Spatial heterogeneity in the Mediterranean Biodiversity Hotspot affects barcoding accuracy of its freshwater fishes.
Incomplete knowledge of biodiversity remains a stumbling block for conservation planning and even occurs within globally important Biodiversity Hotspots (BH). Although technical advances have boosted the power of molecular biodiversity assessments, the link between DNA sequences and species and the analytics to discriminate entities remain crucial. Here, we present an analysis of the first DNA barcode library for the freshwater fish fauna of the Mediterranean BH (526 spp.), with virtually complete species coverage (498 spp., 98% extant species). In order to build an identification system supporting conservation, we compared species determination by taxonomists to multiple clustering analyses of DNA barcodes for 3165 specimens. The congruence of barcode clusters with morphological determination was strongly dependent on the method of cluster delineation, but was highest with the general mixed Yule-coalescent (GMYC) model-based approach (83% of all species recovered as GMYC entity). Overall, genetic morphological discontinuities suggest the existence of up to 64 previously unrecognized candidate species. We found reduced identification accuracy when using the entire DNA-barcode database, compared with analyses on databases for individual river catchments. This scale effect has important implications for barcoding assessments and suggests that fairly simple identification pipelines provide sufficient resolution in local applications. We calculated Evolutionarily Distinct and Globally Endangered scores in order to identify candidate species for conservation priority and argue that the evolutionary content of barcode data can be used to detect priority species for future IUCN assessments. We show that large-scale barcoding inventories of complex biotas are feasible and contribute directly to the evaluation of conservation priorities.


Radio Frequency Tunable Oscillator Device Based on a SmB_{6} Microcrystal.
Radio frequency tunable oscillators are vital electronic components for signal generation, characterization, and processing. They are often constructed with a resonant circuit and a "negative" resistor, such as a Gunn diode, involving complex structure and large footprints. Here we report that a piece of SmB_{6}, 100 μm in size, works as a current-controlled oscillator in the 30 MHz frequency range. SmB_{6} is a strongly correlated Kondo insulator that was recently found to have a robust surface state likely to be protected by the topology of its electronics structure. We exploit its nonlinear dynamics, and demonstrate large ac voltage outputs with frequencies from 20 Hz to 30 MHz by adjusting a small dc bias current. The behaviors of these oscillators agree well with a theoretical model describing the thermal and electronic dynamics of coupled surface and bulk states. With reduced crystal size we anticipate the device to work at higher frequencies, even in the THz regime. This type of oscillator might be realized in other materials with a metallic surface and a semiconducting bulk.


Posttraumatic stress disorder among women after the war in Sarajevo: a rationale for genetic study.
An exposure to extreme trauma events leads to posttraumatic stress disorder (PTSD) in up to 14-50% of war survivors. Recent findings suggest that genetic factors could play a certain role in PTSD development. In order to illustrate this possibility, we present results of a pilot study on gender specific sample of Sarajevo civilians immediately after the war cessation. During the period 1992-1995, Sarajevo civilians experienced continuous life threatening events with a great risk of developing PTSD in such conditions. Our study included 100 women adjusted to same socio-demographic characteristics. All women were interviewed using Harvard Trauma Questionnaire (HTQ) and divided into two groups (domestic and returnees) according to exposure length to extreme war life events of six or forty-three months. Above 50% of total analysed sample fulfilled criteria for PTSD. Regarding duration in trauma exposure no significant difference between these two groups were found. The only significant predictor found was physical abuse (p>0.01) that still cannot explain why some women develop PTSD while others not. Several years after the war, PTSD frequencies are decreased and disorder became chronic and more severe. However, the PTSD prevalence remains high when compared to general population rates. Therefore, Sarajevo population being exposed for almost four years to extreme war life events represents unique model for comparative research on PTSD etiology within the light of latest findings in molecular genetics of PTSD.


Multicenter Initial Guidance on Use of Antivirals for Children With Coronavirus Disease 2019/Severe Acute Respiratory Syndrome Coronavirus 2.
Although coronavirus disease 2019 (COVID-19) is mild in nearly all children, a small proportion of pediatric patients develop severe or critical illness. Guidance is therefore needed regarding use of agents with potential activity against severe acute respiratory syndrome coronavirus 2 in pediatrics. A panel of pediatric infectious diseases physicians and pharmacists from 18 geographically diverse North American institutions was convened. Through a series of teleconferences and web-based surveys, a set of guidance statements was developed and refined based on review of best available evidence and expert opinion. Given the typically mild course of pediatric COVID-19, supportive care alone is suggested for the overwhelming majority of cases. The panel suggests a decision-making framework for antiviral therapy that weighs risks and benefits based on disease severity as indicated by respiratory support needs, with consideration on a case-by-case basis of potential pediatric risk factors for disease progression. If an antiviral is used, the panel suggests remdesivir as the preferred agent. Hydroxychloroquine could be considered for patients who are not candidates for remdesivir or when remdesivir is not available. Antivirals should preferably be used as part of a clinical trial if available. Antiviral therapy for COVID-19 is not necessary for the great majority of pediatric patients. For those rare cases of severe or critical disease, this guidance offers an approach for decision-making regarding antivirals, informed by available data. As evidence continues to evolve rapidly, the need for updates to the guidance is anticipated.


Optical excitation energies, Stokes shift, and spin-splitting of C24H72Si14.
As an initial step toward the synthesis and characterization of sila-diamondoids, such as sila-adamantane (Si(10)H(16),T(d)), the synthesis of a fourfold silylated sila-adamantane molecule (C(24)H(72)Si(14),T(d)) has been reported in literature [Fischer et al., Science 310, 825 (2005)]. We present the electronic structure, ionization energies, quasiparticle gap, and the excitation energies for the Si(14)(CH(3))(24) and the exact silicon analog of adamantane Si(10)H(16) obtained at the all-electron level using the delta-self-consistent-field and transitional state methods within two different density functional models: (i) Perdew-Burke-Ernzerhof generalized gradient approximation and (ii) fully analytic density functional (ADFT) implementation with atom dependent potential. The ADFT is designed so that molecules separate into atoms having exact atomic energies. The calculations within the two models agree well, to within 0.25 eV for optical excitations. The effect of structural relaxation in the presence of electron-hole-pair excitations is examined to obtain its contribution to the luminescence Stokes shift. The spin-influence on exciton energies is also determined. Our calculations indicate overall decrease in the absorption, emission, quasiparticle, and highest occupied molecular orbital-lowest unoccupied molecular orbital gaps, ionization energies, Stokes shift, and exciton binding energy when passivating hydrogens in the Si(10)H(16) are replaced with electron donating groups such as methyl (Me) and trimehylsilyl (-Si(Me)(3)).


Methotrexate in pediatric osteosarcoma: response and toxicity in relation to genetic polymorphisms and dihydrofolate reductase and reduced folate carrier 1 expression.
To determine the influence of the genotype and the level of expression of different enzymes involved in folate metabolism on the response to and toxicity of high-dose methotrexate treatment in pediatric osteosarcomas. DHFR and Reduced folate carrier 1 (RFC1) semiquantitative expression was analyzed in 34 primary and metastatic osteosarcoma tissues by real-time polymerase chain reaction. The following polymorphisms were also analyzed in peripheral blood from 96 children with osteosarcoma and 110 control subjects: C677T, A1298C (MTHFR), G80A (RFC1), A2756G (MTR), C1420T (SHMT), the 28bp-repeat polymorphism, and 1494del6 of the TYMS gene. Treatment toxicity was scored after each cycle according to criteria from the World Health Organization. DHFR and RFC1 expression was lower in initial osteosarcoma biopsy specimens than in metastases (P = .024 and P = .041, respectively). RFC1 expression was moderately decreased in samples with poor histologic response to preoperative treatment (P = .053). Patients with osteosarcoma with G3/G4 hematologic toxicity were more frequently TT than CT/CC for C677T/MTHFR (P = .023) and GG for A2756G/MTR (P = .048 and P = .057 for gastrointestinal and hematologic toxicity, respectively). The role of C677T/MTHFR and A2756G/MTR on chemotherapy-induced toxicity should be further investigated in pediatric osteosarcomas receiving high-dose methotrexate. Altered expression of DHFR and RFC1 is a feasible mechanism by which osteosarcoma cells become resistant to methotrexate.


Bedbug infestations in the news: a picture of an emerging public health problem in the United States.
Bedbug (Cimex lectularis) infestations have become a major complaint in all but three states in the United States. Increasing infestations have also been reported in Asia, Australia, Europe, and Canada. Newspaper articles often herald the onset of a new health issue for the public, and they can be used for epidemiological tracking of increasing populations of irritating and potentially serious vectoring pests. This news coverage model is useful in validating the typical spread of a new contagious entity, or in predicting the waves of public reaction to the spread of a new pest or health problem. The authors' analysis covers the years 2001 to 2006, showing the incidence of bedbug news coverage from the East Coast, the Midwest, the South, and the West in the United States. Rural and urban examples are included in a four-year time frame starting with news in the Northeast and ending with an attempt to eradicate the pest from the state of Hawaii.


