Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells.
The transfer of somatic cell nuclei into oocytes can give rise to pluripotent stem cells that are consistently equivalent to embryonic stem cells, holding promise for autologous cell replacement therapy. Although methods to induce pluripotent stem cells from somatic cells by transcription factors are widely used in basic research, numerous differences between induced pluripotent stem cells and embryonic stem cells have been reported, potentially affecting their clinical use. Because of the therapeutic potential of diploid embryonic stem-cell lines derived from adult cells of diseased human subjects, we have systematically investigated the parameters affecting efficiency of blastocyst development and stem-cell derivation. Here we show that improvements to the oocyte activation protocol, including the use of both kinase and translation inhibitors, and cell culture in the presence of histone deacetylase inhibitors, promote development to the blastocyst stage. Developmental efficiency varied between oocyte donors, and was inversely related to the number of days of hormonal stimulation required for oocyte maturation, whereas the daily dose of gonadotropin or the total number of metaphase II oocytes retrieved did not affect developmental outcome. Because the use of concentrated Sendai virus for cell fusion induced an increase in intracellular calcium concentration, causing premature oocyte activation, we used diluted Sendai virus in calcium-free medium. Using this modified nuclear transfer protocol, we derived diploid pluripotent stem-cell lines from somatic cells of a newborn and, for the first time, an adult, a female with type 1 diabetes.


Different Sensitivity of Control and MICU1- and MICU2-Ablated Trypanosoma cruzi Mitochondrial Calcium Uniporter Complex to Ruthenium-Based Inhibitors.
The mitochondrial Ca2+ uptake in trypanosomatids shares biochemical characteristics with that of animals. However, the composition of the mitochondrial Ca2+ uniporter complex (MCUC) in these parasites is quite peculiar, suggesting lineage-specific adaptations. In this work, we compared the inhibitory activity of ruthenium red (RuRed) and Ru360, the most commonly used MCUC inhibitors, with that of the recently described inhibitor Ru265, on Trypanosoma cruzi, the agent of Chagas disease. Ru265 was more potent than Ru360 and RuRed in inhibiting mitochondrial Ca2+ transport in permeabilized cells. When dose-response effects were investigated, an increase in sensitivity for Ru360 and Ru265 was observed in TcMICU1-KO and TcMICU2-KO cells as compared with control cells. In the presence of RuRed, a significant increase in sensitivity was observed only in TcMICU2-KO cells. However, application of Ru265 to intact cells did not affect growth and respiration of epimastigotes, mitochondrial Ca2+ uptake in Rhod-2-labeled intact cells, or attachment to host cells and infection by trypomastigotes, suggesting a low permeability for this compound in trypanosomes.


Molecular basis of the allosteric mechanism of cAMP in the regulatory PKA subunit.
The second messenger cyclic Adenosine MonoPosphate (cAMP) mediates many biological process by interacting with structurally conserved nucleotide binding domains (cNBD's). Here, we use molecular dynamics simulations on RIIbeta-PKA, one of the best characterized members of the cNBD family, in presence and absence of cAMP. The results of our calculations are fully consistent with the available experimental data and suggest that the key factor of the cAMP allosteric mechanism in cNBDS's is the increased flexibility of the protein upon ligand release along with a mechanical coupling between helical segments. In addition, our calculations provide a rationale for the experimentally observed cAMP selective binding to PKA.


The influence of an artificial playing surface on injury risk and perceptions of muscle soreness in elite Rugby Union.
This prospective cohort study investigated the influence of an artificial playing surface on injury risk and perceptions of muscle soreness in elite English Premiership Rugby Union players. Time loss (from 39.5 matches) and abrasion (from 27 matches) injury risk was compared between matches played on artificial turf and natural grass. Muscle soreness was reported over the 4 days following one match played on each surface by 95 visiting players (i.e., normally play on natural grass surfaces). There was a likely trivial difference in the overall injury burden relating to time-loss injuries between playing surfaces [rate ratio = 1.01, 90% confidence interval (CI): 0.73-1.38]. Abrasions were substantially more common on artificial turf (rate ratio = 7.92, 90% CI: 4.39-14.28), although the majority of these were minor and only two resulted in any reported time loss. Muscle soreness was consistently higher over the 4 days following a match on artificial turf in comparison with natural grass, although the magnitude of this effect was small (effect sizes ranging from 0.26 to 0.40). These results suggest that overall injury risk is similar for the two playing surfaces, but further surveillance is required before inferences regarding specific injury diagnoses and smaller differences in overall injury risk can be made.


Chronic Obstructive Pulmonary Disease Increases the Risk of Hip Fracture: A Nationwide Population-Based Cohort Study.
Hip fractures can lead to functional disability and high mortality rates among elderly patients. The aim of this study was to investigate whether chronic obstructive pulmonary disease (COPD) is a risk factor for hip fracture. A retrospective population-based 4-year cohort study was conducted using case-control matched analysis of data from the Taiwan Longitudinal Health Insurance Database 2005 (LHID2005). Patients with a diagnosis of COPD between January 1, 2004 and December 31, 2007 were enrolled. A 2-stage approach and data from the National Health Interview Survey 2005 were applied to adjust for missing confounders in the LHID2005 cohort. Hazard ratios (HRs) and adjusted HRs were estimated hip fracture risk for the COPD. We enrolled 16,239 patients in the COPD cohort and 48,747 (1:3) patients in non-COPD cohort. The hip fracture incidences were 649 per 100,000 person-years in the study cohort and 369 per 100,000 person-years in non-COPD cohort. The hip fracture HR during the follow-up period was 1.78 (P < 0.001) and the adjusted hip fracture HR was 1.57 (P < 0.001) after adjustment for covariates by using the 2-stage approach method. Patients with COPD were at hip fracture risk and fracture-prevention strategies are essential for better quality of care.


Differential regulation of the expression of interleukin-2 receptor gamma-chain during the in vitro differentiation of human myeloid cells.
The common gamma-chain (gamma c) is a shared component of cell-surface receptors for the interleukins- 2, -4 and -7, and possibly others. We studied its expression in cells and cell lines of myeloid origin and found ubiquitous presence of gamma c mRNA in all cells examined. Differential regulation of gamma c expression was observed in myeloid cell lines induced to differentiate in vitro. In K-562 erythromyeloid cells, a sharp rise in the levels of gamma c mRNA and protein accompanied megakaryocytic, but not erythroid, differentiation. Surface binding of interleukin-2, as well as the transcripts for cognate receptor chains, were scarcely detectable in K-562 cells, whereas a significant increase in the binding of granulocyte-macrophage colony-stimulating factor specifically occurred during their megakaryocytic maturation. Our data indicate that expression of gamma c is a common feature of human myeloid cells, and suggest that its expression may be a requirement for human myelopoiesis.


Tumor necrosis factor-alpha: a promising therapeutic target for asthma?
Asthma is a disease that encompasses a variety of features including airway smooth muscle abnormalities, airway inflammation, and structural changes in the airway. Historically, it has been classified depending on the severity of the disease, the frequency of symptoms, and the level of treatment required to control them. Severe or refractory asthma accounts for approximately 10% of the patient population with asthma and for about 30% of the healthcare costs of this disease. It is often associated with conditions that might lead to activation of innate immunity in the lung, and it has been suggested that some of the features of severe asthma might be due to upregulation of the tumor necrosis factor-alpha (TNFalpha) pathway. In support of this, studies have shown that severe asthma is associated with an increased presence of TNFalpha within the airway and an increase in TNFalpha expression on peripheral blood mononuclear cells. Moreover, TNFalpha has the ability to induce several of the pro-inflammatory changes associated with severe asthma. Interest in the role of TNFalpha in severe asthma has increased following reports that antagonism with etanercept or infliximab is associated with improvement in asthma control in patients with severe asthma. In this article, we discuss the biology, function, and clinical effects of TNFalpha with particular reference to severe asthma.


Synthesis of magnesium complexes of ionic liquids with highly coordinating anions.
Magnesium(ii) complexes, [Mg2+(hfac-)3][Cation+], were prepared as solids from hydrophobic hexafluoroacetylacetonate ionic liquids ([Cation+][hfac-] ILs) and Mg(Tf2N)2. 1-Butyl-3-methylimidazolium ([C4mim]), N-butylpyridinium ([C4Pyr]), N-butyl-N-methylpiperidinium ([C4Pip]), N-hexyl-N-methylmorpholinium ([C6Morp]) and N-butyl-N-methylpyrrolidinium ([C4pyrr]) were used as cationic cores. The [C4Pip][hfac], [C4Pyr][hfac] and [C6Morp][hfac] ILs were prepared for the first time. New Mg(ii) complexes, [C4mim][Mg(hfac)3], [C4Pip][Mg(hfac)3], [C4Pyr][Mg(hfac)3], [C6Morp][Mg(hfac)3] and [C4Pyrr][Mg(hfac)3], were obtained from the [hfac] based ILs. The crystal structures of the novel Mg(ii) complexes show the coordination of three [hfac] anions to the Mg2+ ion through the two oxygen atoms of each [hfac] anion.


The Undiagnosed Diseases Network International: Five years and more!
Undiagnosed rare diseases (URDs) account for a significant portion of the overall rare disease burden, depending upon the country. Hence, URDs represent an unmet medical need. A specific challenge posed by the ensemble of the URD patient cohort is the heterogeneity of its composition; the group, indeed, includes very rare, still unidentified conditions as well as clinical variants of recognized rare diseases. Exact disease recognition requires new approaches that cut across national and institutional boundaries, may need the implementation of methods new to diagnostics, and embrace clinical care and research. To address these issues, the Undiagnosed Diseases Network International (UDNI) was established in 2014, with the major aims of providing diagnoses to patients, implementing additional diagnostic tools, and fostering research on novel diseases, their mechanisms, and their pathways. The UDNI involves centres with internationally recognized expertise, and its scientific resources and know-how aim to fill the knowledge gaps that impede diagnosis, in particularly for ultra-rare diseases. Consequently, the UDNI fosters the translation of research into medical practice, aided by active patient involvement. The goals of the UDNI are to work collaboratively and at an international scale to: 1) provide diagnoses for individuals who have conditions that have eluded diagnosis by clinical experts; 2) gain insights into the etiology and pathogenesis of novel diseases; 3) contribute to standards of diagnosing unsolved patients; and 4) share the results of UDNI research in a timely manner and as broadly as possible.


