had no identifiable contraindications. Indigenous patients are over-represented at a younger age demonstrating the alarming rate of disease burden in NT's young Indigenous population. Generally, ACEI/ARBs were underutilised compared to β-blockers with renal impairment being the primary contraindication. There is a need to develop processes to further improve the use of heart failure medications and setting up a HF database could be the first step in progress.


TNF-alpha modulates expression of the tissue transglutaminase gene in liver cells.
One of several postulated roles for tissue transglutaminase (tTG) is the stabilization and assembly of extracellular matrix via peptide cross-linking. We previously determined that tTG activity increased in an animal model of hepatic fibrogenesis and in human liver disease. To further study the role of tTG in liver disease, we initiated investigations into the effect of a proinflammatory mediator, tumor necrosis factor (TNF)-alpha, on tTG activity in cultured liver cells. Treatment of human Hep G2 cells with 1 ng/ml TNF-alpha increased [14C]putrescine cross-linking to cellular proteins. An increase in tTG mRNA content was observed 1 h after addition of TNF-alpha, and levels of tTG mRNA remained elevated after 24 h. Hep G2 cells, transiently transfected with a luciferase reporter containing 1.67 kb of the human tTG promoter, showed an increase in reporter activity after addition of TNF-alpha. Gel shift experiments using nuclear extracts from TNF-alpha-treated cells and oligonucleotides containing the tTG nuclear factor (NF)-kappa B motif revealed increased binding, concordant with mRNA data. Transient transfections with a truncated reporter construct lacking the tTG NF-kappa B sequence showed an attenuated response to TNF-alpha treatment. Similar responses were seen in stably transfected HeLa cells. Primary hepatocytes isolated from a transgenic mouse line containing the mouse tTG promoter driving the beta-galactosidase reporter, show similar time-dependent increases in promoter activity when treated with TNF-alpha. Furthermore, Hep G2 cells are incapable of upmodulating tTG promoter reporter activity in the presence of TNF-alpha when those cells overexpress a transdominant, negative mutant NF-kappa B subunit. Because TNF-alpha expression is upregulated in hepatic inflammation, the data suggest TNF-alpha-mediated increases in tTG expression may play an important role in the process of hepatic fibrogenesis.


Prospective associations between sugar-sweetened beverage intakes and cardiometabolic risk factors in adolescents.
High sugar-sweetened beverage (SSB) consumption is associated with cardiometabolic disturbances in adults, but this relation is relatively unexplored in children and adolescents. We tested the hypothesis that higher SSB intakes are associated with increases in cardiometabolic risk factors between 14 and 17 y of age. Data were provided by 1433 adolescent offspring from the Western Australian Pregnancy Cohort (Raine) Study. At 14 and 17 y of age, SSB intakes were estimated by using a food-frequency questionnaire; body mass index (BMI), waist circumference, blood pressure, fasting serum lipids, glucose, and insulin were measured, and overall cardiometabolic risk was estimated. Prospective associations between cardiovascular disease risk factors and SSB intake were examined with adjustment for age, pubertal stage, physical fitness, socioeconomic status, and major dietary patterns. The average SSB intake in consumers (89%) was 335 g/d or 1.3 servings/d. Girls who moved into the top tertile of SSB consumption (>1.3 servings/d) between 14 and 17 y of age had increases in BMI (3.8%; 95% CI: 1.8%, 5.7%), increased overweight and obesity risk (OR: 4.8, 95% CI: 2.1, 11.4), and greater overall cardiometabolic risk (OR: 3.2; 95% CI: 1.6, 6.2) (all P-trend ≤ 0.001). Girls and boys who moved into the top tertile of SSB intake showed increases in triglycerides (7.0-8.4%; P-trend ≤ 0.03), and boys showed reductions in HDL cholesterol (-3.1%; 95% CI: -6.2%, 0.1%; P-trend < 0.04) independent of BMI. Some associations were attenuated after adjustment for major dietary patterns. Increased SSB intake may be an important predictor of cardiometabolic risk in young people, independent of weight status.


Clinical Care Redesign to Improve Value for Trigger Finger Release: A Before-and-After Quality Improvement Study.
Background: Trigger finger release (TFR) is a commonly performed procedure. However, there is great variation in the setting, care pathway, anesthetic, and cost. We compared the institutional cost for isolated TFR before and after redesigning our clinical care pathway. Methods: Total direct cost to the health system (excluding the surgeon and anesthesiology costs) and time spent by the patient at the surgery center were collected for 1 hand surgeon's procedures at an ambulatory surgery center over a 3-year period. We implemented a redesigned pathway that altered phases of care and anesthetic use by transitioning from intravenous (IV) sedation to wide awake local anesthesia with no tourniquet. Cost data were reported as percentage change in the median and compared both pre- to post-implementation and with 2 control surgeons using the traditional pathway within the same center. Power analysis was based on prior work on a carpal tunnel pathway. Significance was defined by a P-value < .05. Results: Ten TFRs (90% local with IV sedation) and 44 TFRs (89% local alone) were performed pre- and post-implementation, respectively. From pre- to post-implementation, the study surgeon's total direct cost decreased by 18%, while the control surgeons decreased by 2%. Median time spent at the surgery center decreased by 41 minutes post-implementation with significantly shorter setup time in the operating room (OR), total time in the OR, and time spent in recovery prior to discharge. Conclusions: Redesigning the care pathway for TFR led to a decrease in institutional cost and patient time spent at the surgery center.


This "Ethical Trap" Is for Roboticists, Not Robots: On the Issue of Artificial Agent Ethical Decision-Making.
In this paper we address the question of when a researcher is justified in describing his or her artificial agent as demonstrating ethical decision-making. The paper is motivated by the amount of research being done that attempts to imbue artificial agents with expertise in ethical decision-making. It seems clear that computing systems make decisions, in that they make choices between different options; and there is scholarship in philosophy that addresses the distinction between ethical decision-making and general decision-making. Essentially, the qualitative difference between ethical decisions and general decisions is that ethical decisions must be part of the process of developing ethical expertise within an agent. We use this distinction in examining publicity surrounding a particular experiment in which a simulated robot attempted to safeguard simulated humans from falling into a hole. We conclude that any suggestions that this simulated robot was making ethical decisions were misleading.


Spectral sensitivity measurements reveal partial success in restoring missing rod function with gene therapy.
Restored rod visual function after gene therapy can be established unequivocally by demonstrating that, after dark adaptation, spectral sensitivity has the shape characteristic of rods and that this shape collapses to a cone-like shape before rods have recovered after an intense bleach. We used these tests to assess retinal function in eight young adults and children with early-onset severe retinal dystrophy from Phase II of a clinical gene-therapy trial for RPE65 deficiency that involved the subretinal delivery of a recombinant adeno-associated viral vector carrying RPE65. We found substantial improvements in rod sensitivity in two participants: dark-adapted spectral sensitivity was rod-like after treatment and was cone-like before rods had recovered after a bleach. After 40 min of dark adaptation, one participant showed up to 1,000-fold sensitivity improvements 4 months after treatment and the second up to 100-fold improvements 6 months after treatment. The dark-adapted spectral sensitivities of the other six participants remained cone-like and showed little improvement in sensitivity.


Guidelines for uniform reporting of body fluid biomarker studies in neurologic disorders.
The aim of these guidelines is to make the process of reporting body fluid biomarker studies in neurologic disorders more uniform and transparent, in line with existing standards for reporting research in other biomedical areas. Although biomarkers have been around for decades, there are concerns over the high attrition rate of promising candidate biomarkers at later phases of development. BioMS-eu consortium, a collaborative network working toward improving the quality of biomarker research in neurologic disorders, discussed the merits of standardizing the reporting of body fluid biomarker research. A checklist of items integrating the results of other published guidances, literature, conferences, regulatory opinion, and personal expertise was created to ultimately form a structured summary guidance incorporating the key features. The summary guidance is comprised of a 10-point uniform reporting format ranging from introduction, materials and methods, through to results and discussion. Each item is discussed in detail in the guidance report. To enhance the future development of body fluid biomarkers, it will be important to standardize the reporting of studies. This guideline by the BioMS-eu consortium is aimed at setting a standard for the reporting of future body fluid biomarker research studies in neurologic disorders. We anticipate that following these guidelines will help to accelerate the selection of biomarkers for clinical development.


