Milk A1 and A2 peptides and diabetes.
Food-derived peptides, specifically those derived from milk, may adversely affect health by increasing the risk of insulin-dependent diabetes. This position is based on the relationship of type 1 diabetes (T1D) and the consumption of variants A1 and B β-casein from cow's milk. It appears that β-casomorphin-7 (BCM-7) from β-casein may function as an immunosuppressant and impair tolerance to dietary antigens in the gut immune system, which, in turn, may contribute to the onset of T1D. There are thirteen genetic variants of β-casein in dairy cattle. Among those variants are A1, A2, and B, which are also found in human milk. The amino acid sequences of β-casomorphins among these bovine variants and those found in human milk are similar, often differing only by a single amino acid. In vitro studies indicate BCM-7 can be produced from A1 and B during typical digestive processes; however, BCM-7 is not a product of A2 digestion. Evidence from several epidemiological studies and animal models does not support the association of milk proteins, even proteins in breast milk, and the development of T1D. Ecological data, primarily based on A1/ A2 variations among livestock breeds, do not demonstrate causation, even among countries where there is considerable dairy consumption.


Late Pleistocene shell midden microstratigraphy indicates a complex history of human-environment interactions in the uplands of North Vietnam.
North Vietnam is situated on a major route of Pleistocene hominin dispersal in East Asia, and the area's karstic caves preserve many prehistoric shell middens. Fossil and genomic evidence suggest a complex human history in this region and more widely across Southeast Asia and southern China, but related archaeological investigations are hampered by challenging site stratigraphies. Recent investigations of shell middens in other geographical settings have revealed the microstratigraphic complexity of these anthropogenic deposits. But caves promote distinctive site formation processes, while tropical climates may catalyse geomorphic and diagenetic changes. These environmental factors complicate the interpretation of North Vietnam's shell middens and constraining their effects upon the formation, preservation and destruction of these sites is critical to understanding the archaeology of this region. We examine two archaeological cave sites, dated to the Late Pleistocene and located in the limestone uplands surrounding the Hanoi Basin. Each contains multiple shell midden layers associated with prehistoric occupation and burials. Using thin-section micromorphology (microstratigraphy), we reconstruct the depositional and post-depositional histories of these sites, presenting a geoarchaeological framework of interpretation that is applicable to shell middens in mainland Southeast Asia and tropical zones more widely. This work represents a further step towards improving our understanding of prehistoric human dispersals and adaptations in this region. This article is part of the theme issue 'Tropical forests in the deep human past'.


Abnormal intranuclear and cytoplasmic formations associated with a chemically induced, transplantable chicken sarcoma.
Thirty GRCH/15 tumors (a 1, 2, 5, 6-dibenzanthracene-induced chicken sarcoma) were examined in the light and the electron microscope. Associated with the sarcoma were two types of abnormal intranuclear lesions, one in the form of a vacuole, the other as an aggregate containing glycogen. In the electron microscope, one type of lesion observed showed an organized microfibrillar structure. Abnormal cytoplasmic formations occurred as massed clusters of thread-like or tubular material, which gave rise to small bodies with concentric shell structure; similar bodies were found associated with vacuoles.


Boosting cortical activity at Beta-band frequencies slows movement in humans.
Neurons have a striking tendency to engage in oscillatory activities. One important type of oscillatory activity prevalent in the motor system occurs in the beta frequency band, at about 20 Hz. It is manifest during the maintenance of tonic contractions and is suppressed prior to and during voluntary movement. This and other correlative evidence suggests that beta activity might promote tonic contraction, while impairing motor processing related to new movements. Hence, bursts of beta activity in the cortex are associated with a strengthening of the motor effects of sensory feedback during tonic contraction and with reductions in the velocity of voluntary movements. Moreover, beta activity is increased when movement has to be resisted or voluntarily suppressed. Here we use imperceptible transcranial alternating-current stimulation to entrain cortical activity at 20 Hz in healthy subjects and show that this slows voluntary movement. The present findings are the first direct evidence of causality between any physiological oscillatory brain activity and concurrent motor behavior in the healthy human and help explain how the exaggerated beta activity found in Parkinson's disease can lead to motor slowing in this illness.


Manganese-enhanced magnetic resonance imaging detects declining pancreatic β-cell mass in a cyclophosphamide-accelerated mouse model of type 1 diabetes.
Currently, there is no ideal noninvasive method to quantify the progressive loss of pancreatic β-cell mass (BCM) that occurs in type 1 diabetes. Magnetic resonance imaging has detected gross differences in BCM between healthy and diabetic mice using the contrast agent manganese, which labels functional β-cells and increases the water proton relaxation rate (R1), but its ability to measure gradations in BCM during disease progression is unknown. Our objective was to test the hypothesis that measurements of the manganese-enhanced pancreatic R1 could detect decreasing BCM in a mouse model of type 1 diabetes. We used cyclophosphamide-accelerated BDC2.5 T-cell receptor transgenic nonobese diabetic mice, which experience development of type 1 diabetes during a 7-day time period after cyclophosphamide injection, whereas transgene-negative mice do not. We measured the manganese-enhanced pancreatic R1 before cyclophosphamide injection (day 0) and on days 3, 4, 5, and 7 afterward. Pancreatic R1 remained constant in transgene-negative mice and decreased stepwise day-to-day in transgene-positive mice, mirroring their loss of BCM, confirmed by pancreatic insulin measurements and histology. Changes in R1 in transgene-positive mice occurred before elevations in blood glucose, a clinical indicator of diabetes, suggesting potential for early noninvasive detection of changes in functional BCM.


Rapid Synaptogenesis in the Nucleus Accumbens Is Induced by a Single Cocaine Administration and Stabilized by Mitogen-Activated Protein Kinase Interacting Kinase-1 Activity.
Repeated cocaine exposure produces new spine formation in striatal projection neurons (SPNs) of the nucleus accumbens. However, an acute exposure to cocaine can trigger long-lasting synaptic plasticity in SPNs leading to behavioral alterations. This raises the intriguing question as to whether a single administration of cocaine could enduringly modify striatal connectivity. A three-dimensional morphometric analysis of presynaptic glutamatergic boutons and dendritic spines was performed on SPNs 1 hour and 1 week after a single cocaine administration. Time-lapse two-photon microscopy in adult slices was used to determine the precise molecular-events sequence responsible for the rapid spine formation. A single injection triggered a rapid synaptogenesis and persistent increase in glutamatergic connectivity in SPNs from the shell part of the nucleus accumbens, specifically. Synapse formation occurred through clustered growth of active spines contacting pre-existing axonal boutons. Spine growth required extracellular signal-regulated kinase activation, while spine stabilization involved transcription-independent protein synthesis driven by mitogen-activated protein kinase interacting kinase-1, downstream from extracellular signal-regulated kinase. The maintenance of new spines driven by mitogen-activated protein kinase interacting kinase-1 was essential for long-term connectivity changes induced by cocaine in vivo. Our study originally demonstrates that a single administration of cocaine is able to induce stable synaptic rewiring in the nucleus accumbens, which will likely influence responses to subsequent drug exposure. It also unravels a new functional role for cocaine-induced extracellular signal-regulated kinase pathway independently of nuclear targets. Finally, it reveals that mitogen-activated protein kinase interacting kinase-1 has a pivotal role in cocaine-induced connectivity.


Understanding Bland Altman analysis.
In a contemporary clinical laboratory it is very common to have to assess the agreement between two quantitative methods of measurement. The correct statistical approach to assess this degree of agreement is not obvious. Correlation and regression studies are frequently proposed. However, correlation studies the relationship between one variable and another, not the differences, and it is not recommended as a method for assessing the comparability between methods. In 1983 Altman and Bland (B&A) proposed an alternative analysis, based on the quantification of the agreement between two quantitative measurements by studying the mean difference and constructing limits of agreement. The B&A plot analysis is a simple way to evaluate a bias between the mean differences, and to estimate an agreement interval, within which 95% of the differences of the second method, compared to the first one, fall. Data can be analyzed both as unit differences plot and as percentage differences plot. The B&A plot method only defines the intervals of agreements, it does not say whether those limits are acceptable or not. Acceptable limits must be defined a priori, based on clinical necessity, biological considerations or other goals. The aim of this article is to provide guidance on the use and interpretation of Bland Altman analysis in method comparison studies.


Pathways involved in targeting and secretion of a lysosomal enzyme in Dictyostelium discoideum.
In Dictyostelium discoideum, the lysosomal enzyme alpha-mannosidase is first synthesized as an N-glycosylated precursor of Mr 140,000. After a 20-30-min lag period, up to 30% of the precursor molecules are rapidly secreted, whereas the rest remain cellular and are proteolytically processed (t 1/2 = 8 min) to mature subunits of Mr 58,000 and 60,000. The secreted precursor is modified more extensively than the cellular form, as is revealed by differences in size, charge, and sensitivity to endoglycosidase H. Subcellular fractionation has shown that, following synthesis in the rough endoplasmic reticulum, the precursor is transported to a low density membrane fraction that contains Golgi membranes. Proteolytic processing takes place in these vesicles, since newly cleaved mature enzyme, but no precursor, co-fractionates with lysosomes. Under conditions that disrupt vesicular membranes, the precursor remains associated with the membrane fraction, whereas the newly processed mature enzyme is soluble. Proteolytic cleavage of the precursor thus coincides with the release of the mature enzyme into the lumen of a lysosomal compartment. These findings suggest a possible mechanism for lysosomal targeting that involves the specific association of enzyme precursors with Golgi membranes.


Functional requirement for human pitrilysin metallopeptidase 1 arginine 183, mutated in amyloidogenic neuropathy.
Here we report the enzymologic characterization of recombinant human pitrilysin metallopeptidase 1 (Pitrm1) and derivative mutants including the arginine-to-glutamine substitution mutant Pitrm1 R183Q, which has been implicated in inherited amyloidogenic neuropathy. Recombinant Pitrm1 R183Q was readily expressed in and purified from Escherichia coli, but was less active than the recombinant wild-type enzyme against recombinant amyloid beta-peptide (Aβ 1-40). A novel fluorogenic substrate derived from the reported Aβ 1-40 core peptide cleavage sequence, Mca-KLVFFAEDK-(Dnp)-OH, was synthesized and applied to real-time kinetic study of Pitrm1 and derivative mutants including Pitrm1 R183Q. The Pitrm1 R183Q mutant exhibited significantly decreased rate of fluorogenic peptide hydrolysis, yet retained similar binding affinity by comparison with the wild-type enzyme. Targeted mutagenic analysis revealed a functional requirement for uncharged or electropositive residues in place of Pitrm1 R183. Residue R183 is positioned within an N-terminal strand-loop-strand motif that is conserved among M16C, but not M16A or M16B family metallopeptidases. Truncation analysis revealed that this strand-loop-strand motif inclusive of residue R183 is essential Pitrm1 function. A requirement for charged residues within 4.5 Å of residue R183 was demonstrated, and Pitrm1 R183Q was found to exhibit increased sensitivity to heat inactivation. Our findings indicate that charge sharing in the vicinity of Pitrm1 R183 is critical to enzyme activity, providing potential insight into a molecular basis of Pitrm1 dysfunction.


