Taube nuss is a novel gene essential for the survival of pluripotent cells of early mouse embryos.
The cells of the inner cell mass constitute the pluripotent cell population of the early embryo. They have the potential to form all of the tissues of the embryo proper and some extra-embryonic tissues. They can be considered a transient stem cell population for the whole of the embryo, and stem cells maintaining the same capacity can be isolated from these cells. We have isolated, characterised and mutated a novel gene, taube nuss (Tbn), that is essential for the survival of this important cell population. The taube nuss protein sequence (TBN) was highly conserved between human, mouse, Xenopus laevis, Drosophila melanogaster, Caenorhabditis elegans and Arabidopsis thaliana, particularly in a domain that is not present in any published proteins, showing that TBN is the founding member of a completely new class of proteins with an important function in development. The Tbn gene was expressed ubiquitously as early as E2. 5 and throughout embryonic development. It was also expressed in adult brain with slightly higher levels in the hippocampus. The Tbn mutant embryos developed normally to the blastocyst stage and contained inner cell masses. They hatched from the zonae pellucidae, implanted and induced decidual reactions, but failed to develop beyond E4.0. At this time the trophoblast cells were viable, but inner cell masses were not detectable. At E3.75, massive TUNEL-positive DNA degradation and chromatin condensation were visible within the inner cell masses, whereas the cell membranes where intact. Caspase 3 was expressed in these cells. In vitro, the inner cell mass of mutant embryos failed to proliferate and died after a short period in culture. These results indicate that the novel protein, taube nuss, is necessary for the survival of the inner cell mass cells and that inner cell mass cells died of apoptosis in the absence of the taube nuss protein. As cell pruning by apoptosis is a recognised developmental process at this stage of development, the taube nuss protein may be one of the factors regulating the extent of programmed cell death at this time point.


TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction.
The telomeric shelterin protein telomeric repeat-binding factor 2 (TRF2) recruits origin recognition complex (ORC) proteins, the foundational building blocks of DNA replication origins, to telomeres. We seek to determine whether TRF2-recruited ORC proteins give rise to functional origins in telomere repeat tracts. We find that reduction of telomeric recruitment of ORC2 by expression of an ORC interaction-defective TRF2 mutant significantly reduces telomeric initiation events in human cells. This reduction in initiation events is accompanied by telomere repeat loss, telomere aberrations and dysfunction. We demonstrate that telomeric origins are activated by induced replication stress to provide a key rescue mechanism for completing compromised telomere replication. Importantly, our studies also indicate that the chromatin remodeler SNF2H promotes telomeric initiation events by providing access for ORC2. Collectively, our findings reveal that active recruitment of ORC by TRF2 leads to formation of functional origins, providing an important mechanism for avoiding telomere dysfunction and rescuing challenged telomere replication.


Coping with myocardial infarction: evaluation of a coping questionnaire.
The negative effects of emotional distress on the recovery following myocardial infarction make it important to study coping strategies in this situation. The present study aimed to evaluate the psychometric properties and the validity of a 10 dimensions questionnaire labelled The General Coping Questionnaire (GCQ). The structure of the questionnaire was based on a previous interview study with 26 persons with different diseases. The 10 dimensions are called self-trust, problem-reducing actions, change of values, social trust, minimization, fatalism, resignation, protest, isolation and intrusion. The present study comprised 114 first-time myocardial infarction patients (37 women, 77 men). Five months after myocardial infarction, they answered questions about health-related quality of life, health complaints, sense of coherence and the GCQ. A multi-trait/multi-item analysis showed good item-scale convergent and discriminatory validity when the GCQ was reduced from 47 to 40 items. In conclusion, the results showed that the 40-item GCQ is a well-structured and reliable questionnaire for measuring coping strategies in myocardial infarction patients.


The double polymethylmethacrylate filter (DELETE system) in the removal of light chains in chronic dialysis patients with multiple myeloma.
Multiple myeloma (MM) is still one of the most common haematological diseases and is associated with a poor prognosis. It is frequently worsened by acute kidney failure that, in turn, aggravates the risk of death. In the past few years, the idea has made headway that the removal of free light chains (FLC) by means of extracorporeal blood purification systems may facilitate the recovery of renal function. Up to now, many different extracorporeal techniques have been put forward in FLC removal, such as plasma exchange, dialysis with super-flux filters, and adsorption by means of cartridge of resins. In this paper, we illustrate the use of polymethylmethacrylate (PMMA) dialysis membranes with a high adsorptive capacity (Toray BK-F; Toray Industries, Inc., Tokyo, Japan). We have evaluated light chain removal by means of an original dialysis procedure using a double-filter circuit made of PMMA working in sequential dialysis (DELETE system). The system provides satisfactory results in terms of FLC removal and, at the same time, ensures an adequate dialysis treatment (Kt/V >1.5) with significant reduction in urea, creatinine, and β2-microglobulin. The dual PMMA filter system combines an acceptable cost/efficiency ratio when compared with other methods and constitutes a concrete prospect in FLC removal. Its preferential setting of use is in patients with MM or with monoclonal gammopathies, who are on chronic dialysis and maintain high circulating levels of FLC.


Potential drug-drug interactions in pediatric outpatient prescriptions for newborns and infants.
To surveyed the quantities, types, and related information of potential drug-drug interactions (DDIs) and estimate the off-label use percentage of pediatric outpatient prescriptions for newborns and infants from the National Health Insurance Research Database (NHIRD) of Taiwan. Adverse drug reactions (ADR) may cause morbidity and mortality, potential drug-drug interactions (DDI) increase the probability of ADR. Research on ADR and DDI in infants is of particular urgency and importance but the related profiles in these individuals are not well known. All prescriptions written by physicians in 2000 were analyzed to identify potential DDIs among drugs appearing on the same prescription sheet. Of a total of 150.6 million prescription sheets, with 669.5 million prescriptions registered in the NHIRD of Taiwan, six million (3.99%) prescription sheets were for 2.1 million infants with 19.4 million (2.85%) prescriptions. There were 672,020 potential DDIs in this category, accounting for 3.53% per prescription; an estimated one DDI in every three patients. The interactions between aspirin and aluminum/magnesium hydroxide were most common (4.42%). Of the most significant drug-drug interactions, the interaction of digoxin with furosemide ranked first (20.14%), followed by the interactions of cisapride with furosemide and erythromycin (6.02% and 4.85%, respectively). The interactions of acetaminophen and anti-cholinergic agents comprised most types of drug-drug interactions (6.62%). Although the prevalence rates of DDIs are low, life-threatening interactions may develop. Physicians must be reminded of the potential DDIs when prescribing medications for newborns and infants.


Differences in Postmating Transcriptional Responses between Conspecific and Heterospecific Matings in Drosophila.
In many animal species, females undergo physiological and behavioral changes after mating. Some of these changes are driven by male-derived seminal fluid proteins and are critical for fertilization success. Unfortunately, our understanding of the molecular interplay between female and male reproductive proteins remains inadequate. Here, we analyze the postmating response in a Drosophila species that has evolved strong gametic incompatibility with its sister species; Drosophila novamexicana females produce only ∼1% fertilized eggs in crosses with Drosophila americana males, compared to ∼98% produced in within-species crosses. This incompatibility is likely caused by mismatched male and female reproductive molecules. In this study, we use short-read RNA sequencing to examine the evolutionary dynamics of female reproductive genes and the postmating transcriptome response in crosses within and between species. First, we found that most female reproductive tract genes are slow-evolving compared to the genome average. Second, postmating responses in con- and heterospecific matings are largely congruent, but heterospecific matings induce expression of additional stress-response genes. Some of those are immunity genes that are activated by the Imd pathway. We also identify several genes in the JAK/STAT signaling pathway that are induced in heterospecific, but not conspecific mating. While this immune response was most pronounced in the female reproductive tract, we also detect it in the female head and ovaries. These results show that the female's postmating transcriptome-level response is determined in part by the genotype of the male, and that divergence in male reproductive genes and/or traits can have immunogenic effects on females.


A comparison of the Oxford shoulder score and shoulder pain and disability index: factor structure in the context of a large randomized controlled trial.
To explore and compare the factor structure of the 12-item Oxford shoulder score (OSS) and 13-item shoulder pain and disability index (SPADI). Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) of data from 660 patients attending 46 hospitals in the UK. Complete OSS and SPADI data were available for 648 (98.2%) and 628 (95.2%) participants, respectively. For both instruments, either one or two factors were indicated, depending on the extraction method. On EFA, most OSS items loaded saliently on either of two "Pain" (4 items) and "Function" (8 items) factors, although some items cross-loaded. Cronbach's alphas were 0.75, 0.90, and 0.91 for "Pain" and "Function" subscales, and all 12 OSS items, respectively. CFA suggested marginally better fit for two factors, with neither one- nor two-factor models rejected. EFA indicated two factors for the SPADI, with three of the eight "Disability" items contributing to an 8-item "Pain factor", with 2 items within the 5-item "Disability factor" cross-loading. Cronbach's alpha was 0.87 and 0.93 for the original 5- and 8-item pain and disability scales; 0.94 for all 13 SPADI items, respectively. CFA suggested marginally better fit for the two-factor (original conceptualization) model of the SPADI, with neither one- nor two-factor models rejected. EFA and CFA demonstrated that, in addition to single summary scales usage, separate information on pain and self-reported disability/function can be extracted in a meaningful way, as subscales, from both the OSS and the SPADI. This information can help researchers in choosing primary study endpoints appropriately.


Extracorporeal photopheresis: past, present, and future.
Extracorporeal photopheresis (ECP) is a leukapheresis-based therapy that uses 8-methoxypsoralen and ultraviolet A irradiation. Used alone or in combination with biological agents, ECP is an established and effective therapy for advanced cutaneous T-cell lymphoma. ECP has also shown promising efficacy in a number of other severe and difficult-to-treat conditions, including systemic sclerosis, graft-versus-host disease, prevention and treatment of rejection in solid organ transplantation, and Crohn disease. Furthermore, the use of ECP in some of these conditions may allow a significant reduction in the use of systemic steroids and other immunosuppressants, reducing long-term morbidity and mortality. The accumulated experience shows ECP to be well tolerated, with no clinically significant side effects. Progress is also being made in the search for understanding of the mechanisms of action of ECP, which will ultimately facilitate improvements in the use of this therapy.