Contributions of specificity protein-1 and steroidogenic factor 1 to Adcy4 expression in Y1 mouse adrenal cells.
The type 4 adenylyl cyclase, Adcy4, is the least abundant of five different adenylyl cyclase isoforms expressed in the Y1 mouse adrenocortical cell line and is deficient in a Y1 mutant with impaired steroidogenic factor 1 (SF1) activity. This study examines the contributions of SF1 and other DNA promoter/regulatory elements to Adcy4 expression in the Y1 cell line and its derivative Adcy4-deficient mutant. Primer extension and in silico analyses indicate that Adcy4 transcription initiates from multiple sites just downstream of a GC-rich sequence. Luciferase reporter gene assays identify a 124-bp sequence, situated 19 bp upstream of the major transcription start site and highly conserved among several mammalian species, as the major determinant of Adcy4 expression in Y1 cells and as a site of compromised activity in the Adcy4-deficient mutant. EMSAs using competitor nucleotides and specific antibodies indicate that this conserved region contains three specificity protein (Sp)-1/Sp3-binding sites and one SF1-binding site. As determined by site-specific mutagenesis, the 5'-most Sp1/Sp3-site enhances promoter activity, whereas the middle Sp1/Sp3 and SF1 sites each repress Adcy4 promoter activity. In the Adcy4-deficient mutant, mutating the SF1 site restores Adcy4 promoter activity and knocking down SF1 with small interfering RNAs increases Adcy4 expression, confirming the contribution of SF1 to the mutant phenotype. These studies demonstrate roles for Sp1/Sp3 and SF1 in Adcy4 expression in Y1 cells and establish a repressor function for SF1 in certain promoter contexts.


Cognitive functioning and driving simulator performance in middle-aged and older adults with HIV.
Nearly half of people living with HIV experience cognitive deficits that may impact instrumental activities of daily living. As the number of people aging with HIV increases, concerns mount that disease-related cognitive deficits may be compounded by age-related deficits, which may further compromise everyday functions such as driving. In this cross-sectional pilot study, during a 2.5-hour visit, 26 middle-aged and older adults (40 + years) were administered demographic, health, psychosocial, and driving habits questionnaires; cognitive assessments; and driving simulator tests. Although CD4+ T lymphocyte count and viral load were unrelated to driving performance, older age was related to poorer driving. Furthermore, poorer visual speed of processing performance (i.e., useful field of view) was related to poorer driving performance (e.g., average gross reaction time). Mixed findings were observed between driving performance and cognitive function on self-reported driving habits of participants. Implications for these findings on nursing practice and research are posited.


Partial agonist activity of the progesterone receptor antagonist RU486 mediated by an amino-terminal domain coactivator and phosphorylation of serine400.
Jun dimerization protein-2 (JDP-2) is a progesterone receptor (PR) coregulatory protein that acts by inducing structure and transcriptional activity in the disordered amino-terminal domain (NTD) of PR. JDP-2 can also potentiate the partial agonist activity of the PR antagonist RU486 by mechanisms that have not been defined. Functional mutagenesis experiments revealed that a subregion of the NTD (amino acids 323-427) was required for the partial agonist activity of RU486 induced by PR interaction with JDP-2. However, this subregion was not required for JDP-2 enhancement of the activity of progestin agonists. Mutation of phosphorylation sites within this region of the NTD showed that phosphorylation of serine 400 was required for the partial agonist activity of RU486 stimulated by JDP-2, but was not required for activity of hormone agonist, either in the presence or absence of JDP-2. Cyclin-dependent kinase 2 (Cdk2)/cyclin A is a novel PR coregulator that binds the NTD and acts by phosphorylating steroid receptor coactivator-1 and modulating steroid receptor coactivator-1 interaction with PR. Cdk2/cyclin A also potentiated the partial agonist activity of RU486; however, phosphorylation of serine 400 was not required, indicating that JDP-2 and Cdk2/cyclin A act by distinct mechanisms. We conclude that PR bound to RU486 and associated with JDP-2 adopts an active conformation in a subregion of the NTD requiring phosphorylation of serine 400 that is distinct from that promoted by progestin agonists. These data underscore the structural flexibility of the NTD of PR, and the ability of steroid ligands together with interacting proteins to affect the conformation and activity of the NTD.


Amperometric biosensor based on glucose dehydrogenase and plasma-polymerized thin films.
A novel design is described for an amperometric biosensor based on NAD(P)-dependent glucose dehydrogenase (GDH) combined with a plasma-polymerized thin film (PPF). The GDH is sandwiched between several nanometer thick acetonitrile PPFs on a sputtered gold electrode (PPF/GDH/PPF/Au). The lower PPF layer plays the role as an interface between enzyme and electrode because it is extremely thin, adheres well to the substrate (electrode), has a flat surface and a highly-crosslinked network structure, and is hydrophilic in nature. The upper PPF layer (overcoating) was directly deposited on immobilized GDH. The optimized amperometric biosensor characteristics covered 2.5-26 mM glucose concentration at +0.6 V of applied potential; the least-squares slope was 320 nA mM(-1) cm(-2) and the correlation coefficient was 0.990. Unlike conventional wet-chemical processes that are incompatible with mass production techniques, this dry-chemistry procedure has great potential for enabling high-throughput production of bioelectronic devices.


Modular Solid-Phase Synthesis of Antiprotozoal Barnesin Derivatives.
Here, we applied and optimized a solid support (SP)-based Horner-Wadsworth-Emmons reagent to prepare SP-bound vinylogous amino acids. Subsequent SP-based peptide synthesis, global deprotection, and chemical modifications yielded 14 lipodipeptides carrying vinylogous amino acids, including the natural product barnesin A (1). Biological evaluation revealed that several synthesized derivatives show micromolar to nanomolar inhibitory activity against papain-like cysteine proteases, human cathepsin L, and rhodesain.


Evaluation of a combined supported computer education and employment training program for persons with psychiatric disabilities.
Meaningful work is described as one of the functional indicators of healing and growth beyond the disability and is seen as critical in recovering a personal sense of worth and value. We describe a supported education-supported employment program which focused on teaching computer, recovery and work skills. A program evaluation was implemented on four consecutive classes of this program. Four classes with a convenience sample of sixty-one students were involved in the evaluation over years. The program utilized a one group pretest, posttest design, with repeated measures over time. Following the 10-month classroom training phase, students entered a 2-month internship to give them computer office work experience. Students were interviewed quarterly using standardized assessments involving work and other subjective outcomes. Results suggest that overall the students experienced a positive change in work status and income and a decrease in mental health services utilization. In addition, non-vocational outcomes, specifically self-esteem and empowerment improved. The program represents a successful integration of supported education and supported employment program models.


Quantification of Volume Mismatch of Acetabulum and Femoral Head in Developmental Dysplasia of Hip.
The sustained subluxation or dislocation of the femoral head over time does not permit normal development of acetabulum and results in predictable pattern of acetabular growth disturbance that is termed hip dysplasia. The primary aim of this study is to analyze and quantify the volume mismatch between acetabulum and femoral head of affected side as compared to normal hip. A prospective observational study was conducted by including isolated untreated unilateral idiopathic developmental dysplasia of hip (DDH). After routine clinical and radiographic examination, computed tomography (CT) of both hips was done with pre-determined radiation dosage within safe limits for the pediatric age group in 18 patients of median age 2 years (range 1-5 years). A significant difference was noted between acetabular index (p<0.001), acetabular volume (p<0.001), femoral head volume (p<0.001), and acetabular anterior sectoral angle (p=0.002) of the affected and the normal hips. As compared to the normal side, the acetabulum is 2.6 times smaller than the normal side and femoral epiphysis volume by 3.8 times. A significant negative correlation (r=-0.66, p=0.04) was noted between posterior acetabular sectoral angle and acetabular volume of affected hip. CT is an important investigation in evaluation of late-presenting DDH. The absence of femoral head in its orthotopic location affects the volume of acetabulum as well as that of femoral head. The abnormality of the volume of acetabulum which is seen as related to the dysplasia should be studied and assessed in detail in a child of late-presenting DDH. This would guide us toward the coverage defect and type of osteotomy to be performed.


Extensional flow of blood analog solutions in microfluidic devices.
In this study, we show the importance of extensional rheology, in addition to the shear rheology, in the choice of blood analog solutions intended to be used in vitro for mimicking the microcirculatory system. For this purpose, we compare the flow of a Newtonian fluid and two well-established viscoelastic blood analog polymer solutions through microfluidic channels containing both hyperbolic and abrupt contractions∕expansions. The hyperbolic shape was selected in order to impose a nearly constant strain rate at the centerline of the microchannels and achieve a quasihomogeneous and strong extensional flow often found in features of the human microcirculatory system such as stenoses. The two blood analog fluids used are aqueous solutions of a polyacrylamide (125 ppm w∕w) and of a xanthan gum (500 ppm w∕w), which were characterized rheologically in steady-shear flow using a rotational rheometer and in extension using a capillary breakup extensional rheometer (CaBER). Both blood analogs exhibit a shear-thinning behavior similar to that of whole human blood, but their relaxation times, obtained from CaBER experiments, are substantially different (by one order of magnitude). Visualizations of the flow patterns using streak photography, measurements of the velocity field using microparticle image velocimetry, and pressure-drop measurements were carried out experimentally for a wide range of flow rates. The experimental results were also compared with the numerical simulations of the flow of a Newtonian fluid and a generalized Newtonian fluid with shear-thinning behavior. Our results show that the flow patterns of the two blood analog solutions are considerably different, despite their similar shear rheology. Furthermore, we demonstrate that the elastic properties of the fluid have a major impact on the flow characteristics, with the polyacrylamide solution exhibiting a much stronger elastic character. As such, these properties must be taken into account in the choice or development of analog fluids that are adequate to replicate blood behavior at the microscale.