Corneal collagen cross-linking (CXL) for the treatment of melting keratitis in cats and dogs: a pilot study.
UV-A/riboflavin cross-linking (CXL) of corneal collagen fibers is an established, highly promising therapy for corneal melting in physician-based ophthalmology. A prospective pilot study was conducted to demonstrate proof of principle of this novel method for the treatment of melting corneal ulcers in dogs and cats. After obtaining owner consent, CXL was performed in three cats and three dogs with corneal melting, which either affected the entire corneal surface or was resistant to conventional antibiotic and anticollagenolytic therapy, and affected parts or all of the corneal surface. Medical therapy was continued in all patients. The available follow-up ranged from 2 to 22.5 months and involved slit-lamp examination, fluorescein staining, and photographic documentation during all rechecks. Surgical stabilization of the cornea was not necessary in any case, because progression of corneal melting was arrested in all cases within 1-20 days of CXL treatment. Corneal re-epithelization occurred within 7-40 days in all eyes. At 40 days after CXL, all eyes presented a quiescent corneal state without signs of active inflammation and with beginning scar formation. The complications observed in three of the six animals included a corneal sequestrum, superficial corneal stromal pigmentation, and bullous keratopathy. This study shows the feasibility of CXL to treat progressive corneal melting in veterinary patients. CXL may represent a cost-efficient and safe alternative therapy in the treatment for corneal melting in veterinary ophthalmology. More investigations comparing the effectivity and complication rate of CXL to those of standard medical treatment are necessary.


Graph Curvature for Differentiating Cancer Networks.
Cellular interactions can be modeled as complex dynamical systems represented by weighted graphs. The functionality of such networks, including measures of robustness, reliability, performance, and efficiency, are intrinsically tied to the topology and geometry of the underlying graph. Utilizing recently proposed geometric notions of curvature on weighted graphs, we investigate the features of gene co-expression networks derived from large-scale genomic studies of cancer. We find that the curvature of these networks reliably distinguishes between cancer and normal samples, with cancer networks exhibiting higher curvature than their normal counterparts. We establish a quantitative relationship between our findings and prior investigations of network entropy. Furthermore, we demonstrate how our approach yields additional, non-trivial pair-wise (i.e. gene-gene) interactions which may be disrupted in cancer samples. The mathematical formulation of our approach yields an exact solution to calculating pair-wise changes in curvature which was computationally infeasible using prior methods. As such, our findings lay the foundation for an analytical approach to studying complex biological networks.


Recombinant influenza A viruses as vaccine vectors.
Various viruses, including poxviruses, adenoviruses and vesicular stomatitis virus, have been considered as vaccine vectors for the delivery of antigens of interest in the development of vaccines against newly emerging pathogens. Here, we review results that have been obtained with influenza A viruses (IAV) as vaccine vectors. With the advent of reverse genetics technology, IAV-based recombinant vaccine candidates have been constructed that induce protective immunity to a variety of different pathogens of interest, including West Nile virus, Plasmodium falciparum and respiratory syncytial virus. The various cloning strategies to produce effective and attenuated, safe to use IAV-based viral vectors are discussed. It was concluded that IAV-based vector system has several advantages and holds promise for further development.


[Analysis of single nucleotide polymorphisms (SNPs)].
Single nucleotide polymorphisms (SNPs) are DNA sequence variations occurring when a single nucleotide in the genome differs in paired chromosomes. Some SNPs in the coding region change the amino acid sequence of a protein, and others in the coding region do not affect the protein sequence. SNPs outside the coding region may also affect transcription factor binding, gene splicing, or mRNA degradation. With or without such impacts on the biological function of gene products, SNPs are strongly useful as markers to examine linkage disequilibrium and to explore genetic polymorphisms in the research of population genetics and medical science. Knowledge about SNPs is expected to help in identifying susceptibility genes for complex diseases, drug susceptibility genes, and histocompatibility genes for transfusion or transplantation. Recent breakthroughs in DNA technology, such as high-throughput universal SNP microarrays and next-generation sequencers, have made it possible to carry out comprehensive analyses of DNA at the whole genome level, resulting in the implementation of a genome-wide association study(GWAS). In GWAS, SNPs are used as high-solution markers in gene mapping related to diseases versus normal traits. The outcomes of GWAS provide a number of supportive findings for the promotion of personalized medicine based on genome informatics. This article presents a review of SNP analysis with respect to its scientific significance and current progress.


A metabolic regulon reveals early and late acting enzymes in neuroactive Lycopodium alkaloid biosynthesis.
Plants synthesize many diverse small molecules that affect function of the mammalian central nervous system, making them crucial sources of therapeutics for neurological disorders. A notable portion of neuroactive phytochemicals are lysine-derived alkaloids, but the mechanisms by which plants produce these compounds have remained largely unexplored. To better understand how plants synthesize these metabolites, we focused on biosynthesis of the Lycopodium alkaloids that are produced by club mosses, a clade of plants used traditionally as herbal medicines. Hundreds of Lycopodium alkaloids have been described, including huperzine A (HupA), an acetylcholine esterase inhibitor that has generated interest as a treatment for the symptoms of Alzheimer's disease. Through combined metabolomic profiling and transcriptomics, we have identified a developmentally controlled set of biosynthetic genes, or potential regulon, for the Lycopodium alkaloids. The discovery of this putative regulon facilitated the biosynthetic reconstitution and functional characterization of six enzymes that act in the initiation and conclusion of HupA biosynthesis. This includes a type III polyketide synthase that catalyzes a crucial imine-polyketide condensation, as well as three Fe(II)/2-oxoglutarate-dependent dioxygenase (2OGD) enzymes that catalyze transformations (pyridone ring-forming desaturation, piperidine ring cleavage, and redox-neutral isomerization) within downstream HupA biosynthesis. Our results expand the diversity of known chemical transformations catalyzed by 2OGDs and provide mechanistic insight into the function of noncanonical type III PKS enzymes that generate plant alkaloid scaffolds. These data offer insight into the chemical logic of Lys-derived alkaloid biosynthesis and demonstrate the tightly coordinated coexpression of secondary metabolic genes for the biosynthesis of medicinal alkaloids.


Hesperidin exerts antidepressant-like effects in acute and chronic treatments in mice: possible role of l-arginine-NO-cGMP pathway and BDNF levels.
Hesperidin (4'-methoxy-7-O-rutinosyl-3',5-dihydroxyflavanone), a naturally occurring flavanone glycoside, was previously shown to produce an antidepressant-like effect with modultation of the serotonergic 5-HT1A and kappa-opioid receptors. In this study, the signaling mechanisms underlying their antidepressant-like effects were further evaluated by investigating in acute and chronic treatments. Results showed that chronic treatment of hesperidin or hesperitin (0.1, 0.3 and 1mg/kg, intraperitoneal, i.p.) have an antidepressant-like effect in the mouse tail suspension test (TST) without modified the locomotor activity in the open field test. Pretreatment with l-arginine (a nitric oxide (NO) precursor), sildenafil (a phosphodiesterase 5 inhibitor) or S-nitroso-N-acetyl-penicillamine (a NO donor) significantly reversed the reduction in immobility time elicited by acute treatment with hesperidin (0.3mg/kg) in the TST. Hesperidin (0.01mg/kg, a sub-effective dose in acute treatment) produced an additive antidepressant-like effect with N(G)-nitro-l-arginine (an inhibitor of nitric oxide synthase (NOS)) or 7-nitroindazole (a neuronal NOS inhibitor) in the TST. Pretreatment of animals with methylene blue (an inhibitor of NOS/soluble guanylate cyclase (sGC)) or ODQ (a specific inhibitor sGS) caused an additive effect with hesperidin in the TST. Hesperidin in the acute (1mg/kg) and chronic (0.1, 0.3 and 1mg/kg) treatments caused a significant decrease in nitrate/nitrite (NOX) levels in the hippocampus of mice. Chronic treatment with hesperidin (0.3 and 1mg/kg) also resulted in an increase in hippocampal brain-derived neurotrophic factor (BDNF) levels. These results demonstrated that the antidepressant-like effect of hesperidin is likely mediated by inhibition of l-arginine-NO-cGMP pathway and by increased of the BDNF levels in hippocampus.


Are women ready to do HIV test?
To identify the factors that influence the stage of change with regards to HIV testing in women (16- 45-years-old) in Westmoreland using the trans-theoretical model (TTM) of behaviour change. A structured interview-assisted questionnaire was administered to 372 pregnant and non-pregnant respondents in urban and rural areas of Westmoreland after random selection of four public health facilities. The trans-theoretical model which suggests that behaviour change process moves through five stages from pre-contemplation to maintenance was used to evaluate readiness for HIV testing. Most pregnant women who tested previously were at the preparation stage (78.5%) while non-pregnant women who tested previously were at contemplation (68.5%). The significant predictors of being in the action or maintenance stage among pregnant women was being 20- 24-years-old, experiencing a first pregnancy and being exposed to counselling. For women who had never tested, preparation was significantly associated with being in an unstable union (non-pregnant). No significant association was found for non-pregnant, previously tested females or for pregnant women who had never tested. The majority of women lacked self-efficacy as they were unable to maintain the behaviour and did not recognize its importance in the absence of pregnancy. Interventions are needed to target non-pregnant women, especially teenagers, women over 25-years old and women in unions. Integration of testing services into all aspects of primary healthcare, established testing protocols and simultaneous marketing to selected at-risk groups will increase the uptake of HIV testing opportunities and contribute to the control of this epidemic.