Oral pre-exposure prophylaxis by anti-retrovirals raltegravir and maraviroc protects against HIV-1 vaginal transmission in a humanized mouse model.
Sexual HIV-1 transmission by vaginal route is the most predominant mode of viral transmission, resulting in millions of new infections every year. In the absence of an effective vaccine, there is an urgent need to develop other alternative methods of pre-exposure prophylaxis (PrEP). Many novel drugs that are currently approved for clinical use also show great potential to prevent viral sexual transmission when administered systemically. A small animal model that permits rapid preclinical evaluation of potential candidates for their systemic PrEP efficacy will greatly enhance progress in this area of investigation. We have previously shown that RAG-hu humanized mouse model permits HIV-1 mucosal transmission via both vaginal and rectal routes and displays CD4 T cell loss typical to that seen in the human. Thus far systemic PrEP studies have been primarily limited to RT inhibitors exemplified by tenofovir and emtricitabine. In these proof-of-concept studies we evaluated two new classes of clinically approved drugs with different modes of action namely, an integrase inhibitor raltegravir and a CCR5 inhibitor maraviroc as potential systemically administered chemo-prophylactics. Our results showed that oral administration of either of these drugs fully protects against vaginal HIV-1 challenge in the RAG-hu mouse model. Based on these results both these drugs show great promise for further development as orally administered PrEPs.


Comparison of Control of Clostridium difficile Infection in Six English Hospitals Using Whole-Genome Sequencing.
Variation in Clostridium difficile infection (CDI) rates between healthcare institutions suggests overall incidence could be reduced if the lowest rates could be achieved more widely. We used whole-genome sequencing (WGS) of consecutive C. difficile isolates from 6 English hospitals over 1 year (2013-14) to compare infection control performance. Fecal samples with a positive initial screen for C. difficile were sequenced. Within each hospital, we estimated the proportion of cases plausibly acquired from previous cases. Overall, 851/971 (87.6%) sequenced samples contained toxin genes, and 451 (46.4%) were fecal-toxin-positive. Of 652 potentially toxigenic isolates >90-days after the study started, 128 (20%, 95% confidence interval [CI] 17-23%) were genetically linked (within ≤2 single nucleotide polymorphisms) to a prior patient's isolate from the previous 90 days. Hospital 2 had the fewest linked isolates, 7/105 (7%, 3-13%), hospital 1, 9/70 (13%, 6-23%), and hospitals 3-6 had similar proportions of linked isolates (22-26%) (P ≤ .002 comparing hospital-2 vs 3-6). Results were similar adjusting for locally circulating ribotypes. Adjusting for hospital, ribotype-027 had the highest proportion of linked isolates (57%, 95% CI 29-81%). Fecal-toxin-positive and toxin-negative patients were similarly likely to be a potential transmission donor, OR = 1.01 (0.68-1.49). There was no association between the estimated proportion of linked cases and testing rates. WGS can be used as a novel surveillance tool to identify varying rates of C. difficile transmission between institutions and therefore to allow targeted efforts to reduce CDI incidence.


Altered expression of GFLO, the Gesneriaceae homologue of FLORICAULA/LEAFY, is associated with the transition to bulbil formation in Titanotrichum oldhamii.
Titanotrichum oldhamii inflorescences switch from flower to bulbil production at the end of the flowering season. The structure of the bulbiliferous shoots resembles the abnormal meristematic organization of the Antirrhinum mutant, floricaula. Gesneriaceae- FLORICAULA (GFLO) is thus a candidate gene in the regulation of bulbil formation. To investigate this hypothesis, part of the GFLO gene (between the second and third exon) was isolated using degenerate primers designed in regions conserved between Antirrhinum, Nicotiana and Arabidopsis, followed by genome walking to obtain the complete gene and flanking sequences. RT-PCR results showed that the GFLO homologue is strongly expressed in inflorescence apical meristems and young flowers. However, in meristems that had switched to bulbil formation, GFLO transcription was greatly reduced. The down-regulation of GFLO in bulbil primordia indicates that this gene is connected to, or part of, the bulbil-flower regulatory pathway. Phylogenetic analysis confirms the orthology of GFLO and FLO, and indicates that the gene may be useful for phylogenetic reconstruction at the genus or family level.


Leukocyte endogenous mediator fails to alter protein dynamics in a model of liver dysfunction.
This study examines the effect of a purified leukocyte-derived endogenous mediator (LEM) on protein metabolism during liver dysfunction and after sham surgery and the role of a nonsteroidal, antiinflammatory drug (indomethacin sodium trihydrate) in modifying this response. Febrile response, protein kinetics, urinary end products of protein metabolism, and plasma acute phase protein levels were studied in rats given a pyrogenic dose of LEM or saline solution, and these same indicators were studied after the administration of the same dose of LEM plus 2 mg/kg indomethacin 3 weeks after a portacaval shunt (PCS) or sham operation. In both surgical groups given LEM, a maximum of 1.1 degrees C fever was observed. LEM increased protein turnover and urinary excretion of nitrogen and urea in sham-operated rats but not in the PCS animals. Administration of indomethacin decreased the plasma oxidation of L[1-14C]leucine and prevented the increased excretion of total nitrogen and urea in sham-operated animals treated with LEM. PCS animals showed a constant excretion of nitrogen and urea independent of treatment. alpha 1-Acid glycoprotein levels increased significantly in sham-operated animals treated with LEM but not in the PCS group until indomethacin was added. The coadministration of LEM and indomethacin in shams also enhanced the levels of alpha 2-macroglobulin and alpha 1-acid glycoprotein over values found with LEM alone. These findings confirm the catabolic effect of LEM in normal animals and identify the essential role of the liver in the acute phase response. The data also suggest that indomethacin may modify the acute phase response by reducing plasma amino acid oxidation as well as enhancing the levels of some specific acute phase proteins.


Receptor protein tyrosine phosphatase micro: measuring where to stick.
We review here recent results on the structure and function of a receptor protein tyrosine phosphatase, RPTPmicro. In addition to their intercellular catalytic domains which bear the phosphatase activity, the RPTPs are cell-surface-receptor-type molecules and in many cases have large extracellular regions. What role can these extracellular regions play in function? For RPTPmicro, the extracellular region is known to mediate homophilic adhesion. Sequence analysis indicates that it comprises six domains: an N-terminal MAM (meprin/A5/micro), one immunoglobulin-like domain and four fibronectin type III (FN) repeats. We have determined the crystal structure of the entire extracellular region for RPTPmicro in the form of a functional adhesion dimer. The physical characteristics and dimensions of the adhesion dimer suggest a mechanism by which the location of this phosphatase can be influenced by cell-cell spacings.


Adult Lifetime Diet Quality and Physical Performance in Older Age: Findings From a British Birth Cohort.
Current evidence that links "healthier" dietary patterns to better measured physical performance is mainly from older populations; little is known about the role of earlier diet. We examined adult diet quality in relation to physical performance at age 60-64 years. Diet quality was defined using principal component analysis of dietary data collected at age 36, 43, 53, and 60-64. Throughout adulthood, diets of higher quality were characterized by higher consumption of fruit, vegetables, and wholegrain bread. Diet quality scores calculated at each age indicated compliance with this pattern. Physical performance was assessed using chair rise, timed-up-and-go, and standing balance tests at age 60-64. The analysis sample included 969 men and women. In gender-adjusted analyses, higher diet quality at each age was associated with better measured physical performance (all p < .01 for each test), although some associations were attenuated after adjustment for covariates. Diet quality scores were highly correlated in adulthood (0.44 ≤ r ≤ 0.67). However, conditional models showed that higher diet quality at age 60-64 (than expected from scores at younger ages), was associated with faster chair rise speed and with longer standing balance time (adjusted: 0.08 [95% CI: 0.02, 0.15] and 0.07 [0.01, 0.14] SD increase in chair rise speed and balance time, respectively, per SD increase in conditional diet quality; both p < .05). Higher diet quality across adulthood is associated with better physical performance in older age. Current diet quality may be particularly important for physical performance, suggesting potential for improvements in diet in early older age.


Nuclear reprogramming: kinetics of cell cycle and metabolic progression as determinants of success.
Establishment of totipotency after somatic cell nuclear transfer (NT) requires not only reprogramming of gene expression, but also conversion of the cell cycle from quiescence to the precisely timed sequence of embryonic cleavage. Inadequate adaptation of the somatic nucleus to the embryonic cell cycle regime may lay the foundation for NT embryo failure and their reported lower cell counts. We combined bright field and fluorescence imaging of histone H(2b)-GFP expressing mouse embryos, to record cell divisions up to the blastocyst stage. This allowed us to quantitatively analyze cleavage kinetics of cloned embryos and revealed an extended and inconstant duration of the second and third cell cycles compared to fertilized controls generated by intracytoplasmic sperm injection (ICSI). Compared to fertilized embryos, slow and fast cleaving NT embryos presented similar rates of errors in M phase, but were considerably less tolerant to mitotic errors and underwent cleavage arrest. Although NT embryos vary substantially in their speed of cell cycle progression, transcriptome analysis did not detect systematic differences between fast and slow NT embryos. Profiling of amino acid turnover during pre-implantation development revealed that NT embryos consume lower amounts of amino acids, in particular arginine, than fertilized embryos until morula stage. An increased arginine supplementation enhanced development to blastocyst and increased embryo cell numbers. We conclude that a cell cycle delay, which is independent of pluripotency marker reactivation, and metabolic restraints reduce cell counts of NT embryos and impede their development.