In vivo depletion of CD11c+ dendritic cells abrogates priming of CD8+ T cells by exogenous cell-associated antigens.
Cytotoxic T lymphocytes (CTL) respond to antigenic peptides presented on MHC class I molecules. On most cells, these peptides are exclusively of endogenous, cytosolic origin. Bone marrow-derived antigen-presenting cells, however, harbor a unique pathway for MHC I presentation of exogenous antigens. This mechanism permits cross-presentation of pathogen-infected cells and the priming of CTL responses against intracellular microbial infections. Here, we report a novel diphtheria toxin-based system that allows the inducible, short-term ablation of dendritic cells (DC) in vivo. We show that in vivo DC are required to cross-prime CTL precursors. Our results thus define a unique in vivo role of DC, i.e., the sensitization of the immune system for cell-associated antigens. DC-depleted mice fail to mount CTL responses to infection with the intracellular bacterium Listeria monocytogenes and the rodent malaria parasite Plasmodium yoelii.


The role of cognitions in short-term temporal changes in dental fear among Australian adults.
To determine if temporal changes in dental fear over a 4-month period are predicted by changed cognitive vulnerability-related perceptions of going to the dentist. Australian adults (n = 484) completed mailed questionnaires at baseline and follow-up, containing measures of dental fear, cognitive vulnerability-related perceptions, dental services received, and possible aversive experiences during the study period. Change in dental fear was the main outcome measure, categorized as decreased (Fear- ), unchanged (Fear0 ), and increased (Fear+ ). Across the study period, 15.5 percent of people had Fear- , 73.4 percent had Fear0 , and 11.1 percent had Fear+ . In a multinomial logistic regression, after controlling for participant gender, income, time since last dental visit at baseline, dental fear at baseline and experiencing an aversive event, increased vulnerability-related perceptions were significantly associated with Fear+ (OR = 2.83, P < 0.001) while decreased vulnerability-related perceptions were associated with Fear- (OR = 0.17, P < 0.001). This study found, across a relatively short 4-month period, that increased vulnerability-related perceptions of visiting the dentist predicted increased dental fear while decreased vulnerability-related perceptions predicted decreased dental fear. More appropriate study designs, such as longitudinal designs, and longer follow-up periods are needed to determine the possible causal nature of these associations.


Effectiveness and Safety of Vedolizumab in Anti-TNF-Naïve Patients With Inflammatory Bowel Disease-A Multicenter Retrospective European Study.
Vedolizumab (VDZ) is effective for treatment of ulcerative colitis (UC) and Crohn's disease (CD). In GEMINI trials, anti-tumor necrosis factor (anti-TNF)-naïve patients had a superior response compared with anti-TNF-exposed patients. In real-world experience (RWE), the number of included anti-TNF-naïve patients was low. We aimed to evaluate the effectiveness and safety of VDZ in anti-TNF-naïve patients in an RWE setting. This retrospective multicenter European pooled cohort study included consecutive active anti-TNF-naïve IBD patients treated with VDZ. The primary end point was clinical response at week 14. Patients with follow-up beyond week 14 and those discontinuing VDZ at any time were included for maintenance outcomes analysis. Since January 2015, 184 anti-TNF-naïve patients from 23 centers initiated VDZ treatment (Crohn's disease [CD], 50; ulcerative colitis [UC], 134). In CD, 42/50 (82%) patients responded by week 14 and 32 (64%) were in clinical remission; 26/50 (52%) achieved corticosteroid-free remission (CSFR). At last follow-up (44 weeks; interquartile range [IQR], 30-52 weeks), 27/35 (77.1%) patients with available data responded to treatment; 24/35 (68.6%) were in clinical remission, 21/35 (60%) were in CSFR. For UC, 116/134 (79.1%) responded to treatment by week 14, including 53 (39.5%) in clinical remission; 49/134 (36.6%) achieved CSFR. At last follow-up (42.5 weeks; IQR, 30-52 weeks), 79/103 (76.7%) patients responded to treatment, 69/103 (67.0%) were in remission, and 61/103 (59.2%) were in CSFR. Adverse effects were reported in 20 (11%) of the patients, leading to treatment discontinuation in 6 (3.3%). VDZ is similarly effective in ant-TNF-naïve CD and UC patients. The efficacy is higher than reported in anti-TNF-experienced patients and is comparable to that of anti-TNF biologics in this population.


Slow protein fluctuations explain the emergence of growth phenotypes and persistence in clonal bacterial populations.
One of the most challenging problems in microbiology is to understand how a small fraction of microbes that resists killing by antibiotics can emerge in a population of genetically identical cells, the phenomenon known as persistence or drug tolerance. Its characteristic signature is the biphasic kill curve, whereby microbes exposed to a bactericidal agent are initially killed very rapidly but then much more slowly. Here we relate this problem to the more general problem of understanding the emergence of distinct growth phenotypes in clonal populations. We address the problem mathematically by adopting the framework of the phenomenon of so-called weak ergodicity breaking, well known in dynamical physical systems, which we extend to the biological context. We show analytically and by direct stochastic simulations that distinct growth phenotypes can emerge as a consequence of slow-down of stochastic fluctuations in the expression of a gene controlling growth rate. In the regime of fast gene transcription, the system is ergodic, the growth rate distribution is unimodal, and accounts for one phenotype only. In contrast, at slow transcription and fast translation, weakly non-ergodic components emerge, the population distribution of growth rates becomes bimodal, and two distinct growth phenotypes are identified. When coupled to the well-established growth rate dependence of antibiotic killing, this model describes the observed fast and slow killing phases, and reproduces much of the phenomenology of bacterial persistence. The model has major implications for efforts to develop control strategies for persistent infections.


Patterns of RSA and observed distress during the still-face paradigm predict later attachment, compliance and behavior problems: A person-centered approach.
This study examined profiles of infant observed distress and physiological regulation indexed by respiratory sinus arrhythmia (RSA) levels during the still-face paradigm at 6 months using a person-centered approach. Mothers and infants (N = 206) participated in the study when infants were 6 months, 1 year, and 2 years old. Attachment was assessed at 1 year via the Strange Situation. Children's compliance behaviors were assessed at 2 years during a toy clean-up task. Mothers reported children's behavior problems at 4.5 years. Latent profile analysis yielded four profiles: highly distressed, but regulating; over-regulated; resilient to distress; and under-regulated. Infants in the "resilient to distress" profile characterized by high RSA levels and low negative affect exhibited the most adaptive outcomes such as lower attachment avoidance, higher compliance, and lower behavior problems. Therefore, this study highlights the importance of considering patterns of behavioral and physiological indicators of infant emotionality together for understanding adaptive functioning.


Pericytes resident in postnatal skeletal muscle differentiate into muscle fibres and generate satellite cells.
Skeletal muscle fibres form by fusion of mesoderm progenitors called myoblasts. After birth, muscle fibres do not increase in number but continue to grow in size because of fusion of satellite cells, the postnatal myogenic cells, responsible for muscle growth and regeneration. Numerous studies suggest that, on transplantation, non-myogenic cells also may contribute to muscle regeneration. However, there is currently no evidence that such a contribution represents a natural developmental option of these non-myogenic cells, rather than a consequence of experimental manipulation resulting in cell fusion. Here we show that pericytes, transgenically labelled with an inducible Alkaline Phosphatase CreERT2, but not endothelial cells, fuse with developing myofibres and enter the satellite cell compartment during unperturbed postnatal development. This contribution increases significantly during acute injury or in chronically regenerating dystrophic muscle. These data show that pericytes, resident in small vessels of skeletal muscle, contribute to its growth and regeneration during postnatal life.


Selective Activation of Shoulder, Trunk, and Arm Muscles: A Comparative Analysis of Different Push-Up Variants.
The push-up is a widely used exercise for upper limb strengthening that can be performed with many variants. A comprehensive analysis of muscle activation during the ascendant phase (AP) and descendant phase (DP) in different variants could be useful for trainers and rehabilitators. To obtain information on the effect of different push-up variants on the electromyography (EMG) of a large sample of upper limb muscles and to investigate the role of the trunk and abdomen muscles during the AP and DP. Cross-sectional study. University laboratory. Eight healthy, young volunteers without a history of upper extremity or spine injury. Participants performed a set of 10 repetitions for each push-up variant: standard, wide, narrow, forward (FP), and backward (BP). Surface EMG of 12 selected muscles and kinematics data were synchronously recorded to describe the AP and DP. Mean EMG activity of the following muscles was analyzed: serratus anterior, deltoideus anterior, erector spinae, latissimus dorsi, rectus abdominis, triceps brachii caput longus, triceps brachii caput lateralis, obliquus externus abdominis, pectoralis major sternal head, pectoralis major clavicular head, trapezius transversalis, and biceps brachii. The triceps brachii and pectoralis major exhibited greater activation during the narrow-base variant. The highest activation of abdomen and back muscles was recorded for the FP and BP variants. The DP demonstrated the least electrical activity across all muscles, with less marked differences for the abdominal and erector spinae muscles because of their role as stabilizers. Based on these findings, we suggest the narrow-base variant to emphasize triceps and pectoralis activity and the BP variant for total upper body strength conditioning. The FP and BP variants should be implemented carefully in participants with low back pain because of the greater activation of abdominal and back muscles.


Structure and functional role of dynein's microtubule-binding domain.
Dynein motors move various cargos along microtubules within the cytoplasm and power the beating of cilia and flagella. An unusual feature of dynein is that its microtubule-binding domain (MTBD) is separated from its ring-shaped AAA+ adenosine triphosphatase (ATPase) domain by a 15-nanometer coiled-coil stalk. We report the crystal structure of the mouse cytoplasmic dynein MTBD and a portion of the coiled coil, which supports a mechanism by which the ATPase domain and MTBD may communicate through a shift in the heptad registry of the coiled coil. Surprisingly, functional data suggest that the MTBD, and not the ATPase domain, is the main determinant of the direction of dynein motility.