Acupuncture applied as a sensory discrimination training tool decreases movement-related pain in patients with chronic low back pain more than acupuncture alone: a randomised cross-over experiment.
High-quality clinical evidence suggests that although acupuncture appears superior to usual care in the management of chronic low back pain, there is little meaningful difference between true and sham acupuncture. This suggests that the benefits of acupuncture are mediated by the placebo response. An alternative explanation is that sham acupuncture is an active treatment and shares a mechanism of action with traditionally applied acupuncture. One plausible candidate for this mechanism is improvement in self-perception mediated through the sensory discrimination-like qualities of acupuncture. We aimed to compare the effects of acupuncture with a sensory discrimination training component to acupuncture without. 25 people with chronic low back pain were enroled in a randomised cross-over experiment. We compared the effect of acupuncture delivered when sensory discrimination is optimised to acupuncture delivered when it is not, on movement-related back pain immediately after each intervention. We found that the average pain intensity after participants had received acupuncture with sensory discrimination training (2.8±2.5) was less than when they received acupuncture without sensory discrimination training (3.6±2.0). This difference was statistically significant (after adjustment; mean difference=-0.8, 95% CI -1.4 to -0.3; p=0.011). Our findings are consistent with the idea that acupuncture may offer specific benefit that is not dependent on precisely where the needles are inserted so much as that the patient attends to where they are inserted. If so, the location of the needles might be better focused on the painful area and the need for penetration of the skin may be mitigated.


Phorbol 12-myristate 13-acetate triggers the protein kinase A-mediated phosphorylation and activation of the PDE4D5 cAMP phosphodiesterase in human aortic smooth muscle cells through a route involving extracellular signal regulated kinase (ERK).
Phosphodiesterase 4D5 is the sole PDE4D cAMP phosphodiesterase isoform expressed in human aortic smooth muscle cells (HASMC). Phorbol 12-myristate 13-acetate (PMA) challenge of HASMC rapidly activated PDE4D5 through a process ablated by the mitogen-activated protein kinase kinase inhibitor PD98059. PMA elicited an inhibitory effect on PDE4D5 activity in HASMC treated with the cyclooxygenase (COX) inhibitor indomethacin, the COX-2 selective inhibitor NS-398, the phospholipase A(2) inhibitor quinacrine, and the cAMP-dependent protein kinase A (PKA) inhibitor H89. PMA challenge of COS-1 cells elicited the rapid inhibition and phosphorylation of both recombinant and endogenous PDE4D5 in a manner ablated by PD98059 and not seen in S651A mutant PDE4D5. PMA promoted the generation of PGE(2) in the medium of HASMC and caused activation of both extracellular signal-regulated kinase (ERK) and PKA through a process ablated by indomethacin, NS-398, quinacrine, and PD98059. Exogenous prostaglandin (PG) E(2) increased cAMP levels and activated PKA in HASMC. COX-2 was expressed in HASMC but not in COS-1 cells. Forskolin challenge of COS-1 cells activated PDE4D5 by causing the PKA-mediated phosphorylation of Ser126 as detected using a novel phosphospecific antiserum. PMA challenge of HASMC elicited phosphorylation of the stimulatory PKA-specific phosphorylation site, Ser126 in PDE4D5 in a manner ablated by PD98059, indomethacin, and H89. We propose that, in HASMC, PMA activates PDE4D5 through an ERK-controlled autocrine mechanism. This involves PGE(2) generation, which causes activation of adenylyl cyclase, allowing PKA to elicit net activation of PDE4D5 by phosphorylation at Ser126.


Sea-change or change challenge? Health information access in developing countries: The U.S. National Library of Medicine experience.
Health professionals in developing countries want access to information to help them make changes in health care and contribute to medical research. However, they face challenges of technology limitations, lack of training, and, on the village level, culture and language. This report focuses on the U.S. National Library of Medicine experience with access: for the international medical/scientific community to health information which has been published by researchers in developing countries; for scientists and clinicians in developing countries to their own literature and to that of their colleagues around the world; for medical librarians who are a critical conduit for students, faculty, researchers, and, increasingly, the general public; and for the front line workers at the health center in the village at the end of the line. The fundamental question of whether or not information communication technology can make a difference in access and subsequently in health is illustrated by an anecdote regarding an early intervention in Africa in 1992. From that point, we examine programs to improve access involving malaria researchers, medical journal editors, librarians, and medical students working with local health center staff in the village. Although access is a reality, the positive change in health that the information technology intervention might produce often remains a mirage. Information and technology are not static elements in the equation for better access. They must function together, creating a dialectic in which they transform and inform one another and those whom their combination touches.


Trauma exposure and endothelial function among midlife women.
Trauma is a potent exposure that can have implications for health. However, little research has considered whether trauma exposure is related to endothelial function, a key process in the pathophysiology of cardiovascular disease (CVD). We tested whether exposure to traumatic experiences was related to poorer endothelial function among midlife women, independent of CVD risk factors, demographic factors, psychosocial factors, or a history of childhood abuse. In all, 272 nonsmoking perimenopausal and postmenopausal women aged 40 to 60 years without clinical CVD completed the Brief Trauma Questionnaire, the Child Trauma Questionnaire, physical measures, a blood draw, and a brachial ultrasound for assessment of brachial artery flow-mediated dilation (FMD). Relations between trauma and FMD were tested in linear regression models controlling for baseline vessel diameter, demographics, depression/anxiety, CVD risk factors, health behaviors, and, additionally, a history of childhood abuse. Over 60% of the sample had at least one traumatic exposure, and 18% had three or more exposures. A greater number of traumatic exposures was associated with lower FMD, indicating poorer endothelial function in multivariable models (beta, β [standard error, SE] -1.05 [0.40], P = 0.01). Relations between trauma exposure and FMD were particularly pronounced for three or more trauma exposures (b [SE] -1.90 [0.71], P = 0.008, relative to no exposures, multivariable). A greater number of traumatic exposures were associated with poorer endothelial function. Relations were not explained by demographics, CVD risk factors, mood/anxiety, or a by history of childhood abuse. Women with greater exposure to trauma over life maybe at elevated CVD risk.


Students' individual interest in physical education: Development and validation of a questionnaire.
Identifying the specificity of students' individual interest in physical education is necessary to capture the different facets of this construct. In contrast to existing questionnaires in education which assess the multidimensionality of individual interest, the current scale used in physical education consists of a single-item measurement which rates students' individual interest for multiple physical activities taught during lessons. Even if this single-item rating provides a basis for classifying interest relative to others, it does not provide information about the nature and internal components of individual interest. Therefore, the purpose of this study was to develop a valid and reliable questionnaire measuring the multidimensionality of students' individual interest in physical education. Four stages were followed: (1) the development of a preliminary version based on the findings of a literature review on individual interest and a committee process validation; (2) a first study conducted on 481 secondary school students to establish the factor structure, and internal reliability of this questionnaire; (3) a second study based on 243 secondary school students to confirm its factor structure and to test its temporal stability; and finally, (4) a third study with 253 students to test its concurrent validity. The final form of the questionnaire consists of a three-factor structure based on 14 items that measure students' individual interest in physical education: positive affect and willingness to reengage (five items), stored utility value (four items), and stored attainment value and knowledge-seeking intentions (five items).


Promotion of lymphocyte egress into blood and lymph by distinct sources of sphingosine-1-phosphate.
Lymphocytes require sphingosine-1-phosphate (S1P) receptor-1 to exit lymphoid organs, but the source(s) of extracellular S1P and whether S1P directly promotes egress are unknown. By using mice in which the two kinases that generate S1P were conditionally ablated, we find that plasma S1P is mainly hematopoietic in origin, with erythrocytes a major contributor, whereas lymph S1P is from a distinct radiation-resistant source. Lymphocyte egress from thymus and secondary lymphoid organs was markedly reduced in kinase-deficient mice. Restoration of S1P to plasma rescued egress to blood but not lymph, and the rescue required lymphocyte expression of S1P-receptor-1. Thus, separate sources provide S1P to plasma and lymph to help lymphocytes exit the low-S1P environment of lymphoid organs. Disruption of compartmentalized S1P signaling is a plausible mechanism by which S1P-receptor-1 agonists function as immunosuppressives.


Scoping studies: advancing the methodology.
Scoping studies are an increasingly popular approach to reviewing health research evidence. In 2005, Arksey and O'Malley published the first methodological framework for conducting scoping studies. While this framework provides an excellent foundation for scoping study methodology, further clarifying and enhancing this framework will help support the consistency with which authors undertake and report scoping studies and may encourage researchers and clinicians to engage in this process. We build upon our experiences conducting three scoping studies using the Arksey and O'Malley methodology to propose recommendations that clarify and enhance each stage of the framework. Recommendations include: clarifying and linking the purpose and research question (stage one); balancing feasibility with breadth and comprehensiveness of the scoping process (stage two); using an iterative team approach to selecting studies (stage three) and extracting data (stage four); incorporating a numerical summary and qualitative thematic analysis, reporting results, and considering the implications of study findings to policy, practice, or research (stage five); and incorporating consultation with stakeholders as a required knowledge translation component of scoping study methodology (stage six). Lastly, we propose additional considerations for scoping study methodology in order to support the advancement, application and relevance of scoping studies in health research. Specific recommendations to clarify and enhance this methodology are outlined for each stage of the Arksey and O'Malley framework. Continued debate and development about scoping study methodology will help to maximize the usefulness and rigor of scoping study findings within healthcare research and practice.