Long-term issues in the treatment of sleep disorders.
Insomnia is a disorder characterized by chronic sleep disturbance associated with daytime disability or distress, such as memory impairment and fatigue, that occurs despite adequate opportunity for sleep. Insomnia may present as difficulty falling/staying asleep or as sleep that is nonrestorative. Studies show a strong correlation between insomnia and impaired quality of life. Pain conditions and depression are commonly associated with insomnia, either as secondary or comorbid conditions. In addition, a greater incidence of anxiety, alcohol and drug dependence, and cardiovascular disease is found in people with insomnia. Data indicate insomnia results from over-engaged arousal systems. Insomnia patients experience increased metabolic rate, body temperature, and heart rate, and elevated levels of norepinephrine and catecholamines. Pharmacologic options for the treatment of insomnia include benzodiazepine hypnotics, a selective melatonin receptor agonist, and sedating antidepressants. However, insomnia may be best treated with cognitive-behavioral therapy and instruction in good sleep hygiene, either alone or in concert with pharmacologic agents. Studies on the effects of insomnia treatment use variable methodologies or do not publish negative results, and there are currently no studies of treatment focusing on morbidity. Further research is necessary to better understand the effects of insomnia therapies on medical and psychiatric disorders. In this Clinical Information Supplement, Thomas Roth, PhD, describes the nature of insomnia and its pathophysiology. Next, Andrew D. Krystal, MD, MS, reviews morbidities associated with insomnia. Finally, Joseph A. Lieberman III, MD, MPH, provides an overview of therapeutics utilized in patients with insomnia, including behavioral therapies and pharmacologic options.


Authorship bias in violence risk assessment? A systematic review and meta-analysis.
Various financial and non-financial conflicts of interests have been shown to influence the reporting of research findings, particularly in clinical medicine. In this study, we examine whether this extends to prognostic instruments designed to assess violence risk. Such instruments have increasingly become a routine part of clinical practice in mental health and criminal justice settings. The present meta-analysis investigated whether an authorship effect exists in the violence risk assessment literature by comparing predictive accuracy outcomes in studies where the individuals who designed these instruments were study authors with independent investigations. A systematic search from 1966 to 2011 was conducted using PsycINFO, EMBASE, MEDLINE, and US National Criminal Justice Reference Service Abstracts to identify predictive validity studies for the nine most commonly used risk assessment tools. Tabular data from 83 studies comprising 104 samples was collected, information on two-thirds of which was received directly from study authors for the review. Random effects subgroup analysis and metaregression were used to explore evidence of an authorship effect. We found a substantial and statistically significant authorship effect. Overall, studies authored by tool designers reported predictive validity findings around two times higher those of investigations reported by independent authors (DOR=6.22 [95% CI=4.68-8.26] in designers' studies vs. DOR=3.08 [95% CI=2.45-3.88] in independent studies). As there was evidence of an authorship effect, we also examined disclosure rates. None of the 25 studies where tool designers or translators were also study authors published a conflict of interest statement to that effect, despite a number of journals requiring that potential conflicts be disclosed. The field of risk assessment would benefit from routine disclosure and registration of research studies. The extent to which similar conflict of interests exists in those developing risk assessment guidelines and providing expert testimony needs clarification.


Characterising bone material composition and structure in the ovariectomized (OVX) rat model of osteoporosis.
The ovariectomized (OVX) rat model is well established in investigations of osteoporosis and osteoporotic therapies. Advent of techniques such as Fourier-transform infrared (FTIR) spectroscopy and small angle X-ray scattering (SAXS) facilitate characterization of bone composition and mineral structure, respectively, which are key determinants of bone strength. Limited publications exist on the implementation of these techniques in the OVX rat model. At 12 weeks of age, female Sprague-Dawley rats were either sham-operated (n = 6) or ovariectomized (n = 6) and sacrificed 18 weeks later. L2 lumbar vertebrae and proximal tibiae were assessed by µCT, FTIR and SAXS. Presence of extensive trabecular deterioration in the µCT data confirmed the onset of osteoporosis. FTIR compositional parameters were determined including measures of degree of mineralization, crystallinity, collagen maturity and acid phosphate content. Mineral crystal thickness was determined from the SAXS data using two approaches available in literature. Compositionally, a decline in the heterogeneity of acid phosphate content was observed while measures of crystallinity and collagen maturity remained unaltered. Using an iterative curve fitting method, OVX-induced increases in the mineral crystal thickness of 3.8 and 7.8 % (p < 0.05) were noted in the trabecular of the vertebra and tibia, respectively. In conclusion, implementation of FTIR and SAXS techniques in the OVX rat model, identified no significant compositional changes while substantiating thickening of the mineral crystals as a general structural feature of OVX-induced osteoporosis in rats.


Maternal Vegetable and Fruit Consumption during Pregnancy and Its Effects on Infant Gut Microbiome.
Maternal nutrition intake during pregnancy may affect the mother-to-child transmission of bacteria, resulting in gut microflora changes in the offspring, with long-term health consequences in later life. Longitudinal human studies are lacking, as only a small amount of studies showing the effect of nutrition intake during pregnancy on the gut microbiome of infants have been performed, and these studies have been mainly conducted on animals. This pilot study explores the effects of high or low fruit and vegetable gestational intake on the infant microbiome. We enrolled pregnant women with a complete 3-day dietary record and received postpartum follow-up. The 16S rRNA gene sequence was used to characterize the infant gut microbiome at 2 months (n = 39). Principal coordinate analysis ordination revealed that the infant gut microbiome clustered differently for high and low maternal fruit and vegetable consumption (p < 0.001). The linear discriminant analysis effect size and feature selection identified 6 and 17 taxa from both the high and low fruit and vegetable consumption groups. Among the 23 abundant taxa, we observed that six maternal intake nutrients were associated with nine taxa (e.g., Erysipelatoclostridium, Isobaculum, Lachnospiraceae, Betaproteobacteria, Burkholderiaceae, Sutterella, Clostridia, Clostridiales, and Lachnoclostridium). The amount of gestational fruit and vegetable consumption is associated with distinct changes in the infant gut microbiome at 2 months of age. Therefore, strategies involving increased fruit and vegetable consumption during pregnancy should be employed for modifying the gut microbiome early in life.


Fluorescent microspheres are reliable for serial bone blood flow measurements.
The fluorescent microsphere method is one of the current techniques to determine regional blood flow in various organs. The purpose of this study was to examine the suitability of fluorescent microspheres for serial measurement of regional bone blood flow. Six anesthetized female New Zealand rabbits received five left ventricular injections of fluorescent microspheres in 20-minute intervals. To test the precision of the measurement two types of fluorescent microspheres were injected simultaneously at the first and last injections. Blood flow was calculated in the kidneys, lungs, brain, femurs, and tibias after measuring the fluorescence intensity in each reference blood and tissue sample. Comparison of blood-flow values obtained by simultaneously injected microspheres showed an excellent correlation and a minimal percentage difference at the first and last injections, indicating valid measurements of regional bone blood flow. No significant differences were observed when comparing blood flow in the corresponding regions of bones on the right side and left side. Mean blood flow in the femur and tibia significantly increased at the fourth injection whereas flow distribution within the femur and tibia essentially remained unchanged throughout the experiment. Comparison of blood flow values obtained by simultaneously injected microspheres showed moderate agreement for the kidneys and lungs at the last injections. Because this finding might be attributable to disturbances of microcirculation caused by accumulation of spheres in high-flow organs, the increase in regional bone blood flow observed in our experiments has to be interpreted carefully. This study showed that bone blood flow can be determined reliably in anesthetized rabbits by as many as three serial injections of fluorescent microspheres.


Mental health stigma and barriers to care in World Trade Center responders: Results from a large, population-based health monitoring cohort.
Nearly 20 years after the terrorist attacks of September 11, 2001, multiple studies have documented the adverse mental consequences among World Trade Center (WTC) rescue, recovery, and clean-up workers. However, scarce research has examined mental health stigma and barriers to care in WTC-exposed individuals, and no known study has examined whether rates of endorsement may differ between police and "nontraditional" responders, the latter comprising a heterogeneous group of workers and volunteers. To identify the prevalence and correlates of mental health stigma and barriers to care in WTC responders. Mental health stigma and barriers to care and their correlates were examined in 6,777 police and 6,272 nontraditional WTC responders. Nontraditional responders endorsed more stigma or barriers to care concerns than police responders. Within a subsample who screened positive for a psychiatric disorder, police were more likely than nontraditional responders to endorse "concerns that negative job consequences might result" (17.9% vs. 9.1%), while nontraditional responders were more likely to endorse "I don't know where to go to find counseling services" (18.4% vs.6.6%). Within this subsample, mental health service need and more severe WTC-related posttraumatic stress disorder symptoms were associated with increased likelihood of endorsing stigma or barriers; pre-9/11 psychiatric history and non-Hispanic Black race/ethnicity were associated with lower likelihood of endorsing stigma or barriers. Results of this study underscore the burden of mental health stigma and barriers to care in WTC responders, and highlight the need for targeted interventions to address these concerns and promote mental healthcare utilization in this population.


Hydrophobic surfactant-associated proteins: electrophoretic and immunologic analyses and cellular localization in human lung.
Pulmonary surfactant isolated from a number of animal species contains a protein of molecular weight 38,000 and several very hydrophobic proteins soluble in solvents often used to extract lipids. Although found to be intimately associated with surfactant lipids, localization of the very hydrophobic proteins to alveolar epithelial type II cells, the cells involved in the synthesis and secretion of pulmonary surfactant, has not been demonstrated. Hydrophobic proteins were extracted along with lipids from human surfactant, and, after extraction with deoxycholate, were used to raise an antiserum. The antiserum was characterized by the immunoblotting ("Western") technique, using blots of pulmonary surfactant, and of hydrophobic proteins prepared by deoxycholate extraction and by Sephadex LH-20 chromatography. The antiserum shows reactivity to a 6.5- to 18-kDa surfactant-associated hydrophobic protein, which appears to be the major component of deoxycholate extracted proteins, and has Phe as the N-terminal amino acid. In addition, 28-kDa, 40-kDa, 50-kDa, and 70-kDa bands (chemically unreduced) are seen in human pulmonary surfactant. Whether these bands represent precursors of the major surfactant-associated hydrophobic proteins, different proteolytic cleavage products of the precursor protein, distinct proteins, or associated forms of the same protein is not yet clear. When used in the immunoperoxidase staining of human lung sections, the antiserum yielded a staining pattern identical to that obtained by the use of an antiserum to the 35-kDa surfactant protein. The number, size, and location of the stained cells are consistent with their being the alveolar epithelial type II cells.