Maternal Supplementation With Krill Oil During Breastfeeding and Long-Chain Polyunsaturated Fatty Acids (LCPUFAs) Composition of Human Milk: A Feasibility Study.
Background: Docosahexaenoic acid (DHA) is a major constituent of neuronal and retinal membranes and plays a crucial role in brain and visual development within the first months of life. Dietary intakes are fundamental to provide neonates with adequate DHA supply; hence, maternal supplementation might represent a useful strategy to implement DHA contents in breast milk (BM), with possible benefits on neonatal neurodevelopment. Antarctic krill is a small crustacean rich in highly available phospholipid-bound DHA. This pilot study aimed to evaluate whether maternal supplementation with krill oil during breastfeeding increases long-chain polyunsaturated fatty acids (LCPUFAs) BM contents. Methods: Mothers of infants admitted to the Neonatal Intensive Care Unit were enrolled in this open, randomized-controlled study between 4 and 6 weeks after delivery and randomly allocated in 2 groups. Group 1 received an oral krill oil-based supplement providing 250 mg/day of DHA and 70 mg/day of eicosapentaenoic acid (EPA) for 30 days; group 2 served as control. BM samples from both groups were collected at baseline (T0) and day 30 (T1) and underwent a qualitative analysis of LCPUFAs composition by gas chromatography/mass spectrometry. Results: Sixteen breastfeeding women were included. Of these, 8 received krill-oil supplementation and 8 were randomized to the control group. Baseline percentage values of DHA (%DHA), arachidonic acid (%AA), and EPA (%EPA) did not differ between groups. A significant increase in %DHA (T0: median 0.23% [IQR 0.19;0.38], T1:0.42% [0.32;0.49], p 0.012) and %EPA (T0: median 0.10% [IQR 0.04;0.11], T1:0.11% [0.04;0.15], p 0.036) and a significant reduction in %AA (T0: median 0.48% [IQR 0.42;0.75], T1:0.43% [0.38;0.61], p 0.017) between T0 and T1 occurred in Group 1, whereas no difference was seen in Group 2. Consistently, a significant between-group difference was observed in percentage changes from baseline of DHA (Δ%DHA, group 1: median 64.2% [IQR 27.5;134.6], group 2: -7.8% [-12.1;-3.13], p 0.025) and EPA (Δ%EPA, group 1: median 39% [IQR 15.7;73.4]; group 2: -25.62% [-32.7;-3.4], p 0.035). Conclusions: Oral krill oil supplementation effectively increases DHA and EPA contents in BM. Potential benefits of this strategy on brain and visual development in breastfed preterm neonates deserve further evaluation in targeted studies. Clinical Trial Registration: www.ClinicalTrials.gov, identifier NCT03583502.


Socio-demographic variations in moves to institutional care 1991-2001: a record linkage study from England and Wales.
Only a minority of older people in England and Wales live in institutional care, but among the older of the old, this minority is large. Disability is the major driver of admissions, but socio-demographic factors are also relevant. Understanding more about the influence of these is important for planning by long-term care. To investigate effects of socio-demographic factors, including housing tenure, household type, marital status, and number of children, on the proportions of elderly people who made a transition from living in the community in 1991 to living in institutional care in 2001. Nationally representative record linkage study including 36,650 people aged 65 years and over, living in the community in England and Wales in 1991, who were still alive in 2001. Nineteen thousand women aged 75-89 years in 2001 were included in additional analyses of effects of parity (number of children borne). Bi-variate and multivariate analyses of variations in sample proportions, who by 2001 were resident in institutional care. 4.3% of men and 9.3% of women in the surviving sample then aged 75 years and over, were in institutional care in 2001. Older age, living in rented accommodation, living alone in 1991 and being unmarried in 2001, as well as long-term illness, were associated with higher proportions making this transition. Women had higher risks than men. Childless women aged 64-79 years in 1991 had a 25% higher risk than women with children of being in institutional care in 2001. Socio-demographic factors continue to influence risks of entry to institutional care in England and Wales.


Neuraminidase inhibitor susceptibility testing in human influenza viruses: a laboratory surveillance perspective.
Neuraminidase inhibitors (NAIs) are vital in managing seasonal and pandemic influenza infections. NAI susceptibilities of virus isolates (n = 5540) collected during the 2008-2009 influenza season were assessed in the chemiluminescent neuraminidase inhibition (NI) assay. Box-and-whisker plot analyses of log-transformed IC(50)s were performed for each virus type/subtype and NAI to identify outliers which were characterized based on a statistical cutoff of IC(50) >3 interquartile ranges (IQR) from the 75(th) percentile. Among 1533 seasonal H1N1 viruses tested, 1431 (93.3%) were outliers for oseltamivir; they all harbored the H275Y mutation in the neuraminidase (NA) and were reported as oseltamivir-resistant. Only 15 (0.7%) of pandemic 2009 H1N1 viruses tested (n = 2259) were resistant to oseltamivir. All influenza A(H3N2) (n = 834) and B (n = 914) viruses were sensitive to oseltamivir, except for one A(H3N2) and one B virus, with D151V and D197E (D198E in N2 numbering) mutations in the NA, respectively. All viruses tested were sensitive to zanamivir, except for six seasonal A(H1N1) and several A(H3N2) outliers (n = 22) which exhibited cell culture induced mutations at residue D151 of the NA. A subset of viruses (n = 1058) tested for peramivir were sensitive to the drug, with exception of H275Y variants that exhibited reduced susceptibility to this NAI. This study summarizes baseline susceptibility patterns of seasonal and pandemic influenza viruses, and seeks to contribute towards criteria for defining NAI resistance.


Screw-Dislocation-Induced Strengthening-Toughening Mechanisms in Complex Layered Materials: The Case Study of Tobermorite.
Nanoscale defects such as dislocations have a profound impact on the physics of crystalline materials. Understanding and characterizing the motion of screw dislocation and its corresponding effects on the mechanical properties of complex low-symmetry materials has long been a challenge. Herein, we focus on triclinic tobermorite, as a model system and a crystalline analogue of layered hydrated cement, and report for the first time how the motion of screw dislocation can influence the strengthening-toughening relationship, imparting brittle-to-ductile transitions. By applying shear loading in tobermorite systems with single and dipole screw dislocations, we observe dislocation jogs around the dislocation core, which increases the yield shear stress and the work-of-fracture when the dislocation lines are along the [100] and [010] directions. Our results demonstrate that the dislocation core acts as a bottleneck for the initial straight gliding to induce intralaminar gliding, which consequently leads to a significant improvement in the mechanical properties. Together, the fundamental knowledge gained in this work on the role of the motion of the dislocation core on the mechanical properties provides an improved understanding of deformation mechanisms in cementitious materials and other complex layered systems, providing new hypotheses and design guidelines for the development of strong, ductile, and tough materials.


Finite element analysis of a floating microstimulator.
Analytical solutions for voltage fields in a volume conductor are available only for ideal electrodes with radially symmetric contacts and infinitely extending substrates. Practical electrodes for neural stimulation may have asymmetric contacts and finite substrate dimensions and hence deviate from the ideal geometries. For instance, it needs to be determined if the analytical solutions are adequate for simulations of narrow shank electrodes where the substrate width is comparable to the size of the contacts. As an extension to this problem, a "floating" stimulator can be envisioned where the substrate would be finite in all directions. The question then becomes how small this floating stimulator can be made before its stimulation strength is compromised by the decrease in the medium impedance between the contacts as the contacts are approaching each other. We used finite element modeling to solve the voltage and current profiles generated by these radially asymmetric electrode geometries in a volume conductor. The simulation results suggest that both the substrate size and the bipolar contact separation influence the voltage field when these parameters are as small as a few times the contact size. Both of these effects are larger for increasing elevations from the contact surface, and even stronger for floating electrodes (finite substrate in all directions) than the shank-type electrodes. Location of the contacts on the floating electrode also plays a role in determining the voltage field. The voltage field for any device size and current, and any specific resistance of the volume conductor can be predicted from these results so long as the aspect ratios are preserved.


Functional interplay between the cell cycle and cell phenotypes.
Cell cycle distribution of adherent cells is typically assessed using flow cytometry, which precludes the measurements of many cell properties and their cycle phase in the same environment. Here we develop and validate a microscopy system to quantitatively analyze the cell-cycle phase of thousands of adherent cells and their associated cell properties simultaneously. This assay demonstrates that population-averaged cell phenotypes can be written as a linear combination of cell-cycle fractions and phase-dependent phenotypes. By perturbing the cell cycle through inhibition of cell-cycle regulators or changing nuclear morphology by depletion of structural proteins, our results reveal that cell cycle regulators and structural proteins can significantly interfere with each other's prima facie functions. This study introduces a high-throughput method to simultaneously measure the cell cycle and phenotypes at single-cell resolution, which reveals a complex functional interplay between the cell cycle and cell phenotypes.


RNA probes detect nucleotide sequence homology between members of two different nairovirus serogroups.
Cloned cDNA derived from the small (S) and medium (M) genomic RNA segments of Dugbe (DUG) virus, isolate ArD44313, a member of the Nairobi sheep disease (NSD) serogroup of nairoviruses (family, Bunyaviridae) was used to prepare 32P-labelled DNA and RNA probes. The S and M segments of six isolates of DUG virus all hybridised to both DNA and RNA probes, although the M segment of isolate KT281/75 reacted only weakly. Of nine other nairoviruses tested, representing all the six other serogroups within the Nairovirus genus, none hybridised to the DNA probes. However, under conditions of low stringency, the DUG S and M RNA probes hybridised to the respective S and M segments of Ganjam (GAN) virus (another member of the NSD serogroup). The DUG S RNA probe also hybridised to the S segments of Crimean-Congo haemorrhagic fever (CCHF) virus and Hazara (HAZ) virus (members of the CCHF serogroup). The indicated sequence relationships between DUG, GAN, CCHF and HAZ viruses show that the NSD serogroup is more closely related to members of the CCHF serogroup than it is to nairoviruses of the other five serogroups.


Nonhost diversity and density reduce the strength of parasitoid-host interactions.
The presence of nonprey or nonhosts is known to reduce the strength of consumer- resource interactions by increasing the consumer's effort needed to find its resource. These interference effects can have a stabilizing effect on consumer-resource dynamics, but have also been invoked to explain parasitoid extinctions. To understand how nonhosts affect parasitoids, we manipulated the density and diversity of nonhost aphids using experimental host-parasitoid communities and tested how this affects parasitation efficiency of two aphid parasitoid species. To further study the behavioral response of parasitoids to nonhosts, we tested for changes in parasitoid time allocation in relation to their host-finding strategies. The proportion of successful attacks (attack rate) in both parasitoid species was reduced by the presence of nonhosts. The parasitoid Aphidius megourae was strongly affected by increasing nonhost diversity with the attack rate dropping from 0.39 without nonhosts to 0.05 with high diversity of nonhosts, while Lysiphlebus fabarum responded less strongly, but in a more pronounced way to an increase in nonhost density. Our experiments further showed that increasing nonhost diversity caused host searching and attacking activity levels to fall in A. megourae, but not in L. fabarum, and that A. megourae changed its behavior after a period of time in the presence of nonhosts by increasing its time spent resting. This study shows that nonhost density and diversity in the environment are crucial determinants for the strength of consumer-resource interactions. Their impact upon a consumer's efficiency strongly depends on its host/prey finding strategy as demonstrated by the different responses for the two parasitoid species. We discuss that these trait-mediated indirect interactions between host and nonhost species are important for community stability, acting either stabilizing or destabilizing depending on the level of nonhost density or diversity present.