Increasing lysine content of waxy maize through introgression of opaque-2 and opaque-16 genes using molecular assisted and biochemical development.
The low lysine content of waxy maize cannot meet the nutritional requirements of humans, livestock, or poultry. In the present study, the high-lysine genes o2 and o16 were backcrossed into wx lines using the maize high-lysine inbreds TAIXI19 (o2o2) and QCL3021 (o16o16) as donors and the waxy maize inbred line QCL5019 (wxwx) as a receptor. In the triple-cross F1, backcross, and inbred generations, the SSR markers phi027 and phi112 within the wx and o2 genes and the SSR marker umc1121 linked to the o16 gene were used for foreground selection. Background selection of the whole-genome SSR markers was performed for the selected individuals. The grain lysine content was determined using the dye-binding lysine method. The waxiness of the grain was determined with the I2-KI staining and dual-wavelength spectrophotometric analysis. The BC2F2 generation included 7 plants of genotype wxwxo2o2O16_, 19 plants of genotype wxwxo16o16O2_, and 3 plants of genotype wxwxo2o2o16o16. In these seeds, the average amylopectin content was 96.67%, 96.87%, and 96.62%, respectively, which is similar to that of QCL5019. The average lysine content was 0.555%, 0.380%, and 0.616%, respectively, representing increases of 75.1%, 19.9%, 94.3%, respectively, over QCL5019. The average genetic background recovery rate of the BC2F3 families was 95.3%, 94.3%, 94.2%, respectively. Among these 3 wxwxo2o2O16O16 families, 4 wxwxo2o2O16o16 families, and 3 wxwxo2o2o16o16 families, the longest imported parent donor fragment was 113.35 cM and the shortest fragment was 11.75 cM. No significant differences in lysine content were found between the BC2F4 seeds and the BC2F3 seeds in these 10 families. This allowed us to increase the lysine content of waxy corn and produce seeds with excellent nutritional characteristics suitable for human consumption, animal feed, and food processing. This may be of significance in the breeding of high-quality corn and in improvement of the nutrition of humans, livestock, and poultry.


Microbial community composition and glyphosate degraders of two soils under the influence of temperature, total organic carbon and pH.
Glyphosate can be degraded by soil microorganisms rapidly and is impacted by temperature and soil properties. Enhanced temperature and total organic carbon (TOC) as well as reduced pH increased the rate of 13C315N-glyphosate conversion to CO2 and biogenic non-extractable residues (bioNERs) in a Haplic Chernozem (Muskus et al., 2019) and in a Humic Cambisol (Muskus et al., 2020). To date; however, the combined effect of temperature and TOC or pH on microbial community composition and glyphosate degraders in these two soils has not been investigated. Phospholipid fatty acid [PLFA] biomarker analysis combined with 13C labeling was employed to investigate the effect of two soil properties (pH, TOC) and of three temperatures (10 °C, 20 °C, 30 °C) on soil microorganisms. Before incubation, the properties of a Haplic Chernozem and a Humic Cambisol were adjusted to obtain five treatments: (a) Control (Haplic Chernozem: 2.1% TOC and pH 6.6; Humic Cambisol: 3% TOC and pH 7.0), (b) 3% TOC (Haplic Chernozem) or 4% TOC (Humic Cambisol), (c) 4% TOC (Haplic Chernozem) or 5% TOC (Humic Cambisol), (d) pH 6.0 (Haplic Chernozem) or pH 6.5 (Humic Cambisol), and (e) pH 5.5 for both soils. All treatments were amended with 50 mg kg-1 glyphosate and incubated at 10 °C, 20 °C or 30 °C. We observed an increase in respiration, microbial biomass and glyphosate mineralization with incubation temperature. Although respiration and microbial biomass in the Humic Cambisol was higher, the microorganisms in the Haplic Chernozem were more active in glyphosate degradation. Increased TOC shifted the microbiome and the 13C-glyphosate degraders towards Gram-positive bacteria in both soils. However, the abundance of 13C-PLFAs indicative for the starvation of Gram-negative bacteria increased with increasing TOC or decreasing pH at higher temperatures. Gram-negative bacteria thus may have been involved in earlier stages of glyphosate degradation.


Psychological adjustment after breast cancer: a systematic review of longitudinal studies.
Breast cancer (BC) can be a traumatic and stressful experience for women, but there are wide-ranging differences in the ways in which women respond and adapt to BC. This systematic review examines which sociodemographic, disease-related, and psychosocial factors near diagnosis predict later psychological adjustment to BC. Database searches were conducted in 9 different health-related databases from 2000 to December 2015 using relevant search terms. Full-text, peer-reviewed articles in English that analyzed potential predictors of psychological adjustment in longitudinal studies were considered for inclusion. Of 1780 abstracts, 41 studies fulfilled inclusion criteria. Consistent sociodemographic and disease-related variable predictors of adjustment were income, fatigue, cancer stage, and physical functioning. Psychosocial factors, particularly optimism and trait anxiety, as well as perceived social support, coping strategies, and initial levels of psychological functioning, were found to be predictive of later depressive and anxiety symptoms, psychological distress, and quality of life for women with BC, in predictable ways. Other psychosocial variables, such as cognitive and body image factors, predicted psychological adjustment but were explored only by a few studies. The majority of studies showed a significant relationship between psychosocial factors and psychological adjustment. These results point to specific sociodemographic, disease-related, and psychosocial factors that can help to identify women at the time of diagnosis who are at risk for long-term psychological challenges so they can be referred for psychological support that targets their specific needs and can improve their quality of life and mood and decrease indicators of anxiety, depression, and psychological distress.


Ozone inactivation of Cryptosporidium parvum in demand-free phosphate buffer determined by in vitro excystation and animal infectivity.
Inactivation of Cryptosporidium parvum oocysts by ozone was performed in ozone demand-free 0.05 M phosphate buffer (pH 6.9) in bench-scale batch reactors at 7 and 22 degrees C. Ozone was added to each trial from a concentrated stock solution for contact times ranging from 5 to 15 min. The viability of the control and treated oocysts was determined by using in vitro excystation and infection in neonatal CD-1 mice. It was found that excystation consistently underestimated inactivation when compared with animal infectivity (P < or = 0.05). As inactivations increased, the difference between excystation and infectivity also increased. The inactivation kinetics of C. parvum by ozone deviated from the simple first-order Chick-Watson model and was better described by a nonlinear Hom model. The use of the Hom model for predicting inactivation resulted in a family of unique concentration and time values for each inactivation level rather than the simple CT product of the Chick-Watson model.


Epigenomic signature of adrenoleukodystrophy predicts compromised oligodendrocyte differentiation.
Epigenomic changes may either cause disease or modulate its expressivity, adding a layer of complexity to mendelian diseases. X-linked adrenoleukodystrophy (X-ALD) is a rare neurometabolic condition exhibiting discordant phenotypes, ranging from a childhood cerebral inflammatory demyelination (cALD) to an adult-onset mild axonopathy in spinal cords (AMN). The AMN form may occur with superimposed inflammatory brain demyelination (cAMN). All patients harbor loss of function mutations in the ABCD1 peroxisomal transporter of very-long chain fatty acids. The factors that account for the lack of genotype-phenotype correlation, even within the same family, remain largely unknown. To gain insight into this matter, here we compared the genome-wide DNA methylation profiles of morphologically intact frontal white matter areas of children affected by cALD with adult cAMN patients, including male controls in the same age group. We identified a common methylomic signature between the two phenotypes, comprising (i) hypermethylation of genes harboring the H3K27me3 mark at promoter regions, (ii) hypermethylation of genes with major roles in oligodendrocyte differentiation such as MBP, CNP, MOG and PLP1 and (iii) hypomethylation of immune-associated genes such as IFITM1 and CD59. Moreover, we found increased hypermethylation in CpGs of genes involved in oligodendrocyte differentiation, and also in genes with H3K27me3 marks in their promoter regions in cALD compared with cAMN, correlating with transcriptional and translational changes. Further, using a penalized logistic regression model, we identified the combined methylation levels of SPG20, UNC45A and COL9A3 and also, the combined expression levels of ID4 and MYRF to be good markers capable of discriminating childhood from adult inflammatory phenotypes. We thus propose the hypothesis that an epigenetically controlled, altered transcriptional program may drive an impaired oligodendrocyte differentiation and aberrant immune activation in X-ALD patients. These results shed light into disease pathomechanisms and uncover putative biomarkers of interest for prognosis and phenotypic stratification.


[Analysis of heat loss using inhalation agents in rats subjected to laparotomy and increased intra-abdominal pressure, using digital infrared thermal image].
Anesthesiology involves the handling of situations inherent to anesthetic and surgical procedures which lead to patients thermal homeostasis unbalance, with noxious physiological effects. This study aimed at qualifying and quantifying thermal redistribution in rats subjected to inhalation anesthesia, during induction and in surgical situations of laparotomy and increased intra-abdominal pressure. The study involved 90 rats, submitted to inhalation anesthesia, which were distributed in three groups: halothane; isoflurane; sevoflurane. Each group was divided in subgroups: I - control; II - median laparotomy with bowel exposure; III - 15 mmHg Increase in intra-abdominal pressure. Heat loss was measured by an esophageal probe and infrared thermal image. There were no significant differences among inhalation anesthetics regarding heat loss between groups I and II. In group III, there was a difference between isoflurane and sevoflurane and isoflurane was responsible for the highest heat loss.


Survey study of members of the Canadian Orthopaedic Association on the natural history and treatment of anterior cruciate ligament injury.
To describe current preferences and opinions of members of the Canadian Orthopaedic Association (COA) pertaining to anterior cruciate ligament (ACL) reconstruction. Survey study. All orthopedic surgeon members of the COA residing in Canada were invited to participate. The primary measure was a survey completed via an Internet-based survey manager. It was composed of 30 questions on the natural history of ACL-injured knees, and surgical and postsurgical treatment choices. Two hundred eighty-three surgeons (50%) responded to the survey. One hundred forty-four performed ACL reconstruction in the past year. In terms of natural history, the only area of agreement was that hamstring and quadricep strength affects function in ACL-deficient knees (92%). A majority of surgeons indicated preference for hamstring autograft (73%), transtibial versus anteromedial portal for establishment of the femoral tunnel (70% vs 28%), and promotion of full weight bearing and range of motion immediately after surgery (72% and 75%, respectively). The most frequent surgeon-reported complication was tunnel widening (10%). A greater proportion of high-volume surgeons permitted earlier return to sport (P < 0.008). In the rapidly evolving area of ACL reconstruction, no recent surveys of opinions and preferences of Canadian orthopedic surgeons have been published. In addition to providing information from a Canadian perspective, the findings from this study will allow surgeons to evaluate a range of treatment decisions based on the general opinions of their colleagues and also highlights areas of dissimilarity that can be targeted for more extensive research.