Predictors of health service use by family caregivers of persons with multimorbidity.
Informal caregivers of patients with multiple chronic conditions are socially good, promoting the sustainability of a large part of home care provision. However, this very demanding activity causes health problems that increase their own need for health services. This study analyses the use of health services by informal carers, comparing it with the use made by the general population with similar characteristics. Cross-sectional analytical study carried out in the Malaga-Valle Guadalhorce Primary Health Care District (Spain). Healthcare demand and perceived health were measured in the family caregivers, compared to the general population. Strobe Statement for observational studies has been used to strength the report of the results. Final sample consisted of 314 family caregivers together with a subsample of 2.290 non-caregivers taken from data of the National Health Survey. This subsample was paired by gender with our sample. Formal caregivers make fewer annual visits to the health services, with respect to the general population, regardless of the perceived level of health. The difference of the means between those who perceive their health as very poor was 0.11 (95% CI: 0.01 to 0.20) consultations with the family doctor, 0.21 (95% CI: 0.15 to 0.26) consultations with medical specialists and 1.70 (95% CI: 1.52 to 1.87) emergency room attention. Three independent factors were identified that predispose to the increased use of health services: background of greater education achievement (OR 8.13, 95% CI: 1.30 to 50.68), non-cohabitation with the care recipient (OR 3.57, 95% CI: 1.16 to 11.11) and a more positive physical quality of life component (OR 1.06; 95% CI: 1.03 to 1.09). Intrinsic components of the caregiver reveal their independent relationship with the provision of informal care and the use of health services. A broader vision is needed for the factors that influence the health of these caregivers to develop multipurpose interventions and improve the consistency and effectiveness of the health services offered to the caregiver.


Linking neural and clinical measures of glaucoma with diffusion magnetic resonance imaging (dMRI).
To link optic nerve (ON) structural properties to clinical markers of glaucoma using advanced, semi-automated diffusion magnetic resonance imaging (dMRI) tractography in human glaucoma patients. We characterized optic neuropathy in patients with unilateral advanced-stage glaucoma (n = 6) using probabilistic dMRI tractography and compared their results to those in healthy controls (n = 6). We successfully identified the ONs of glaucoma patients based on dMRI in all patients and confirmed that dMRI measures of the ONs correlated with clinical markers of glaucoma severity. Specifically, we found reduced fractional anisotropy (FA) in the ONs of eyes with advanced, as compared to mild, glaucoma (F(1,10) = 55.474, p < 0.0001, FDR < 0.0005). Furthermore, by comparing the ratios of ON FA in glaucoma patients to those of healthy controls (n = 6), we determined that this difference was beyond that expected from normal anatomical variation (F(1,9) = 20.276, p < 0. 005). Finally, we linked the dMRI measures of ON FA to standard clinical glaucoma measures. ON vertical cup-to-disc ratio (vCD) predicted ON FA (F(1,10) = 11.061, p < 0.01, R2 = 0.66), retinal nerve fiber layer thickness (RNFL) predicted ON FA (F(1,10) = 11.477, p < 0.01, R2 = 0.63) and ON FA predicted perceptual deficits (visual field index [VFI]) (F(1,10) = 15.308, p < 0.005, R2 = 0.52). We describe semi-automated methods to detect glaucoma-related structural changes using dMRI and confirm that they correlate with clinical measures of glaucoma.


High-fat Diet Increases Phospholipid Peroxidation in the Liver of Mature Fischer 344 Rats.
Phospholipid peroxidation is considered to be involved in the pathophysiology of various diseases. While dietary antioxidants are believed to help prevent these diseases via inhibition of phospholipid peroxidation, further evaluation is needed to prove this hypothesis. For this, it is crucial to establish an animal model with accelerated phospholipid peroxidation. In this study, we hypothesized that a combination of aging and high-fat diet feeding may accelerate phospholipid peroxidation in vivo. High-fat diets were fed to mature and juvenile Fischer 344 rats for 12 weeks. The mature rats in particular accumulated body fat and liver phosphatidylcholine hydroperoxide (PCOOH). Interestingly, the increase in PCOOH levels was abrogated by the co-administration of antioxidants to mature rats. This may be attributed to factors including the decrease in body fat, functions of vitamin E, and/or the involvement of antioxidant-related genes, each caused by antioxidant administration. These results indicate that the high-fat diet-fed aging animal model may be suitable for investigation of the relationship between phospholipid peroxidation, oxidative stress-related diseases, and dietary antioxidants.


Vitamin C Deficiency, High-Sensitivity C-Reactive Protein, and Cardiac Event-Free Survival in Patients With Heart Failure.
Vitamin C is related to lower levels of high-sensitivity C-reactive protein (hsCRP), an inflammatory biomarker that predicts cardiovascular disease. Whether vitamin C deficiency is associated with hsCRP and cardiac events in heart failure (HF) patients has not been examined. The aim of this study is to determine the relationships among vitamin C intake, serum levels of hsCRP, and cardiac events. A total of 200 HF patients completed a 3-day food diary to determine vitamin C deficiency and provided blood to measure serum levels of hsCRP. Patients were followed for 2 years to obtain data on cardiac event-free survival. Moderation analyses with hierarchical logistic and Cox regressions were used for the data analysis. Seventy-eight patients (39%) had vitamin C deficiency and 100 (50%) had an hsCRP level higher than 3 mg/L. Vitamin C deficiency was associated with an hsCRP level higher than 3 mg/L in the hierarchical logistic regression (odds ratio, 2.40; 95% confidence interval, [1.13-5.10]; P = .023). Vitamin C deficiency (hazard ratio, 1.68; 95% CI, 1.05-2.69, P = .029) and hsCRP level higher than 3 mg/L (hazard ratio, 1.79; 95% CI, 1.07-3.01; P = .027) predicted shorter cardiac event-free survival in hierarchical Cox regression. The interaction of hsCRP level higher than 3 mg/L and vitamin C deficiency produced a 2.3-fold higher risk for cardiac events (P = .002) in moderation analysis. Higher level of hsCRP predicted shorter cardiac event-free survival only in patients with vitamin C deficiency (P = .027), but not in those with vitamin C adequacy. Vitamin C deficiency moderated the relationship between inflammation and cardiac events in patients with HF. Future study is required to determine whether adequate intake of vitamin C could play a protective role against the impact of inflammation on cardiac events in HF patients.


The relationship between preoperative general mental health and postoperative quality of life in minimally invasive lumbar spine surgery.
In assessing poor lumbar surgery outcomes, researchers continue to investigate psychosocial predictors of patient postoperative quality of life. This is the first study of its kind to investigate this relationship in an exclusively minimally invasive patient sample. To determine the association between preoperative mental health and postoperative patient-centered outcomes in patients undergoing minimally invasive lumbar surgery. In 83 adults undergoing single-level minimally invasive lumbar spine surgery, Pearson correlation and partial correlation analyses were conducted between all demographic and clinical baseline variables and Oswestry Disability Index (ODI), Visual Analogue Scale (VAS), and 36-item Short-Form Health Survey Version 2.0 (SF-36v2) scores at 6 to 12 months postoperatively. SF-36v2 mental component summary scores (MCS) were used to assess pre- and postoperative general mental health. Post hoc analysis consisted of Pearson correlations between baseline SF-36v2, ODI, and VAS scores, and an identical set of correlations at outcomes. Preoperative MCS showed no significant association with outcomes VAS, ODI, or physical component summary scores. Baseline disability correlated significantly and more strongly with baseline MCS (P < .001, r = -0.40) than baseline pain levels (VAS back not significant, VAS leg P = .015, r = 0.27). Outcomes disability correlated significantly and more strongly with outcome back and leg pain levels (P < .001, r = 0.60 and 0.66) than outcome MCS (P = .031, r = -0.24). In a patient sample with mental health scores comparable to the population mean, there is no relationship between preoperative general mental health and postoperative patient-centered outcomes. Surgeons should consider the dynamic relationships between patient disability, mental health, and pain levels in assessing quality of life at different time points.


Microcapsules through polymer complexation. Part 3: Encapsulation and culture of human Burkitt lymphoma cells in vitro.
Methacrylic acid (MAA) based polyelectrolytes were complexed with protonated or quaternized dimethylaminoethyl methacrylate (DMAEMA) containing polyelectrolytes to form microcapsules in vitro. Anchorage independent human Burkitt lymphoma (Raji) cells were successfully cultured in the presence of dissolved MAA containing polymer. Capsule morphology was investigated by light microscopy and by scanning electron microscopy (SEM). Capsules based on quaternized DMAEMA containing polymer were found to be more stable than capsules containing protonated DMAEMA functionality. Raji cells were successfully encapsulated in both systems and divided to confluence; thereafter sufficient pressure was exerted to burst open the capsules. Cells released from these capsules appeared to suffer no discernible trauma and were successfully isolated and subcultured to confluence.