The K box, a conserved 3' UTR sequence motif, negatively regulates accumulation of enhancer of split complex transcripts.
Cell-cell interactions mediated by the Notch receptor play an essential role in the development of the Drosophila adult peripheral nervous system (PNS). Transcriptional activation of multiple genes of the Enhancer of split Complex [E(spl)-C] is a key intracellular response to Notch receptor activity. Here we report that most E(spl)-C genes contain a novel sequence motif, the K box (TGTGAT), in their 3' untranslated regions (3' UTRs). We present three lines of evidence that demonstrate the importance of this element in the post-transcriptional regulation of E(spl)-C genes. First, K box sequences are specifically conserved in the orthologs of two structurally distinct E(spl)-C genes (m4 and m8) from a distantly related Drosophila species. Second, the wild-type m8 3' UTR strongly reduces accumulation of heterologous transcripts in vivo, an activity that requires its K box sequences. Finally, m8 genomic DNA transgenes lacking these motifs cause mild gain-of-function PNS defects and can partially phenocopy the genetic interaction of E(spl)D with Notchspl. Although E(spl)-C genes are expressed in temporally and spatially specific patterns, we find that K box-mediated regulation is ubiquitous, implying that other targets of this activity may exist. In support of this, we present sequence analyses that implicate genes of the iroquois Complex (Iro-C) and engrailed as additional targets of K box-mediated regulation.


Vigabatrin caused rapidly progressive deterioration in two cases with early myoclonic encephalopathy associated with nonketotic hyperglycinemia.
Vigabatrin, a structural analogue of gamma-aminobutyric acid (GABA), is used for the treatment of generalized and partial seizures in infants. The drug inhibits the GABA transaminase and elevates the GABA concentration in the brain. Here we present the vigabatrin experience in two patients with early myoclonic encephalopathy owing to nonketotic hyperglycinemia (glycine encephalopathy). Both patients had early infantile seizures characterized by fragmentary myoclonic jerks associated with burst-suppression pattern on electroencephalography. Nonketotic hyperglycinemia was diagnosed with elevated cerebrospinal fluid and plasma glycine levels. The seizures were initially thought to be infantile spasms, and vigabatrin (50 mg /kg/day) was started for the treatment of seizures. Rapidly progressive deterioration was noticed after a few days. Acute encephalopathy associated with sleepiness and respiratory failure developed. Vigabatrin produced acute encephalopathy, which regressed in a few days after vigabatrin was stopped in the first patient. However, in the second case, despite the discontinuation of vigabatrin, there was no recovery of general conditions. Our observations in two cases indicate the risk of using vigabatrin in patients with nonketotic hyperglycinemia. The elevated GABA concentration in the brain can enhance the encephalopathy, together with the elevated levels of glycine. (J Child Neurol 2006;21:82-84).


Identification of Angiotensin I-Converting Enzyme-Inhibitory and Anticoagulant Peptides from Enzymatic Hydrolysates of Chicken Combs and Wattles.
Peptides from protein hydrolysate of a mixture of chicken combs and wattles (CCWs) were obtained through enzymatic hydrolysis, and their anticoagulant and inhibitory effects on angiotensin I-converting enzyme (ACE) were investigated. The protein hydrolysate exhibited anticoagulant capacity by the intrinsic pathway (activated partial thromboplastin time) and potent ACE-inhibitory activity. The peptides were sequenced by LC-MS to identify those with higher inhibitory potential. From the pool of sequenced peptides, the following three peptides were selected and synthesized based on their low molecular weight and the presence of amino acids with ACE-inhibitory potential at the C-terminus: peptide I (APGLPGPR), peptide II (Piro-GPPGPT), and peptide III (FPGPPGP). Peptide III (FPGPPGP) showed the highest ACE-inhibitory capacity among the peptides selected. In conclusion, a peptide (FPGPPGP) of unknown sequence was identified as having potent ACE-inhibitory capacity. This peptide originated from unconventional hydrolysates from poultry slaughter waste, including combs and wattles.


Effect of nocturnal melatonin intake on cellular damage and recovery from repeated sprint performance during an intensive training schedule.
An optimal recovery between training sessions is of similar if not greater importance as the training content and program of the training, itself. One of the most used strategies for improving recovery is the ingestion of supplements. The present study aimed to evaluate the effect of 5 mg oral melatonin supplementation on the recovery from repeated sprint (RSA) of performance and biochemical responses (i.e. oxidative stress, leukocytosis cellular damage) after an intensive training camp (TC). Twenty soccer players performed an RSA test before and after an intensive six-day TC associated with nocturnal melatonin (n = 10) or placebo (n = 10) ingestion. Resting and post-RSA test blood samples were obtained before and after the TC. Compared to placebo, melatonin intake decreased resting oxidative stress markers (i.e, advanced oxidation protein products), leukocytosis (i.e. white blood cells (WBC), neutrophils (NE)) and biomarkers of cellular damage (i.e. creatine kinase (CK)). It also lowered post-exercise leukocytosis (i.e. WBC, NE, lymphocytes (LY), monocytes (MO)) and biomarkers of cellular damage (i.e. CK, aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT)) and raised the activity of the main antioxidant enzymes (i.e. glutathione peroxidase (GPx), glutathione reductase (GR)). In addition, compared to placebo, melatonin reduced the deterioration of the best and total time during the RSA test after the TC. In conclusion, nocturnal melatonin supplementation during an intensive TC alleviated oxidative stress, leukocytosis and cellular damage and improved recovery of RSA performance in soccer players.


Thermal activation of the co-chaperonins GroES and gp31 probed by mass spectrometry.
Many biological active proteins are assembled in protein complexes. Understanding the (dis)assembly of such complexes is therefore of major interest. Here we use mass spectrometry to monitor the disassembly induced by thermal activation of the heptameric co-chaperonins GroES and gp31. We use native electrospray ionization mass spectrometry (ESI-MS) on a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer to monitor the stoichiometry of the chaperonins. A thermally controlled electrospray setup was employed to analyze conformational and stoichiometric changes of the chaperonins at varying temperature. The native ESI-MS data agreed well with data obtained from fluorescence spectroscopy as the measured thermal dissociation temperatures of the complexes were in good agreement. Furthermore, we observed that thermal denaturing of GroES and gp31 proceeds via intermediate steps of all oligomeric forms, with no evidence of a transiently stable unfolded heptamer. We also evaluated the thermal dissociation of the chaperonins in the gas phase using infrared multiphoton dissociation (IRMPD) for thermal activation. Using gas-phase activation the smaller (2-4) oligomers were not detected, only down to the pentamer, whereafter the complex seemed to dissociate completely. These results demonstrate clearly that conformational changes of GroES and gp31 due to heating in solution and in the gas phase are significantly different.


BM88/Cend1 regulates stimuli-induced intracellular calcium mobilization.
In neurogenesis, little is known about signal transduction pathways upstream of gene expression however, mounting evidence suggests that calcium release from internal stores plays a critical role. We have previously demonstrated that BM88 is a neuronal lineage-specific regulator of cell cycle exit and differentiation; we now report a link between BM88 and calcium signaling. Calcium imaging experiments revealed that P2Y-induced calcium mobilization is diminished in mouse neuroblastoma Neuro 2a cells stably transfected with BM88 (N2A-BM88 cells) as compared with N2A cells or N2A cells differentiated with retinoic acid. This effect is not restricted to N2A cells but is also observed in HeLa cells that are transiently transfected with BM88, indicating that cells of both neural and non-neural origin respond similarly. Further, activation of P2Y1 but not purinergic P2X receptors induces proliferation of N2A and to a lesser extent of N2A-BM88 cells. Conversely, knockdown of BM88 facilitates N2A cell proliferation both under stimulating and non-stimulating conditions. Importantly, N2A-BM88 cells are less susceptible to apoptosis triggered by C2-ceramide and exhibit reduced C2-ceramide-induced intracellular calcium release. Higher calcium uptake from mitochondria and/or lower calcium levels inside the endoplasmic reticulum may explain the reduced calcium mobilization in response to BM88. Overall, our data reveal a novel signaling mechanism by which BM88 interferes with calcium release from inositol 1,4,5-trisphosphate-sensitive stores and exerts anti-proliferative and anti-apoptotic functions.


Penile fracture: retrospective analysis of our case history with long-term assessment of the erectile and sexological outcome.
To review the cases of patients with suspected penile fracture and asses erectile and sexological outcomes. from 1987 to 2013 presented to the Urology Clinic of Trieste and at the AIED of Pordenone a total of 41 cases that were divided into two groups according to the timing of treatment: 18 patients with anamnestic diagnosis of penile fracture treated nonimmediately and 23 patients treated immediately after the trauma. For all patients we evaluated the type of treatment adopted, the occurrence of complications and reoperations and the follow-up. The erectile function was also evaluated through the IIEF, as well as the psychological impact of the trauma on social and sex life, using a psycho- sexological questionnaire. Among patients treated immediately after the trauma 14 were subjected to surgery. About a year after surgery, penile curvature was reported in 1 patient, pain in 3 patients, urinary disorders in 1 patient, while none reported erectile dysfunction (ED). Out of these, only 3 patients underwent reoperation. Among those treated conservatively 1 patient reported curvature, 1 patient reported pain and none reported ED. Among patients who were admitted at a later date, 14 reported curvature and 4 reported pain whereas urinary disorders were reported in 1 and ED in 4 patients. From a psychological point of view, the trauma caused in most cases a fear of new trauma and of repercussions on erectile function and sensitivity. The diagnosis is mainly clinical; however, radiological investigation is essential to confirm the diagnosis, assess the site and extent of the trauma and possible urethral involvement, so as to plan the most appropriate treatment. In addition, immediate treatment leads to better long-term results, with a lower incidence of ED and penile curvature. Psychologically, penile trauma intensifies the fear of reoccurrence; it decreases, however, with the passage of time.