The spleen of patients with myelofibrosis harbors defective mesenchymal stromal cells.
Splenic hematopoiesis is a major feature in the course of myelofibrosis (MF). In fact, the spleen of patients with MF contains malignant hematopoietic stem cells retaining a complete differentiation program, suggesting both a pivotal role of the spleen in maintaining the disease and a tight regulation of hematopoiesis by the splenic microenvironment, in particular by mesenchymal stromal cells (MSCs). Little is known about splenic MSCs (Sp-MSCs), both in normal and in pathological context. In this work, we have in vitro expanded and characterized Sp-MSCs from 25 patients with MF and 13 healthy subjects (HS). They shared similar phenotype, growth kinetics, and differentiation capacity. However, MF Sp-MSCs expressed significant lower levels of nestin, and favored megakaryocyte (Mk) differentiation in vitro at a larger extent than their normal counterpart. Moreover, they showed a significant upregulation of matrix metalloprotease 2 (MMP2) and fibronectin 1 (FN1) genes both at mRNA expression and at protein level, and, finally, developed genetic abnormalities which were never detected in HS-derived Sp-MSCs. Our data point toward the existence of a defective splenic niche in patients with MF that could be responsible of some pathological features of the disease, including the increased trafficking of CD34+ cells and the expansion of the megakaryocytic lineage.


Model of associations between psychosocial variables and health-outcome measures of adolescents with IDDM.
The purpose of this study was to develop a model that describes the contributions of key psychosocial variables to the health outcome of adolescents with insulin-dependent diabetes mellitus (IDDM). Subjects were 93 adolescents with IDDM and their parents. Health-outcome measures included adherence and metabolic control (HbA1c). Psychosocial variables included adolescent age, chronic life stress, social competence, family relations, and family knowledge about IDDM. Multiple regression analyses showed that adherence (P less than .029) and stress (P less than .052) were directly related to metabolic control and that knowledge about IDDM (P less than .029), family relations (P less than .099), and adolescent age (P less than .086) had direct effects on adherence. Combined, the independent variables accounted for 14.5% of the variance in predicting HbA1c and 18.5% of the variance in predicting adherence. In general, these findings are consistent with extant theory. The direct link between stress and metabolic control, however, contrasts with the current view that psychosocial variables affect metabolic control indirectly through their influence on adherence behavior. The methodological limitations of the findings are noted, directions for future research are suggested, and the implications for clinical interventions are described.


Rapid detection of drug resistance and mutational patterns of extensively drug-resistant strains by a novel GenoType® MTBDRsl assay.
The emergence of extensively drug-resistant tuberculosis (XDR-TB) is a major concern in the India. The burden of XDR-TB is increasing due to inadequate monitoring, lack of proper diagnosis, and treatment. The GenoType ® Mycobacterium tuberculosis drug resistance second line (MTBDRsl) assay is a novel line probe assay used for the rapid detection of mutational patterns conferring resistance to XDR-TB. The aim of this study was to study the rapid detection of drug resistance and mutational patterns of the XDR-TB by a novel GenoType ® MTBDRsl assay. We evaluated 98 multidrug-resistant (MDR) M. tuberculosis isolates for second line drugs susceptibility testing by 1% proportion method (BacT/ALERT 3D system) and GenoType ® MTBDRsl assay for rapid detection of conferring drug resistance to XDR-TB. A total of seven (17.4%) were identified as XDR-TB by using standard phenotypic method. The concordance between phenotypic and GenoType ® MTBDRsl assay was 91.7-100% for different antibiotics. The sensitivity and specificity of the MTBDRsl assay were 100% and 100% for aminoglycosides; 100% and 100% for fluoroquinolones; 91.7% and 100% for ethambutol. The most frequent mutations and patterns were gyrA MUT1 (A90V) in seven (41.2%) and gyrA + WT1-3 + MUT1 in four (23.5%); rrs MUT1 (A1401G) in 11 (64.7%), and rrs WT1-2 + MUT1 in eight (47.1%); and embB MUT1B (M306V) in 11 (64.7%) strains. These data suggest that the GenoType ® MTBDRsl assay is rapid, novel test for detection of resistance to second line anti-tubercular drugs. This assay provides additional information about the frequency and mutational patterns responsible for XDR-TB resistance.


HELICAL COMPUTERIZED TOMOGRAPHY CAN MEASURE SUBCUTANEOUS, VISCERAL AND TOTAL FAT AREAS?
Abdominal obesity or android obesity, that is, the increase in adipose tissue in the abdominal region, is considered a risk factor for several morbidities. Different ways of quantifying it have been proposed, one method is the measurement of the abdominal fat area by computed tomography. To establish correspondence between the groups defined by degree of obesity in relation to the total, subcutaneous and visceral fat area. Cross-sectional observational study carried out through the analysis of tomographic examinations. Horos v3.3.5 medical image visualization software was used, with abdominal tomography in a single cut including the L4 vertebral body and the umbilical scar, to obtain the areas of total, visceral and subcutaneous fat. Of the 40 patients, 10 had grade II obesity, 23 grade III and 7 superobese. The amount of total fat showed an increase in relation to the degree of obesity. Visceral fat did not show significant differences between the degrees of obesity, but the data showed a lower average in the group of obesity grade II. The area of subcutaneous fat, as well as total fat, showed an increase in its measurements, according to the progression of the patients' BMI, but there was no statistical significance in this difference between the groups of grade II and super-obese individuals. The area of total and subcutaneous fat showed an increase in its measurements according to the progression of the BMI groups, which did not happen with visceral fat.


Attachment of Burkholderia pseudomallei to pharyngeal epithelial cells: a highly pathogenic bacteria with low attachment ability.
Respiratory infections are initiated by the attachment of bacteria to pharyngeal epithelial cells. We studied the attachment of Burkholderia pseudomallei to pharyngeal epithelial cells. After one, two, three, and four washes, there were 22.6+/-8.9, 15.7+/-7.0, 6.8+/-3.1, and 4.6+/-1.1 (mean+/-SD) attached bacteria/cell, respectively. If the bacterial concentration was maintained at 1 X 10(8) colony-forming units (cfu)/ml and three washes were done, at concentrations of 2.5 x 10(4), 5 X 10(4), and 1 x 10(5) cells/ml there were 9.9+/-3.6, 3.3+/-0.8, and 2.5+/-1.1 attached bacteria/cell, respectively. If the cell concentration was kept at 2.5 x 10(4) cells/ml and three washes were done, at bacterial concentrations of 1 x 10(5), 1 X 10(6), 1 X 10(7), 1 x 10(8), and 1 x 10(9) cfu/ml, there were 0.3+/-0.3, 0.6+/-0.6, 1.0+/-0.2, 5.1+/-2.3, and 9.6+/-1.9 attached bacteria/cell, respectively. There were 4.8+/-1.9, 5.5+/-2.5, 5.6+/-1.9, and 6.4+/-2.6 attached bacteria/cell at 0, 30, 120, and 240 min of incubation, respectively. Pharyngeal cells from 10 persons (seven men and three women, mean+/-SD age = 30.7+/-8.1 years, 12 experiments with a single isolate) showed that there were 7.8+/-4.3 attached bacteria/cell. It was found that the efficiency of attachment of this bacteria was very low (7.0+/-3.3 bacteria/cell). Electron microscopy revealed that there were no fimbriae but a thin capsular polysaccharide layer on the surface of B. pseudomallei. Attachment to pharyngeal epithelial cells appeared to be mediated by this structure.


Correlation of Trichoscopic Findings in Androgenetic Alopecia and the Disease Severity.
Androgenetic alopecia (AGA) refers to the appearance of the common nonscarring progressive patterned loss of terminal hair on the frontal scalp and/or vertex of the scalp in both men and women, seen with increasing age in genetically predisposed individuals. Until recently, a scalp biopsy was the only objective tool to diagnose and monitor the disease severity. Trichoscopy of scalp is a new noninvasive technique applied to facilitate the diagnosis of hair and scalp disorders using a manual or video dermatoscope. We found a significant difference in some of the variables such as brown peripilar sign (BPPS), white peripilar sign (WPPS), focal atrichia which may aid in the diagnosis of early and late stages of both male and female AGA along with its clinical correlation. No significant difference in the occipital area was found in all AGA patients. This study aims to study the trichoscopic findings of AGA and to correlate their relationship with disease severity in our tertiary care hospital. This was a prospective, observational study. A total of 91 patients (66 males and 25 females) of the age group between 18 and 70 years, were included in the study at the outpatient department of dermatology in 1 year. Each patient underwent a detailed general physical, systemic, and dermatological examination. The diagnosis of AGA was based on clinical grounds. The type of hair loss in each patient was recorded. Trichoscopic evaluation and capture of trichoscopic images was performed using an eScope Oitez Digital Microscope. In accordance with the Helsinki Declaration of 1975 (revised in 2000), the study was approved by Ethical and Scientific Research committees of Care Institute of Medical Sciences, Hyderabad. Statistical analysis was carried out with R-studio. Statistical significance in the difference in the outcome variables between the stages was assessed by Fisher's exact test. The statistical test was considered statistically significant at P < 0.05. A positive correlation between clinical and trichoscopic findings with respect to disease severity was seen in some of the variables in our study. Both male and female AGA patients have hair shaft thickness heterogeneity as the most common feature. BPPS is seen in early grades of AGA (P < 0.01); WPPS and focal atrichia are seen in later grades of AGA (P < 0.01). Scalp honeycomb pigmentation was most commonly seen in all stages and is not correlated to the disease severity of AGA. As trichoscopy can reveal early variations in hair follicle diameter long before hair loss becomes clinically visible and has the advantage of examining larger areas in a relatively short duration makes it a practical choice for clinic set up. It adds new easily recognizable images for visual teledermatology. Besides, the easy documentation allows the doctor and patient to view the video graphics images simultaneously and helps in evaluating a therapeutic response by comparing it with pre-treatment images.