Healthcare utilization, medical costs and mortality associated with malnutrition in patients with chronic obstructive pulmonary disease: a matched cohort study.
Objective: Although disease-related malnutrition has prognostic implications for patients with chronic obstructive pulmonary disease (COPD), its health-economic impact and clinical burdens are uncertain. We conducted a population-level study to investigate these questions. Methods: We excerpted data relevant to malnutrition, prolonged mechanical ventilation and medications from claims by 1,197,098 patients which were consistent with COPD and registered by the Taiwan National Health Insurance Administration between 2009 and 2013. These patients were separated into cohorts with or without respiratory failure requiring long-term mechanical ventilation, and each cohort was divided to compare cases who developed malnutrition after their first diagnosis consistent with COPD, versus non-malnourished propensity-score matched controls. Results: The prevalence of malnutrition was 3.8% overall (10,259/287,000 non-ventilator-dependent; 1198/15,829 ventilator-dependent). Propensity-score matched non-ventilator-dependent patients who became malnourished (N = 10,242) had comparatively more hospitalizations, emergency room and outpatient visits, longer hospitalization (all p < .01), and higher mortality (HR = 2.26, 95% CI 2.18-2.34) than non-malnourished controls (N = 40,968). Malnourished ventilator-dependent patients (N = 1197) had higher rates of hospitalization, emergency room and outpatient visits, but shorter hospitalization (all p < .001) and lower mortality (HR = 0.85, 95% CI 0.80-0.93) than matched non-malnourished controls (N = 4788). Total medical expenditure on malnourished non-ventilator-dependent COPD patients was 75% higher than controls (p < .001), whereas malnourished ventilator-dependent patients had total costs 7% lower than controls (p < .001). Conclusions: Malnourishment among COPD patients who were not dependent on mechanical ventilation was associated with greater healthcare resource utilization and higher aggregate medical costs.


Feasibility and effects of preventive home visits for at-risk older people: design of a randomized controlled trial.
The search for preventive methods to mitigate functional decline and unwanted relocation by older adults living in the community is important. Preventive home visit (PHV) models use infrequent but regular visits to older adults by trained practitioners with the goal of maintaining function and quality of life. Evidence about PHV efficacy is mixed but generally supportive. Yet interventions have rarely combined a comprehensive (biopsychosocial) occupational therapy intervention protocol with a home visit to older adults. There is a particular need in the USA to create and examine such a protocol. The study is a single-blind randomized controlled pilot trial designed to assess the feasibility, and to obtain preliminary efficacy estimates, of an intervention consisting of preventive home visits to community-dwelling older adults. An occupational therapy-based preventive home visit (PHV) intervention was developed and is being implemented and evaluated using a repeated measures design. We recruited a sample of 110 from a population of older adults (75+) who were screened and found to be at-risk for functional decline. Participants are currently living in the community (not in assisted living or a skilled nursing facility) in one of three central North Carolina counties. After consent, participants were randomly assigned into experimental and comparison groups. The experimental group receives the intervention 4 times over a 12 month follow-up period while the comparison group receives a minimal intervention of mailed printed materials. Pre- and post-intervention measures are being gathered by questionnaires administered face-to-face by a treatment-blinded research associate. Key outcome measures include functional ability, participation, life satisfaction, self-rated health, and depression. Additional information is collected from participants in the experimental group during the intervention to assess the feasibility of the intervention and potential modifiers. Fidelity is being addressed and measured across several domains. Feasibility indications to date are positive. Although the protocol has some limitations, we expect to learn enough about the intervention, delivery and effects to support a larger trial with a more stringent design and enhanced statistical power. ClinicalTrials.gov ID NCT00985283.


Long-term safety of radiotherapy and breast cancer laterality in older survivors.
Although adjuvant radiotherapy (RT) following surgery for breast cancer improves overall survival, controversy exists about its long-term adverse impact on cardiovascular health in older survivors. To determine whether incident cardiovascular disease (CVD) is associated with RT and whether tumor laterality modifies this association. Women aged 65+ years diagnosed with stage I and II breast cancer between 1990 and 1994 were identified from three health plans. Women were followed through CVD outcomes, health plan disenrollment, death, or study end (December 31, 2004). The main independent variable was RT use. Adjusted HRs and 95% CIs were estimated using Cox proportional hazards models with time-dependent tamoxifen and RT use status. We adjusted for age, race, stage, estrogen receptor/progesterone receptor, hypertension, and diabetes. In the full cohort (N = 806), RT was not associated with greater risk of CVD (maximum follow-up was 14 years). However, within the RT-exposed group (N = 340), women treated for left-side breast cancer had a significant increased risk of CVD outcomes (HR = 1.53, 95% CI: 1.06-2.21) compared with women with right-sided tumors. Laterality is critical to understanding the effect of RT on CVD. Studies of more contemporary cohorts of women treated with RT should incorporate this variable to determine whether the risk persists with refinements in the dosing and delivery of RT. As some irradiation to the heart is unavoidable even with refined modern RT techniques, continued effort is required to minimize such exposures, especially in older women with left-sided tumors.


A systematic review of conservative treatment of carpal tunnel syndrome.
To assess the effectiveness of conservative therapy in carpal tunnel syndrome. A computer-aided search of MEDLINE and the Cochrane Collaboration was conducted for randomized controlled trials (RCTs) from January 1985 to May 2006. RCTs were included if: (1) the patients, with clinically and electrophysiologically confirmed carpal tunnel syndrome, had not previously undergone surgical release, (2) the efficacy of one or more conservative treatment options was evaluated, (3) the study was designed as a randomized controlled trial. Two reviewers independently selected the studies and performed data extraction using a standardized form. In order to assess the methodological quality, the criteria list of the Cochrane Back Review Group for systematic reviews was applied. The different treatment methods were grouped (local injections, oral therapies, physical therapies, therapeutic exercises and splints). Thirty-three RCTs were included in the review. The studies were analysed to determine the strength of the available evidence for the efficacy of the treatment. Our review shows that: (1) locally injected steroids produce a significant but temporary improvement, (2) vitamin B6 is ineffective, (3) steroids are better than non-steroidal anti-inflammatory drugs (NSAIDs) and diuretics, but they can produce side-effects, (4) ultrasound is effective while laser therapy shows variable results, (5) exercise therapy is not effective, (6) splints are effective, especially if used full-time. There is: (1) strong evidence (level 1) on efficacy of local and oral steroids; (2) moderate evidence (level 2) that vitamin B6 is ineffective and splints are effective and (3) limited or conflicting evidence (level 3) that NSAIDs, diuretics, yoga, laser and ultrasound are effective whereas exercise therapy and botulinum toxin B injection are ineffective.


Imposing genetic diversity.
The idea that a world in which everyone was born "perfect" would be a world in which something valuable was missing often comes up in debates about the ethics of technologies of prenatal testing and preimplantation genetic diagnosis (PGD). This thought plays an important role in the "disability critique" of prenatal testing. However, the idea that human genetic variation is an important good with significant benefits for society at large is also embraced by a wide range of figures writing in the bioethics literature, including some who are notoriously hostile to the idea that we should not select against disability. By developing a number of thought experiments wherein we are to contemplate increasing genetic diversity from a lower baseline in order to secure this value, I argue that this powerful intuition is more problematic than is generally recognized, especially where the price of diversity is the well-being of particular individuals.


Profiling Proteinic Changes Induced by Vildagliptin Treatment in a Mouse Lung Transplantation Model: The Role of Kininogen-1.
BACKGROUND This study investigated the protective effects of pharmaceutical CD26/dipeptidylpeptidase-4 (CD26/DPP-4) inhibitor in lung transplantation (LTx). Changes in protein expression associated with the treatment were screened and identified to evaluate the role of kininogen-1 in early-term ischemia/reperfusion (I/R) injury after LTx. MATERIAL AND METHODS Orthotopic single LTx was performed in syngeneic C57BL/6 mice, with a pharmaceutical CD26/DPP-4 inhibitor (vildagliptin, subcutaneous injection, 10 mg/kg, every 12 h) administered to the investigational group. All donors were perfused and preserved with low potassium dextran (LPD). Grafts were harvested at 60 h post-transplantation after 8 h of cold ischemia. Myeloperoxidase activity and wet/dry weight ratio were measured, followed by histopathological examination. Proteins were separated, analyzed, and identified using proteomics and database searches. The target proteins were validated by Western blot. Immunohistochemical studies were performed in the same lung specimen locus. RESULTS Investigational group (IN) versus control group (CON) comparison showed decreased myeloperoxidase enzymatic activity, as well as decreased edema and interstitial-alveolar inflammation. Proteomics results revealed 78 spots with significant differences in abundance between the 2 groups. Fifteen proteins were identified. Kininogen-1 was up-regulated in CON and down-regulated in IN, with contrasting results for the heat shock protein 70. Immunohistochemical results revealed significantly different staining with kininogen-1 in alveolar macrophages and inflammatory cells. CONCLUSIONS Combined vildagliptin and LPD significantly ameliorated I/R injury after LTx. This treatment may change local pulmonary protein levels. Moreover, proper application of proteins such as kininogen-1 may enhance the protective effects against I/R injury during transplantation.


Bioimpedance and NIR for Non-invasive Assessment of Blood Glucose.
Sixteen volunteers each drank 700 ml sugar-containing soft drink during two successive periods and the blood sugar was measured at 10 min intervals together with electrical impedance spectroscopy and near infrared spectroscopy (NIR). A maximum correlation of 0.46 was found for the electrical measurements but no clear separation between low and high blood glucose levels were found in the NIR measurements. The latter was attributed to the experimental design where the NIR probe was removed from the skin between each measurement.


Rationally designed trimetallic Prussian blue analogues on LDH/Ni foam for high performance supercapacitors.
Rational design of a Prussian blue analogue (PBA)@Ni-Co layered double hydroxide (NiCo-LDH) nanocomposite electrode material is vitally important for synthesizing high-performance supercapacitor electrodes. In this work, such nanocomposite electrode materials were successfully fabricated by a facile hydrothermal method. Firstly, three-dimensional (3D) regulated NiCo-LDH nanosheets with high interlayer space were grown on nickel foam under mild synthetic conditions. Then these nanosheets as a precursor were in situ converted into the target PBA@NiCo-LDH/NF nanocomposite electrode by a facile thermal ion-exchange reaction with potassium ferricyanide (K3[Fe(CN)6]). A series of PBA@NiCo-LDH/NF nanocomposite electrodes were fabricated with different ratios of Ni and Co and reaction temperatures. Their structures and morphologies were characterized by X-ray diffraction (XRD), FT-IR and scanning electron microscopy (SEM). Electrochemical investigation reveals that the firstname.lastname@example.com electrode exhibits the best electrochemical performance with an area specific capacitance of 2004.26 mF cm-2 at 1 mA cm-2, which is much higher (about three times) than the properties of each single component. All results demonstrate that (1) high-performance composite electrodes can be effectively fabricated and (2) fabrication of such composites is highly necessary and important.