Attitudes of emergency department patients and visitors regarding emergency exception from informed consent in resuscitation research, community consultation, and public notification.
To assess public views on emergency exception to informed consent in resuscitation research, public awareness of such studies, and effective methods of community consultation and public notification. A face-to-face survey was conducted in two academic Level I trauma center emergency departments (EDs) in Oregon and Minnesota from June through August 2001. Five hundred thirty people completed the survey, with an 82% response rate. The mean age of the respondents was 41 years (range 18-95) with a standard deviation of 14.5; 46% were female and 64% white. Most (88%) believed that research subjects should be informed prior to being enrolled, while 49% believed enrolling patients without prior consent in an emergency situation would be acceptable and 70% (369) would not object to be entered into such a study without providing prospective informed consent. Informing and consulting the community as a substitute for patient consent in emergency research was thought to be reasonable by 45% of the respondents. Most respondents would prefer to be informed about a study using emergency exception from informed consent by radio and television media (42%). Two hundred fifty-eight respondents (49%) stated they would attend a community meeting; the less educated were more likely to attend than those with college degrees (OR = 0.53; 95% CI = 0.33 to 0.85, p = 0.008). However, only 5% knew of ongoing studies in their community using emergency exception from informed consent. Most respondents disagreed with foregoing prospective informed consent for research participation even in emergency situations; however, many would be willing to participate in studies using emergency exception from informed consent. Most respondents would not attend community meetings, and would prefer to rely upon the media for information. Very few were aware of emergency exception from informed consent studies in their community. This suggests that current methods of community notification may not be effective.


Assessment of multi-pulse laser-induced damage threshold of metallic mirrors for Thomson scattering system.
Multi-pulse laser-induced damage threshold (LIDT) was experimentally investigated up to ~10(6) pulses for Cu, Ag mirrors. The surface roughness and the hardness dependence on the LIDT were also examined. The LIDT of OFHC-Cu decreased with the pulse number and was 1.0 J/cm(2) at 1.8 × 10(6) pulses. The expected LIDT of cutting Ag at 10(7) pulses was the highest; Ag mirror would be one of the best choices for ITER Thomson scattering system. For the roughness and hardness, material dependences of LIDT are discussed with experimental results.


The Food and Beverage Occurrence of Furfuryl Alcohol and Myrcene-Two Emerging Potential Human Carcinogens?
For decades, compounds present in foods and beverages have been implicated in the etiology of human cancers. The World Health Organization (WHO) International Agency for Research on Cancer (IARC) continues to classify such agents regarding their potential carcinogenicity in humans based on new evidence from animal and human studies. Furfuryl alcohol and β-myrcene are potential human carcinogens due to be evaluated. The major source of furfuryl alcohol in foods is thermal processing and ageing of alcoholic beverages, while β-myrcene occurs naturally as a constituent of the essential oils of plants such as hops, lemongrass, and derived products. This study aimed to summarize the occurrence of furfuryl alcohol and β-myrcene in foods and beverages using literature review data. Additionally, results of furfuryl alcohol occurrence from our own nuclear magnetic resonance (NMR) analysis are included. The highest content of furfuryl alcohol was found in coffee beans (>100 mg/kg) and in some fish products (about 10 mg/kg), while among beverages, wines contained between 1 and 10 mg/L, with 8 mg/L in pineapple juice. The content of β-myrcene was highest in hops. In conclusion, the data about the occurrence of the two agents is currently judged as insufficient for exposure and risk assessment. The results of this study point out the food and beverage groups that may be considered for future monitoring of furfuryl alcohol and β-myrcene.


Lactobacillus reuteri strains protect epithelial barrier integrity of IPEC-J2 monolayers from the detrimental effect of enterotoxigenic Escherichia coli.
Lactobacillus reuteri is an inhabitant of the gastrointestinal (GI) tract of mammals and birds and several strains of this species are known to be effective probiotics. The mechanisms by which L. reuteri confers its health-promoting effects are far from being fully understood, but protection of the mucosal barrier is thought to be important. Leaky gut is a state of abnormal intestinal permeability with implications for the pathophysiology of various gastrointestinal disorders. Enterotoxigenic Escherichia coli (ETEC) can invade the intestinal mucosa and induce changes in barrier function by producing enterotoxin or by direct invasion of the intestinal epithelium. Our hypothesis was that L. reuteri can protect the mucosal barrier, and the goal of the study was to challenge this hypothesis by monitoring the protective effect of L. reuteri strains on epithelial dysfunction caused by ETEC. Using an infection model based on the porcine intestinal cell line IPEC-J2, it was demonstrated that pretreatment of the cells with human-derived L. reuteri strains (ATCC PTA 6475, DSM 17938 and 1563F) and a rat strain (R2LC) reduced the detrimental effect of ETEC in a dose-dependent manner, as monitored by permeability of FITC-dextran and transepithelial electrical resistance (TEER). Moreover, the results revealed that ETEC upregulated proinflammatory cytokines IL-6 and TNFα and decreased expression of the shorter isoform of ZO-1 (187 kDa) and E-cadherin. In contrast, pretreatment with L. reuteri DSM 17938 and 1563F downregulated expression of IL-6 and TNFα, and led to an increase in production of the longer isoform of ZO-1 (195 kDa) and maintained E-cadherin expression. Interestingly, expression of ZO-1 (187 kDa) was preserved only when the infected cells were pretreated with strain 1563F. These findings demonstrate that L. reuteri strains exert a protective effect against ETEC-induced mucosal integrity disruption.


Effects of yoga in patients with paroxysmal atrial fibrillation - a randomized controlled study.
Patients with atrial fibrillation often have an impaired quality of life (QoL). Practising yoga may decrease stress and have positive effects on mental and physical health. The aim of this study was to investigate whether yoga can improve QoL and decrease blood pressure and heart rate in patients with paroxysmal atrial fibrillation (PAF). In this pilot study, 80 patients diagnosed with PAF were randomized to standard treatment (control group, n=40) or standard treatment in combination with yoga (yoga group, n=40) during a 12-week period. QoL, blood pressure and heart rate were evaluated at baseline and at the end of the study (12 (+2) weeks). EuroQoL-5D (EQ-5D) Visual Analogue Scale (VAS) and the two dimensions in Short-Form Health Survey (SF-36) were used to evaluate QoL. At baseline there was a significant difference in QoL between the groups in EQ-5D VAS- scale ( p=0.02) and SF-36 mental health score ( p<0.001) in which the control group had higher scores. At the end of the study, the yoga group averaged higher SF-36 mental health scores. There was a significant difference between the two groups ( p=0.016), but no differences in EQ-5D VAS- scale and physiological health score was seen between the two groups. At the end of the study, the yoga group had significantly lower heart rate ( p=0.024) and systolic ( p=0.033) and diastolic blood pressure ( p<0.001) compared to the control group. Yoga with light movements and deep breathing may lead to improved QoL, lower blood pressure and lower heart rate in patients with PAF compared to a control group. Yoga could be a complementary treatment method to standard therapy.


STEPS: efficient simulation of stochastic reaction-diffusion models in realistic morphologies.
Models of cellular molecular systems are built from components such as biochemical reactions (including interactions between ligands and membrane-bound proteins), conformational changes and active and passive transport. A discrete, stochastic description of the kinetics is often essential to capture the behavior of the system accurately. Where spatial effects play a prominent role the complex morphology of cells may have to be represented, along with aspects such as chemical localization and diffusion. This high level of detail makes efficiency a particularly important consideration for software that is designed to simulate such systems. We describe STEPS, a stochastic reaction-diffusion simulator developed with an emphasis on simulating biochemical signaling pathways accurately and efficiently. STEPS supports all the above-mentioned features, and well-validated support for SBML allows many existing biochemical models to be imported reliably. Complex boundaries can be represented accurately in externally generated 3D tetrahedral meshes imported by STEPS. The powerful Python interface facilitates model construction and simulation control. STEPS implements the composition and rejection method, a variation of the Gillespie SSA, supporting diffusion between tetrahedral elements within an efficient search and update engine. Additional support for well-mixed conditions and for deterministic model solution is implemented. Solver accuracy is confirmed with an original and extensive validation set consisting of isolated reaction, diffusion and reaction-diffusion systems. Accuracy imposes upper and lower limits on tetrahedron sizes, which are described in detail. By comparing to Smoldyn, we show how the voxel-based approach in STEPS is often faster than particle-based methods, with increasing advantage in larger systems, and by comparing to MesoRD we show the efficiency of the STEPS implementation. STEPS simulates models of cellular reaction-diffusion systems with complex boundaries with high accuracy and high performance in C/C++, controlled by a powerful and user-friendly Python interface. STEPS is free for use and is available at http://steps.sourceforge.net/


Association between a functional polymorphism in the serotonin transporter gene and diarrhoea predominant irritable bowel syndrome in women.
Serotonin (5-hydroxtryptamine, 5-HT) is an important factor in gut function, playing key roles in intestinal peristalsis and secretion, and in sensory signalling in the brain-gut axis. Removal from its sites of action is mediated by a specific protein called the serotonin reuptake transporter (SERT or 5-HTT). Polymorphisms in the promoter region of the SERT gene have effects on transcriptional activity, resulting in altered 5-HT reuptake efficiency. It has been speculated that such functional polymorphisms may underlie disturbance in gut function in individuals suffering with disorders such as irritable bowel syndrome (IBS). The aim of this study was to assess the potential association between SERT polymorphisms and the diarrhoea predominant IBS (dIBS) phenotype. A total of 194 North American Caucasian female dIBS patients and 448 female Caucasian controls were subjected to genotyping. Leucocyte DNA of all subjects was analysed by polymerase chain reaction based technologies for nine SERT polymorphisms, including the insertion/deletion polymorphism in the promoter (SERT-P) and the variable tandem repeat in intron 2. Statistical analysis was performed to assess association of any SERT polymorphism allele with the dIBS phenotype. A strong genotypic association was observed between the SERT-P deletion/deletion genotype and the dIBS phenotype (p = 3.07x10(-5); n = 194). None of the other polymorphisms analysed was significantly associated with the presence of disease. Significant association was observed between dIBS and the SERT-P deletion/deletion genotype, suggesting that the serotonin transporter is a potential candidate gene for dIBS in women.


Light generation and harvesting in a van der Waals heterostructure.
Two-dimensional (2D) materials are a new type of materials under intense study because of their interesting physical properties and wide range of potential applications from nanoelectronics to sensing and photonics. Monolayers of semiconducting transition metal dichalcogenides MoS2 or WSe2 have been proposed as promising channel materials for field-effect transistors. Their high mechanical flexibility, stability, and quality coupled with potentially inexpensive production methods offer potential advantages compared to organic and crystalline bulk semiconductors. Due to quantum mechanical confinement, the band gap in monolayer MoS2 is direct in nature, leading to a strong interaction with light that can be exploited for building phototransistors and ultrasensitive photodetectors. Here, we report on the realization of light-emitting diodes based on vertical heterojunctions composed of n-type monolayer MoS2 and p-type silicon. Careful interface engineering allows us to realize diodes showing rectification and light emission from the entire surface of the heterojunction. Electroluminescence spectra show clear signs of direct excitons related to the optical transitions between the conduction and valence bands. Our p-n diodes can also operate as solar cells, with typical external quantum efficiency exceeding 4%. Our work opens up the way to more sophisticated optoelectronic devices such as lasers and heterostructure solar cells based on hybrids of 2D semiconductors and silicon.