The down-regulation of miR-125b in chronic lymphocytic leukemias leads to metabolic adaptation of cells to a transformed state.
MiR-125b-1 maps at 11q24, a chromosomal region close to the epicenter of 11q23 deletions in chronic lymphocytic leukemias (CLLs). Our results establish that both aggressive and indolent CLL patients show reduced expression of miR-125b. Overexpression of miR-125b in CLL-derived cell lines resulted in the repression of many transcripts encoding enzymes implicated in cell metabolism. Metabolomics analyses showed that miR-125b overexpression modulated glucose, glutathione, lipid, and glycerolipid metabolism. Changes on the same metabolic pathways also were observed in CLLs. We furthermore analyzed the expression of some of miR-125b-target transcripts that are potentially involved in the aforementioned metabolic pathways and defined a miR-125b-dependent CLL metabolism-related transcript signature. Thus, miR-125b acts as a master regulator for the adaptation of cell metabolism to a transformed state. MiR-125b and miR-125b-dependent metabolites therefore warrant further investigation as possible novel therapeutic approaches for patients with CLL.


A recommended "minimum data set" framework for SD-OCT retinal image acquisition and analysis from the Atlas of Retinal Imaging in Alzheimer's Study (ARIAS).
We propose a minimum data set framework for the acquisition and analysis of retinal images for the development of retinal Alzheimer's disease (AD) biomarkers. Our goal is to describe methodology that will increase concordance across laboratories, so that the broader research community is able to cross-validate findings in parallel, accumulate large databases with normative data across the cognitive aging spectrum, and progress the application of this technology from the discovery stage to the validation stage in the search for sensitive and specific retinal biomarkers in AD. The proposed minimum data set framework is based on the Atlas of Retinal Imaging Study (ARIAS), an ongoing, longitudinal, multi-site observational cohort study. However, the ARIAS protocol has been edited and refined with the expertise of all co-authors, representing 16 institutions, and research groups from three countries, as a first step to address a pressing need identified by experts in neuroscience, neurology, optometry, and ophthalmology at the Retinal Imaging in Alzheimer's Disease (RIAD) conference, convened by the Alzheimer's Association and held in Washington, DC, in May 2019. Our framework delineates specific imaging protocols and methods of analysis for imaging structural changes in retinal neuronal layers, with optional add-on procedures of fundus autofluorescence to examine beta-amyloid accumulation and optical coherence tomography angiography to examine AD-related changes in the retinal vasculature. This minimum data set represents a first step toward the standardization of retinal imaging data acquisition and analysis in cognitive aging and AD. A standardized approach is essential to move from discovery to validation, and to examine which retinal AD biomarkers may be more sensitive and specific for the different stages of the disease severity spectrum. This approach has worked for other biomarkers in the AD field, such as magnetic resonance imaging; amyloid positron emission tomography; and, more recently, blood proteomics. Potential context of use for retinal AD biomarkers is discussed.


Differential effect of cross-linking the CD98 heavy chain on fusion and amino acid transport in the human placental trophoblast (BeWo) cell line.
CD98 (otherwise known as 4F2) is an integral membrane protein with multiple functions including amino acid transport, integrin activation, cell fusion and cell activation. The molecular mechanisms coordinating these multiple functions remain unclear. We have studied CD98 heavy chain (hc) function in a human placental trophoblast cell line (BeWo). We show that cross-linking of CD98hc by incubation of cells in the presence of functional monoclonal antibodies causes cellular re-distribution of the protein from the cytoplasm to the plasma membrane as measured by flow cytometry, western blotting and quantitative immuno-electron microscopy. The latter technique also indicated that CD98hc is trafficked between cell surface and cytoplasmic pools in vesicles. Increased cell surface CD98 correlates with increased cellular fusion in BeWo cells. In addition, we show reduced LAT 1 surface expression and neutral amino acid transport in the presence of the CD98 mabs. The results thus suggest that the function of CD98 in cell fusion is distinct from its role in cellular nutrient delivery.


Biological implications of longevity in dairy cows: 2. Changes in methane emissions and efficiency with age.
Previous studies indicated that absolute CH4 emissions and CH4 yield might increase and that milk production efficiency might decrease with age in cattle. Both would make strategies to increase longevity in dairy cattle less attractive. These aspects were experimentally determined in Brown Swiss cattle distributed continuously across a large age range. Thirty lactating dairy cows (876-3,648 d of age) received diets consisting of hay, corn silage, and grass pellets supplemented with 0 or 5kg of concentrate per day. Twelve heifers (199-778 d of age) received hay only. Cows and heifers were members of herds subjected to the 2 different feeding regimens (with or without concentrate) for the past 10 yr. Methane emissions were measured individually for 2 d in open-circuit respiration chambers, followed by quantifying individual feed intake and milk yield over 8 d. Additional data on digestibility, rumination time, and passage time of feed of all experimental animals were available. Regression analyses were applied to evaluate effects of age and feeding regimen. Body weight, milk yield, and the hay proportion of forage dry matter intake were considered as covariates. Methane emissions per unit of intake, body weight, and milk yield were significantly related to age. Their development in the cows with age was characterized by an increase to maximum at around 2,000 d of age, followed by a decline. This response was not accompanied by corresponding age-related changes in intake, chewing activity, digesta passage time, and digestibility of organic matter, which would have explained shifts in CH4. However, fiber digestibility showed a similar change with age as methane emissions, resulting in quite stable methane emissions per unit of digestible fiber. As expected, methane emissions intensity per unit of milk produced was greater by 8% without concentrate than with concentrate, but no difference was noted in the response to age when the animals were subjected to different feeding regimens. The efficiency of milk production was only marginally influenced by age and diet, and no different response was observed for age in the 2 dietary regimens. In conclusion, life cycle analyses of milk production systems focusing on longevity should consider changing methane yields with age in addition to the variation in environmental costs for replacements of culled cows.


Remote ischemic preconditioning protects neurocognitive function of rats following cerebral hypoperfusion.
Protection of remote ischemic preconditioning on neurocognitive function caused by bilateral common carotid artery occlusion has been investigated in rats. Thirty-six male Sprague-Dawley rats were divided into 3 groups - control group (Group C, n=12), bilateral carotid arteries occlusion group (Group B, n=12) and remote ischemic precondition group (Group P, n=12). In Group P, remote ischemic preconditioning (RIPC) was performed on the right femoral artery with 3 cycles (10 min) of occlusion/perfusion. After 3 cycles of preconditioning, bilateral carotid arteries were occluded immediately for 60 min. In Group B, ischemic insults were conducted without RIPC. Sham surgeries were performed in Group C. Evaluation of memory and learning capacity was performed on days 5-8 after surgery by Morris water maze testing of spatial learning capacity (n=6 for each group). Apoptosis of cells in the hippocampus region was determined by TUNEL tests and Bcl-2 at this region was determined by ELISA 24 h and 9 days after vessel occlusion (n=6 for each group). Neurocognitive tests showed that latency time was significantly longer in Group B than in Group P on day 7 (p=0.016) and day 8 (p=0.036). Moreover, frequency of platform crossings was significant less in group B than in the other 2 groups on day 9. Bcl-2 level was significantly increased in the hippocampal region of rats in Group P on days 1 and 9 after vessel occlusion. TUNEL test showed that apoptosis could be observed at 24 h after occlusion in Group B, but not in Group P and Group C. No apoptosis was observed on day 9. Our results suggest that RIPC can protect neurocognitive function of rats after bilateral carotid occlusions, and that Bcl-2 may play an important role in this protective effect.


Human activated T lymphocytes modulate IDO expression in tumors through Th1/Th2 balance.
Previous cancer vaccination approaches have shown some efficiency in generating measurable immune responses, but they have rarely led to tumor regression. It is therefore possible that tumors emerge with the capacity to down-regulate immune counterparts, through the local production of immunosuppressive molecules, such as IDO. Although it is known that IDO exerts suppressive effects on T cell functions, the mechanisms of IDO regulation in tumor cells remain to be characterized. Here, we demonstrate that activated T cells can induce functional IDO expression in breast and kidney tumor cell lines, and that this is partly attributable to IFN-gamma. Moreover, we found that IL-13, a Th2 cytokine, has a negative modulatory effect on IDO expression. Furthermore, we report IDO expression in the majority of breast and kidney carcinoma samples, with infiltration of activated Th1-polarized T cells in human tumors. These findings demonstrate complex control of immune activity within tumors. Future immune therapeutic interventions should thus include strategies to counteract these negative mechanisms.


PARP-1 controls NK cell recruitment to the site of viral infection.
The activation and recruitment of NK cells to the site of viral infection are crucial for virus control. However, it remains largely unknown what controls the recruitment of the activated NK cells to the infection site. In a model of intraperitoneal infection with vaccinia virus (VV), we showed that poly(ADP-ribose) polymerase-1 (PARP-1), a sensor of DNA damage, is critical for NK cell recruitment to the site of infection and viral control in vivo. We further demonstrated that PARP-1 promotes the production of CCL2 and that the CCL2-CCR2 axis is essential for NK cell recruitment to the infection site. In addition, we demonstrated that peritoneal macrophages are the main producer of PARP-1-dependent CCL2 secretion. Mechanistically, PARP-1 functions as a regulator of NF-κB by promoting its nuclear translocation and binding to its response sequences in macrophages upon VV infection. Taken together, our results reveal a potentially previously unknown role for PARP-1-dependent CCL2 production in NK cell migration and viral control and may provide important insights into the design of effective NK cell-based therapies for viral infections and cancer.


Enhanced transmission through squeezed modes in a self-cladding magnonic waveguide.
We study spin-wave propagation in 360-nm wide Ni(80)Fe(20) nanowires using all-electrical spin-wave spectroscopy. Creating a zigzag-like magnetization state, we find enhanced spin-wave transmission compared to the states of more homogeneous magnetization. Micromagnetic simulations show that the spin waves propagate in narrow channels, which in particular, are remotely positioned from the edges. The internal channels reflect field-controlled self-cladding. Interestingly, rotation of the magnetic field at a specific value is found to vary the propagation velocity without changing the eigenfrequency. This opens the perspective of the velocity modulation transistor following a concept known from semiconductor electronics.