Deleting the IF1-like ζ subunit from Paracoccus denitrificans ATP synthase is not sufficient to activate ATP hydrolysis.
In oxidative phosphorylation, ATP synthases interconvert two forms of free energy: they are driven by the proton-motive force across an energy-transducing membrane to synthesize ATP and displace the ADP/ATP ratio from equilibrium. For thermodynamically efficient energy conversion they must be reversible catalysts. However, in many species ATP synthases are unidirectional catalysts (their rates of ATP hydrolysis are negligible), and in others mechanisms have evolved to regulate or minimize hydrolysis. Unidirectional catalysis by Paracoccus denitrificans ATP synthase has been attributed to its unique ζ subunit, which is structurally analogous to the mammalian inhibitor protein IF1 Here, we used homologous recombination to delete the ζ subunit from the P. denitrificans genome, and compared ATP synthesis and hydrolysis by the wild-type and knockout enzymes in inverted membrane vesicles and the F1-ATPase subcomplex. ATP synthesis was not affected by loss of the ζ subunit, and the rate of ATP hydrolysis increased by less than twofold, remaining negligible in comparison with the rates of the Escherichia coli and mammalian enzymes. Therefore, deleting the P. denitrificans ζ subunit is not sufficient to activate ATP hydrolysis. We close by considering our conclusions in the light of reversible catalysis and regulation in ATP synthase enzymes.


Elevated circulating amyloid concentrations in obesity and diabetes promote vascular dysfunction.
Diabetes, obesity, and Alzheimer's disease (AD) are associated with vascular complications and impaired nitric oxide (NO) production. Furthermore, increased β-site amyloid precursor protein-cleaving (APP-cleaving) enzyme 1 (BACE1), APP, and β-amyloid (Aβ) are linked with vascular disease development and increased BACE1 and Aβ accompany hyperglycemia and hyperlipidemia. However, the causal relationship between obesity and diabetes, increased Aβ, and vascular dysfunction is unclear. We report that diet-induced obesity (DIO) in mice increased plasma and vascular Aβ42 that correlated with decreased NO bioavailability, endothelial dysfunction, and increased blood pressure. Genetic or pharmacological reduction of BACE1 activity and Aβ42 prevented and reversed, respectively, these outcomes. In contrast, expression of human mutant APP in mice or Aβ42 infusion into control diet-fed mice to mimic obese levels impaired NO production, vascular relaxation, and raised blood pressure. In humans, increased plasma Aβ42 correlated with diabetes and endothelial dysfunction. Mechanistically, higher Aβ42 reduced endothelial NO synthase (eNOS), cyclic GMP (cGMP), and protein kinase G (PKG) activity independently of diet, whereas endothelin-1 was increased by diet and Aβ42. Lowering Aβ42 reversed the DIO deficit in the eNOS/cGMP/PKG pathway and decreased endothelin-1. Our findings suggest that BACE1 inhibitors may have therapeutic value in the treatment of vascular disease associated with diabetes.


Dorsal Lingual Surface and Halitosis: a Morphological Point of View.
The interest in the study of the tongue papillary niches and the related biofilm has increased in recent years because they form a suitable source of periodontal microorganisms and are associated with development of halitosis. Tongue dorsum structure represents a factor favoring a particular and complex bacterial biofilm where periodontal pathogens are frequently found. The aim of this preliminary study was to associate the tongue papillary structure with the biofilm causing halitosis by means of a new clinical protocol. In this study, one subject affected by oral malodor was selected and included. A photograph of lingual dorsum was taken to spot the areas with visible lingual coating. A tongue dorsum impression was obtained, divided and cut with a blade in six parts, according to Winkel Tongue Coated Index by means of the 2-step double-mix impression technique. The contours of the six parts were observed by the stereomicroscope LEICA LED2000 and analyzed by ImageJ software. The results showed that the depth of papillae was associated with visible presence of the tongue biofilm and indirectly correlated with halitosis in patients.. The morphological papillary structure of the tongue dorsal surface influences the presence of the tongue biofilm. The presented protocol can be further considered in clinical application for a correct diagnosis and a personalized treatment of halitosis.


Establishment of an automated patch-clamp platform for electrophysiological and pharmacological evaluation of hiPSC-CMs.
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have evolved into widely used and reliable cell sources for modeling cardiovascular channelopathies and for drug safety pharmacology. However, the electrophysiological and pharmacological applications of hiPSC-CMs are hampered by manual patch-clamp technique, which is labor-intensive and generates low output. The automated patch-clamp technique is showing potential to overcome this problem. Here, we describe a new dissociation method, with which we can harvest a vast number of single relaxed hiPSC-CMs with smooth membrane suited for automated patch-clamp. Using the automated whole-cell patch-clamp technology, we report a high success rate for cell capture and whole-cell access (around 70%). We are able to identify and record several currents and paced action potentials (APs) with different success rates, including Na+ current (INa), L-type Ca2+ current (ICaL), two specific K+ currents, the transient outward K+ current (Ito) and the inward rectifier K+ current (IK1). Moreover, we successfully applied dynamic current-clamp to virtually increase IK1 for AP recordings. Our study suggests that automated patch-clamp technology could be used to investigate the relevant ionic currents and APs in hiPSC-CMs. The combination of automated patch-clamp and hiPSC-CM technologies promises a wide range of applications in the future.


Inverted translational control of eukaryotic gene expression by ribosome collisions.
The canonical model of eukaryotic translation posits that efficient translation initiation increases protein expression and mRNA stability. Contrary to this model, we find that increasing initiation rate can decrease both protein expression and stability of certain mRNAs in the budding yeast Saccharomyces cerevisiae. These mRNAs encode a stretch of polybasic residues that cause ribosome stalling. Our computational modeling predicts that the observed decrease in gene expression at high initiation rates occurs when ribosome collisions at stalls stimulate abortive termination of the leading ribosome or cause endonucleolytic mRNA cleavage. Consistent with this prediction, the collision-associated quality-control factors Asc1 and Hel2 (orthologs of human RACK1 and ZNF598, respectively) decrease gene expression from stall-containing mRNAs only at high initiation rates. Remarkably, hundreds of S. cerevisiae mRNAs that contain ribosome stall sequences also exhibit lower translation efficiency. We propose that inefficient translation initiation allows these stall-containing endogenous mRNAs to escape collision-stimulated reduction in gene expression.


Accurate characterization of the IFITM locus using MiSeq and PacBio sequencing shows genetic variation in Galliformes.
Interferon inducible transmembrane (IFITM) proteins are effectors of the immune system widely characterized for their role in restricting infection by diverse enveloped and non-enveloped viruses. The chicken IFITM (chIFITM) genes are clustered on chromosome 5 and to date four genes have been annotated, namely chIFITM1, chIFITM3, chIFITM5 and chIFITM10. However, due to poor assembly of this locus in the Gallus Gallus v4 genome, accurate characterization has so far proven problematic. Recently, a new chicken reference genome assembly Gallus Gallus v5 was generated using Sanger, 454, Illumina and PacBio sequencing technologies identifying considerable differences in the chIFITM locus over the previous genome releases. We re-sequenced the locus using both Illumina MiSeq and PacBio RS II sequencing technologies and we mapped RNA-seq data from the European Nucleotide Archive (ENA) to this finalized chIFITM locus. Using SureSelect probes capture probes designed to the finalized chIFITM locus, we sequenced the locus of a different chicken breed, namely a White Leghorn, and a turkey. We confirmed the Gallus Gallus v5 consensus except for two insertions of 5 and 1 base pair within the chIFITM3 and B4GALNT4 genes, respectively, and a single base pair deletion within the B4GALNT4 gene. The pull down revealed a single amino acid substitution of A63V in the CIL domain of IFITM2 compared to Red Jungle fowl and 13, 13 and 11 differences between IFITM1, 2 and 3 of chickens and turkeys, respectively. RNA-seq shows chIFITM2 and chIFITM3 expression in numerous tissue types of different chicken breeds and avian cell lines, while the expression of the putative chIFITM1 is limited to the testis, caecum and ileum tissues. Locus resequencing using these capture probes and RNA-seq based expression analysis will allow the further characterization of genetic diversity within Galliformes.


Bisubstrate-Type Chemical Probes Identify GRP94 as a Potential Target of Cytosine-Containing Adenosine Analogs.
We synthesized affinity-based chemical probes of cytosine-adenosine bisubstrate analogs and identified several potential targets by proteomic analysis. The validation of the proteomic analysis identified the chemical probe as a specific inhibitor of glucose-regulated protein 94 (GRP94), a potential drug target for several types of cancers. Therefore, as a result of the use of bisubstrate-type chemical probes and a chemical-biology methodology, this work opens the way to the development of a new family of GRP94 inhibitors that could potentially be of therapeutic interest.


Effects of Newcastle disease virus vaccine antibodies on the shedding and transmission of challenge viruses.
Different genotypes of avian paramyxovirus serotype-1 virus (APMV-1) circulate in many parts of the world. Traditionally, Newcastle disease virus (NDV) is recognized as having two major divisions represented by classes I and II, with class II being further divided into sixteen genotypes. Although all NDV are members of APMV-1 and are of one serotype, antigenic and genetic diversity is observed between the different genotypes. Reports of vaccine failure from many countries and reports by our lab on the reduced ability of classical vaccines to significantly decrease viral replication and shedding have created renewed interest in developing vaccines formulated with genotypes homologous to the virulent NDV (vNDV) circulating in the field. We assessed how the amount and specificity of humoral antibodies induced by inactivated vaccines affected viral replication, clinical protection and evaluated how non-homologous (heterologous) antibody levels induced by live NDV vaccines relate to transmission of vNDV. In an experimental setting, all inactivated NDV vaccines protected birds from morbidity and mortality, but higher and more specific levels of antibodies were required to significantly decrease viral replication. It was possible to significantly decrease viral replication and shedding with high levels of antibodies and those levels could be more easily reached with vaccines formulated with NDV of the same genotype as the challenge viruses. However, when the levels of heterologous antibodies were sufficiently high, it was possible to prevent transmission. As the level of humoral antibodies increase in vaccinated birds, the number of infected birds and the amount of vNDV shed decreased. Thus, in an experimental setting the effective levels of humoral antibodies could be increased by (1) increasing the homology of the vaccine to the challenge virus, or (2) allowing optimal time for the development of the immune response.