Milnacipran: a unique antidepressant?
Tricyclic antidepressants (TCAs) are among the most effective antidepressants available, although their poor tolerance at usual recommended doses and toxicity in overdose make them difficult to use. While selective serotonin reuptake inhibitors (SSRIs) are better tolerated than TCAs, they have their own specific problems, such as the aggravation of sexual dysfunction, interaction with coadministered drugs, and for many, a discontinuation syndrome. In addition, some of them appear to be less effective than TCAs in more severely depressed patients. Increasing evidence of the importance of norepinephrine in the etiology of depression has led to the development of a new generation of antidepressants, the serotonin and norepinephrine reuptake inhibitors (SNRIs). Milnacipran, one of the pioneer SNRIs, was designed from theoretic considerations to be more effective than SSRIs and better tolerated than TCAs, and with a simple pharmacokinetic profile. Milnacipran has the most balanced potency ratio for reuptake inhibition of the two neurotransmitters compared with other SNRIs (1:1.6 for milnacipran, 1:10 for duloxetine, and 1:30 for venlafaxine), and in some studies milnacipran has been shown to inhibit norepinephrine uptake with greater potency than serotonin (2.2:1). Clinical studies have shown that milnacipran has efficacy comparable with the TCAs and is superior to SSRIs in severe depression. In addition, milnacipran is well tolerated, with a low potential for pharmacokinetic drug-drug interactions. Milnacipran is a first-line therapy suitable for most depressed patients. It is frequently successful when other treatments fail for reasons of efficacy or tolerability.


Hemorrhagic shock hemodynamic and metabolic behavior: experimental study in dogs.
Different experimental models have been used to illustrate hemodynamic and metabolic changes during hemorrhagic shock. This study aimed at observing hemodynamic and metabolic behaviors during a sequential and progressive model of hemorrhagic shock in dogs to determine which indices are earlier changed. The study involved 13 dogs under total intravenous anesthesia with sodium pentobarbital, standard ventilation and previously splenectomized. Animals were not hydrated and bleeding velocity was dictated by animals blood pressure. Evaluated parameters were: hemodynamic (heart rate - HR, mean blood pressure - MBP, systemic vascular resistance index - SVRI, stroke index - SI, cardiac index - CI, shock index - i.Shock, left ventricular stroke work index - LVSWI, pulmonary capillary pressure - PCP, central venous pressure - CVP); and metabolic (mixed venous saturation - SvO2, venous oxygen pressure - PvO2, oxygen transport - O2T, oxygen consumption - VO2, oxygen extraction - TEO2, serum lactate). Results and parameters were evaluated in 6 different moments: M1 - control, and the other moments corresponding to gradual 10% decrease in calculated volume for each animal. Hemorrhage has determined significant heart rate decrease in M6 only; MBP, CI, i.Shock, LVSWI decrease in all studied moments; mild CVP and PCP change in all moments; PVO2 and SVO2 decrease in all moments; O2T decrease, VO2 stabilization and TEO2 increase in all moments; shock index was increased from M1 to M3, was decreased in M4 and again increased until M6; SVRI was increased from M1 to M4, remained unchanged in M5 and has significantly decreased in M6; lactate has increased as from M5 and M6. Mean blood pressure, heart rate, central venous pressure and pulmonary capillary pressure have not reflected real volume status of dogs in our experimental model, oxygen transport, consumption and extraction rate are useful parameters to determine hemorrhagic shock reversibility and prognosis.


Serial human passage of simian immunodeficiency virus by unsterile injections and the emergence of epidemic human immunodeficiency virus in Africa.
There is compelling evidence that both human immunodeficiency virus (HIV) types emerged from two dissimilar simian immunodeficiency viruses (SIVs) in separate geographical regions of Africa. Each of the two HIVs has its own simian progenitor and specific genetic precursor, and all of the primates that carry these SIVs have been in close contact with humans for thousands of years without the emergence of epidemic HIV. To date no plausible mechanism has been identified to account for the sudden emergence in the mid-20th century of these epidemic HIVs. In this study we examine the conditions needed for SIV to complete the genetic transition from individual human SIV infections to epidemic HIV in humans. The genetic distance from SIV to HIV and the mutational activity needed to achieve this degree of adaptation to human hosts is placed within a mathematical model to estimate the probabilities of SIV completing this transition within a single SIV-infected human host. We found that the emergence of even one epidemic HIV strain, following a single human exposure to SIV, was very unlikely. And the probability of four or more such transitions (i.e. HIV-1 groups M, O and HIV-2 subtypes A and B) occurring in a brief period is vanishingly small. We conclude that SIV cannot become a zoonosis, but requires adaptive mutations to become HIV. Some modern event must have aided in the transition of SIV to HIV. Our research indicates that serial passage of partially adapted SIV between humans could produce the series of cumulative mutations sufficient for the emergence of epidemic HIV strains. We examined the rapid growth of unsterile injections in Africa beginning in the 1950s as a biologically plausible event capable of greatly increasing serial human passage of SIV and generating HIV by a series of multiple genetic transitions. We conclude that increased unsterile injecting in Africa during the period 1950-1970 provided the agent for SIV human infections to emerge as epidemic HIV in the modern era.


Activity-dependent regulation of potassium currents in an identified neuron of the stomatogastric ganglion of the crab Cancer borealis.
Identified neurons of the stomatogastric ganglion of the crab Cancer borealis were voltage-clamped, and the current densities of three K+ currents were measured. The current densities of each of the three K+ currents varied twofold to fivefold in inferior cardiac (IC) neurons from different animals. Conventionally, this degree of variability has been attributed to experimental artifacts. Instead, we suggest that it reflects a natural variability that may be related to an underlying process of plasticity. First, we found that there is no fixed ratio among the three K+ currents. Second, we found that several hours of stimulation with depolarizing current pulses (0.5 sec duration at 1 Hz) altered the current density of the Ca2+-dependent outward current, IK(Ca), and the transient outward current, IA. This stimulation paradigm mimics the normal pattern of activity for these neurons. The effect of stimulation on the IA current density was eliminated when Ca2+ influx was blocked by extracellular Cd2+. In contrast, the K+ current densities of the lateral pyloric (LP) neuron were unaffected by the same pattern of stimulation, and the currents expressed by both the IC and the LP neurons were insensitive to hyperpolarizing pulses at the same frequency. We conclude that the conductance densities expressed by neurons may vary continually depending on the recent history of electrical activity in the preparation, and that intracellular Ca2+ may play a role in the processes by which activity influences the regulation of current densities in neurons.


Reduced virulence of an extensively drug-resistant outbreak strain of Mycobacterium tuberculosis in a murine model.
Bacterial drug resistance is often associated with a fitness cost. Large outbreaks of multidrug-resistant (MDR) and extensively drug-resistant (XDR) TB have been described that predominately affect persons with HIV infection. We obtained four closely-related Mycobacterium tuberculosis strains (genotype F15/LAM4/KZN) from an outbreak in KwaZulu-Natal (KZN), South Africa, including drug-sensitive, MDR, and XDR clinical isolates. We compared the virulence of these strains in a murine model of aerosol M. tuberculosis infection for four phenotypes: (1) competitive in vivo growth in lung and spleen, (2) non-competitive in vivo growth in lung and spleen, (3) murine survival time, and (4) lung pathology. When mixtures of sensitive, MDR, and XDR KZN strains were aerosolized (competitive model), lung CFUs were similar at 60 days after infection, and spleen CFUs were ordered as follows: sensitive > MDR > XDR. When individual strains were aerosolized (non-competitive model), modest differences in lung and spleen CFUs were observed with the same ordering. C57BL/6, C3H/FeJ, and SCID mice all survived longer after infection with MDR as compared to sensitive strains. SCID mice infected with an XDR strain survived longer than those infected with MDR or sensitive strains. Lung pathology was reduced after XDR TB infection compared to sensitive or MDR TB infection. In summary, increasing degrees of drug resistance were associated with decreasing murine virulence in this collection of KZN strains as measured by all four virulence phenotypes. The predominance of HIV-infected patients in MDR and XDR TB outbreaks may be explained by decreased virulence of these strains in humans.


Chinese Medicine for Coronavirus Disease 2019 (COVID-19): A GRADE-Assessed Systematic Review and Meta-Analysis.
Coronavirus disease 2019 (COVID-19) has caused enormous public health and socioeconomic burden globally. This study aims to evaluate the efficacy and safety of Chinese medicine (CM) against COVID-19. Eleven databases were searched on April 30, 2021, and 52 studies were included. The RoB 2.0, ROBINS-I, and GRADE tools were employed to assess the risks and evidence grades. The findings with moderate certainty in GRADE showed that compared with routine treatment (RT), Lianhua Qingwen granules (LHQW) adjunctive to RT showed significantly improved efficacy rate (relative risk (RR) = 1.19, 95% confidence interval (CI): [1.09, 1.31]), febrile score (standard mean difference (SMD) = -1.21, 95% CI: [-1.43, -0.99]), and computerized tomography (CT) lung images (RR = 1.23, 95% CI: [1.10, 1.38]); Qingfei Paidu decoction (QFPD) plus RT significantly shortened the length of hospital stay (SMD = -1.83, 95% CI: [-2.18, -1.48]); Feiyan Yihao formula (FYYH) plus RT significantly improved the clinical efficacy rate (RR = 1.07, 95% CI: [1, 1.15]), febrile time (SMD = -0.02, 95% CI: [-0.23, 0.19]), and time to negative PCR test for COVID-19 (SMD = -0.72, 95% CI: [-0.94, -0.51]). Adjunctive effects of CM with lower certainty of evidence were found, including the improvements of symptoms, laboratory findings, and mortality. No or mild adverse events were observed in most of the studies. In conclusion, the current evidence indicates that CM formulae, particularly LHQW, QFPD, and FYYH, have adjunctive effects on the standard treatment of COVID-19.