Epidemiology of Taenia saginata taeniosis/cysticercosis: a systematic review of the distribution in central and western Asia and the Caucasus.
The zoonotic parasite Taenia saginata transmits between humans, the definitive host (causing taeniosis), and bovines as the intermediate host (causing cysticercosis). Central and western Asia and the Caucasus have large cattle populations and beef consumption is widespread. However, an overview of the extent of human T. saginata infection and bovine cysticercosis is lacking. This review aims to summarize the distribution of T. saginata in this region. A systematic review was conducted, that gathered published and grey literature, and official data concerning T. saginata taeniosis and bovine cysticercosis in central and western Asia and the Caucasus published between January 1st, 1990 and December 31st, 2018. Where no data were available for a country within this period, published data from 1985-1990 were also accessed. From 10,786 articles initially scanned, we retrieved 98 full-text articles from which data were extracted. In addition, two unpublished datasets were provided on the incidence of human taeniosis. Data for human taeniosis and bovine cysticercosis were found for all countries except Turkmenistan. Human taeniosis prevalence varied from undetected to over 5.3%, with regional variations. Where bovine cysticercosis was detected, prevalences varied from case reports to 25%. The public health burden of T. saginata is assumed to be small as the parasite is of low pathogenicity to humans. However, this review indicates that infection continues to be widespread and this may result in a large economic burden, due to the resources utilized in meat inspection and condemnation or processing with subsequent downgrading of infected carcasses.


Sex estimation by femur in modern Thai population.
Sex estimation is an important step of postmortem investigation and the femur is a useful bone for sex estimation by using metric analysis method. Even though there have been a reported sex estimation method by using femur in Thais, the temporal change related to time and anthropological data need to be renewed. Thus the aim of this study is to re-evaluate sex estimation by femur in Thais. 97 adult male and 103 female femora were random chosen from Forensic osteology research center and 6 measurements were applied tend to. To compare with previous Thai data, mid shaft diameter to increase but femoral head and epicondylar breadth to stabilize and when tested previous discriminant function by vertical head diameter and epicondalar breadth, the accuracy of prediction was lower than previous report. From the new data, epicondalar breadth is the best variable for distinguishing male and female at 88.7 percent of accuracy, following by transverse and vertical head diameter at 86.7 percent and femoral neck diameter at 81.7 percent of accuracy. Multivariate discriminant analysis indicated transverse head diameter and epicondylar breadth performed highest rate of accuracy at 89.7 percent. The percent of accuracy of femur was close to previous reported sex estimation by talus and calcaneus in Thai population. Thus, for especially in case of lower limb remain, which absence of pelvis.


Touch and surgical division of the anterior quadrant of the spinal cord.
An investigation was carried out to determine whether tactile sensibility was affected by anterolateral cordotomy. There were 65 patients who had cordotomies for painful forms of cancer. Thirty eight had necropsy examination with histological investigation of the spinal cord. No form of mechanoreception was removed in any of the 65 patients and in the majority no forms of tactile sensibility were altered by division of the pathways in the anterolateral and anterior columns. In no case was graphaesthesia affected. Knowledge of joint position and movement and awareness of vibration was normal in 62 of the 65 patients. But information carried by these anterolateral pathways does reach neural levels of consciousness, for with total lesions of the posterior columns, previously reported, touch and pressure are still felt. Itch was removed by division of the anterolateral pathways. Although the posterior columns are essential for discrimination in mechanoreception, discrimination may be disturbed by lesions of the anterolateral pathways, notably two-point discrimination. The evidence on the pathways essential for conveying impulses giving rise to tickle was inconclusive.


Salt tolerance in Solanum pennellii: antioxidant response and related QTL.
Excessive soil salinity is an important problem for agriculture, however, salt tolerance is a complex trait that is not easily bred into plants. Exposure of cultivated tomato to salt stress has been reported to result in increased antioxidant content and activity. Salt tolerance of the related wild species, Solanum pennellii, has also been associated with similar changes in antioxidants. In this work, S. lycopersicum M82, S. pennellii LA716 and a S. pennellii introgression line (IL) population were evaluated for growth and their levels of antioxidant activity (total water-soluble antioxidant activity), major antioxidant compounds (phenolic and flavonoid contents) and antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase and peroxidase) under both control and salt stress (150 mM NaCl) conditions. These data were then used to identify quantitative trait loci (QTL) responsible for controlling the antioxidant parameters under both stress and nonstress conditions. Under control conditions, cultivated tomato had higher levels of all antioxidants (except superoxide dismutase) than S. pennellii. However, under salt stress, the wild species showed greater induction of all antioxidants except peroxidase. The ILs showed diverse responses to salinity and proved very useful for the identification of QTL. Thus, 125 loci for antioxidant content under control and salt conditions were detected. Eleven of the total antioxidant activity and phenolic content QTL matched loci identified in an independent study using the same population, thereby reinforcing the validity of the loci. In addition, the growth responses of the ILs were evaluated to identify lines with favorable growth and antioxidant profiles. Plants have a complex antioxidant response when placed under salt stress. Some loci control antioxidant content under all conditions while others are responsible for antioxidant content only under saline or nonsaline conditions. The localization of QTL for these traits and the identification of lines with specific antioxidant and growth responses may be useful for breeding potentially salt tolerant tomato cultivars having higher antioxidant levels under nonstress and salt stress conditions.


Changes induced by fasting and dietetic obesity in thermogenic parameters of rat brown adipose tissue mitochondrial subpopulations.
The effects of starvation on the thermogenic parameters of three different mitochondrial subpopulations in brown adipose tissue (BAT) of both post-cafeteria obese and lean rats were investigated. Tissue from different BAT depots from fed and 24 h starved rats were collected, pooled and three mitochondrial subpopulations were isolated by differential centrifugation; the M1 fraction (1000 g), the M3 fraction (3000 g) and the M15 fraction (15,000 g). Thermogenic parameters were measured in the three mitochondrial subtypes, and uncoupling protein (UCP) mRNA was determined in BAT. The results showed that starvation induced a decrease in mitochondrial turnover in BAT from both lean and obese rats. Moreover, a selective net loss of UCP from the lightest mitochondrial fraction (M15) in lean rats, with a concomitant reduction of UCP mRNA was observed. The reductions did not occur in obese rats and, as a result, a change in UCP distribution between the mitochondrial subpopulations was produced, with an increase in the M1 mitochondrial subtype. The lack of response of UCP to starvation observed in BAT of obese rats compared with the decrease seen in lean animals, is a consequence of a different mitochondrial subpopulation composition and/or a different response of a particular subpopulation to starvation.


Staple Food Item Availability among Small Retailers in Providence, RI.
Inventory requirements for authorized Supplemental Nutrition Assistance Program (SNAP) retailers have undergone several revisions to increase the availability of healthful foods. A proposed rule of 84 staple food items was not implemented due to concerns that stores would not withstand this expansion, resulting in a final rule requiring 36 items. This study used the Food Access Research Atlas data to characterize food provisions in 30 small retailers in areas with high and low proportions of SNAP and racial minority residents in Providence, Rhode Island (RI). Stores were assessed with an audit instrument to tally variety, perishability, and depth of stock of four staple food categories. Descriptive, analysis of variance, and chi-square analyses were performed. Across stores, 80% were compliant with the final rule, but 66.7% would need to expand their offerings to meet the proposed rule. Mean dairy variety was lowest among all categories (p < 0.05). Most stores met the perishability (92.3%) and depth-of-stock requirements (96.1%) under both rules. No difference was detected between areas with high and low proportions of SNAP and racial minority residents. Future expansion of requirements may increase healthful food availability without imposing undue burdens on retailers in Providence, RI, excluding increased requirements for dairy variety.


Tyrosine Kinase Inhibitors for the Treatment of Chronic-Phase Chronic Myeloid Leukemia: Long-Term Patient Care and Management.
Several tyrosine kinase inhibitors (TKIs) are now approved for the treatment of chronic myeloid leukemia in chronic phase. The efficacy of these drugs has been repeatedly demonstrated, as has their tolerability in most patients. However, late and chronic toxicities become an important issue for many patients facing long-term TKI exposure. For patients on long-term imatinib, gastrointestinal events, fluid retention, muscle cramps, fatigue, and hepatotoxicity are among the most common and most clinically relevant adverse events (AEs). A few of these have also emerged as important AEs with some of the newer TKIs. Distinct long-term toxicity concerns have emerged for dasatinib (pleural effusion, pulmonary hypertension, headache, and dyspnea) and nilotinib (rash, headache, myalgia, alopecia, and hyperglycemia), whereas due to the recent approval of bosutinib and ponatinib, their long-term toxicity profiles have not been fully characterized. Clinical experience with each of these drugs is accumulating, and ensuring proper adherence and monitoring for potential AEs is essential for effective treatment.