Hematopoietic Cell Transplantation for Myelodysplastic Syndromes.
Allogeneic hematopoietic cell transplantation (HCT) offers the only potential cure for patients with myelodysplastic syndromes (MDS). However, with current approaches to HCT, many older patients with comorbidities are poor HCT candidates, and treatment-related morbidity and mortality may offset benefit for patients with lower-risk disease. Consequently, selection of patients with MDS for HCT should take into consideration disease risk category including mutational status, HCT comorbidity index, functional status, donor options, and available institutional resources. Formal geriatric assessment may further guide use of HCT and, if HCT is chosen, selection of conditioning intensity. Patients with higher-risk MDS should be considered for HCT at the time of diagnosis, whereas expectant nontransplant management is more appropriate for those with lower-risk disease. A high blast burden at the time of HCT increases the risk of subsequent relapse; however, the role of pretransplant cytoreductive therapy and the regimen of choice remain controversial. Patients with MDS younger than 65 years and with an HCT comorbidity index ≤ 4 may benefit from more intense conditioning regimens. The presence of complex or monosomal karyotype or mutations in TP53, DNMT3A, or other genes identify patients with poorer outcomes following HCT. Patients with TP53 mutations have particularly poor survival, and should be enrolled in clinical trials whenever possible. Several important HCT studies are ongoing and will better define the role of HCT in MDS as well as the value of pretransplant cytoreductive therapy or post-transplant relapse-prevention strategies. Given the apparent underuse of HCT in eligible patients and low enrollment in MDS HCT clinical trials to date, timely referral of patients with MDS to such trials and HCT programs is critical.


Large-scale proteomics and phosphoproteomics of urinary exosomes.
Normal human urine contains large numbers of exosomes, which are 40- to 100-nm vesicles that originate as the internal vesicles in multivesicular bodies from every renal epithelial cell type facing the urinary space. Here, we used LC-MS/MS to profile the proteome of human urinary exosomes. Overall, the analysis identified 1132 proteins unambiguously, including 177 that are represented on the Online Mendelian Inheritance in Man database of disease-related genes, suggesting that exosome analysis is a potential approach to discover urinary biomarkers. We extended the proteomic analysis to phosphoproteomic profiling using neutral loss scanning, and this yielded multiple novel phosphorylation sites, including serine-811 in the thiazide-sensitive Na-Cl co-transporter, NCC. To demonstrate the potential use of exosome analysis to identify a genetic renal disease, we carried out immunoblotting of exosomes from urine samples of patients with a clinical diagnosis of Bartter syndrome type I, showing an absence of the sodium-potassium-chloride co-transporter 2, NKCC2. The proteomic data are publicly accessible at http://dir.nhlbi.nih.gov/papers/lkem/exosome/.


Polarization variations induced by high hydrostatic pressures in the isolated frog skin as related to the effects on passive ionic permeability and active Na+ transport.
The effects of a wide range of hydrostatic pressures (from 50 to 1000 kg/cm2) have been investigated on the spontaneous potential difference (PD), the short-circuit current (SCC) and the activity of the membrane ATPases of the isolated abdominal skin from the frog Rana temporaria L. Two types of variations in PD are induced by pressure changes: short and transient potential variations which appear to be related to the pressure change (compression and decompression) and lasting variations which persist as long as pressure is applied and whose nature appears to be related to the pressure magnitude. Long-lasting potential changes have particularly been investigated. At pressures lower than 500 kg/cm2, the skin potential increases while a pressure over 500-600 kg/cm2 induces a depolarization. Both variations consecutively occur at 500 +/- 100 kg/cm2. These effects of pressure have been shown to be reversible up to about 800 kg/cm2. The question of the origin of the potential changes is discussed and it is proposed that the lasting hyperpolarization results from an effect on the passive permeabilities to Na+, K+ and Cl- ions inducing in turm a secondary readjustment (stimulation) of the Na+ active transport while the depolarization at high pressures reflects a direct inhibition of the Na+ pump. These interpretations are supported by experimental data on the effects of pressure on the short-circuit current and on the activity of the skin (Na+ + K+) ATPase.


Rebamipide Attenuates Mandibular Condylar Degeneration in a Murine Model of TMJ-OA by Mediating a Chondroprotective Effect and by Downregulating RANKL-Mediated Osteoclastogenesis.
Temporomandibular joint osteoarthritis (TMJ-OA) is characterized by progressive degradation of cartilage and changes in subchondral bone. It is also one of the most serious subgroups of temporomandibular disorders. Rebamipide is a gastroprotective agent that is currently used for the treatment of gastritis and gastric ulcers. It scavenges reactive oxygen radicals and has exhibited anti-inflammatory potential. The aim of this study was to investigate the impact of rebamipide both in vivo and in vitro on the development of cartilage degeneration and osteoclast activity in an experimental murine model of TMJ-OA, and to explore its mode of action. Oral administration of rebamipide (0.6 mg/kg and 6 mg/kg) was initiated 24 h after TMJ-OA was induced, and was maintained daily for four weeks. Rebamipide treatment was found to attenuate cartilage degeneration, to reduce the number of apoptotic cells, and to decrease the expression levels of matrix metalloproteinase-13 (MMP-13) and inducible nitric oxide synthase (iNOS) in TMJ-OA cartilage in a dose-dependent manner. Rebamipide also suppressed the activation of transcription factors (e.g., NF-κB, NFATc1) and mitogen-activated protein kinases (MAPK) by receptor activator of nuclear factor kappa-B ligand (RANKL) to inhibit the differentiation of osteoclastic precursors, and disrupted the formation of actin rings in mature osteoclasts. Together, these results demonstrate the inhibitory effects of rebamipide on cartilage degradation in experimentally induced TMJ-OA. Furthermore, suppression of oxidative damage, restoration of extracellular matrix homeostasis of articular chondrocytes, and reduced subchondral bone loss as a result of blocked osteoclast activation suggest that rebamipide is a potential therapeutic strategy for TMJ-OA.


Neoadjuvant immune checkpoint blockade in high-risk resectable melanoma.
Preclinical studies suggest that treatment with neoadjuvant immune checkpoint blockade is associated with enhanced survival and antigen-specific T cell responses compared with adjuvant treatment1; however, optimal regimens have not been defined. Here we report results from a randomized phase 2 study of neoadjuvant nivolumab versus combined ipilimumab with nivolumab in 23 patients with high-risk resectable melanoma ( NCT02519322 ). RECIST overall response rates (ORR), pathologic complete response rates (pCR), treatment-related adverse events (trAEs) and immune correlates of response were assessed. Treatment with combined ipilimumab and nivolumab yielded high response rates (RECIST ORR 73%, pCR 45%) but substantial toxicity (73% grade 3 trAEs), whereas treatment with nivolumab monotherapy yielded modest responses (ORR 25%, pCR 25%) and low toxicity (8% grade 3 trAEs). Immune correlates of response were identified, demonstrating higher lymphoid infiltrates in responders to both therapies and a more clonal and diverse T cell infiltrate in responders to nivolumab monotherapy. These results describe the feasibility of neoadjuvant immune checkpoint blockade in melanoma and emphasize the need for additional studies to optimize treatment regimens and to validate putative biomarkers.


Effect of hypoxia and reoxygenation on the activity of transglutaminase in brain of newborn piglets.
The transglutaminase activity in five regions of the brain of newborn piglets was measured and the effects of hypoxia and posthypoxic period on this activity evaluated. Enzyme activity was measured in homogenates from cortex, hippocampus, striatum, thalamus and midbrain. The control activities were 7.2, 6.2, 6.0, 5.7 and 4.6 pmol/mg protein/min, respectively. The activities at the end of an 18 min period of hypoxia induced by an FiO2 of 9% were not significantly different from control activities. By 3 h after the hypoxic episode, however, the transglutaminase activities were significantly above control levels in all five regions of the brain. Measurements of the kinetic constants of tranglutaminase indicated that increases in enzyme activity were associated with an increase in Vmax with no significant change in the apparent affinity of the enzyme for the substrate, putrescine. The increased activity of transglutaminase during the posthypoxic period, with no changes immediately after hypoxia, suggest that the increases could be due to increased enzyme synthesis rather than activation of existing enzyme. The rise in transglutaminase activity subsequent to a hypoxic episode may contribute significantly to the long-term disturbances in cellular metabolism in the immature brain induced by hypoxic episodes.


Long-range atmospheric infrasound propagation from subsurface sources.
In seismology and ocean acoustics, the interface with the atmosphere is typically represented as a free surface. Similarly, these interfaces are considered as a rigid surface for infrasound propagation. This implies that seismic or acoustic waves are not transmitted into the atmosphere from subsurface sources, and vice versa. Nevertheless, infrasound generated by subsurface sources has been observed. In this work, seismo-acoustic modeling of infrasound propagation from underwater and underground sources will be presented. The fast field program (FFP) is used to model the seismo-acoustic coupling between the solid earth, the ocean, and the atmosphere under the variation of source and media parameters. The FFP model allows for a detailed analysis of the seismo-acoustic coupling mechanisms in frequency-wavenumber space. A thorough analysis of the coupling mechanisms reveals that evanescent wave coupling and leaky surface waves are the main energy contributors to long-range infrasound propagation. Moreover, it is found that source depth affects the relative amplitude of the tropospheric and stratospheric phases, which allows for source depth estimation in the future.