Evaluation of nicotine in tobacco-free-nicotine commercial products.
Recently, a variety of new tobacco-free-nicotine, TFN, products have been commercialized as e-liquids. Tobacco-derived nicotine contains predominantly (S)-(-)-nicotine, whereas TFN products may not. The TFN products are said to be cleaner, purer substances, devoid of toxic components that come from the tobacco extraction process. A variety of commercial tobacco and TFN products were analyzed to identify the presence and composition of each nicotine enantiomer. A rapid and effective enantiomeric separation of nicotine has been developed using a modified macrocyclic glycopeptide bonded to superficially porous particles. The enantiomeric assay can be completed in <2 min with high resolution and accuracy using high performance liquid chromatography with electrospray ionization mass spectrometry. The results of this study suggest the need for pharmacological studies of (R)-(+)-nicotine, which is present in much greater quantities in commercial TFN products compared to commercial tobacco-derived products. Such studies are required by the FDA for new enantiomeric pharmacological products. Copyright © 2016 John Wiley & Sons, Ltd.


The Americleft Project: Comparison of Ratings Using Two-Dimensional Versus Three-Dimensional Images for Evaluation of Nasolabial Appearance in Patients With Unilateral Cleft Lip and Palate.
This study was conducted to determine if nasolabial appearance is rated with comparable results and reliability on 3-dimensional stereophotogrammetric facial images versus standard clinical photographs (2-dimensional). Twenty-seven consecutively treated patients with repaired complete unilateral cleft lip and palate were selected. Six trained and calibrated raters assessed cropped 2- and 3-dimensional facial images. Nasolabial profile, nasolabial frontal, and vermillion border esthetics were rated with the 5-point scale described by Asher-McDade using the modified Q-sort method. Cropped 3-dimensional images were available for viewing by each rater, allowing for complete rotational control for viewing the images from all aspects. Two- and three-dimensional ratings were done separately and repeated the next day.Interrater reliability scores were good for 2-dimensional (κ = 0.607-0.710) and fair to good for 3-dimensional imaging (κ = 0.374-0.769). Intrarater reliability was good to very good for 2-dimensional (κ = 0.749-0.836) and moderate to good for 3-dimensional imaging (κ = 0.554-0.855). Bland-Altman analysis showed satisfactory agreement of 2- and 3-dimensional scores for nasolabial profile and nasolabial frontal, but more systematic error occurred in the assessment of vermillion border.Although 3-dimensional images may be perceived as more representative of a direct clinical facial evaluation, their use for subjective rating of nasolabial aesthetics was not more reliable than 2-dimensional images in this study. Conventional 2-dimensional images provide acceptable reliability while being readily accessible for most cleft palate centers.


Sialic acid catabolism drives intestinal inflammation and microbial dysbiosis in mice.
Rapid shifts in microbial composition frequently occur during intestinal inflammation, but the mechanisms underlying such changes remain elusive. Here we demonstrate that an increased caecal sialidase activity is critical in conferring a growth advantage for some bacteria including Escherichia coli (E. coli) during intestinal inflammation in mice. This sialidase activity originates among others from Bacteroides vulgatus, whose intestinal levels expand after dextran sulphate sodium administration. Increased sialidase activity mediates the release of sialic acid from intestinal tissue, which promotes the outgrowth of E. coli during inflammation. The outburst of E. coli likely exacerbates the inflammatory response by stimulating the production of pro-inflammatory cytokines by intestinal dendritic cells. Oral administration of a sialidase inhibitor and low levels of intestinal α2,3-linked sialic acid decrease E. coli outgrowth and the severity of colitis in mice. Regulation of sialic acid catabolism opens new perspectives for the treatment of intestinal inflammation as manifested by E. coli dysbiosis.


Emerging viral diseases of tomato crops.
Viral diseases are an important limiting factor in many crop production systems. Because antiviral products are not available, control strategies rely on genetic resistance or hygienic measures to prevent viral diseases, or on eradication of diseased crops to control such diseases. Increasing international travel and trade of plant materials enhances the risk of introducing new viruses and their vectors into production systems. In addition, changing climate conditions can contribute to a successful spread of newly introduced viruses or their vectors and establishment of these organisms in areas that were previously unfavorable. Tomato is economically the most important vegetable crop worldwide and many viruses infecting tomato have been described, while new viral diseases keep emerging. Pepino mosaic virus is a rapidly emerging virus which has established itself as one of the most important viral diseases in tomato production worldwide over recent years. Begomovirus species and other whitefly-transmitted viruses are invading into new areas, and several recently described new viruses such as Tomato torrado virus and new Tospovirus species are rapidly spreading over large geographic areas. In this article, emerging viruses of tomato crops are discussed.


Holographic spatial walk-off polarizer and its application to a 4-port polarization-independent optical circulator.
Based on the coupled-wave theory, a holographic spatial walk-off polarizer (HSWP) is designed. This HSWP is a transmission-type phase volume holographic grating on a substrate and its optical recording geometry can be derived from Chen's corrected methodology with a desired reconstruction condition. A pair of fabricated HSWPs with the splitting angle of 60 degrees is applied to assemble a new type of 4-port polarizationindependent optical circulator. The operating principles and the characteristics of the proposed HSWP and the prototype optical circulator are discussed.


Acute Ultraviolet Light Exposure and Post-Resistance Exercise Serum Testosterone: A Pilot Study in Older Men.
Testosterone deficiency is linked to a multitude of conditions including depression, sexual dysfunction, and cognitive impairment. Although synthetic testosterone-boosting pharmaceuticals have gained wide use, recent data suggests that vitamin D, ingested orally, may positively impact serum testosterone. Furthermore, evidence tying ultraviolet (UV) light exposure to endogenous vitamin D synthesis presents an intriguing possibility for naturally enhancing serum testosterone. This investigation sought to determine the effect of acute UV light exposure on resistance exercise-induced acute testosteronemia and vitamin D in older men. Six older adult men of varying activity levels (age 62 ± 1.79 yrs., height 179.92 ± 1.12 cm., body mass 83.79 ± 3.12 kg., BMI 25.95 ± 1.15 kg/m2) participated in two testing sessions separated by one week: 1) Resistance exercise followed by standard passive recovery (RERC) and 2) RE plus UV light exposure during the first 10-minutes of RE passive recovery (RERC-UV). The RE protocol was identical in both sessions and consisted of four sets of 10RM on leg press, chest press, and back row with 1-minute of rest between sets followed by 30-minutes of post-RE passive recovery. Serum testosterone and vitamin D were measured preand post-RE in 5-minute increments during the 30-minute recovery. Analysis of variance revealed neither RE or RERC-UV significantly affected serum testosterone or vitamin D. These findings suggest that acute UV light exposure may be insufficient to positively impact serum testosterone and vitamin D following a single bout of RE in older adult men.


Host range evolution is not driven by the optimization of larval performance: the case of Lycaeides melissa (Lepidoptera: Lycaenidae) and the colonization of alfalfa.
Herbivorous insects that have recently incorporated novel hosts into their diet provide unique opportunities for understanding factors that promote or constrain the evolution of niche breadth. Lycaeides melissa has colonized both cultivated and feral alfalfa (Medicago sativa) throughout much of North America within the past 200 years. We investigated the quality of the novel host as a resource for juvenile development, and asked if the novel host is a preferred host for oviposition relative to a native host (Astragalus canadensis). Larval-performance and oviposition-preference were examined using L. melissa individuals from a population associated with both M. sativa and A. canadensis, and oviposition-preference was also examined in another population associated exclusively with M. sativa. In addition, we investigated the effects of M. sativa and A. canadensis flowers on both preference and performance. Only one of the hosts, M. sativa, has flowers that are accessible to nectaring butterflies, and we hypothesized that the presence of flowers could affect female behavior. We find that the novel host is a relatively poor larval resource: adults that were reared as larvae on M. sativa were roughly one-third the size of adults that were reared on the native host, A. canadensis. The native host, Astragalus canadensis, is the preferred host in choice experiments involving only foliage. However, when flowers were included in preference assays, the native and novel hosts received similar numbers of eggs. Thus, the presence of flowers on hosts in the field might influence the utilization of a novel and inferior larval resource. These results are consistent with a model in which host shifts are driven by adult behavior that does not directly optimize larval performance.


Activity-dormancy transition in the cambial meristem involves stage-specific modulation of auxin response in hybrid aspen.
The molecular basis of short-day-induced growth cessation and dormancy in the meristems of perennial plants (e.g., forest trees growing in temperate and high-latitude regions) is poorly understood. Using global transcript profiling, we show distinct stage-specific alterations in auxin responsiveness of the transcriptome in the stem tissues during short-day-induced growth cessation and both the transition to and establishment of dormancy in the cambial meristem of hybrid aspen trees. This stage-specific modulation of auxin signaling appears to be controlled via distinct mechanisms. Whereas the induction of growth cessation in the cambium could involve induction of repressor auxin response factors (ARFs) and down-regulation of activator ARFs, dormancy is associated with perturbation of the activity of the SKP-Cullin-F-box(TIR) (SCF(TIR)) complex, leading to potential stabilization of repressor auxin (AUX)/indole-3-acetic acid (IAA) proteins. Although the role of hormones, such as abscisic acid (ABA) and gibberellic acid (GA), in growth cessation and dormancy is well established, our data now implicate auxin in this process. Importantly, in contrast to most developmental processes in which regulation by auxin involves changes in cellular auxin contents, day-length-regulated induction of cambial growth cessation and dormancy involves changes in auxin responses rather than auxin content.