Effect of the herbicide atrazine and its metabolite DACT on bovine sperm quality.
Atrazine (ATZ), one of the most extensively used herbicides, is considered a ubiquitous environmental contaminant. ATZ is a known endocrine disruptor, and deleterious effects on reproductive function have been shown, even at low, ecologically relevant doses (0.1-3μg/L). Once it enters the body, ATZ is metabolized to various metabolites, which are further detected in the urine, serum and tissues. In mammals, the major ATZ metabolite is diaminochlorotriazine (DACT). The current study focuses on direct effects of low doses of ATZ and DACT on bovine sperm isolated from ejaculates or epididymis compartments (head, body and tail). Sperm were incubated under capacitation conditions with or without 0.1-10μM ATZ or 1-100μM DACT. The integrity and functionality of sperm membranes (plasma, acrosomal and mitochondrial) were examined simultaneously by fluorescence staining at 0, 2 and 4h of incubation. Acrosome reaction (AR) was induced by Ca++ ionophore, after capacitation. The findings indicated that both ATZ and DACT adversely affect sperm, expressed by damaged sperm membranes. ATZ had a prominent effect on epididymal-tail sperm, expressed as disruption of all examined membranes, mostly at low (0.1 or 1μM) concentrations; pseudo-AR and that induced by Ca++ ionophore were both affected by exposure to 0.1μM ATZ (P<0.05 and P<0.00004, respectively). A similar pattern was documented for sperm isolated from ejaculates (P<0.002 and P<0.001, respectively). ΔYm was affected by ATZ in sperm isolated from the epididymis tail (1μM, P<0.0009), but not in that isolated from ejaculates. DACT reduced sperm viability at all examined concentrations and in all fractions. DACT at 1μM impaired ΔΨm in sperm isolated from the epididymis tail and ejaculate (P<0.005). DACT at 100μM did not induce pseudo-AR in sperm isolated from the ejaculate, but did in sperm isolated from the epididymis tail (P<0.05). Induction of AR by Ca++ ionophore was impaired in sperm isolated from ejaculate and exposed to 10 or 100μM DACT (P<0.05) and in sperm isolated from the epididymis tail and exposed to 1, 10 or 100μM DACT (P<0.0004). These findings reveal the harmful effect of exposure to ATZ and DACT, mainly at low ecologically relevant doses, on sperm viability, AR and mitochondrial function. We conclude that sperm at advanced stages of spermatogenesis, through its passage and storage in the epididymis compartments as well as in the ejaculate, is sensitive to herbicide. The results suggest that ATZ- or DACT-induced disruptions of sperm membranes might impair sperm fertilization competence.


Abnormal strong burn-in degradation of highly efficient polymer solar cells caused by spinodal donor-acceptor demixing.
The performance of organic solar cells is determined by the delicate, meticulously optimized bulk-heterojunction microstructure, which consists of finely mixed and relatively separated donor/acceptor regions. Here we demonstrate an abnormal strong burn-in degradation in highly efficient polymer solar cells caused by spinodal demixing of the donor and acceptor phases, which dramatically reduces charge generation and can be attributed to the inherently low miscibility of both materials. Even though the microstructure can be kinetically tuned for achieving high-performance, the inherently low miscibility of donor and acceptor leads to spontaneous phase separation in the solid state, even at room temperature and in the dark. A theoretical calculation of the molecular parameters and construction of the spinodal phase diagrams highlight molecular incompatibilities between the donor and acceptor as a dominant mechanism for burn-in degradation, which is to date the major short-time loss reducing the performance and stability of organic solar cells.


Nature of the nonequilibrium phase transition in the non-Markovian driven Dicke model.
The Dicke model famously exhibits a phase transition to a superradiant phase with a macroscopic population of photons and is realized in multiple settings in open quantum systems. In this paper, we study a variant of the Dicke model where the cavity mode is lossy due to the coupling to a Markovian environment while the atomic mode is coupled to a colored bath. We analytically investigate this model by inspecting its low-frequency behavior via the Schwinger-Keldysh field theory and carefully examine the nature of the corresponding superradiant phase transition. Integrating out the fast modes, we can identify a simple effective theory allowing us to derive analytical expressions for various critical exponents including the dynamical exponent. We find excellent agreement with previous numerical results when the non-Markovian bath is at zero temperature; however, contrary to these studies, our low-frequency approach reveals that the same exponents govern the critical behavior when the colored bath is at finite temperature unless the chemical potential is zero. Furthermore, we show that the superradiant phase transition is classical in nature, while it is genuinely nonequilibrium. We derive a fractional Langevin equation and conjecture the associated fractional Fokker-Planck equation that captures the system's long-time memory as well as its nonequilibrium behavior. Finally, we consider finite-size effects at the phase transition and identify the finite-size scaling exponents, unlocking a rich behavior in both statics and dynamics of the photonic and atomic observables.


Compromised genomic integrity impedes muscle growth after Atrx inactivation.
ATR-X syndrome is a severe intellectual disability disorder caused by mutations in the ATRX gene. Many ancillary clinical features are attributed to CNS deficiencies, yet most patients have muscle hypotonia, delayed ambulation, or kyphosis, pointing to an underlying skeletal muscle defect. Here, we identified a cell-intrinsic requirement for Atrx in postnatal muscle growth and regeneration in mice. Mice with skeletal muscle-specific Atrx conditional knockout (Atrx cKO mice) were viable, but by 3 weeks of age presented hallmarks of underdeveloped musculature, including kyphosis, 20% reduction in body mass, and 34% reduction in muscle fiber caliber. Atrx cKO mice also demonstrated a marked regeneration deficit that was not due to fewer resident satellite cells or their inability to terminally differentiate. However, activation of Atrx-null satellite cells from isolated muscle fibers resulted in a 9-fold reduction in myoblast expansion, caused by delayed progression through mid to late S phase. While in S phase, Atrx colocalized specifically to late-replicating chromatin, and its loss resulted in rampant signs of genomic instability. These observations support a model in which Atrx maintains chromatin integrity during the rapid developmental growth of a tissue.


The EMBARC European Bronchiectasis Registry: protocol for an international observational study.
Bronchiectasis is one of the most neglected diseases in respiratory medicine. There are no approved therapies and few large-scale, representative epidemiological studies. The EMBARC (European Multicentre Bronchiectasis Audit and Research Collaboration) registry is a prospective, pan-European observational study of patients with bronchiectasis. The inclusion criterion is a primary clinical diagnosis of bronchiectasis consisting of: 1) a clinical history consistent with bronchiectasis; and 2) computed tomography demonstrating bronchiectasis. Core exclusion criteria are: 1) bronchiectasis due to known cystic fibrosis; 2) age <18 years; and 3) patients who are unable or unwilling to provide informed consent. The study aims to enrol 1000 patients by April 2016 across at least 20 European countries, and 10 000 patients by March 2020. Patients will undergo a comprehensive baseline assessment and will be followed up annually for up to 5 years with the goal of providing high-quality longitudinal data on outcomes, treatment patterns and quality of life. Data from the registry will be available in the form of annual reports. and will be disseminated in conference presentations and peer-reviewed publications. The European Bronchiectasis Registry aims to make a major contribution to understanding the natural history of the disease, as well as guiding evidence-based decision making and facilitating large randomised controlled trials.


Fluoro-edenite and carbon nanotubes: The health impact of 'asbestos-like' fibres.
Several decades have passed since Wagner et al demonstrated a causal link between asbestos fibre inhalation and the development of pleural mesothelioma in 1960. It was later suggested that pleural plaques are a benign consequence of exposure to these fibres. Most recently, a significant association between exposure to asbestos and cancer diagnosed at various sites, such as the peritoneum, stomach, pharynx, colon and ovaries has been demonstrated. The great concerns about public health that arose from the scientific evidence presented above have led to the banning of asbestos in several countries. Over the years, the suspicion that particles with a high aspect ratio may have asbestos-like pathogenicity has been supported by increasing evidence. Natural occurring minerals, as well as man-made fibres, have proven capable of inducing either chronic inflammation of serous membranes, or, in some cases, the development of peritoneal and pleural mesothelioma. The pathogenic role of both fluoro-edenite and carbon nanotubes, two 'asbestos-like' fibres is summarized and discussed in this review. The data presented herein support the notion that occupational exposure to these two types of fibre contributes to the development of different types of cancer.


A proton-coupled folate transporter mutation causing hereditary folate malabsorption locks the protein in an inward-open conformation.
The proton-coupled folate transporter (PCFT, SLC46A1) is required for folate intestinal absorption and transport across the choroid plexus. Recent work has identified a F392V mutation causing hereditary folate malabsorption. However, the residue properties responsible for this loss of function remains unknown. Using site-directed mutagenesis, we observed complete loss of function with charged (Lys, Asp, and Glu) and polar (Thr, Ser, and Gln) Phe-392 substitutions and minimal function with some neutral substitutions; however, F392M retained full function. Using the substituted-cysteine accessibility method (with N-biotinyl aminoethyl methanethiosulfonate labeling), Phe-392 mutations causing loss of function, although preserving membrane expression and trafficking, also resulted in loss of accessibility of the substituted cysteine in P314C-PCFT located within the aqueous translocation pathway. F392V function and accessibility of the P314C cysteine were restored by insertion of a G305L (suppressor) mutation. A S196L mutation localized in proximity to Gly-305 by homology modeling was inactive. However, when inserted into the inactive F392V scaffold, function was restored (mutually compensatory mutations), as was accessibility of the P314C cysteine residue. Reduced function, documented with F392H PCFT, was due to a 15-fold decrease in methotrexate influx Vmax, accompanied by a decreased influx Kt (4.5-fold) and Ki (3-fold). The data indicate that Phe-392 is required for rapid oscillation of the carrier among its conformational states and suggest that this is achieved by dampening affinity of the protein for its folate substrates. F392V and other inactivating Phe-392 PCFT mutations lock the protein in its inward-open conformation. Reach (length) and hydrophobicity of Phe-392 appear to be features required for full activity.