Impact and Duration of Brief Surgeon-Delivered Smoking Cessation Advice on Attitudes Regarding Nicotine Dependence and Tobacco Harms for Patients with Peripheral Arterial Disease.
Despite the recognized benefits of smoking cessation, many clinicians question if a brief smoking cessation intervention can help dedicated smokers with peripheral arterial disease understand nicotine dependence and harms related to smoking. We investigated the impact and durability of a multimodal smoking cessation intervention on patient attitudes regarding nicotine dependence and the health effects of smoking. We conducted a pilot cluster-randomized trial of a brief smoking cessation intervention at 8 vascular surgery practices between September 1, 2014 and August 31, 2015. Compared with control sites, patients at intervention sites received protocolized brief cessation counseling, medications, and referrals to a quitline. After their clinic visit and again at 3 months, participants completed a brief survey about patient attitudes regarding nicotine dependence and the health effects of smoking. Responses to questions were analyzed using chi-squared test and Student's t-test. All trial participants (n = 156) complete the initial survey, and 75 (45%) participants completed the follow-up survey. Intervention and control patients both reported a greater than 30-pack-year history (80% vs. 90%, P = 0.07) and previous failed quit attempts (77% vs. 78%, P = 0.8). Compared with usual care, patients in the intervention group were more likely to describe hearing advice to quit from their surgeon (98% vs. 77%, P < 0.001), and expressed "a lot" or "some" interest in quitting (95.4% vs. 85.7%, P = 0.05). Patients in the intervention group were also more likely to acknowledge their addictive behaviors, consistently scoring higher on question bank items regarding nicotine addiction (52.9 vs. 48.0, P = 0.006) and the negative health effects of smoking (scaled score 56.6 vs. 50.6, P = 0.001). When resurveyed 3 months after intervention, patients in the intervention group had larger declines in nicotine dependence and health effect domains, suggesting durable impact of the intervention on patient attitudes regarding nicotine addiction and smoking harms. Brief smoking cessation counseling by a vascular surgeon increases patient interest in smoking cessation and awareness of smoking harms, and this effect was durable 3 months after intervention. This evidence suggests that even brief counseling within a surgical clinic has the potential to impact patient desire to quit.


Nucleic acid-based antiviral and gene therapy of chronic hepatitis B infection.
Persistent hepatitis B virus (HBV) infection often leads to the development of chronic hepatitis, cirrhosis and hepatocellular carcinoma. There is a need to develop new antiviral approaches for the treatment of this disease. We have explored various nucleic acid-based strategies designed to inhibit HBV replication including: the use of antisense RNA and DNA constructs, DNA-based immunization techniques to stimulate broad-based cellular immune responses with particular emphasis on the generation of cytotoxic lymphocyte (CTL) activity to viral structural proteins, hammerhead ribozymes to cleave HBV pregenomic RNA in vitro and dominant negative HBV core mutant proteins as inhibitors of nucleocapsid formation within cells. In order to optimize these antiviral effects, various novel expression vectors have been developed to deliver such DNA constructs to cells. For example, adenoviral vectors carrying genes that encode for dominant negative proteins have been employed to transfect hepatocytes in vitro and in vivo. In addition, plasmid vectors have been produced to promote expression of HBV structural genes following injection into muscle cells as a means to stimulate the host's cellular and humoral immune response in the context of histocompatibility antigen (HLA) class I and II antigen presentation. These experimental approaches may have important implications for the generation of efficient antiviral effects during chronic HBV infection.


Misophonia and comorbid psychiatric symptoms: a preliminary study of clinical findings.
Objective: Misophonia is a neurophysiological disorder, phenotypically characterized by heightened autonomic nervous system arousal which is accompanied by a high magnitude of emotional reactivity to repetitive and pattern-based auditory stimuli. This study identifies the prevalence of psychiatric symptoms in misophonia sufferers, the association between severity of misophonia and psychiatric symptoms, and the association between misophonia severity and gender. Methods: Fifty-two misophonia sufferers, 30 females (mean age = 40.93 ± 15.29) and 22 males (mean age = 51.18 ± 15.91) were recruited in our study and they were diagnosed according the criteria proposed by Schröder et al. The participants were evaluated by the A-MISO-S for the severity of misophonia and the MINI to assess the presence of psychiatric symptoms. Results: The most common comorbid symptoms reported by the misophonia patients were respectively PTSD (N = 8, 15.38%), OCD (N = 6, 11.53%), MDD (N = 5, 9.61%), and anorexia (N = 5, 9.61%). Misophonia severity was associated with the symptoms of MDD, OCD, and PTSD as well as anorexia. There was an indication of a significant difference between men and women in the severity of misophonic symptoms. Conclusion: Our findings highlight the importance of recognizing psychiatric comorbidity among misophonia sufferers. The presence of these varying psychiatric disorders' features in individuals with misophonia suggests that while misophonia has unique clinical characteristics with an underlying neurophysiological mechanism, may be associated with psychiatric symptoms. Therefore, when assessing individuals with misophonia symptoms, it is important to screen for psychiatric symptoms. This will assist researchers and clinicians to better understand the nature of the symptoms and how they may be interacting and ultimately allocating the most effective therapeutic strategies.


Irrational exuberance and neural crash warning signals during endogenous experimental market bubbles.
Groups of humans routinely misassign value to complex future events, especially in settings involving the exchange of resources. If properly structured, experimental markets can act as excellent probes of human group-level valuation mechanisms during pathological overvaluations--price bubbles. The connection between the behavioral and neural underpinnings of such phenomena has been absent, in part due to a lack of enabling technology. We used a multisubject functional MRI paradigm to measure neural activity in human subjects participating in experimental asset markets in which endogenous price bubbles formed and crashed. Although many ideas exist about how and why such bubbles may form and how to identify them, our experiment provided a window on the connection between neural responses and behavioral acts (buying and selling) that created the bubbles. We show that aggregate neural activity in the nucleus accumbens (NAcc) tracks the price bubble and that NAcc activity aggregated within a market predicts future price changes and crashes. Furthermore, the lowest-earning subjects express a stronger tendency to buy as a function of measured NAcc activity. Conversely, we report a signal in the anterior insular cortex in the highest earners that precedes the impending price peak, is associated with a higher propensity to sell in high earners, and that may represent a neural early warning signal in these subjects. Such markets could be a model system to understand neural and behavior mechanisms in other settings where emergent group-level activity exhibits mistaken belief or valuation.


Effect of U-73,122, an inhibitor of phospholipase C, on actions of parathyroid hormone in opossum kidney cells.
The relative roles of the adenylate cyclase-protein kinase A system (AC-PKA), the phospholipase C-protein kinase C system (PLC-PKC), and increases in cytosolic calcium in mediating the final actions of parathyroid hormone (PTH) remain ill defined. Although an important role for the PLC-PKC system in the regulation of phosphate transport in response to PTH has been suggested, previous studies from our laboratory and others, in OK cells, have emphasized the major role of AC-PKA. The present studies were designed to dissociate the second messengers for PTH by using an inhibitor of PLC (U-73,122). Studies were performed in confluent cultures of OK cells with and without preincubation with U-73,122 (1 microM). This inhibitor did not alter adenosine 3',5'-cyclic monophosphate (cAMP) production or the activation of PKA in response to PTH. Preincubation with U-73,122, however, totally abolished PTH-stimulated increases in diglyceride mass, consistent with inhibition of PLC. Activation of particulate PKC was then examined in response to PTH in the absence and presence of U-73,122. Although PTH resulted in an increase in particulate PKC activity in control cultures, this effect was abolished in the presence of U-73,122 and actually decreased significantly. Therefore, having documented marked attenuation of PLC-PKC, we next examined the effects of PTH on phosphate transport. Basal phosphate uptake was not altered by 1 microM U-73,122. Dose-response curves of the inhibition of phosphate transport in response to PTH were identical in the presence or absence of U-73,122. Thus inhibition of PLC and PKC activities did not alter the effects of PTH on phosphate transport.(ABSTRACT TRUNCATED AT 250 WORDS)


Severe bone demineralisation is associated with higher mortality in children with cystic fibrosis.
Decreased bone mineral density (BMD) is an emerging problem for clinicians who care for children with Cystic fibrosis (CF). The aim of this study was to determine prevalence and assess risk factors for reduced BMD in our adolescent population with CF. All bone densitometry scans (n=99) performed on children (n=79) with a mean age 13.6 (10-19.2) years over a 7 year period (2000-2007) were reviewed. Patient records were reviewed for correlating clinical data. Low BMD is frequently present in adults and children with variable reports (36-66%). In our study, BMD expressed as z score of L2-L4 spine was reduced in a total of 50% children with a preponderance of males. Bone demineralization was strongly associated with increasing age (p=0.03), diminished lung function (p=0.027), reduced body mass index (p=0.001) and treatment with oral corticosteroids (p=0.02).


Human immunodeficiency virus type-1 episomal cDNA in semen.
Episomal 2-long terminal repeat (LTR) HIV-1 cDNA, a by-product of HIV-1 infection, is used in clinical trials as a marker for ongoing viral replication. It would be useful to employ 2-LTR cDNA to monitor cryptic HIV-1 infection in the genital tract of men on antiretroviral therapy (ART) to predict the evolution of sexually transmissible drug-resistant HIV-1, but studies thus far have failed to detect this marker in semen. The objectives of this study were: 1) to use a technique that maximizes DNA recovery from HIV-1 infected white blood cells in semen to determine if episomal 2-LTR cDNA is detectable in semen of ART-naïve men with other evidence of genital tract HIV-1 infection, and 2) to compare levels of HIV-1 2-LTR cDNA, RNA, and proviral DNA in semen from HIV-1+ men on ART. Using a somatic cell DNA extraction technique, 2-LTR cDNA was detected by PCR/ELISA in 4 out of 8 semen samples from ART-naïve men selected for other signs of seminal HIV-1 infection (positive controls). Southern blot and DNA sequencing confirmed that the amplified sequences were HIV-1 2-LTR cDNA; copy numbers ranged from 55 to 504 copies/sample. Two semen samples from a cohort of 22 HIV-1-infected men on dual nucleoside therapy, one with and one without detectable seminal HIV-1 RNA, were 2-LTR cDNA positive (336 and 8,560 copies/sample). Following addition of indinavir to the therapy regimen, no semen samples from 21 men with controlled peripheral and seminal viral loads were 2-LTR cDNA positive at 1 and 6 month time points, despite the persistence of HIV-1 proviral DNA+ semen cells and seminal cytomegalovirus (CMV) shedding in some cases. However, one individual who failed indinavir therapy and later developed distinct protease inhibitor (PI) drug resistance mutations in semen, maintained elevated levels of HIV-1 RNA and 2-LTR cDNA in semen. 2-LTR HIV-1 cDNA is detectable in semen of HIV-1-infected men. Two men on ART had 2-LTR HIV-1 cDNA in semen, suggesting that this marker may prove to be useful to monitor HIV-1 infection in the genital tract of men on ART to predict the evolution of drug resistance mutations in semen.