Citrus aurantium L. essential oil exhibits anxiolytic-like activity mediated by 5-HT(1A)-receptors and reduces cholesterol after repeated oral treatment.
The current treatments for anxiety disorders and depression have multiple adverse effects in addition to a delayed onset of action, which has prompted efforts to find new substances with potential activity in these disorders. Citrus aurantium was chosen based on ethnopharmacological data because traditional medicine refers to the Citrus genus as useful in diminishing the symptoms of anxiety or insomnia, and C. aurantium has more recently been proposed as an adjuvant for antidepressants. In the present work, we investigated the biological activity underlying the anxiolytic and antidepressant effects of C. aurantium essential oil (EO), the putative mechanism of the anxiolytic-like effect, and the neurochemical changes in specific brain structures of mice after acute treatment. We also monitored the mice for possible signs of toxicity after a 14-day treatment. The anxiolytic-like activity of the EO was investigated in a light/dark box, and the antidepressant activity was investigated in a forced swim test. Flumazenil, a competitive antagonist of benzodiazepine binding, and the selective 5-HT(1A) receptor antagonist WAY100635 were used in the experimental procedures to determine the mechanism of action of the EO. To exclude false positive results due to motor impairment, the mice were submitted to the rotarod test. The data suggest that the anxiolytic-like activity observed in the light/dark box procedure after acute (5 mg/kg) or 14-day repeated (1 mg/kg/day) dosing was mediated by the serotonergic system (5-HT(1A) receptors). Acute treatment with the EO showed no activity in the forced swim test, which is sensitive to antidepressants. A neurochemical evaluation showed no alterations in neurotransmitter levels in the cortex, the striatum, the pons, and the hypothalamus. Furthermore, no locomotor impairment or signs of toxicity or biochemical changes, except a reduction in cholesterol levels, were observed after treatment with the EO. This work contributes to a better understanding of the biological activity of C. aurantium EO by characterizing the mechanism of action underlying its anxiolytic-like activity.


Cyclosporin-A differentially affects apoptosis during in vivo rat thymocyte maturation.
Maturation arrest and interference with selection are two well-documented effects of cyclosporin-A (CsA) on the thymus. We recently hypothesized that these effects are related and owing to the reduced T-cell receptor (TCR)-CD3 complex-mediated signal transduction in thymocytes upon CsA treatment. In this hypothesis, the maturation arrest is the result of the additional depletion of thymocytes that normally survive by positive selection, whereas the impaired self-tolerance induction is caused by an increased survival of thymocytes that normally undergo negative selection. In this view, it is anticipated that CsA differentially affects thymocyte apoptosis during in vivo thymocyte maturation. Indeed, we report in this study a strong increase in apoptotic cells in the thymic cortex on in situ analysis. Simultaneously, the number of apoptotic cells had decreased at the cortico-medullary zone which is held to be the site for negative selection. Rapamycin (Rapa) also interferes with thymocyte maturation by inhibiting cytokine-driven proliferation. Hence, Rapa preferentially affects the early maturational stages of thymocyte development and is considered not to alter thymocyte selection and subsequent apoptotic events. Indeed, the number of apoptotic events appears not to be altered. However, possibly owing to the decrease in cortical macrophages, the apoptotic cells revealed an atypical enumeration around blood vessels. Taken together, our results favour the hypothesis that the dominant effect of CsA on the thymus is the reduction of the TCR-CD3 complex-mediated signal transduction in thymocytes upon interaction with stromal cells. Furthermore, the preferential localization of apoptotic cells next to blood vessels upon Rapa administration may indicate that endothelial cells are a back-up system for the removal of apoptotic cells.


Structural and biochemical characterization of human orphan DHRS10 reveals a novel cytosolic enzyme with steroid dehydrogenase activity.
To this day, a significant proportion of the human genome remains devoid of functional characterization. In this study, we present evidence that the previously functionally uncharacterized product of the human DHRS10 gene is endowed with 17beta-HSD (17beta-hydroxysteroid dehydrogenase) activity. 17beta-HSD enzymes are primarily involved in the metabolism of steroids at the C-17 position and also of other substrates such as fatty acids, prostaglandins and xenobiotics. In vitro, DHRS10 converts NAD+ into NADH in the presence of oestradiol, testosterone and 5-androstene-3beta,17beta-diol. Furthermore, the product of oestradiol oxidation, oestrone, was identified in intact cells transfected with a construct plasmid encoding the DHRS10 protein. In situ fluorescence hybridization studies have revealed the cytoplasmic localization of DHRS10. Along with tissue expression data, this suggests a role for DHRS10 in the local inactivation of steroids in the central nervous system and placenta. The crystal structure of the DHRS10 apoenzyme exhibits secondary structure of the SDR (short-chain dehydrogenase/reductase) family: a Rossmann-fold with variable loops surrounding the active site. It also reveals a broad and deep active site cleft into which NAD+ and oestradiol can be docked in a catalytically competent orientation.


Listener Modeling and Context-Aware Music Recommendation Based on Country Archetypes.
Music preferences are strongly shaped by the cultural and socio-economic background of the listener, which is reflected, to a considerable extent, in country-specific music listening profiles. Previous work has already identified several country-specific differences in the popularity distribution of music artists listened to. In particular, what constitutes the "music mainstream" strongly varies between countries. To complement and extend these results, the article at hand delivers the following major contributions: First, using state-of-the-art unsupervized learning techniques, we identify and thoroughly investigate (1) country profiles of music preferences on the fine-grained level of music tracks (in contrast to earlier work that relied on music preferences on the artist level) and (2) country archetypes that subsume countries sharing similar patterns of listening preferences. Second, we formulate four user models that leverage the user's country information on music preferences. Among others, we propose a user modeling approach to describe a music listener as a vector of similarities over the identified country clusters or archetypes. Third, we propose a context-aware music recommendation system that leverages implicit user feedback, where context is defined via the four user models. More precisely, it is a multi-layer generative model based on a variational autoencoder, in which contextual features can influence recommendations through a gating mechanism. Fourth, we thoroughly evaluate the proposed recommendation system and user models on a real-world corpus of more than one billion listening records of users around the world (out of which we use 369 million in our experiments) and show its merits vis-à-vis state-of-the-art algorithms that do not exploit this type of context information.


Discovery of an inhibitor of insulin-like growth factor 1 receptor activation: implications for cellular potency and selectivity over insulin receptor.
Insulin-like growth factor 1 receptor (IGF-1R) is an attractive target for anti-cancer therapy due to its anti-apoptotic effect on tumor cells, but inhibition of insulin receptor (IR) may have undesired metabolic consequences. The primary sequences of the ATP substrate-binding sites of these receptors are identical and the crystal structures of the activated kinase domains are correspondingly similar. Thus, most small-molecule inhibitors described to date are equally potent against the activated kinase domains of IGF-1R and IR. In contrast, the non-phosphorylated kinase domains of these receptors have several structural features that may accommodate differences in binding affinity for kinase inhibitors. We used a cell-based assay measuring IGF-1R autophosphorylation as an inhibitor screen, and identified a potent purine derivative that is selective compared to IR. Surprisingly, the compound is a weak inhibitor of the activated IGF-1R tyrosine kinase domain. Biochemical and structural studies are presented that indicate the compound preferentially binds to the ATP site of non-phosphorylated IGF-1R compared to phosphorylated IGF-1R. The potential selectivity and potency advantages of this binding mode are discussed.


Current mass spectrometry strategies for the analysis of pesticides and their metabolites in food and water matrices.
Analysis of pesticides and their metabolites in food and water matrices continues to be an active research area closely related to food safety and environmental issues. This review discusses the most widely applied mass spectrometric (MS) approaches to pesticide residues analysis over the last few years. The main techniques for sample preparation remain solvent extraction and solid-phase extraction. The QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) approach is being increasingly used for the development of multi-class pesticide residues methods in various sample matrices. MS detectors-triple quadrupole (QqQ), ion-trap (IT), quadrupole linear ion trap (QqLIT), time-of-flight (TOF), and quadrupole time-of-flight (QqTOF)-have been established as powerful analytical tools sharing a primary role in the detection/quantification and/or identification/confirmation of pesticides and their metabolites. Recent developments in analytical instrumentation have enabled coupling of ultra-performance liquid chromatography (UPLC) and fast gas chromatography (GC) with MS detectors, and faster analysis for a greater number of pesticides. The newly developed "ambient-ionization" MS techniques (e.g., desorption electrospray ionization, DESI, and direct analysis in real time, DART) hyphenated with high-resolution MS platforms without liquid chromatography separation, and sometimes with minimum pre-treatment, have shown potential for pesticide residue screening. The recently introduced Orbitrap mass spectrometers can provide high resolving power and mass accuracy, to tackle complex analytical problems involved in pesticide residue analysis.


Authenticity at Work: A Matter of Fit?
Authenticity at work refers to the extent to which a worker feels in touch with their true self while at work. At first sight this concept seems to overlap with the concept of person-environment (P-E) fit, that is, the degree to which an individual experiences good fit with their work environment. Drawing on a sample of 867 Dutch gifted workers, structural equation modeling was used to investigate (i) whether authenticity at work and P-E fit can be distinguished, and (ii) how authenticity at work and P-E fit were associated with employee well-being. As expected, confirmatory factor analysis revealed that authenticity at work and P-E fit were distinct from each other. Moreover, the mediated effect of authenticity at work was stronger for two negative forms of well-being (burnout and boredom) than for two positive forms of well-being (work engagement and job satisfaction). The theoretical and practical implications of these findings are discussed, especially focusing on the distinction between authenticity and P-E fit.


Does a tree-like phylogeny only exist at the tips in the prokaryotes?
The extent to which prokaryotic evolution has been influenced by horizontal gene transfer (HGT) and therefore might be more of a network than a tree is unclear. Here we use supertree methods to ask whether a definitive prokaryotic phylogenetic tree exists and whether it can be confidently inferred using orthologous genes. We analysed an 11-taxon dataset spanning the deepest divisions of prokaryotic relationships, a 10-taxon dataset spanning the relatively recent gamma-proteobacteria and a 61-taxon dataset spanning both, using species for which complete genomes are available. Congruence among gene trees spanning deep relationships is not better than random. By contrast, a strong, almost perfect phylogenetic signal exists in gamma-proteobacterial genes. Deep-level prokaryotic relationships are difficult to infer because of signal erosion, systematic bias, hidden paralogy and/or HGT. Our results do not preclude levels of HGT that would be inconsistent with the notion of a prokaryotic phylogeny. This approach will help decide the extent to which we can say that there is a prokaryotic phylogeny and where in the phylogeny a cohesive genomic signal exists.