Carbamazepine-Responsive Chorea in a Toddler with Semilobar Holoprosencephaly: Case Report.
Holoprosencephaly (HPE) is a central nervous system malformation defined by incomplete separation of the prosencephalon in two hemispheres and determines a broad spectrum of clinical presentations based on extension of non-separation. A 1 year and 8 months' old girl with semilobar HPE and 18p deletion syndrome was admitted to our hospital due to viral bronchiolitis. During hospitalization, she started generalized choreic movements, with face dyskinesia and without any identified aggravating factors. Haloperidol, clonazepam, and valproic acid did not achieve an attenuation of the movement disorder. Significant symptom relief was obtained with the use of trihexyphenidyl, with reduced amplitude and frequency of movements, but hyperthermia compromised its use. Control of chorea with no important side effects was only achieved after the introduction of carbamazepine. Despite significant morbidity, there are few cases described in the literature of chorea and movement disorders in HPE and no effective treatment strategies described. Carbamazepine is an antiepileptic drug that stabilizes voltage-gated sodium channels and is the most effective treatment for paroxysmal kinesigenic dyskinesia. Although it has been used successfully in the treatment of different movement disorders, few therapeutic trials have been reported. The mechanism by which carbamazepine alleviates chorea is still unknown but may be justified through the blocking of post-synaptic dopamine receptors and stimulation of cholinergic pathways.


Molecular cloning and immunolocalization of a novel vertebrate trp homologue from Xenopus.
We report the sequence, structure and distribution of a novel transient receptor potential (trp) homologue from Xenopus, Xtrp, determined by screening an oocyte cDNA library. On the basis of sequence similarity and predicted structure, Xtrp appears to be a homologue of mammalian trp1 proteins. Two polyclonal antibodies raised against distinct regions of the Xtrp sequence revealed Xtrp expression in various Xenopus tissues, and the localization of Xtrp at the plasma membrane of Xenopus oocytes and HeLa cells. Since capacitative calcium entry into Xenopus oocytes has been shown previously to be substantially inhibited by trp1 antisense oligonucleotides [Tomita, Kaneko, Funayama, Kondo, Satoh and Akaike (1998) Neurosci. Lett. 248, 195-198] we suggest that Xtrp may underlie capacitative calcium entry in Xenopus tissues.


Effect of oral administration of nicotinamide mononucleotide on clinical parameters and nicotinamide metabolite levels in healthy Japanese men.
Recent studies have revealed that decline in cellular nicotinamide adenine dinucleotide (NAD+) levels causes aging-related disorders and therapeutic approaches increasing cellular NAD+ prevent these disorders in animal models. The administration of nicotinamide mononucleotide (NMN) has been shown to mitigate aging-related dysfunctions. However, the safety of NMN in humans have remained unclear. We, therefore, conducted a clinical trial to investigate the safety of single NMN administration in 10 healthy men. A single-arm non-randomized intervention was conducted by single oral administration of 100, 250, and 500 mg NMN. Clinical findings and parameters, and the pharmacokinetics of NMN metabolites were investigated for 5 h after each intervention. Ophthalmic examination and sleep quality assessment were also conducted before and after the intervention. The single oral administrations of NMN did not cause any significant clinical symptoms or changes in heart rate, blood pressure, oxygen saturation, and body temperature. Laboratory analysis results did not show significant changes, except for increases in serum bilirubin levels and decreases in serum creatinine, chloride, and blood glucose levels within the normal ranges, independent of the dose of NMN. Results of ophthalmic examination and sleep quality score showed no differences before and after the intervention. Plasma concentrations of N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-5-carboxamide were significantly increased dose-dependently by NMN administration. The single oral administration of NMN was safe and effectively metabolized in healthy men without causing any significant deleterious effects. Thus, the oral administration of NMN was found to be feasible, implicating a potential therapeutic strategy to mitigate aging-related disorders in humans.


Free radicals mediate actions of oxyhemoglobin on cerebrovascular smooth muscle cells.
Single smooth muscle cells were isolated from the basilar artery of the rat by enzymatic dispersion. The membrane properties of the cells were assessed using the patch-electrode voltage-clamp technique, and cell viability was monitored using fluorescein diacetate uptake. Exposure of the cells to oxyhemoglobin (5 microM) resulted in 1) contraction, 2) the appearance of membrane blebs, 3) an increase in the outward potassium currents, 4) a decrease in the membrane resistance, and 5) cell death. In contrast, no effect of oxyhemoglobin on cultured murine neuroblastoma cells was observed. Methemoglobin (100 microM) had no effects on the smooth muscle cells. Catalase (300 units/ml) or dimethyl sulfoxide (0.5%) protected against the effects of oxyhemoglobin; superoxide dismutase (100-1,000 units/ml) provided only partial protection. Exposure of the cells to superoxide anions generated by xanthine (1 mM) plus xanthine oxidase (10 units/l) or to hydrogen peroxide (500 microM) caused an increase in the outward potassium currents without affecting membrane resistance. Generation of hydroxyl radicals by metal ions plus hydrogen peroxide caused the same effects as oxyhemoglobin, that is, an increase in the potassium currents, followed by a decrease in the membrane resistance and cell death. In conclusion, it appears that oxyhemoglobin exerts its effects on vascular smooth muscle cells by the generation of free radicals, chiefly hydroxyl radicals.


The Kruppel-like factor 6 genotype is associated with fibrosis in nonalcoholic fatty liver disease.
Although nonalcoholic fatty liver disease (NAFLD) is increasingly common, only a minority of affected individuals develop fibrotic liver disease. Based on its role in liver growth and repair, we explored whether Kruppel-like factor 6 (KLF6) plays a role in NAFLD progression. KLF6 expression in 31 fibrosis scored NAFLD liver biopsy specimens was assessed by real-time polymerase chain reaction. Transfected minigene constructs were used to study the effect of a polymorphism, KLF6-IVS1-27G>A, that promotes KLF6 alternative splicing in vitro. We genotyped KLF6-IVS1-27G>A in 3 groups of patients (UK group 1, n = 306; Italian group 2, n = 109; trio group 3, n = 61 children and parents). KLF6 expression was increased in association with increased steatosis, inflammation, and fibrosis in NAFLD livers. KLF6-IVS1-27G>A promoted alternative splicing of KLF6 and abrogated the up-regulation of both alpha-smooth muscle actin and collagen 1 in LX-2 cells. Group 1 genotyping identified KLF6-IVS1-27G>A in 44 of 306 (14.4%) patients. Notably, KLF6-IVS1-27G>A was associated significantly with milder NAFLD, with only 25% having more advanced fibrosis compared with 45% of wild-type (wt) individuals. This trend was confirmed in group 2. A linear regression analysis including all 415 patients, adjusted for age, sex, body mass index, and blood glucose level, confirmed that presence of the wt KLF6 allele was an independent predictor of fibrotic NAFLD. Furthermore, we have shown preferential transmission of the wt allele to children with fibrotic NAFLD. We report a functional polymorphism in the KLF6 gene associated with advanced NAFLD and believe further study of KLF6 may enhance our understanding of this disease process.


Full body illusion is associated with widespread skin temperature reduction.
A central feature of our consciousness is the experience of the self as a unified entity residing in a physical body, termed bodily self-consciousness. This phenomenon includes aspects such as the sense of owning a body (also known as body ownership) and has been suggested to arise from the integration of sensory signals from the body. Several studies have shown that temporally synchronous tactile stimulation of the real body and visual stimulation of a fake or virtual body can induce changes in bodily self-consciousness, typically resulting in a sense of illusory ownership over the fake body. The present study assessed the effect of anatomical congruency of visuo-tactile stimulation on bodily self-consciousness. A virtual body was presented and temporally synchronous visuo-tactile stroking was applied simultaneously to the participants' body and to the virtual body. We manipulated the anatomical locations of the visuo-tactile stroking (i.e., on the back, on the leg), resulting in congruent stroking (stroking was felt and seen on the back or the leg) or incongruent stroking (i.e., stroking was felt on the leg and seen on the back). We measured self-identification with the virtual body and self-location as well as skin temperature. Illusory self-identification with the avatar as well as changes in self-location were experienced in the congruent stroking conditions. Participants showed a decrease in skin temperature across several body locations during congruent stimulation. These data establish that the full-body illusion (FBI) alters bodily self-consciousness and instigates widespread physiological changes in the participant's body.