Aging enhances cognitive biases to friends but not the self.
We measured changes in self and friend biases in perceptual matching in young and older participants. Participants learned associations between neutral geometric shapes and three personal labels (You, Friend, or Stranger), representing themselves, their named best friend, and a stranger not corresponding to anyone they knew. They then responded whether the shapes and labels matched or mismatched. In addition, participants reported the perceived personal distance between themselves, their best friend, and a stranger. Relative to young participants, older adults showed an increased bias toward matching their friends over strangers, whereas the bias toward the self over friends tended to decrease. Equivalent results occurred for a perceived personal distance measure, and, on measures of perceptual sensitivity with older participants, the personal distance between friends and strangers correlated with the friend bias in matching. These results indicate that the social bias toward a familiar best friend increases with age and modulates perceptual matching.


Efficacy of Nonsteroidal Anti-inflammatory Drug Prophylaxis for Heterotrophic Ossification in Hip Arthroscopy: A Systematic Review.
The purpose of this systematic review was to investigate the efficacy of nonsteroidal anti-inflammatory drug (NSAID) prophylaxis for preventing heterotopic ossification (HO) in the setting of hip arthroscopy. A systematic search was performed in duplicate for studies comparing the use of NSAID prophylaxis for HO in the setting of hip arthroscopy until March 2015. Study parameters--including sample size, incidence of HO, adverse effects, and level of symptoms--were obtained. Furthermore, the level of evidence of studies was collected and quality assessment was performed. The difference in incidence as well as pooled odds ratios were calculated and analyzed to compare no prophylaxis versus NSAID prophylaxis. This systematic review identified 5 studies, consisting of 1,662 patients, investigating NSAID prophylaxis in hip arthroscopy. HO was diagnosed with the use of postoperative hip radiographs at follow-up, with 95% of cases classified using the Brooker classification. The incidence of HO was 13.4% without NSAID prophylaxis and 3.3% with NSAID prophylaxis. Pooled odds ratios from the prospective studies were 0.07 (95% confidence interval [CI], 0.02 to 0.28; P = .0002; I(2) = 0%), showing with statistical significance that NSAID prophylaxis decreased the incidence of HO. The retrospective data similarly showed pooled odds ratios of 0.03 (95% CI, 0.00 to 1.43); P = .08; I(2) = 84%), although it was not statistically significant. Most of the patients who experienced HO in both groups were not reported to be symptomatic. Adverse effects and compliance were not consistently reported. The available orthopaedic literature suggests that the incidence of postoperative HO may be decreased with the use of NSAID prophylaxis in hip arthroscopy. However, the evidence is unclear regarding NSAID drug regimen choice, drug compliance, and adverse effects. Level III, systematic review of Level I, Level II, and Level III studies.


Pre- and postjunctional actions of neuropeptide Y and related peptides.
The effects of neuropeptide Y (NPY) and related peptide fragments on blood pressure and vagal action at the heart were compared in the anaesthetized rat. A change in vagal action was taken as a measure of presynaptic activity and a change in blood pressure was taken as a measure of postsynaptic activity. NPY, NPY-(13-36), PYY-(13-36), des-Ser22-NPY-(13-36) and a stabilized 13-36 analogue of NPY (ANA NPY) all exerted pressor actions and attenuated vagal action at the heart. The maximum vagal inhibitory or presynaptic action in order of potency was NPY, ANA-NPY, PYY-(13-36) significantly greater than NPY-(13-36), des-Ser22-NPY-(13-36). The order of potency for the half time of this effect was NPY, ANA-NPY significantly longer than PYY-(13-36) and NPY-(13-36), which were significantly longer than des Ser22-NPY-(13-36). For the pressor or postsynaptic effects, NPY increased blood pressure significantly more and for a longer duration than all the 13-36 fragments, which were not demonstrably different in this respect. These results are consistent with the proposal that there are two populations of NPY receptors. The C-terminal flanking peptide of NPY (CPON) and desamido-NPY had no effect on either vagal action at the heart or on blood pressure.


Imminent fracture risk.
The clinical significance of osteoporosis is in the occurrence of fractures and re-fractures. The main risk factor of sustaining a fracture is a previous one, but a recent fracture is a better fracture risk factor than fracture history. The role of the recency of fracture has been shown for both vertebral and non-vertebral fracture risk. This imminent risk is explained by both bone-related factors (underlying osteoporosis) and fall-related factors (including those related to postfracture care). Such a short-term increased risk has been shown also in patients initiating corticosteroids and in frail osteoporotic subjects with central nervous system (CNS) diseases or drugs targeting CNS, and thus a high risk of falls. Patients with an imminent (i.e. 2 years) risk of fracture or refracture should be identified in priority in order to receive an immediate treatment and a program of fall prevention.


Adherence to and invasion of human intestinal epithelial cells by Campylobacter jejuni and Campylobacter coli isolates from retail meat products.
The abilities of 34 Campylobacter jejuni and 9 Campylobacter coli isolates recovered from retail meats to adhere to and invade human intestinal epithelial T84 cells were examined and compared with those of a well-characterized human clinical strain, C. jejuni 81-176, to better assess the pathogenic potential of these meat isolates. The meat isolates exhibited a wide range of adherence and invasion abilities; a few of the isolates adhered to and invaded T84 cells almost as well as did C. jejuni 81-176. There was a significant correlation between the adherence ability and the invasion ability of the Campylobacter isolates. The presence of eight putative virulence genes in these Campylobacter isolates that are potentially responsible for adherence and invasion or that encode cytolethal distending toxin was determined using PCR. All Campylobacter isolates possessed flaA, cadF, pldA, cdtA, cdtB, and cdtC, and most (91%) also contained the ciaB gene. However, the virB11 gene, carried by virulence plasmid pVir, was absent in almost all the Campylobacter isolates. Our findings indicated that C. jejuni and C. coli present in retail meat were diverse in their ability to adhere to and invade human intestinal epithelial cells and that the putative virulence genes were widespread among the Campylobacter isolates. Thus, despite of the presence of the putative virulence genes, only some but not all Campylobacter strains isolated from retail meat can effectively invade human intestinal epithelial cells in vitro.


Comparison of human cardiac gene expression profiles in paired samples of right atrium and left ventricle collected in vivo.
Studies of expressed genes in human heart provide insight into both physiological and pathophysiological mechanisms. This is of importance for extended understanding of cardiac function as well as development of new therapeutic drugs. Heart tissue for gene expression studies is generally hard to obtain, particularly from the ventricles. Since different parts of the heart have different functions, expression profiles should likely differ between these parts. The aim of the study was therefore to compare the global gene expression in cardiac tissue from the more accessible auricula of the right atrium to expression in tissue from the left ventricle. Tissue samples were collected from five men undergoing aortic valve replacement or coronary artery bypass grafting. Global gene expression analysis identified 542 genes as differentially expressed between the samples extracted from these two locations, corresponding to ~2% of the genes covered by the microarray; 416 genes were identified as abundantly expressed in right atrium, and 126 genes were abundantly expressed in left ventricle. Further analysis of the differentially expressed genes according to available annotations, information from curated pathways and known protein interactions, showed that genes with higher expression in the ventricle were mainly associated with contractile work of the heart. Transcription in biopsies from the auricula of the right atrium on the other hand indicated a wider area of functions, including immunity and defense. In conclusion, our results suggest that biopsies from the auricula of the right atrium may be suitable for various genetic studies, but not studies directly related to muscle work.


Homing of human B cells to lymphoid organs and B-cell lymphoma engraftment are controlled by cell adhesion molecule JAM-C.
Junctional adhesion molecule C (JAM-C) is expressed by vascular endothelium and human but not mouse B lymphocytes. The level of JAM-C expression defines B-cell differentiation stages and allows the classification of marginal zone-derived (JAM-C-positive) and germinal center-derived (JAM-C-negative) B-cell lymphomas. In the present study, we investigated the role of JAM-C in homing of human B cells, using a xenogeneic nonobese diabetic/severe combined immunodeficient mouse model. Treatment with anti-JAM-C antibodies in short-term experiments reduced migration of normal and malignant JAM-C-expressing B cells to bone marrow, lymph nodes, and spleen. Blocking homing to the spleen is remarkable, as most other antiadhesion antibodies reduce homing of B cells only to bone marrow and lymph nodes. Long-term administration of anti-JAM-C antibodies prevented engraftment of JAM-Cpos lymphoma cells in bone marrow, spleen, and lymph nodes of mice. Plasmon resonance studies identified JAM-B as the major ligand for JAM-C, whereas homotypic JAM-C interactions remained at background levels. Accordingly, anti-JAM-C antibodies blocked adhesion of JAM-C-expressing B cells to their ligand JAM-B, and immunofluorescence analysis showed the expression of JAM-B on murine and human lymphatic endothelial cells. Targeting JAM-C could thus constitute a new therapeutic strategy to prevent lymphoma cells from reaching supportive microenvironments not only in the bone marrow and lymph nodes but also in the spleen.