Systematic fluoroscopic and electrical assessment of implantable cardioverter-defibrillator patients implanted with silicone-polyurethane copolymer (Optim™) coated leads.
Serious concerns have been recently raised about the reliability of the silicone-polyurethane copolymer (Optim™) lead insulation system. We sought to identify insulation defects and Optim-lead failures by systematic fluoroscopic and electrical assessment in a prospectively defined cohort of implantable cardioverter-defibrillator (ICD) patients. Between July 2007 and December 2011, 234 patients were implanted with 413 optim-coated leads as part of an ICD system at a single centre. Fluoroscopic screening with high-resolution cine-fluoroscopy at 30 frames per second was offered to all patients. In addition, the electrical integrity of all implanted leads was assessed. Durata, Riata ST Optim, and low-voltage Optim leads were implanted in 199, 26, and 188 cases, respectively. During a total follow-up of 10 036 lead-months, there were 7 Optim-lead failures (defined as electrical malfunction resulting in lead replacement) and 31 deaths; no cases of electrical noises were encountered. The overall incidence of lead failure was 1.2 vs. 0.3 per 100 lead-years, for high- and low-voltage leads, respectively (P = 0.1). One hundred fifty-one patients agreed to undergo fluoroscopy screening; none of the 264 analysed Optim leads were found to have any fluoroscopically visible structural defects after an average of 31 months post-implant. This study represents the first systematic screening of Optim-coated leads in a large unselected cohort of ICD patients. Over a 5-year period few lead failures were observed and normal fluoroscopic appearance was present in all patients.


Stability of end-of-life preferences: a systematic review of the evidence.
Policies and practices that promote advance care planning and advance directive completion implicitly assume that patients' choices for end-of-life (EOL) care are stable over time, even with changes in health status. To systematically evaluate the evidence on the stability of EOL preferences over time and with changes in health status. We searched for longitudinal studies of patients' preferences for EOL care in PubMed, EMBASE, and using citation review. Studies restricted to preferences regarding the place of care at the EOL were excluded. A total of 296 articles were assessed for eligibility, and 59 met inclusion criteria. Twenty-four articles had sufficient data to extract or calculate the percentage of individuals with stable preferences or the percentage of total preferences that were stable over time. In 17 studies (71%) more than 70% of patients' preferences for EOL care were stable over time. Preference stability was generally greater among inpatients and seriously ill outpatients than among older adults without serious illnesses (P < .002). Patients with higher education and who had engaged in advance care planning had greater preference stability, and preferences to forgo therapies were generally more stable than preferences to receive therapies. Among 9 of the 24 studies (38%) assessing changes in health status, no consistent relationship with preference changes was identified. Considerable variability among studies in the methods of preference assessment, the time between assessments, and the definitions of stability preclude meta-analytic estimates of the stability of patients' preferences and the factors influencing these preferences. Although more seriously ill patients and those who engage in advance care planning most commonly have stable preferences for future treatments, further research in real-world settings is needed to confirm the utility of advance care plans for future decision making.


Application of MALDI-TOF MS for the Identification of Food Borne Bacteria.
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently emerged as a powerful tool for the routine identification of clinical isolates. MALDI-TOF MS based identification of bacteria has been shown to be more rapid, accurate and cost-efficient than conventional phenotypic techniques or molecular methods. Rapid and reliable identification of food-associated bacteria is also of crucial importance for food processing and product quality. This review is concerned with the applicability of MALDI-TOF MS for routine identification of foodborne bacteria taking the specific requirements of food microbiological laboratories and the food industry into account. The current state of knowledge including recent findings and new approaches are discussed.


High Figure of Merit in Gallium-Doped Nanostructured n-Type PbTe-xGeTe with Midgap States.
PbTe-based thermoelectric materials are some of the most promising for converting heat into electricity, but their n-type versions still lag in performance the p-type ones. Here, we introduce midgap states and nanoscale precipitates using Ga-doping and GeTe-alloying to considerably improve the performance of n-type PbTe. The GeTe alloying significantly enlarges the energy band gap of PbTe and subsequent Ga doping introduces special midgap states that lead to an increased density of states (DOS) effective mass and enhanced Seebeck coefficients. Moreover, the nucleated Ga2Te3 nanoscale precipitates and off-center discordant Ge atoms in the PbTe matrix cause intense phonon scattering, strongly reducing the thermal conductivity (∼0.65 W m-1 K-1 at 623 K). As a result, a high room-temperature thermoelectric figure of merit ZT ∼ 0.59 and a peak ZTmax of ∼1.47 at 673 K were obtained for the Pb0.98Ga0.02Te-5%GeTe. The ZTavg value that is most relevant for devices is ∼1.27 from 400 to 773 K, the highest recorded value for n-type PbTe.


Evolution of duplicate genes in a tetraploid animal, Xenopus laevis.
To understand the evolution of duplicate genes, we compared rates of nucleotide substitution between 17 pairs of nonallelic duplicated genes in the tetraploid frog Xenopus laevis with rates between the orthologous loci of human and rodent. For all duplicated X. laevis genes, the number of synonymous substitutions per site (dS) was greater than the number of nonsynonymous substitutions per site (dN), indicating that these genes are subject to purifying selection. There was also a significant positive correlation (r = 0.915) between dN for the X. laevis genes and dN for the mammalian genes, suggesting that, at the amino acid level, the X. laevis genes and the mammalian genes are under similar constraints. Results of relative-rate tests showed nearly equal rates of nonsynonymous substitution in each copy of the X. laevis genes; apparently there are similar constraints on both copies. No correlation was found between dS for the X. laevis genes and dS for the mammalian genes. There was a significant positive correlation both between members of pairs of duplicated X. laevis genes (r = 0.951) and between human and rodent orthologues (r = 0.854) with respect to third-position G+C content but no such relationship between the X. laevis genes and either of their mammalian orthologues. The results indicate that both copies of a duplicate gene can be subject to purifying selection and thus support the hypothesis of selection against all genotypes containing a null allele at either of two duplicate loci.


The activating TERT promoter mutation C228T is recurrent in subsets of adrenal tumors.
The telomerase reverse transcriptase gene (TERT) encodes the reverse transcriptase component of the telomerase complex, which is essential for telomere stabilization and cell immortalization. Recent studies have demonstrated a transcriptional activation role for the TERT promoter mutations C228T and C250T in many human cancers, as well as a role in aggressive disease with potential clinical applications. Although telomerase activation is known in adrenal tumors, the underlying mechanisms are not established. We assessed C228T and C250T TERT mutations by direct Sanger sequencing in tumors of the adrenal gland, and further evaluated potential associations with clinical parameters and telomerase activation. A total of 199 tumors were evaluated, including 34 adrenocortical carcinomas (ACC), 47 adrenocortical adenomas (ACA), 105 pheochromocytomas (PCC; ten malignant and 95 benign), and 13 abdominal paragangliomas (PGL; nine malignant and four benign). TERT expression levels were determined by quantitative RT-PCR. The C228T mutation was detected in 4/34 ACCs (12%), but not in any ACA (P=0.028). C228T was also observed in one benign PCC and in one metastatic PGL. The C250T mutation was not observed in any case. In the ACC and PGL groups, TERT mutation-positive cases exhibited TERT expression, indicating telomerase activation; however, since expression was also revealed in TERT WT cases, this could denote additional mechanisms of TERT activation. To conclude, the TERT promoter mutation C228T is a recurrent event associated with TERT expression in ACCs, but rarely occurs in PGL and PCC. The involvement of the TERT gene in ACC represents a novel mutated gene in this entity.


Fluorogen-activating scFv biosensors target surface markers on live cells via streptavidin or single-chain avidin.
Fluorescence biosensors are indispensable tools for understanding protein behavior and function in cells. Recent advancements utilize fluorogen-activating-proteins (FAPs) that form complexes with small organic molecules (fluorogens) and result in their fluorescence activation. The technology has found multiple uses in protein discovery applications; however, the current method of detection requires the expression of FAPs as gene fusion tags in cells-a process that is time- and labor-intensive. In this report, we present an alternate method that utilizes FAPs as affinity reagents. Accordingly, we isolated soluble reagents based on FAP fusions with streptavidin (Strep) or avidin proteins, both highly selective for biotin. When tested in vitro, the reagents displayed bi-functional activity, fluorogen activation, and biotin affinity. For live-cell protein discovery, surface targets were biotinylated via biotin-tagged immunoglobulins or a genetically encoded biotin acceptor peptide. As a result, when the cells were labeled with FAP-Strep or FAP-avidin reagent, the in vivo fluorescence measurements indicated high target specificity, minimal background, and bright signal detection. In summary, we present a novel FAP reagent platform that offers a rapid and efficient approach for cell surface protein detection.


VEGF-induced retinal angiogenic signaling is critically dependent on Ca²⁺ signaling by Ca²⁺/calmodulin-dependent protein kinase II.
The authors conducted an in vitro investigation of the role of Ca(2+)-dependent signaling in vascular endothelial growth factor (VEGF)-induced angiogenesis in the retina. Bovine retinal endothelial cells (BRECs) were stimulated with VEGF in the presence or absence of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM; intracellular Ca(2+) chelator), U73122 (phospholipase C (PLC) inhibitor), xestospongin C (Xe-C), and 2-aminoethoxydiphenyl borate (2APB) (inhibitors of inositol-1,4,5 triphosphate (IP(3)) signaling). Intracellular Ca(2+) concentration ([Ca(2+)](i)) was estimated using fura-2 Ca(2+) microfluorometry, Akt phosphorylation quantified by Western blot analysis, and angiogenic responses assessed using cell migration, proliferation, tubulogenesis, and sprout formation assays. The effects of the Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) inhibitor KN93 were also evaluated on VEGF-induced Akt signaling and angiogenic activity. Stimulation of BRECs with 25 ng/mL VEGF induced a biphasic increase in [Ca(2+)](i), with an initial transient peak followed by a sustained plateau phase. VEGF-induced [Ca(2+)](i) increases were almost completely abolished by pretreating the cells with BAPTA-AM, U73122, Xe-C, or 2APB. These agents also inhibited VEGF-induced phosphorylation of Akt, cell migration, proliferation, tubulogenesis, and sprouting angiogenesis. KN93 was similarly effective at blocking the VEGF-induced activation of Akt and angiogenic responses. VEGF increases [Ca(2+)](i) in BRECs through activation of the PLC-IP(3) signal transduction pathway. VEGF-induced phosphorylation of the proangiogenic protein Akt is critically dependent on this increase in [Ca(2+)](i) and the subsequent activation of CaMKII. Pharmacologic inhibition of Ca(2+)-mediated signaling in retinal endothelial cells blocks VEGF-induced angiogenic responses. These results suggest that the PLC/IP(3)/Ca(2+)/CaMKII signaling pathway may be a rational target for the treatment of angiogenesis-related disorders of the eye.