Online GESS: prediction of miRNA-like off-target effects in large-scale RNAi screen data by seed region analysis.
RNA interference (RNAi) is an effective and important tool used to study gene function. For large-scale screens, RNAi is used to systematically down-regulate genes of interest and analyze their roles in a biological process. However, RNAi is associated with off-target effects (OTEs), including microRNA (miRNA)-like OTEs. The contribution of reagent-specific OTEs to RNAi screen data sets can be significant. In addition, the post-screen validation process is time and labor intensive. Thus, the availability of robust approaches to identify candidate off-targeted transcripts would be beneficial. Significant efforts have been made to eliminate false positive results attributable to sequence-specific OTEs associated with RNAi. These approaches have included improved algorithms for RNAi reagent design, incorporation of chemical modifications into siRNAs, and the use of various bioinformatics strategies to identify possible OTEs in screen results. Genome-wide Enrichment of Seed Sequence matches (GESS) was developed to identify potential off-targeted transcripts in large-scale screen data by seed-region analysis. Here, we introduce a user-friendly web application that provides researchers a relatively quick and easy way to perform GESS analysis on data from human or mouse cell-based screens using short interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs), as well as for Drosophila screens using shRNAs. Online GESS relies on up-to-date transcript sequence annotations for human and mouse genes extracted from NCBI Reference Sequence (RefSeq) and Drosophila genes from FlyBase. The tool also accommodates analysis with user-provided reference sequence files. Online GESS provides a straightforward user interface for genome-wide seed region analysis for human, mouse and Drosophila RNAi screen data. With the tool, users can either use a built-in database or provide a database of transcripts for analysis. This makes it possible to analyze RNAi data from any organism for which the user can provide transcript sequences.


Using Constrained Density Functional Theory to Track Proton Transfers and to Sample Their Associated Free Energy Surface.
Ab initio molecular dynamics (AIMD) and quantum mechanics/molecular mechanics (QM/MM) methods are powerful tools for studying proton solvation, transfer, and transport processes in various environments. However, due to the high computational cost of such methods, achieving sufficient sampling of rare events involving excess proton motion-especially when Grotthuss proton shuttling is involved-usually requires enhanced free energy sampling methods to obtain informative results. Moreover, an appropriate collective variable (CV) that describes the effective position of the net positive charge defect associated with an excess proton is essential both for tracking the trajectory of the defect and for the free energy sampling of the processes associated with the resulting proton transfer and transport. In this work, such a CV is derived from first principles using constrained density functional theory (CDFT). This CV is applicable to a broad array of proton transport and transfer processes as studied via AIMD and QM/MM simulations.


Association Between Influenza Infection and Vaccination During Pregnancy and Risk of Autism Spectrum Disorder.
Maternal infections and fever during pregnancy are associated with increased risk for autism spectrum disorders (ASDs). To our knowledge, no study has investigated the association between influenza vaccination during pregnancy and ASD. To investigate the association between influenza infection and vaccination during pregnancy and ASD risk. This cohort study included 196 929 children born at Kaiser Permanente Northern California from January 1, 2000 to December 31, 2010, at a gestational age of at least 24 weeks. Data on maternal influenza infection and vaccination from conception date to delivery date, obtained from Kaiser Permanente Northern California inpatient and outpatient databases. Influenza infection was defined by the International Classification of Diseases, Ninth Revision, Clinical Modification codes or positive influenza laboratory test results. Clinical diagnoses of ASDs identified by International Classification of Diseases, Ninth Revision, Clinical Modification codes 299.0, 299.8, or 299.9 recorded in Kaiser Permanente Northern California electronic medical records on at least 2 occasions any time from birth through June 2015. Within this cohort of 196 929 children, influenza was diagnosed in 1400 (0.7%) mothers and 45 231 (23%) received an influenza vaccination during pregnancy. The mean (SD) ages of vaccinated and unvaccinated women were 31.6 (5.2) and 30.4 (5.6) years, respectively. A total number of 3101 (1.6%) children were diagnosed with ASD. After adjusting for covariates, we found that maternal influenza infection (adjusted hazard ratio, 1.04; 95% CI, 0.68-1.58) or influenza vaccination (adjusted hazard ratio, 1.10; 95% CI, 1.00-1.21) anytime during pregnancy was not associated with increased ASD risk. In trimester-specific analyses, first-trimester influenza vaccination was the only period associated with increased ASD risk (adjusted hazard ratio, 1.20; 95% CI, 1.04-1.39). However, this association could be due to chance (P = 0.1) if Bonferroni corrected for the multiplicity of hypotheses tested (n = 8). Maternal influenza vaccination in the second or third trimester was not associated with increased ASD risk. There was no association between maternal influenza infection anytime during pregnancy and increased ASD risk. There was a suggestion of increased ASD risk among children whose mothers received an influenza vaccination in their first trimester, but the association was not statistically significant after adjusting for multiple comparisons, indicating that the finding could be due to chance. These findings do not call for changes in vaccine policy or practice, but do suggest the need for additional studies on maternal influenza vaccination and autism.


Serial blood water estimations and in-line blood viscometry: the continuous measurement of blood volume during dialysis procedures.
It has been shown in vitro that serial measurements of blood water during ultrafiltration accurately reflect changing blood volume. It has been shown that minute changes in blood hydration produce detectable changes in blood viscosity. An 'in-line' capillary viscometer has been constructed which can be placed in parallel with an extracorporeal circuit to give a continuous semi-quantitative measure of blood viscosity during ultrafiltration or haemodialysis. By making serial measurements of blood water each 'viscometer curve' can be corrected to permit calculation of blood volume provided that the starting blood volume is known. Blood volume changes of less than 1% can be detected in vitro and provided that blood volume changes solely as a result of the removal or influx of water it can be measured continuously to within an accuracy of 4% for volume changes up to 30% irrespective of starting packed cell volume or blood water.


Retinal vascular calibre changes after intravitreal bevacizumab or dexamethasone implant treatment for diabetic macular oedema.
To compare changes in retinal vascular calibre after 2 years of treatment with intravitreal bevacizumab (BVZ) or dexamethasone implant (DEX) in patients with centre-involving diabetic macular oedema (DMO). At baseline, 88 eyes of 61 patients with DMO were recruited in a prospective, multicentre, randomised, single-masked clinical trial. Of these subjects, 22 BVZ-treated (52%) and 22 DEX-treated (48%) eyes of 34 patients (56%) had gradable retinal photographs at both the baseline and 24-month visits. Retinal vascular calibre was measured from digital fundus photographs and summarised as central retinal artery (CRAE) and vein (CRVE) equivalents in all gradable eyes at baseline and 24 months. At 24 months, 40.9% of BVZ and 45.5% of DEX eyes gained 10 or more letters (p=0.77). There was concurrent reduction in mean central macular thickness, -157.7 μm in BVZ and -192.5 μm in DEX-treated eyes (p=0.40). DEX-treated eyes showed a statistically significant reduction in CRVE compared with BVZ-treated eyes, with a mean change from baseline of -31.78 to +4.34 µm, respectively (p<0.001). CRAE showed a non-statistically significant trend towards reduction over time in DEX-treated eyes compared with BVZ-treated eyes, with a mean change from baseline of -6.09 and +1.66, respectively (p=0.077). DEX had a significant narrowing effect on venular diameter in eyes with DMO not seen with BVZ. The changes in retinal vascular calibre suggest that these agents have a differing actions effects retinal vasculature and thereby suggest a potentially different mechanism of action on reducing DMO. NCT01298076.


Host availability drives distributions of fungal endophytes in the imperilled boreal realm.
Boreal forests represent the world's largest terrestrial biome and provide ecosystem services of global importance. Highly imperilled by climate change, these forests host Earth's greatest phylogenetic diversity of endophytes, a hyperdiverse group of symbionts that are defined by their occurrence within living, symptomless plant and lichen tissues. Endophytes shape the ecological and evolutionary trajectories of plants and are therefore key to the function and resilience of terrestrial ecosystems. A critical step in linking the ecological functions of endophytes with those of their hosts is to understand the distributions of these symbionts at the global scale; however, turnover in host taxa with geography and climate can confound insights into endophyte biogeography. As a result, global drivers of endophyte diversity and distributions are not known. Here, we leverage sampling from phylogenetically diverse boreal plants and lichens across North America and Eurasia to show that host filtering in distinctive environments, rather than turnover with geographical or environmental distance, is the main determinant of the community composition and diversity of endophytes. We reveal the distinctiveness of boreal endophytes relative to soil fungi worldwide and endophytes from diverse temperate biomes, highlighting a high degree of global endemism. Overall, the distributions of endophytes are directly linked to the availability of compatible hosts, highlighting the role of biotic interactions in shaping fungal communities across large spatial scales, and the threat that climate change poses to biological diversity and function in the imperilled boreal realm.


OTA Staffs Up.
In the issue of 25 January, p. 291, Science reported that Representative Charles A. Mosher (R-Ohio), vice chairman of the Technology Assessment Board, would become chairman of the board in January 1975. However, by law, any House member of the board may become the chairman. It is likely that a member of the majority party, in this case a Democrat, will be chosen. Edward Wenk, Jr., is chairman of the Committee on Public Engineering Policy of the National Academy of Engineering.