Rasch Mixture Models for DIF Detection: A Comparison of Old and New Score Specifications.
Rasch mixture models can be a useful tool when checking the assumption of measurement invariance for a single Rasch model. They provide advantages compared to manifest differential item functioning (DIF) tests when the DIF groups are only weakly correlated with the manifest covariates available. Unlike in single Rasch models, estimation of Rasch mixture models is sensitive to the specification of the ability distribution even when the conditional maximum likelihood approach is used. It is demonstrated in a simulation study how differences in ability can influence the latent classes of a Rasch mixture model. If the aim is only DIF detection, it is not of interest to uncover such ability differences as one is only interested in a latent group structure regarding the item difficulties. To avoid any confounding effect of ability differences (or impact), a new score distribution for the Rasch mixture model is introduced here. It ensures the estimation of the Rasch mixture model to be independent of the ability distribution and thus restricts the mixture to be sensitive to latent structure in the item difficulties only. Its usefulness is demonstrated in a simulation study, and its application is illustrated in a study of verbal aggression.


Death of photoreceptors in organotypic retinal explant cultures: implication of rhodopsin accumulation and endoplasmic reticulum stress.
Here we suggest that endoplasmic reticulum (ER)-stress may be induced following aberrant rhodopsin accumulation in photoreceptors in explanted rat retinas. Rhodopsin accumulation was accompanied by increased phosphorylation of pancreatic ER-kinase and eukaryotic initiator factor 2α as well as increased levels of C/EBP homologous protein, glucose-regulated protein 78 and eventually increased cleaved caspase-12 and cleaved caspase-3. Glucose-regulated protein 78, pancreatic ER-kinase, caspase-12 and cleaved caspase-3 were present in photoreceptors, indicating that ER-stress and apoptosis are induced in this cell population. These results suggest that ER-stress and subsequent apoptosis is induced in healthy photoreceptors, presumably by aberrant accumulation of rhodopsin and the phosphorylation of eukaryotic initiator factor 2α. The explant culture system may allow investigations of neuroprotective strategies.


Apj+ Vessels Drive Tumor Growth and Represent a Tractable Therapeutic Target.
Identification of cellular surface markers that distinguish tumorous from normal vasculature is important for the development of tumor vessel-targeted therapy. Here, we show that Apj, a G protein-coupled receptor, is highly enriched in tumor endothelial cells but absent from most endothelial cells of adult tissues in homeostasis. By genetic targeting using Apj-CreER and Apj-DTRGFP-Luciferase, we demonstrated that hypoxia-VEGF signaling drives expansion of Apj+ tumor vessels and that targeting of these vessels, genetically and pharmacologically, remarkably inhibits tumor angiogenesis and restricts tumor growth. These in vivo findings implicate Apj+ vessels as a key driver of pathological angiogenesis and identify Apj+ endothelial cells as an important therapeutic target for the anti-angiogenic treatment of tumors.


Religious orientation and pain management.
Occupational therapists may find pain management less effective than anticipated for the patient's condition. In exploring additional avenues of relief, it is important to look beyond physical agents. The therapist must sort physical, psychological, and spiritual components of pain to provide appropriate intervention or referral. The interface between health care and religion is strongest at the point of confrontation with a life-changing event. Literature addressing the relationship between religious orientation and pain perception differentiates among major religions but not among differences in beliefs of diverse denominational groups. Understanding beliefs about pain may be indispensable to its effective management. Productivity demands necessitate sensitivity to factors other than the physical in order to avoid ineffective treatment. The patient's beliefs can be explored within the contexts of initial evaluation and the course to treatment. Knowledge of the mechanics of pain and treatment modalities can be augmented by awareness of the influence of beliefs on response to treatment.


Circulating thyroglobulin transcytosed by thyroid cells in complexed with secretory components of its endocytic receptor megalin.
After its endocytosis from the colloid, some thyroglobulin (Tg) is transcytosed intact across thyrocytes, accounting in part for its presence in the circulation. We previously showed that megalin (gp330), an endocytic Tg receptor, mediates apical to basolateral Tg transcytosis. Here we investigated whether a portion of megalin remains combined with Tg after its transcytosis, using studies with cultured thyroid cells and in vivo observations. FRTL-5 cells, a rat thyroid cell line, cultured on filters in dual chambers form tight junctions and exhibit features of polarity, with expression of megalin exclusively on the upper (apical) surface. After the addition of unlabeled Tg to the upper chamber and incubation at 37 C, some Tg was transcytosed intact across FRTL-5 cells into the lower chamber. Two antimegalin ectodomain antibodies precipitated transcytosed Tg in fluids collected from the lower chamber. After the addition of Tg to surface-biotinylated FRTL-5 cells, an anti-Tg antibody and the two antimegalin ectodomain antibodies precipitated high molecular mass biotinylated material in fluids collected from the lower chamber, corresponding to much of the megalin ectodomain, as well as smaller amounts of lower molecular mass material. The results indicate that Tg transcytosed across FRTL-5 cells remains complexed with megalin ectodomain components, which we refer to as megalin secretory components. In aminotriazole-treated rats, which develop increased megalin-mediated Tg transcytosis, antimegalin antibodies precipitated some of the Tg in the serum. Tg was also precipitated by antimegalin antibodies in sera from patients with Graves' disease, in which we found increased megalin expression on the apical surface of thyrocytes. In contrast, in thyroidectomized patients with metastatic papillary thyroid carcinoma, in whom Tg is directly secreted by neoplastic thyroid cells into the circulation rather than transcytosed, serum Tg was not precipitated by antimegalin antibodies. The detection of Tg-megalin complexes may help identify the source of serum Tg in patients with thyroid diseases.


Tetrahydrobiopterin protects against guanabenz-mediated inhibition of neuronal nitric-oxide synthase in vitro and in vivo.
It is established that guanabenz inhibits neuronal nitric-oxide (NO) synthase (nNOS) and causes the enhanced proteasomal degradation of nNOS in vivo. Although the time- and NADPH-dependent inhibition of nNOS has been reported in studies where guanabenz was incubated with crude cytosolic preparations of nNOS, the exact mechanism for inhibition is not known. Moreover, even less is known about how the inhibition of nNOS triggers its proteasomal degradation. In the current study, we show, with the use of purified nNOS, that guanabenz treatment leads to the oxidation of tetrahydrobiopterin and formation of a pterin-depleted nNOS, which is not able to form NO. With the use of 14C-labeled guanabenz, we were unable to detect any guanabenz metabolites or guanabenz-nNOS adducts, indicating that reactive intermediates of guanabenz probably do not play a role in the inhibition. Superoxide dismutase, however, prevents the guanabenz-mediated oxidation of tetrahydrobiopterin and inhibition of nNOS, suggesting the role of superoxide as an intermediate. Studies in rats show that administration of tetrahydrobiopterin prevents the inhibition and loss of penile nNOS due to guanabenz, indicating that the loss of tetrahydrobiopterin plays a major role in the effects of guanabenz in vivo. Our findings are consistent with the destabilization and enhanced degradation of nNOS found after tetrahydrobiopterin depletion. These studies suggest that drug-mediated destabilization and subsequent enhanced degradation of protein targets will likely be an important toxicological consideration.


Mutation rate analysis via parent-progeny sequencing of the perennial peach. I. A low rate in woody perennials and a higher mutagenicity in hybrids.
Mutation rates vary between species, between strains within species and between regions within a genome. What are the determinants of these forms of variation? Here, via parent-offspring sequencing of the peach we ask whether (i) woody perennials tend to have lower per unit time mutation rates compared to annuals, and (ii) hybrid strains have high mutation rates. Between a leaf from a low heterozygosity individual, derived from an intraspecific cross, to a leaf of its selfed progeny, the mutation rate is 7.77 × 10-9 point mutations per bp per generation, similar to Arabidopsis thaliana (7.0-7.4 × 10-9 point mutations per bp per generation). This suggests a low per unit time mutation rate as the generation time is much longer in peach. This is supported by our estimate of 9.48 × 10-9 point mutations per bp per generation from a 200-year-old low heterozygosity peach to its progeny. From a more highly heterozygous individual derived from an interspecific cross to its selfed progeny, the mutation rate is 1.38 × 10-8 mutations per site per generation, consistent with raised rates in hybrids. Our data thus suggest that (i) peach has an approximately order of magnitude lower mutation rate per unit time than Arabidopsis, consistent with reports of low evolutionary rates in woody perennials, and (ii) hybridization may, indeed, be associated with increased mutation rates as considered over a century ago.


Melatonin modulates the cadmium-induced expression of MT-2 and MT-1 metallothioneins in three lines of human tumor cells (MCF-7, MDA-MB-231 and HeLa).
Cadmium (Cd) is a human carcinogen present in tobacco smoke and contaminated industrial soils. Metallothioneins (MTs) are intracellular proteins involved in protecting against Cd. The toxic effects of Cd can be modified by compounds able to modulate MTs synthesis. Melatonin has oncostatic properties and has also been shown to counteract the toxic effects of Cd. In this study we examine the possible role of melatonin in Cd-induced expression of several MT isoforms (MT-2A, MT-1X, MT-1F and MT-1E) in three human tumor cell lines (MCF-7, MDA-MB-231 and HeLa). We found that, in all cell types, melatonin increases Cd-induced expression of MT-2A, which is considered to protect against Cd toxicity. As regards MT-1 subtypes, which have been related with cell invasiveness and high histological grade tumors, melatonin caused Cd-induced expression in both breast cancer cell lines to decrease. These effects point towards melatonin's possible role as a preventive agent for carcinogenesis dependent on Cd contamination.


Delivery of Antioxidant and Anti-inflammatory Agents for Tissue Engineered Vascular Grafts.
The treatment of patients with severe coronary and peripheral artery disease represents a significant clinical need, especially for those patients that require a bypass graft and do not have viable veins for autologous grafting. Tissue engineering is being investigated to generate an alternative graft. While tissue engineering requires surgical intervention, the release of pharmacological agents is also an important part of many tissue engineering strategies. Delivery of these agents offers the potential to overcome the major concerns for graft patency and viability. These concerns are related to an extended inflammatory response and its impact on vascular cells such as endothelial cells. This review discusses the drugs that have been released from vascular tissue engineering scaffolds and some of the non-traditional ways that the drugs are presented to the cells. The impact of antioxidant compounds and gasotransmitters, such as nitric oxide and carbon monoxide, are discussed in detail. The application of tissue engineering and drug delivery principles to biodegradable stents is also briefly discussed. Overall, there are scaffold-based drug delivery techniques that have shown promise for vascular tissue engineering, but much of this work is in the early stages and there are still opportunities to incorporate additional drugs to modulate the inflammatory process.