Loss of Drosophila melanogaster TRPA1 Function Affects "Siesta" Behavior but Not Synchronization to Temperature Cycles.
To maintain synchrony with the environment, circadian clocks use a wide range of cycling sensory cues that provide input to the clock (zeitgebers), including environmental temperature cycles (TCs). There is some knowledge about which clock neuronal groups are important for temperature synchronization, but we currently lack knowledge on the temperature receptors and their signaling pathways that feed temperature information to the (neuronal) clock. Since TRPA1 is a well-known thermosensor that functions in a range of temperature-related behaviors, and it is potentially expressed in clock neurons, we set out to test the putative role of TRPA1 in temperature synchronization of the circadian clock. We found that flies lacking TRPA1 are still able to synchronize their behavioral activity to TCs comparable to wild-type flies, both in 16°C : 25°C and 20°C : 29°C TCs. In addition, we found that flies lacking TRPA1 show higher activity levels during the middle of the warm phase of 20°C : 29°C TCs, and we show that this TRPA1-mediated repression of locomotor activity during the "siesta" is caused by a lack of sleep. Based on these data, we conclude that the TRPA1 channel is not required for temperature synchronization in this broad temperature range but instead is required to repress activity during the warm part of the day.


Chemical and biosynthetic evolution of the antimycin-type depsipeptides.
Evolution of natural products, and particularly those resulting from microbial assembly line-like enzymes, such as polyketide (PK) and nonribosomal peptides (NRP), has resulted in a variety of pharmaceutically important and chemically diverse families of molecules. The antimycin-type depsipeptides are one such grouping, with a significant level of diversity and members that have noted activities against key targets governing human cellular apoptosis (e.g. Bcl-xL and GRP78). Chemical variance originates from ring size, with 9-, 12-, 15-, and 18-membered classes, and we show that such distinctions influence their molecular targeting. Further, we present here a systematic interrogation of the chemistry and assembly line evolution of antimycin-type analogues by conducting metabolomic profiling and biosynthetic gene cluster comparative analysis of the depsipeptide assembly lines for each member of the antimycin-group. Natural molecular evolution principles of such studies should assist in artificial re-combinatorializing of PK and NRP assembly lines.


A phase II study (ARCHER 1042) to evaluate prophylactic treatment of dacomitinib-induced dermatologic and gastrointestinal adverse events in advanced non-small-cell lung cancer.
ARCHER 1042, a randomized phase II trial, explored the impact of prophylactic treatment on select dermatologic adverse events of interest (SDAEI), diarrhea, and mucositis associated with dacomitinib, an oral irreversible pan-human epidermal growth factor receptor (HER) inhibitor, in development for advanced non-small-cell lung cancer (NSCLC). Patients with advanced NSCLC treated with dacomitinib were enrolled in two cohorts. Cohort I patients were randomized 1:1 to receive oral doxycycline or placebo (4 weeks). Cohort II patients received oral VSL#3 probiotic plus topical alclometasone. Primary end points for Cohorts I and II were incidence of all grade and grade ≥2 SDAEI in the first 8 weeks of treatment and quality of life (QoL) assessed by the Skindex-16 survey. Additional primary end points for Cohort II were incidence of all grade and grade ≥2 diarrhea and mucositis in the first 8 weeks of treatment; QoL regarding diarrhea and mucositis incidence was assessed by the modified-Oral Mucositis Daily Questionnaire. Cohort I randomized 114 evaluable patients: 56 in the doxycycline arm, 58 in the placebo arm. Cohort II enrolled 59 evaluable patients. Doxycycline significantly reduced the incidence of grade ≥2 SDAEI by 50% (P = 0.016) compared with placebo. The incidence of all grade SDAEI was lower with doxycycline than with placebo but did not reach statistical significance. Doxycycline was associated with less deterioration in QoL compared with placebo. Alclometasone was associated with less deterioration in QoL compared with placebo but did not statistically significantly reduce the incidence of all grade or grade ≥2 SDAEI. VSL#3 did not reduce the incidence of all grade or grade ≥2 diarrhea and did not impact mucositis scores. Doxycycline was effective as a prophylactic treatment for dacomitinib-induced grade ≥2 SDAEI. Both doxycycline and alclometasone reduced the negative impact in patient-reported dermatologic AEs. The probiotic was not effective for preventing diarrhea or mucositis.


Multidisciplinary team functioning and decision making within forensic mental health.
The purpose of this paper is to investigate the operation of multidisciplinary team (MDT) meetings within a forensic hospital in England, UK. Mixed methods, including qualitative face to face interviews with professionals and service users, video observations of MDT meetings and documentary analysis. Data were collected from 142 staff and 30 service users who consented to take part in the research and analysed using the constant comparison technique of grounded theory and ethnography. Decisions taken within MDT meetings are unequally shaped by the professional and personal values and assumptions of those involved, as well as by the power dynamics linked to the knowledge and responsibility of each member of the team. Service users' involvement is marginalised. This is linked to a longstanding tradition of psychiatric paternalism in mental health care. Future research should explore the nuances of interactions between MDT professionals and service users during the meetings, the language used and the approach taken by professionals to enable/empower service user to be actively involved. Clear aims, responsibilities and implementation actions are a pre-requisite to effective MDT working. There is a need to give service users greater responsibility and power regarding their care. While direct (video) observations were very difficult to achieve in secure settings, they enabled unmediated access to how people conducted themselves rather than having to rely only on their subjective accounts (from the interviews).


Cutaneous vascular reactivity is reduced in aging and in heart failure: association with inflammation.
In the present study, we have investigated whether changes in vascular reactivity in congestive heart failure (CHF) patients can be detected in the cutaneous microvessels and whether these changes are due to endothelial dysfunction, are affected by increasing age and related to an ongoing inflammation. The responses to local warming and iontophoretically administered endothelium-dependent and -independent vasodilators were investigated in healthy young adults, healthy elderly adults and elderly adults with CHF. The results were correlated with plasma concentrations of vascular risk factors and markers for endothelial dysfunction and inflammation. The vasorelaxant responses were reduced in the elderly groups and were attenuated further in the CHF group. This group also had increases in levels of several markers associated with inflammation, higher blood glucose and homocysteine levels, a lower low-density lipoprotein-cholesterol and a rise in the concentration of von Willebrand factor, indicating a prothrombotic endothelial function. The severity of the heart failure, measured as the plasma level of brain natriuretic peptide, correlated with the intensity of inflammation and to the changes in vascular risk factors and endothelial function. It is concluded that the reactivity of the cutaneous microvessels is reduced with age, and the presence of CHF causes a further impairment. There is endothelial dysfunction in CHF, but it is uncertain to what extent this contributes to the reduced vasodilatory capacity. The inflammatory response appears central for many of the manifestations of the CHF syndrome.


Interfering with protein-protein interactions: potential for cancer therapy.
Genotype-specific cancer therapy promises to engender the era of personalised medicines in which rapid identification of tumour specific gene mutations coupled to rapid methods for efficacious drug identification will be applied. Aberrant signal transduction via protein-protein interactions is generally difficult to target with small molecules. However, macromolecules (macrodrugs) can be developed that interfere with protein-protein interactions by binding with high affinity and specificity to contact surfaces. Inhibitors of mutant RAS and its effector protein interactions affect cancer by attenuating aberrant RAS-dependent signal transduction and would be effective against mutant RAS in dividing cells of overt tumours and in putative cancer stem cells when they move into cell-cycle. Results with an antibody fragment blocking effector binding to RAS, illustrates that this is sufficient to prevent cancer. While macrodrugs have inherent problems of bio-distribution and delivery to target cells in patients, their efficacy suggests that efforts to achieve the goal of clinical use should be pursued.


Regional Nodal Irradiation in Early-Stage Breast Cancer.
Most women with breast cancer who undergo breast-conserving surgery receive whole-breast irradiation. We examined whether the addition of regional nodal irradiation to whole-breast irradiation improved outcomes. We randomly assigned women with node-positive or high-risk node-negative breast cancer who were treated with breast-conserving surgery and adjuvant systemic therapy to undergo either whole-breast irradiation plus regional nodal irradiation (including internal mammary, supraclavicular, and axillary lymph nodes) (nodal-irradiation group) or whole-breast irradiation alone (control group). The primary outcome was overall survival. Secondary outcomes were disease-free survival, isolated locoregional disease-free survival, and distant disease-free survival. Between March 2000 and February 2007, a total of 1832 women were assigned to the nodal-irradiation group or the control group (916 women in each group). The median follow-up was 9.5 years. At the 10-year follow-up, there was no significant between-group difference in survival, with a rate of 82.8% in the nodal-irradiation group and 81.8% in the control group (hazard ratio, 0.91; 95% confidence interval [CI], 0.72 to 1.13; P=0.38). The rates of disease-free survival were 82.0% in the nodal-irradiation group and 77.0% in the control group (hazard ratio, 0.76; 95% CI, 0.61 to 0.94; P=0.01). Patients in the nodal-irradiation group had higher rates of grade 2 or greater acute pneumonitis (1.2% vs. 0.2%, P=0.01) and lymphedema (8.4% vs. 4.5%, P=0.001). Among women with node-positive or high-risk node-negative breast cancer, the addition of regional nodal irradiation to whole-breast irradiation did not improve overall survival but reduced the rate of breast-cancer recurrence. (Funded by the Canadian Cancer Society Research Institute and others; MA.20 ClinicalTrials.gov number, NCT00005957.).


Insights into strand displacement and processivity from the crystal structure of the protein-primed DNA polymerase of bacteriophage phi29.
The DNA polymerase from phage phi29 is a B family polymerase that initiates replication using a protein as a primer, attaching the first nucleotide of the phage genome to the hydroxyl of a specific serine of the priming protein. The crystal structure of phi29 DNA polymerase determined at 2.2 A resolution provides explanations for its extraordinary processivity and strand displacement activities. Homology modeling suggests that downstream template DNA passes through a tunnel prior to entering the polymerase active site. This tunnel is too small to accommodate double-stranded DNA and requires the separation of template and nontemplate strands. Members of the B family of DNA polymerases that use protein primers contain two sequence insertions: one forms a domain not previously observed in polymerases, while the second resembles the specificity loop of T7 RNA polymerase. The high processivity of phi29 DNA polymerase may be explained by its topological encirclement of both the downstream template and the upstream duplex DNA.