Acousto-optic imaging of a color picture hidden behind a scattering layer.
An imaging technique has been developed to image a color picture hidden behind a 5 mm thick, highly scattering layer with low transmittance of 0.24%. Small vibrations (< 1 µm) were induced in the hidden picture, causing a time-varying speckle pattern on the scattering layer in the front, which is captured by a CCD camera and quantified as speckle contrast difference (SCD). With two lasers at 543 nm and 633 nm, the imaging system raster-scans the front of the scattering layer and the resulting SCD image reveals the color features of the hidden picture.


Direct coupling of normal-phase high-performance liquid chromatography to atmospheric pressure laser ionization fourier transform ion cyclotron resonance mass spectrometry for the characterization of crude oil.
The high complexity of crude oil makes the use of chromatographic separation an important tool especially for sample simplification. The coupling of normal-phase high-performance liquid chromatography (HPLC) using a polar aminocyano column to a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer offers the best attributes of good separation prior to ultrahigh resolution mass spectrometry (MS) detection. Atmospheric pressure laser ionization (APLI) was used as an ionization technique to analyze the nitrogen-containing aromatic compounds in a deasphalted crude oil due to its unique selectivity toward aromatic compounds and also due to its sensitivity. Two main chromatographic peaks were observed during this separation indicating a class-based separation. Mass spectra obtained from fractions were collected along the entire retention time and compared with each other to assign the unique constituents. By coupling the HPLC system directly to the FTICR mass spectrometer, comparable ion and UV chromatograms were obtained, reflecting the scan-to-scan sensitivity of the coupling system. The results show that it is possible to calculate reconstructed class chromatograms (RCC), allowing differences in class composition to be traced along the retention time. As an example, radical and protonated nitrogen species generated by APLI were detected along the retention time which enabled a differentiation between basic and nonbasic species in the same polar peak, thus overcoming the limitation of chromatographic resolution. This report represents the first online LC-FTICR MS coupling in the field of crude oil analysis.


Perception of the chitin oligosaccharides contributes to disease resistance to blast fungus Magnaporthe oryzae in rice.
Chitin is a component of fungal cell walls, and its fragments act as elicitors in many plants. The plasma membrane glycoprotein CEBiP, which possesses LysM domains, is a receptor for the chitin elicitor (CE) in rice. Here, we report that the perception of CE by CEBiP contributes to disease resistance against the rice blast fungus, Magnaporthe oryzae, and that enhanced responses to CE by engineering CEBiP increase disease tolerance. Knockdown of CEBiP expression allowed increased spread of the infection hyphae. To enhance defense responses to CE, we constructed chimeric genes composed of CEBiP and Xa21, which mediate resistance to rice bacterial leaf blight. The expression of either CRXa1 or CRXa3, each of which contains the whole extracellular portion of CEBiP, the whole intracellular domain of XA21, and the transmembrane domain from either CEBiP or XA21, induced cell death accompanied by an increased production of reactive oxygen and nitrogen species after treatment with CE. Rice plants expressing the chimeric receptor exhibited necrotic lesions in response to CE and became more resistant to M. oryzae. Deletion of the first LysM domain in CRXA1 abolished these cellular responses. These results suggest that CEs are produced and recognized through the LysM domain of CEBiP during the interaction between rice and M. oryzae and imply that engineering pattern recognition receptors represents a new strategy for crop protection against fungal diseases.


Hypoperfusion of the Adventitial Vasa Vasorum Develops an Abdominal Aortic Aneurysm.
The aortic wall is perfused by the adventitial vasa vasorum (VV). Tissue hypoxia has previously been observed as a manifestation of enlarged abdominal aortic aneurysms (AAAs). We sought to determine whether hypoperfusion of the adventitial VV could develop AAAs. We created a novel animal model of adventitial VV hypoperfusion with a combination of a polyurethane catheter insertion and a suture ligation of the infrarenal abdominal aorta in rats. VV hypoperfusion caused tissue hypoxia and developed infrarenal AAA, which had similar morphological and pathological characteristics to human AAA. In human AAA tissue, the adventitial VV were stenotic in both small AAAs (30-49 mm in diameter) and in large AAAs (> 50 mm in diameter), with the sac tissue in these AAAs being ischemic and hypoxic. These results indicate that hypoperfusion of adventitial VV has critical effects on the development of infrarenal AAA.


Investigation of trace metals in the blood plasma and scalp hair of gastrointestinal cancer patients in comparison with controls.
Gastrointestinal cancer is one of the most common types of cancer which is predominantly associated with the environmental factors. The carcinogenic processes are linked with the imbalances of trace metals in body fluid and tissues. Trace metals (Cd, Cr, Cu, Fe, Ni, Pb and Zn) are estimated in blood plasma and scalp hair of the cancer patients and controls employing nitric acid-perchloric acid based wet-digestion followed by atomic absorption spectrophotometric method. The mean concentrations of Cd, Cr, Cu and Ni were found to be significantly higher in the plasma of patients compared with the controls, however, appreciably higher concentrations of Fe and Zn were observed in the plasma of controls. The average scalp hair concentrations of Zn, Fe, Pb, Cu and Cd were notably higher in the patients than controls. The correlation study revealed significantly different mutual variations of the trace metals in the plasma and scalp hair of the patients and controls. The apportionment of trace metals in the plasma and scalp hair of the patients and controls was also considerably different. The study revealed that the carcinogenic processes are significantly affecting the trace metal burden and mutual variations in the cancerous patients compared with the controls.


Multiparametric MR assessment of pediatric brain tumors.
MR assessment of pediatric brain tumors has expanded to include physiologic information related to cellular metabolites, hemodynamic and diffusion parameters. The purpose of this study was to investigate the relationship between MR and proton MR spectroscopic imaging in children with primary brain tumors. Twenty-one patients (mean age 9 years) with histologically verified brain tumors underwent conventional MR imaging, hemodynamic MR imaging (HMRI) and proton MR spectroscopic imaging (MRSI). Fourteen patients also had diffusion-weighted MR imaging (DWMRI). Metabolic indices including choline-containing compounds (Cho), total creatine (tCr) and lipids/lactate (L) were derived by proton MRSI, relative cerebral blood volume (rCBV) by HMRI, and apparent tissue water diffusion coefficients (ADC) by DWMRI. Variables were examined by linear regression and correlation as well as by ANOVA. Cho (suggestive of tumor cellularity and proliferative activity) correlated positively with rCBV, while the relationship between Cho and ADC (suggestive of cellular density) was inverse ( P<0.001). The relationship between rCBV and ADC was also inverse ( P=0.004). Cho and lipids (suggestive of necrosis and/or apoptosis) were not significantly correlated ( P=0.51). A positive relationship was found between lipids and ADC ( P=0.002). The relationships between Cho, rCBV, ADC and lipids signify that tumor physiology is influenced by the tumor's physical and chemical environment. Normalized Cho and lipids distinguished high-grade from low-grade tumors ( P<0.05). Multiparametric MR imaging using MRSI, HMRI and DWMRI enhances assessment of brain tumors in children and improves our understanding of tumor physiology while promising to distinguish higher- from lower-malignancy tumors, a distinction that is particularly clinically important among inoperable tumors.


Unique flexibility in energy metabolism allows mycobacteria to combat starvation and hypoxia.
Mycobacteria are a group of obligate aerobes that require oxygen for growth, but paradoxically have the ability to survive and metabolize under hypoxia. The mechanisms responsible for this metabolic plasticity are unknown. Here, we report on the adaptation of Mycobacterium smegmatis to slow growth rate and hypoxia using carbon-limited continuous culture. When M. smegmatis is switched from a 4.6 h to a 69 h doubling time at a constant oxygen saturation of 50%, the cells respond through the down regulation of respiratory chain components and the F1Fo-ATP synthase, consistent with the cells lower demand for energy at a reduced growth rate. This was paralleled by an up regulation of molecular machinery that allowed more efficient energy generation (i.e. Complex I) and the use of alternative electron donors (e.g. hydrogenases and primary dehydrogenases) to maintain the flow of reducing equivalents to the electron transport chain during conditions of severe energy limitation. A hydrogenase mutant showed a 40% reduction in growth yield highlighting the importance of this enzyme in adaptation to low energy supply. Slow growing cells at 50% oxygen saturation subjected to hypoxia (0.6% oxygen saturation) responded by switching on oxygen scavenging cytochrome bd, proton-translocating cytochrome bc1-aa3 supercomplex, another putative hydrogenase, and by substituting NAD+-dependent enzymes with ferredoxin-dependent enzymes thus highlighting a new pattern of mycobacterial adaptation to hypoxia. The expression of ferredoxins and a hydrogenase provides a potential conduit for disposing of and transferring electrons in the absence of exogenous electron acceptors. The use of ferredoxin-dependent enzymes would allow the cell to maintain a high carbon flux through its central carbon metabolism independent of the NAD+/NADH ratio. These data demonstrate the remarkable metabolic plasticity of the mycobacterial cell and provide a new framework for understanding their ability to survive under low energy conditions and hypoxia.