Dietary intake of total, animal, and vegetable protein and risk of type 2 diabetes in the European Prospective Investigation into Cancer and Nutrition (EPIC)-NL study.
Dietary recommendations are focused mainly on relative dietary fat and carbohydrate content in relation to diabetes risk. Meanwhile, high-protein diets may contribute to disturbance of glucose metabolism, but evidence from prospective studies is scarce. We examined the association among dietary total, vegetable, and animal protein intake and diabetes incidence and whether consuming 5 energy % from protein at the expense of 5 energy % from either carbohydrates or fat was associated with diabetes risk. A prospective cohort study was conducted among 38,094 participants of the European Prospective Investigation into Cancer and Nutrition (EPIC)-NL study. Dietary protein intake was measured with a validated food frequency questionnaire. Incident diabetes was verified against medical records. During 10 years of follow-up, 918 incident cases of diabetes were documented. Diabetes risk increased with higher total protein (hazard ratio 2.15 [95% CI 1.77-2.60] highest vs. lowest quartile) and animal protein (2.18 [1.80-2.63]) intake. Adjustment for confounders did not materially change these results. Further adjustment for adiposity measures attenuated the associations. Vegetable protein was not related to diabetes. Consuming 5 energy % from total or animal protein at the expense of 5 energy % from carbohydrates or fat increased diabetes risk. Diets high in animal protein are associated with an increased diabetes risk. Our findings also suggest a similar association for total protein itself instead of only animal sources. Consumption of energy from protein at the expense of energy from either carbohydrates or fat may similarly increase diabetes risk. This finding indicates that accounting for protein content in dietary recommendations for diabetes prevention may be useful.


Molecular investigations into vaginal immunization with HIV gp41 antigenic construct H4A in a quick release solid dosage form.
A robust vaginal immune response is considered essential for an effective prophylactic vaccine that prevents transmission of HIV and other sexually acquired diseases. Considerable attention has recently focused on the potential of vaginally administered vaccines as a means to induce such local immunity. However, the potential for vaccination at this site remains in doubt as the vaginal mucosa is generally considered to have low immune inductive potential. In the current study, we explored for the first time the use of a quick release, freeze-dried, solid dosage system for practical vaginal administration of a protein antigen. These solid dosage forms overcome the common problem associated with leakage and poor retention of vaginally administered antigen solutions. Mice were immunized vaginally with H4A, an HIV gp41 envelope based recombinant protein, using quick release, freeze-dried solid rods, and the immune responses compared to a control group immunized via subcutaneous H4A injection. Vaginally immunized mice failed to elicit robust immune responses. Our detailed investigations, involving cytokine analysis, the stability of H4A in mouse cervicovaginal lavage, and elucidation of the state of H4A protein in the immediate-release dosage form, revealed that antigen instability in vaginal fluid, the state of the antigen in the dosage form, and the cytokine profile induced are all likely to have contributed to the observed lack of immunogenicity. These are important factors affecting vaginal immunization and provide a rational basis for explaining the typically poor and variable elicitation of immunity at this site, despite the presence of immune responsive cells within the vaginal mucosae. In future mucosal vaccine studies, a more explicit focus on antigen stability in the dosage form and the immune potential of available antigen-responsive cells is recommended.


Reflection of successful anticancer drug development processes in the literature.
The development of cancer drugs is time-consuming and expensive. In particular, failures in late-stage clinical trials are a major cost driver for pharmaceutical companies. This puts a high demand on methods that provide insights into the success chances of new potential medicines. In this study, we systematically analyze publication patterns emerging along the drug discovery process of targeted cancer therapies, starting from basic research to drug approval - or failure. We find clear differences in the patterns of approved drugs compared with those that failed in Phase II/III. Feeding these features into a machine learning classifier allows us to predict the approval or failure of a targeted cancer drug significantly better than educated guessing. We believe that these findings could lead to novel measures for supporting decision making in drug development.


Immediate Dental Implant Stabilization in a Canine Model Using a Novel Mineral-Organic Adhesive: 4-Month Results.
This study evaluated a novel injectable, self-setting, osteoconductive, resorbable adhesive that provides immediate implant stabilization. Twenty-six large canines had the mandibular second through fourth premolars and the first molar removed bilaterally. After 3 months, oversized osteotomies were prepared with only the apical 2 mm of the implant engaging native bone. One site had a novel resorbable, self-setting, mineral-organic adhesive (TN-SM) placed around the implant, a second site received bone graft, and a third site received only blood clot. Removal torque, standardized radiography, and histology were used to evaluate implant stability and tissue contact after 24 hours, 10 days, and 4 months. Mean removal torque values after 24 hours were 1.4, 1.3, and 22.2 Ncm for the control, bone graft, and mineral-organic adhesive, respectively. After 10 days, these values were 5.7, 6.2, and 45.7 Ncm and at 4 months increased to 88.7, 77.8, and 104.7 Ncm, respectively. Clinical, radiographic, and histologic evaluations showed a lack of inflammatory reaction. Control defects were initially radiolucent in the coronal area; grafted sites revealed particles in the gap, with both conditions gradually filling with bone over time. At 10 days, histologic evaluation demonstrated excellent biocompatibility and intimate contact of mineral-organic adhesive to both the implant and bone, providing an osseointegration-like bond; control sites revealed no bone contact in the defect area, while the bone-grafted sites revealed unattached graft particles. At 4 months, much of the mineral-organic adhesive was replaced with bone; the control and grafted sites had some bone fill, and many of the defects demonstrated no bone-to-implant contact and were filled with soft tissue or isolated graft particles. The mineral-organic adhesive provides immediate (osseointegration-like) and continued implant stabilization over 4 months in sites lacking primary stability. Experimental sites demonstrated maintenance of crestal bone levels adjacent to the mineral-organic adhesive and soft tissue exclusion without the use of membranes in this canine model. These results demonstrate that this novel mineral-organic adhesive can enable implant osseointegration in a site where insufficient native bone exists to allow immediate implant placement.


Health-related quality of life during bosutinib (SKI-606) therapy in patients with advanced chronic myeloid leukemia after imatinib failure.
The tyrosine kinase inhibitor (TKI) bosutinib has demonstrated activity in patients with advanced phase chronic myeloid leukemia (CML), but effects on health-related quality of life (HRQoL) remain unexplored. This study evaluated HRQoL in advanced CML patients receiving bosutinib in an ongoing phase 2 study following resistance or intolerance to prior imatinib therapy. This analysis included data from 76 accelerated-phase (AP) and 64 blast-phase (BP) patients resistant/intolerant to prior imatinib with or without prior exposure to other TKIs. Patient-reported HRQoL assessments completed at baseline; weeks 4, 8, and 12; every 12 weeks thereafter; and at treatment completion included the Functional Assessment of Cancer Therapy-Leukemia (FACT-Leu); general health status was assessed using the 5-item EuroQol (EQ-5D) instrument and a visual analog scale (VAS). HRQoL at baseline was somewhat worse in BP versus AP CML patients. There was a significant improvement in the mean FACT-Leu Total scale at weeks 24, 36, and 48 in AP CML patients and at weeks 4, 8, 12, 24, 36, 48, and 96 in BP CML patients compared with baseline. EQ-5D Utility scores were stable throughout treatment in AP CML patients but significantly improved versus baseline in BP CML patients at weeks 4, 8, 12, and 36. Mean VAS scores were significantly improved at weeks 8, 36, and 48 in AP CML patients and at weeks 4, 8, 12, 24, 36, and 96 in BP CML patients. The lack of a comparison group limits attribution of improvements in HRQoL specifically to bosutinib treatment; potential bias due to non-ignorable dropout may limit the ability to generalize these findings to situations where durations of therapy exceed the 96-week follow-up duration of the present study. These findings suggest that bosutinib therapy is associated with improved HRQoL in advanced phase CML patients. NCT00261846.


Continuous-wave laser spectrometer automatically aligned and continuously tuned from 11.8 to 16.1 microm by use of diode-laser-pumped difference-frequency generation in GaSe.
We report a fully automated mid-IR difference-frequency spectrometer with a spectral resolution under 70 MHz pumped by a pair of conventional room-temperature 800-900-nm diode lasers. 0.1 microW of tunable cw radiation is produced from incident-diode powers of 120 and 75 mW. The system has computer-controlled beam alignment with compact CCD cameras, motorized mirrors and positioners to obtain 0.01 degrees crystal-angle positioning, 4-microm beam overlap at the nonlinear crystal, and automated diode laser beam collimation. Computer-operated frequency control uses temperature tuning and current tuning of the free-running diode lasers. The system has been demonstrated by successfully scanning, without any human intervention, 64 randomly selected acetylene absorption lines between 12 and 15 microm. Spectral scans of ammonia are also presented. This mid-IR spectrometer is suitable for fully automated spectroscopy of an unlimited list of mid-IR frequencies and has the potential to detect any trace gas that has an acceptable absorption line within the large tuning range.


Heterogeneous responses of temperate-zone amphibian populations to climate change complicates conservation planning.
The pervasive and unabated nature of global amphibian declines suggests common demographic responses to a given driver, and quantification of major drivers and responses could inform broad-scale conservation actions. We explored the influence of climate on demographic parameters (i.e., changes in the probabilities of survival and recruitment) using 31 datasets from temperate zone amphibian populations (North America and Europe) with more than a decade of observations each. There was evidence for an influence of climate on population demographic rates, but the direction and magnitude of responses to climate drivers was highly variable among taxa and among populations within taxa. These results reveal that climate drivers interact with variation in life-history traits and population-specific attributes resulting in a diversity of responses. This heterogeneity complicates the identification of conservation 'rules of thumb' for these taxa, and supports the notion of local focus as the most effective approach to overcome global-scale conservation challenges.


Influence of teichoic acid from S. aureus on metabolic activity of macrophages and cytotoxic activity of splenocytes of mice bearing Lewis lung carcinoma.
To investigate the effect of teichoic acid (TA) from the cell wall of S. aureus on some indices of immunological reactivity in mice bearing Lewis lung carcinoma (LLC). The teichoic acid at the doses of 1, 2 and 4 microg/g of body weight has been administered subcutaneously simultaneously with tumor cells transplantation and in 7 days. The cytotoxic activity of peritoneal macrophages has been assessed by NBT-test. The splenocyte cytotoxic activity against the LLC cells has been tested by flow cytometry. The evaluation of tumor infiltration by lymphoid cells was carried out as well. TA had no significant effect on oxidative metabolism of peritoneal macrophages in tumor bearing mice. Upon TA administration, the cytotoxic activity of splenocytes against the LLC cells has been augmented in a dose-dependent manner (at the TA dose of 4 microg/g, 2-fold decrease of tumor growth and metastasis has been registered) and leads to decreased tumor infiltration by mononuclear cells. TA caused a dose dependent inhibition of growth and metastasis of LLC. It was supposed that TA can influence the tumor grows by activation of splenocytes cytotoxic activity.