Systematic review of the current evidence in the use of postoperative radiotherapy for oral squamous cell carcinoma.
Improved disease-free survival for oral squamous cell carcinoma (SCC) with the use of postoperative radiotherapy (PORT) has to be balanced against the risk of recurrence, the relative morbidity of radiotherapy, reduced options for treatment, and survival with recurrent disease. In the absence of randomised trials, a review of current evidence is timely because of increasing differences in outcome and response to treatment for cancers of the larynx, oropharynx, and oral cavity. From a search of 109 papers, 25 presented relevant data in tabular form, and reported local, regional, and total recurrence, and overall survival. Most data come from non-randomised studies that compared the effects of interventions with previous or historical information. A summary of the results shows local recurrence of 11%, 17%, and 15% for early, late, and all stages after operation alone, compared with 13%, 16%, and 19% after PORT. Regional recurrence is reported as 13%, 12%, and 11% for early, late, and all stages after operation alone compared with 6%, 11%, and 9% after PORT. Overall survival is reported as 76%, 74%, and 77% for operation alone compared with 65%, 62%, and 62% for early, late and all stages of oral SCC, respectively. It is acknowledged that this is a weak level of evidence as patients who have PORT probably have a high pathological-stage of disease. Knowing that PORT increases morbidity and reduces salvage rates and options for treating recurrent disease, this difference in overall survival emphasises the need for randomised studies or a re-evaluation of our current protocols.


A novel secretory tumor necrosis factor-inducible protein (TSG-6) is a member of the family of hyaluronate binding proteins, closely related to the adhesion receptor CD44.
TSG-6 cDNA was isolated by differential screening of a lambda cDNA library prepared from tumor necrosis factor (TNF)-treated human diploid FS-4 fibroblasts. We show that TSG-6 mRNA was not detectable in untreated cells, but became readily induced by TNF in normal human fibroblast lines and in peripheral blood mononuclear cells. In contrast, TSG-6 mRNA was undetectable in either control or TNF-treated human vascular endothelial cells and a variety of tumor-derived or virus-transformed cell lines. The sequence of full-length TSG-6 cDNA revealed one major open reading frame predicting a polypeptide of 277 amino acids, including a typical cleavable signal peptide. The NH2-terminal half of the predicted TSG-6 protein sequence shows a significant homology with a region implicated in hyaluronate binding, present in cartilage link protein, proteoglycan core proteins, and the adhesion receptor CD44. The most extensive sequence homology exists between the predicted TSG-6 protein and CD44. Western blot analysis with an antiserum raised against a TSG-6 fusion protein detected a 39-kD glycoprotein in the supernatants of TNF-treated FS-4 cells and of cells transfected with TSG-6 cDNA. Binding of the TSG-6 protein to hyaluronate was demonstrated by coprecipitation. Our data indicate that the inflammatory cytokine (TNF or IL-1)-inducible, secretory TSG-6 protein is a novel member of the family of hyaluronate binding proteins, possibly involved in cell-cell and cell-matrix interactions during inflammation and tumorigenesis.


Mass Spectrometry-Based Metabolomics Reveals Occupational Exposure to Per- and Polyfluoroalkyl Substances Relates to Oxidative Stress, Fatty Acid β-Oxidation Disorder, and Kidney Injury in a Manufactory in China.
Occupational workers are usually exposed to high levels of per- and polyfluoroalkyl substances (PFASs), placing them under greater health risks compared to the general population. Herein, 40 occupational workers from a factory in China and 52 control subjects from the general population were involved in an investigation on the potential health concerns of occupational exposure to PFASs by mass spectrometry-based metabolomics analysis. The PFAS levels in plasma from both groups were analyzed. Six PFAS congeners (∑6PFASs) were found to be the main components of the 13 detected PFASs, with a geometric mean of 1770 and 22.2 ng mL-1 in occupational workers and the general population, respectively. Metabolic profiles of the plasma samples were acquired using liquid chromatography coupled with orbitrap high-resolution mass spectrometry and gas chromatography-mass spectrometry. The partial least-squares-discriminant analysis model indicated that the plasma metabolic profiles of the two groups could be clearly separated. Differential and correlation analyses were applied to discover potential biomarkers. A total of 14 potential biomarkers were identified, and they were found to be associated with oxidative stress, fatty acid β-oxidation disorder, and kidney injury. The obtained results indicated that the health effects of occupational exposure to PFASs on workers should not be ignored.


Magnetic and electrical characterization of nickel-rich NiFe thin films synthesized by atomic layer deposition and subsequent thermal reduction.
Nickel-rich NiFe thin films (Ni92Fe8, Ni89Fe11 and Ni83Fe17) were prepared by combining atomic layer deposition (ALD) with a subsequent thermal reduction process. In order to obtain Ni x Fe1-x O y films, one ALD supercycle was performed according to the following sequence: m NiCp2/O3, with m = 1, 2 or 3, followed by one FeCp2/O3 cycle. The supercycle was repeated n times. The thermal reduction process in hydrogen atmosphere was investigated by in situ x-ray diffraction studies as a function of temperature. The metallic nickel iron alloy thin films were investigated and characterized with respect to crystallinity, morphology, resistivity, and magnetism. As proof-of-concept magnetic properties of an array of Ni83Fe17, close to the perfect Permalloy stoichiometry, nanotubes and an isolated tube were investigated.


The influence of active site conformations on the hydride transfer step of the thymidylate synthase reaction mechanism.
The hydride transfer from C6 of tetrahydrofolate to the reaction's exocyclic methylene-dUMP intermediate is the rate limiting step in thymidylate synthase (TSase) catalysis. This step has been studied by means of QM/MM molecular dynamics simulations to generate the corresponding free energy surfaces. The use of two different initial X-ray structures has allowed exploring different conformational spaces and the existence of chemical paths with not only different reactivities but also different reaction mechanisms. The results confirm that this chemical conversion takes place preferentially via a concerted mechanism where the hydride transfer is conjugated to thiol-elimination from the product. The findings also confirm the labile character of the substrate-enzyme covalent bond established between the C6 of the nucleotide substrate and a conserved cysteine residue. The calculations also reproduce and rationalize a normal H/T 2° kinetic isotope effect measured for that step. From a computational point of view, the results demonstrate that the use of an incomplete number of coordinates to describe the real reaction coordinate can render biased results.


Are corticosterone levels a good indicator of food availability and reproductive performance in a kittiwake colony?
We evaluated the use of corticosterone to gauge forage availability and predict reproductive performance in black-legged kittiwakes (Rissa tridactyla) breeding in Alaska during 1999 and 2000. We modeled the relationship between baseline levels of corticosterone and a suite of individual and temporal characteristics of the sampled birds. We also provided supplemental food to a sample of pairs and compared their corticosterone levels with that of pairs that were not fed. Corticosterone levels were a good predictor of forage availability in some situations, although inconsistencies between corticosterone levels and reproductive performance of fed and unfed kittiwakes suggested that this was not always the case. In general, higher corticosterone levels were found in birds that lacked breeding experience and in birds sampled shortly after arriving from their wintering grounds. All parameters investigated, however, explained only a small proportion of the variance in corticosterone levels. We also investigated whether corticosterone, supplemental feeding, year of the study, breeding experience, body weight, and sex of a bird were able to predict laying, hatching, and fledging success in kittiwakes. Here, breeding experience, year of the study, and body weight were the best predictors of a bird's performance. Corticosterone level and supplemental feeding were good predictors of kittiwake reproductive performance in some cases. For example, corticosterone levels of birds sampled during the arrival stage reliably predicted laying success, but were less reliable at predicting hatching and fledging success. Counts of active nests with eggs or chicks may be more reliable estimates of the actual productivity of the colony. Supplemental feeding had strong effects on kittiwake productivity when natural forage was poor, but had little effect when natural forage was plentiful.


Nanoscale coherent interface strengthening of Mg alloys.
Structural materials with higher strength and ductility could reduce weight and improve energy efficiency from the ecological and economical viewpoints. However, most classical strengthening strategies, such as refining grain size and forming secondary particles, can effectively hinder dislocation motion but remarkably decrease ductility. In this research, a nanoscale contraction twins-stacking faults (CTWSFs) hierarchical structure was achieved in a model sample of magnesium-lithium (Mg-Li) alloy using an industrial ultrahigh pressure technique. The specific yield strength and elongation of the treated Mg-Li alloy were ∼179 kN m kg-1 and ∼25% at room temperature, respectively. Both of these are the highest values reported so far, even compared to commercial aluminum alloys and steel. Depending on molecular dynamics simulations, it was demonstrated that the basal-plane stacking faults (SFs) attribute to the nucleation of contraction twins (CTWs), but prevented the broadening of the CTWs. The results were confirmed using ex situ transmission electron microscopy. The models also verified that the unique structure of nanoscale coherent boundaries-basal-plane SFs were effective in preventing dislocation motion in a single Mg crystal, which was analogous to twin boundaries. Finding this new CTWSFs structure might provide an alternative perspective for designing more innovative hexagonal close packed (hcp)-type structural materials with superior mechanical properties.


Nitric oxide delays the death of trophic factor-deprived PC12 cells and sympathetic neurons by a cGMP-mediated mechanism.
We have used cultured PC12 cells and rat sympathetic neurons as model systems to examine the regulation of neuronal cell death and survival. Because nitric oxide (NO) may be involved in nerve growth factor (NGF) signaling in PC12 cells, we tested NO-generating compounds for their ability to protect PC12 cells and sympathetic neurons from death after withdrawal of trophic support. Three such agents, S-nitroso-N-acetylpenicillamine (SNAP), diethylenetriamine NO adduct (DETA-NO), and sodium nitroprusside provide (SNP), were found to promote complete short-term survival after removal of serum from naive PC12 cells and of NGF from neuronally differentiated PC12 cells and sympathetic neurons. One major target of NO action is guanylate cyclase, which is activated by nitrosylation of its heme prosthetic group. We observed that inhibition of guanylate cyclase blocks the protective effects of the NO generators on trophic factor-deprived PC12 cells and sympathetic neurons without preventing NGF-induced survival. We also found that permeant cGMP analogs and an inhibitor of cGMP-specific phosphodiesterase enhance cell survival, suggesting that the protective effects of NO are mediated by activation of guanylate cyclase and increased intracellular cGMP. N-Nitro-L-arginine methyl ester, a NO synthase inhibitor, did not block NGF-promoted PC12 cell or sympathetic neuron survival. These findings indicate that like NGF, NO has survival-promoting actions on neurons but that the two agents work by initially independent mechanisms.