Dramatic response to climate change in the Southwest: Robert Whittaker's 1963 Arizona Mountain plant transect revisited.
Models analyzing how Southwestern plant communities will respond to climate change predict that increases in temperature will lead to upward elevational shifts of montane species. We tested this hypothesis by reexamining Robert Whittaker's 1963 plant transect in the Santa Catalina Mountains of southern Arizona, finding that this process is already well underway. Our survey, five decades after Whittaker's, reveals large changes in the elevational ranges of common montane plants, while mean annual rainfall has decreased over the past 20 years, and mean annual temperatures increased 0.25°C/decade from 1949 to 2011 in the Tucson Basin. Although elevational changes in species are individualistic, significant overall upward movement of the lower elevation boundaries, and elevational range contractions, have occurred. This is the first documentation of significant upward shifts of lower elevation range boundaries in Southwestern montane plant species over decadal time, confirming that previous hypotheses are correct in their prediction that mountain communities in the Southwest will be strongly impacted by warming, and that the Southwest is already experiencing a rapid vegetation change.


A comparison of training methods to increase neck muscle strength.
To compare two neck strength training modalities. Neck injury in pilots flying high performance aircraft is a concern in aviation medicine. Strength training may be an effective means to strengthen the neck and decrease injury risk. The cohort consisted of 32 age-height-weight matched participants, divided into two experimental groups; the Multi-Cervical Unit (MCU) and Thera-Band tubing groups (THER), and a control (CTRL) group. Ten weeks of training were undertaken and pre-and post isometric strength testing for all groups was performed on the MCU. Comparisons between the three groups were made using a Kruskal-Wallis test and effect sizes between the MCU and the THER groups and the THER and CTRL groups were also calculated. The MCU group displayed the greatest increase in isometric strength (flexion 64.4%, extension 62.9%, left lateral flexion 53.3%, right lateral flexion 49.1%) and differences were only statistically significant (p<0.05) when compared to the CTRL group. Increases in neck strength for the THER group were lower than that shown in the MCU group (flexion 42.0%, extension 29.9%, left lateral flexion 26.7%, right lateral flexion 24.1%). Moderate to large effect sizes were found between the MCU and THER as well as the THER and CTRL groups. This study demonstrated that the MCU was the most effective training modality to increase isometric cervical muscle strength. Thera-Band tubing did however, produce moderate gains in isometric neck strength.


Comparative studies of thought disorders. I. Mania and schizophrenia.
Thought disorder in 20 manic and 43 schizophrenic patients was examined using the Thought Disorder Index. To ensure the inclusion of patients with unambiguous mania and schizophrenia, the patients met Research Diagnostic Criteria, DSM-III criteria, and Washington University (St Louis) criteria. While the quantity of thought disorder did not differ significantly between the two groups, there were distinct qualitative differences. The thought disorder of manic patients was extravagantly combinatory, usually with humor, flippancy, and playfulness. The thought disorder of schizophrenic patients appeared disorganized, confused, and ideationally fluid, with many peculiar words and phrases.


A Perspective on Polo-Like Kinase-1 Inhibition for the Treatment of Rhabdomyosarcomas.
Rhabdomyosarcomas are the most common pediatric soft tissue sarcoma and are a major cause of death from cancer in young patients requiring new treatment options to improve outcomes. High-risk patients include those with metastatic or relapsed disease and tumors with PAX3-FOXO1 fusion genes that encode a potent transcription factor that drives tumourigenesis through transcriptional reprogramming. Polo-Like Kinase-1 (PLK1) is a serine/threonine kinase that phosphorylates a wide range of target substrates and alters their activity. PLK1 functions as a pleiotropic master regulator of mitosis and regulates DNA replication after stress. Taken together with high levels of expression that correlate with poor outcomes in many cancers, including rhabdomyosarcomas, it is an attractive therapeutic target. This is supported in rhabdomyosarcoma models by characterization of molecular and phenotypic effects of reducing and inhibiting PLK1, including changes to the PAX3-FOXO1 fusion protein. However, as tumor re-growth has been observed, combination strategies are required. Here we review preclinical evidence and consider biological rationale for PLK1 inhibition in combination with drugs that promote apoptosis, interfere with activity of PAX3-FOXO1 and are synergistic with microtubule-destabilizing drugs such as vincristine. The preclinical effects of low doses of the PLK1 inhibitor volasertib in combination with vincristine, which is widely used in rhabdomyosarcoma treatment, show particular promise in light of recent clinical data in the pediatric setting that support achievable volasertib doses predicted to be effective. Further development of novel therapeutic strategies including PLK1 inhibition may ultimately benefit young patients with rhabdomyosarcoma and other cancers.


Leishmania braziliensis Infection Enhances Toll-Like Receptors 2 and 4 Expression and Triggers TNF-α and IL-10 Production in Human Cutaneous Leishmaniasis.
Cutaneous leishmaniasis (CL) caused by infection with Leishmania braziliensis is characterized by an exaggerated inflammatory response that controls the parasite burden, but also contributes to pathology. While myeloid cells are required to eliminate the parasite, recent studies indicate that they may also participate in the inflammatory response driving disease progression. The innate immune response to leishmania is driven in part by the Toll-like receptors (TLRs) TLR2, TLR4, and TLR9. In this study, we used flow cytometric analysis to compare TLR2 and TLR4 expression in monocyte subsets (classical, intermediate, and non-classical) from CL patients and healthy subjects (HS). We also determined if there was an association of either the pro-inflammatory cytokine TNF or the anti-inflammatory cytokine IL-10 with TLR2 or TLR4 expression levels after L. braziliensis infection. In vitro infection with L. braziliensis caused CL monocytes to up-regulate TLR2 and TLR4 expression. We also found that intermediate monocytes expressed the highest levels of TLR2 and TLR4 and that infected monocytes produced more TNF and IL-10 than uninfected monocytes. Finally, while classical and intermediate monocytes were mainly responsible for TNF production, classical monocytes were the main source of IL-10. Collectively, our studies revealed that up-regulated TLR2/4 expression and TNF production by intermediate/inflammatory subsets of monocytes from patients correlates with detrimental outcome of cutaneous leishmaniasis.


Responsible, Inclusive Innovation and the Nano-Divide.
Policy makers from around the world are trying to emulate successful innovation systems in order to support economic growth. At the same time, innovation governance systems are being put in place to ensure a better integration of stakeholder views into the research and development process. In Europe, one of the most prominent and newly emerging governance frameworks is called Responsible Research and Innovation (RRI). This article aims to substantiate the following points: (1) The concept of RRI and the concept of justice can be used to derive similar ethical positions on the nano-divide. (2) Given the ambitious policy aims of RRI (e.g. economic competitiveness enhancer), the concept may be better suited to push for ethical outcomes on access to nanotechnology and its products rather than debates based on justice issues alone. It may thus serve as a mediator concept between those who push solely for competitiveness considerations and those who push solely for justice considerations in nano-technology debates. (3) The descriptive, non-normative Systems of Innovation approaches (see below) should be linked into RRI debates to provide more evidence on whether the approach advocated to achieve responsible and ethical governance of research and innovation (R&I) can indeed deliver on competitiveness (in nano-technology and other fields).


The rheological properties of beta amyloid Langmuir monolayers: Comparative studies with melittin peptide.
We determined the rheological properties of β-amyloid Langmuir films at the air/water interface, a peptide whose interfacial structure is extended β-sheet, and compared them with those of films composed of Melittin (Mel), which adopts an α-helical conformation at neutral pH. To determine the dilatational and shear moduli we evaluated the response of pure peptide monolayers to an oscillatory anisotropic compressive work. Additionally, a micro-rheological characterization was performed by tracking the diffusion of micrometer sized latex beads onto the interface. This technique allowed us the detection of different rheological behaviour between monolayers presenting a low shear response. Monolayers of the β-sheet structure-adopting peptides, such as β-amyloid peptides, exhibited a marked shear (elastic) modulus even at low surface pressures. In contrast, Mel monolayers exhibited negligible shear modulus and the micro-rheological shear response was markedly lower than that observed for either Aβ1-40 or Aβ1-42 amyloid peptides. When Mel monolayers were formed at the interface of an aqueous solution at pH 11, we observed an increase in both the lateral stability and film viscosity as detected by a slower diffusion of the latex beads, in keeping with an increase in β-sheet structure at this high pH (verified by ATR and FT-IR measurements). We suggest that the interactions responsible for the marked response upon shear observed for β-amyloid peptide monolayers are the hydrogen bonds of the β-sheet structure that can form an infinite planar network at the interface. Conversely, α-helical Mel peptide lack of these inter-molecular interactions and, therefore the shear contribution was negligible. We propose that the secondary structure is important for modulating the rheological behavior of short peptide monolayers regardless of the mass density or surface charge at the surface.