[Downregulation of AKR1B10 gene expression in colorectal cancer].
Colorectal cancer is one of the most common cancers in the world. In our work changes of AKR1B1 and AKR1B10 gene expression levels in colorectal tumors were studied. Their potential diagnostic value was previously shown for several other cancer types. These genes encode aldoso reductases, which belong to the aldo-keto reductases superfamily consisting of enzymes capable to reduce numerous aromatic and aliphatic aldehydes and ketones. They are also involved into retinoid metabolism and cancerogenesis. We have carried out comparative analysis of mRNA levels of AKR1B1 and AKR1B10 genes in paired samples of normal and colorectal tumor tissues using RT-PCR and quantitative PCR. We have shown for the first time the decrease of activity of these genes in colorectal carcinomas. Significant reduction of AKR1B10 mRNA level was detected in the most of tumor samples (88%, 65/74) even at the early stages of malignancy, and in more than 60% of cases this downregulation was much higher than 10 folds. The decrease of AKR1B1 mRNA level was shown in 10% of tumors only. Therefore, we have detected quite different mRNA expression patterns in colorectal cancer for these two structurally similar genes. These data could indicate different functional roles of these two genes in colorectum. The significant decrease of AKR1B10 mRNA in most samples of colorectal cancer could be considered as potential diagnostic marker of this type of cancer.


Development of novel EST-SSR markers in the macaúba palm (Acrocomia aculeata) using transcriptome sequencing and cross-species transferability in Arecaceae species.
The macaúba palm is a novel feedstock for oil production suitable for multiple uses, including as biodiesel and in the food and cosmetic industries. As an efficient alternative, the macaúba palm has limited genomic resources, particularly expressed sequence tag (EST) markers. We report a comprehensive set of validated EST-simple sequence repeat (SSR) markers by using transcriptome sequencing, its application in genetic diversity analysis and cross transferability in other palm trees with environmental and economic importance. In this study, a total of 418 EST-SSRs were identified to be unique for one transcript and region; 232 EST-SSRs were selected, with trinucleotide repeats being the most frequent motif, representing 380 (90.9%), followed by composited (4.5%), di- (3.6%), and hexanucleotides (3.6%). A total of 145 EST-SSRs (62.5%) were validated for consistent amplification in seventeen macaúba palm samples, and 100 were determined to be polymorphic with PIC values ranging from 0.25 to 0.77. Genetic diversity analysis was performed with the 20 most informative EST-SSR markers showing a distinct separation of the different groups of macaúba palm. Additionally, these 145 markers were transferred in six other palm species resulting in transferability rates of 99% (144) in Acrocomia intumescens, 98% (143) in Acrocomia totai, 80.7% (117 EST-EST) in African oil palm (Elaeis guineensis) and peach palm (Bactris gasipaes) samples, 70% (102) in the juçara palm (Euterpe edulis) and 71.7% (104) in the hat palm (Sabal causiarum). Analysis of genetic distance showed a high separation in accordance with geographic location, establishing distinct groups by genera. The EST markers identified in our study are a valuable resource and provide a genomic tool for genetic mapping and further genetic studies, as well as evaluation of co-location between QTLs and functionally associated markers.


Stimulation of peripheral blood mononuclear cells from patients with schistosomiasis mansoni by living and fixed schistosomula, and schistosomular membrane extracts and vesicles.
Several antigenic preparations derived from Schistosoma mansoni schistosomula were compared as immunogens in proliferation assays utilizing peripheral blood mononuclear cells (PBMN) from patients with chronic intestinal schistosomiasis mansoni. Living and fixed schistosomula, either irradiated or nonirradiated, and freshly transformed or 24 hr cultured worms were found to induce very similar proliferative responses of patients' PBMN. Two schistosomulum surface membrane preparations were also compared. Membrane vesicles from schistosomula were found to elicit mean proliferative responses similar to those stimulated by a CaCl2-extracted tegument preparation and to do so at lower total protein concentrations. However, the responses to membrane vesicles were somewhat more variable. When saline extracts from schistosomular bodies submitted to these two treatments were used as stimulants in parallel PBMN cultures it was found that vesiculation more effectively removed antigens involved in the stimulation of PBMN. Broken vesicles were less stimulatory than intact ones. This suggests that the manner in which membrane antigens are presented to antigen processing cells is critical. Incubation of schistosomula at 37 degrees C, but not 4 degrees C in medium containing 10% fetal calf serum from 3 to 48 hr decreased the capacity of the membrane to produce vesicles when treated with high salt solution.


Individualizing and optimizing the use of early warning scores in acute medical care for deteriorating hospitalized patients.
While early warning scores (EWS) have the potential to identify physiological deterioration in an acute care setting, the implementation of EWS in clinical practice has yet to be fully realized. The primary aim of this study is to identify optimal patient-centered rapid response team (RRT) activation rules using electronic medical records (EMR)-derived Markovian models. The setting for the observational cohort study included 38,356 adult general floor patients hospitalized in 2011. The national early warning score (NEWS) was used to measure the patient health condition. Chi-square and Kruskal Wallis tests were used to identify statistically significant subpopulations as a function of the admission type (medical or surgical), frailty as measured by the Braden skin score, and history of prior clinical deterioration (RRT, cardiopulmonary arrest, or unscheduled ICU transfer). Statistical tests identified 12 statistically significant subpopulations which differed clinically, as measured by length of stay and time to re-admission (P < .001). The Chi-square test of independence results showed a dependency structure between subsequent states in the embedded Markov chains (P < .001). The SMDP models identified two sets of subpopulation-specific RRT activation rules for each statistically unique subpopulation. Clinical deterioration experience in prior hospitalizations did not change the RRT activation rules. The thresholds differed as a function of admission type and frailty. EWS were used to identify personalized thresholds for RRT activation for statistically significant Markovian patient subpopulations as a function of frailty and admission type. The full potential of EWS for personalizing acute care delivery is yet to be realized.


A transferable coarse-grained model for hydrogen-bonding liquids.
We present here a recent development of a generalized coarse-grained model for use in molecular simulations. In this model, interactions between coarse-grained particles consist of both van der Waals and explicit electrostatic components. As a result, the coarse-grained model offers the transferability that is lacked by most current effective-potential based approaches. The previous center-of-mass framework (P. A. Golubkov and P. Ren, J. Chem. Phys., 2006, 125, 64103) is generalized here to include arbitrary off-center interaction sites for both Gay-Berne and multipoles. The new model has been applied to molecular dynamic simulations of neat methanol liquid. By placing a single point multipole at the oxygen atom rather than at the center of mass of methanol, there is a significant improvement in the ability to capture hydrogen-bonding. The critical issue of transferability of the coarse-grained model is verified on methanol-water mixtures, using parameters derived from neat liquids without any modification. The mixture density and internal energy from coarse-grained molecular dynamics simulations show good agreement with experimental measurements, on a par with what has been obtained from more detailed atomic models. By mapping the dynamics trajectory from the coarse-grained simulation into the all-atom counterpart, we are able to investigate atomic-level structure and interaction. Atomic radial distribution functions of neat methanol, neat water and mixtures compare favorably to experimental measurements. Furthermore, hydrogen-bonded 6- and 7-molecule chains of water and methanol observed in the mixture are in agreement with previous atomic simulations.


Pesticides and child neurodevelopment.
This review summarizes the recent research on pesticide exposure and child neurobehavioral development with a focus on in-utero exposure to organochlorine and organophosphate pesticides. Recent studies on in-utero exposure to the organochlorine pesticide dichlorodiphenyltrichloroethane and its breakdown product, dichlorodiphenyldichloroethene, indicate that exposure is associated with poorer infant (6 months and older) and child neurodevelopment. Yet, the studies differ on the domain of development that is affected. Research on organophosphate pesticide exposure and neurodevelopment is limited but suggests some negative association of exposure and neurodevelopment at certain ages. Two reports agree that increased levels of organophosphate exposure in utero result in greater numbers of abnormal reflexes in neonates and studies in older infants and young children also point to a negative association with development. In young children (2-3 years) two separate studies observed an increase in maternally reported pervasive developmental disorder with increased levels of organophosphate exposure. Given that the literature suggests a link between organochlorine and in-utero pesticide exposure and impaired child neurodevelopment, clinicians should educate parents about prevention of exposure, especially in populations living in agricultural areas or where household use is common.


SU-E-I-45: Measurement of CT Dose to An HDPE Phantom Using Calorimetry: A Feasibility Study.
Radiation dose in CT is traditionally evaluated using an ionization chamber calibrated in terms of air kerma in a phantom of specific dimensions. The radiation absorbed dose, J/kg, can also be realized directly by measuring the temperature rise in the medium. We investigate using this primary method to determine the CT dose at a point (a few mm), using the recently proposed (APMM TG220) high density polyethylene (HDPE) phantom as a medium. The calorimeter detection scheme is adapted from the second generation NIST water calorimeter using sensitive thermistors in a Wheatstone bridge powered by a lock-in amplifier. The temperature sensitivity is about 3 microK. The expected temperature rise in PE is about 0.6 mK per Gy. The thermistor sensors were placed inside a 26 cm dia. × 10 cm HDPE phantom. Two preliminary tests were made: at a linear accelerator with a 6 MV photon beam, and at a 16-slice CT scanner with a 120 kV beam, each with the thermal sensor and with a calibrated ionization chamber. The 6 MV photon beam with 10 on/off cycles at 60 s each yielded the (uncorrected) run-to-run average dose of 3.06 Gy per cycle (sdm 0.3%), about 8% higher than the Result from the ionization chamber (calibrated in terms of absorbed to water). The CT measurements were also made in the middle section of the TG200 30 cm phantom. Twenty consecutive axial scans at 250 mA, which delivers a nominal accumulated dose (CTDIvol) of 705 mGy in 50 s at three axial and three radial locations were measured. The accumulated dose measured by the ionization chamber at the center of the smaller phantom was 347 mGy. The calorimeter data show qualitative tracking of the chamber measurements. Detailed thermal and electrical analysis of the system are planned to obtain quantitative results.


Suffering from care as expressed in the narratives of former patients in somatic wards.
To illuminate patients' experiences of suffering from care, ten former patients in somatic wards narrated a desirable care episode (n = 39) and an undesirable care episode (n = 51). The interviews were analysed using a phenomenological hermeneutic method inspired by Ricoeur's philosophy. Four themes were found: having a good rest, suffering through, searching for autonomy and being cared for by attentive and committed staff. The findings were interpreted in light of Eriksson's description of suffering, which describes three kinds of suffering: 'suffering of life', 'suffering of illness' and 'suffering of caring'. Although not mentioned explicitly, it was evident that cases of suffering from care were indicated in the patients' narratives. The themes were related to the patients' states of health, their experience of the care situation and their descriptions of themselves, and could be understood as related to the acts of the 'drama of suffering' described by Eriksson. The study highlights the need for the patient to find a co-actor in the drama of suffering in order to prevent suffering from care, i.e. prevent hindrance to the patient in her/his struggle against the 'suffering of illness' and the 'suffering of life'. The patients must be seen as the directors of their own dramas of suffering.


Microgel adhesives for wet cellulose: measurements and modeling.
Nanostructured adhesive layers were prepared by adsorbing and/or grafting polyvinylamine (PVAm) onto carboxylated poly(N-isopropylacrylamide) (PNIPAM) microgels that were then assembled between layers of wet oxidized cellulose. The wet delamination force was measured as functions of PVAm content, PVAm molecular weight, coverage (mass adhesive/joint area), and the distribution of carboxyl groups in the PNIPAM microgels. The use of microgels is attractive because simple physical adsorption onto the cellulose surfaces before lamination gives much higher adhesive content and strength compared to the corresponding adsorbed linear PVAm. Wet adhesion increased with PVAm content in the microgels and the quantity of microgels in the joint whereas adhesion was independent of PVAm molecular weight. Physical adsorption of the PVAm onto/into the microgels gave the same adhesion as covalently coupled PVAm. Finally, the roles of microgel diameter, elasticity, and coverage were simulated by a simple peel adhesion model in which the microgels were treated as ideal springs.


Assessing women's preferences towards tests that may reveal uncertain results from prenatal genomic testing: Development of attributes for a discrete choice experiment, using a mixed-methods design.
Prenatal DNA tests, such as chromosomal microarray analysis or exome sequencing, increase the likelihood of receiving a diagnosis when fetal structural anomalies are identified. However, some parents will receive uncertain results such as variants of uncertain significance and secondary findings. We aimed to develop a set of attributes and associated levels for a discrete-choice experiment (DCE) that will examine parents' preferences for tests that may reveal uncertain test results. A two phase mixed-methods approach was used to develop attributes for the DCE. In Phase 1, a "long list" of candidate attributes were identified via two approaches: 1) a systematic review of the literature around parental experiences of uncertainty following prenatal testing; 2) 16 semi-structured interviews with parents who had experienced uncertainty during pregnancy and 25 health professionals who return uncertain prenatal results. In Phase 2, a quantitative scoring exercise with parents prioritised the candidate attributes. Clinically appropriate levels for each attribute were then developed. A final set of five attributes and levels were identified: likelihood of getting a result, reporting of variants of uncertain significance, reporting of secondary findings, time taken to receive results, and who tells you about your result. These attributes will be used in an international DCE study to investigate preferences and differences across countries. This research will inform best practice for professionals supporting parents to manage uncertainty in the prenatal setting.


Hypoxia-inducible factor-1 (HIF-1) is involved in the regulation of hypoxia-stimulated expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and MCP-5 (Ccl12) in astrocytes.
Neuroinflammation has been implicated in various brain pathologies characterized by hypoxia and ischemia. Astroglia play an important role in the initiation and propagation of hypoxia/ischemia-induced inflammation by secreting inflammatory chemokines that attract neutrophils and monocytes into the brain. However, triggers of chemokine up-regulation by hypoxia/ischemia in these cells are poorly understood. Hypoxia-inducible factor-1 (HIF-1) is a dimeric transcriptional factor consisting of HIF-1alpha and HIF-1beta subunits. HIF-1 binds to HIF-1-binding sites in the target genes and activates their transcription. We have recently shown that hypoxia-induced expression of IL-1beta in astrocytes is mediated by HIF-1alpha. In this study, we demonstrate the role of HIF-1alpha in hypoxia-induced up-regulation of inflammatory chemokines, human monocyte chemoattractant protein-1 (MCP-1/CCL2) and mouse MCP-5 (Ccl12), in human and mouse astrocytes, respectively. Primary fetal human astrocytes or mouse astrocytes generated from HIF-1alpha+/+ and HIF-1alpha+/- mice were subjected to hypoxia (<2% oxygen) or 125 muM CoCl2 for 4 h and 6 h, respectively. The expression of HIF-1alpha, MCP-1 and MCP-5 was determined by semi-quantitative RT-PCR, western blot or ELISA. The interaction of HIF-1alpha with a HIF-1-binding DNA sequence was examined by EMSA and supershift assay. HIF-1-binding sequence in the promoter of MCP-1 gene was cloned and transcriptional activation of MCP-1 by HIF-1alpha was analyzed by reporter gene assay. Sequence analyses identified HIF-1-binding sites in the promoters of MCP-1 and MCP-5 genes. Both hypoxia and HIF-1alpha inducer, CoCl2, strongly up-regulated HIF-1alpha expression in astrocytes. Mouse HIF-1alpha+/- astrocytes had lower basal levels of HIF-1alpha and MCP-5 expression. The up-regulation of MCP-5 by hypoxia or CoCl2 in HIF-1alpha+/+ and HIF-1alpha+/- astrocytes was correlated with the levels of HIF-1alpha in cells. Both hypoxia and CoCl2 also up-regulated HIF-1alpha and MCP-1 expression in human astrocytes. EMSA assay demonstrated that HIF-1 activated by either hypoxia or CoCl2 binds to wild-type HIF-1-binding DNA sequence, but not the mutant sequence. Furthermore, reporter gene assay demonstrated that hypoxia markedly activated MCP-1 transcription but not the mutated MCP-1 promoter in transfected astrocytes. These findings suggest that both MCP-1 and MCP-5 are HIF-1 target genes and that HIF-1alpha is involved in transcriptional induction of these two chemokines in astrocytes by hypoxia.


Statistical security for Social Security.
The financial viability of Social Security, the single largest U.S. government program, depends on accurate forecasts of the solvency of its intergenerational trust fund. We begin by detailing information necessary for replicating the Social Security Administration's (SSA's) forecasting procedures, which until now has been unavailable in the public domain. We then offer a way to improve the quality of these procedures via age- and sex-specific mortality forecasts. The most recent SSA mortality forecasts were based on the best available technology at the time, which was a combination of linear extrapolation and qualitative judgments. Unfortunately, linear extrapolation excludes known risk factors and is inconsistent with long-standing demographic patterns, such as the smoothness of age profiles. Modern statistical methods typically outperform even the best qualitative judgments in these contexts. We show how to use such methods, enabling researchers to forecast using far more information, such as the known risk factors of smoking and obesity and known demographic patterns. Including this extra information makes a substantial difference. For example, by improving only mortality forecasting methods, we predict three fewer years of net surplus, $730 billion less in Social Security Trust Funds, and program costs that are 0.66% greater for projected taxable payroll by 2031 compared with SSA projections. More important than specific numerical estimates are the advantages of transparency, replicability, reduction of uncertainty, and what may be the resulting lower vulnerability to the politicization of program forecasts. In addition, by offering with this article software and detailed replication information, we hope to marshal the efforts of the research community to include ever more informative inputs and to continue to reduce uncertainties in Social Security forecasts.


An update on hepatitis C virus.
Chronic hepatitis caused by hepatitis C virus infection is one of the leading causes of liver cirrhosis and hepatocellular carcinoma globally. Over the last few years, there have been significant advances in our understanding of the immunopathogenesis of this infection, as well as revolutionary developments in the therapeutic armamentarium. The introduction of a number of new oral antiviral agents is already delivering major benefits to patients as they are generally well tolerated and are associated with excellent response rates in the majority. Nonetheless, a number of clinical scenarios remain challenging and research is ongoing in these areas. In this update, we review developments in both basic as well as translational research in the field.