Spatiotemporally separated antigen uptake by alveolar dendritic cells and airway presentation to T cells in the lung.
Asthma pathogenesis is focused around conducting airways. The reasons for this focus have been unclear because it has not been possible to track the sites and timing of antigen uptake or subsequent antigen presentation to effector T cells. In this study, we use two-photon microscopy of the lung parenchyma and note accumulation of CD11b(+) dendritic cells (DCs) around the airway after allergen challenge but very limited access of these airway-adjacent DCs to the contents of the airspace. In contrast, we observed prevalent transepithelial uptake of particulate antigens by alveolar DCs. These distinct sites are temporally linked, as early antigen uptake in alveoli gives rise to DC and antigen retention in the airway-adjacent region. Antigen-specific T cells also accumulate in the airway-adjacent region after allergen challenge and are activated by the accumulated DCs. Thus, we propose that later airway hyperreactivity results from selective retention of allergen-presenting DCs and antigen-specific T cells in airway-adjacent interaction zones, not from variation in the abilities of individual DCs to survey the lung.


Voltage sensor of Kv1.2: structural basis of electromechanical coupling.
Voltage-dependent ion channels contain voltage sensors that allow them to switch between nonconductive and conductive states over the narrow range of a few hundredths of a volt. We investigated the mechanism by which these channels sense cell membrane voltage by determining the x-ray crystal structure of a mammalian Shaker family potassium ion (K+) channel. The voltage-dependent K+ channel Kv1.2 grew three-dimensional crystals, with an internal arrangement that left the voltage sensors in an apparently native conformation, allowing us to reach three important conclusions. First, the voltage sensors are essentially independent domains inside the membrane. Second, they perform mechanical work on the pore through the S4-S5 linker helices, which are positioned to constrict or dilate the S6 inner helices of the pore. Third, in the open conformation, two of the four conserved Arg residues on S4 are on a lipid-facing surface and two are buried in the voltage sensor. The structure offers a simple picture of how membrane voltage influences the open probability of the channel.


Activated mTORC1 promotes long-term cone survival in retinitis pigmentosa mice.
Retinitis pigmentosa (RP) is an inherited photoreceptor degenerative disorder that results in blindness. The disease is often caused by mutations in genes that are specific to rod photoreceptors; however, blindness results from the secondary loss of cones by a still unknown mechanism. Here, we demonstrated that the mammalian target of rapamycin complex 1 (mTORC1) is required to slow the progression of cone death during disease and that constitutive activation of mTORC1 in cones is sufficient to maintain cone function and promote long-term cone survival. Activation of mTORC1 in cones enhanced glucose uptake, retention, and utilization, leading to increased levels of the key metabolite NADPH. Moreover, cone death was delayed in the absence of the NADPH-sensitive cell death protease caspase 2, supporting the contribution of reduced NADPH in promoting cone death. Constitutive activation of mTORC1 preserved cones in 2 mouse models of RP, suggesting that the secondary loss of cones is caused mainly by metabolic deficits and is independent of a specific rod-associated mutation. Together, the results of this study address a longstanding question in the field and suggest that activating mTORC1 in cones has therapeutic potential to prolong vision in RP.


Exploring Niches for Short-Season Grain Legumes in Semi-Arid Eastern Kenya - Coping with the Impacts of Climate Variability.
Climate variability is the major risk to agricultural production in semi-arid agroecosystems and the key challenge to sustain farm livelihoods for the 500 million people who inhabit these areas worldwide. Short-season grain legumes have great potential to address this challenge and help to design more resilient and productive farming systems. However, grain legumes display a great diversity and differ widely in growth, development, and resource use efficiency. Three contrasting short season grain legumes common bean (Phaseolus vulgaris L.), cowpea (Vigna unguiculata (L.) Walp.] and lablab [Lablab purpureus (L.) Sweet] were selected to assess their agricultural potential with respect to climate variability and change along the Machakos-Makueni transect in semi-arid Eastern Kenya. This was undertaken using measured data [a water response trial conducted during 2012/13 and 2013/14 in Machakos, Kenya] and simulated data using the Agricultural Production System sIMulator (APSIM). The APSIM crop model was calibrated and validated to simulate growth and development of short-season grain legumes in semi-arid environments. Water use efficiency (WUE) was used as indicator to quantify the production potential. The major traits of adaptation include early flowering and pod and seed set before the onset of terminal drought. Early phenology together with adapted canopy architecture allowed more optimal water use and greater partitioning of dry matter into seed (higher harvest index). While common bean followed a comparatively conservative strategy of minimizing water loss through crop transpiration, the very short development time and compact growth habit limited grain yield to rarely exceed 1,000 kg ha-1. An advantage of this strategy was relatively stable yields independent of in-crop rainfall or season length across the Machakos-Makueni transect. The growth habit of cowpea in contrast minimized water loss through soil evaporation with rapid ground cover and dry matter production, reaching very high grain yields at high potential sites (3,000 kg ha-1) but being highly susceptible to in-season drought. Lablab seemed to be best adapted to dry environments. Its canopy architecture appeared to be best in compromising between the investment in biomass as a prerequisite to accumulate grain yield by minimizing water loss through soil evaporation and crop transpiration. This lead to grain yields of up to 2,000 kg ha-1 at high potential sites and >1,000 kg ha-1 at low potential sites. The variance of observed and simulated WUE was high and no clear dependency on total rainfall alone was observed for all three short-season grain legumes, highlighting that pattern of water use is also important in determining final WUEbiomass and WUEgrain. Mean WUEgrain was lowest for cowpea (1.5-3.5 kggrain ha-1 mm-1) and highest for lablab (5-7 kggrain ha-1 mm-1) reflecting the high susceptibility to drought of cowpea and the good adaptation to dry environments of lablab. Results highlight that, based on specific morphological, phonological, and physiological characteristics, the three short-season grain legumes follow different strategies to cope with climate variability. The climate-smart site-specific utilization of the three legumes offers promising options to design more resilient and productive farming systems in semi-arid Eastern Kenya.


Diffusion kernel-based logistic regression models for protein function prediction.
Assigning functions to unknown proteins is one of the most important problems in proteomics. Several approaches have used protein-protein interaction data to predict protein functions. We previously developed a Markov random field (MRF) based method to infer a protein's functions using protein-protein interaction data and the functional annotations of its protein interaction partners. In the original model, only direct interactions were considered and each function was considered separately. In this study, we develop a new model which extends direct interactions to all neighboring proteins, and one function to multiple functions. The goal is to understand a protein's function based on information on all the neighboring proteins in the interaction network. We first developed a novel kernel logistic regression (KLR) method based on diffusion kernels for protein interaction networks. The diffusion kernels provide means to incorporate all neighbors of proteins in the network. Second, we identified a set of functions that are highly correlated with the function of interest, referred to as the correlated functions, using the chi-square test. Third, the correlated functions were incorporated into our new KLR model. Fourth, we extended our model by incorporating multiple biological data sources such as protein domains, protein complexes, and gene expressions by converting them into networks. We showed that the KLR approach of incorporating all protein neighbors significantly improved the accuracy of protein function predictions over the MRF model. The incorporation of multiple data sets also improved prediction accuracy. The prediction accuracy is comparable to another protein function classifier based on the support vector machine (SVM), using a diffusion kernel. The advantages of the KLR model include its simplicity as well as its ability to explore the contribution of neighbors to the functions of proteins of interest.


The Association Between Age at Initiation of Alcohol Consumption and Type 2 Diabetes Mellitus: A Cohort Study of 0.5 Million Persons in China.
It is well known that alcohol consumption is associated with type 2 diabetes mellitus. However, the association of age at initiation of alcohol consumption and duration of alcohol drinking with type 2 diabetes mellitus among Chinese adults is not fully understood. This study was based on data from the China Kadoorie Biobank, which included 512,712 participants aged 30-79 years who were living in China in 2004-2008. A Cox proportional hazards model was used to estimate the association of AAI and drinking duration with type 2 diabetes. After adjustment for potential covariates, ages at alcohol initiation (AAIs) of 18.1-29.0 years, 29.1-39.0 years, and >39.0 years were associated with 22% (95% confidence interval (CI): 14, 30), 25% (95% CI: 17, 33), and 32% (95% CI: 24, 39) lower hazards of type 2 diabetes compared with abstaining, respectively. Drinking durations of <10.1 years, 10.1-20.0 years, and 20.1-30.0 years were associated with a lower risk of type 2 diabetes, compared with abstaining. Among current (weekly) drinkers, AAI <18.1 years and drinking duration >30.0 years were associated with 18% (95% CI: 4, 33) and 20% (95% CI: 3, 40) higher hazards of type 2 diabetes, compared with AAI 18.1-29.0 years and drinking duration <10.1 years, respectively. In conclusion, late AAI and a short drinking duration were associated with a lower risk of type 2 diabetes in this large prospective cohort study of Chinese adults, but early AAI and long drinking duration were not.


Antioxidants in Alzheimer's disease-vitamin C delivery to a demanding brain.
Levels of several antioxidants and related markers were measured in cerebrospinal fluid (CSF) and plasma of 10 Alzheimer's disease (AD) patients and 10 controls. Daily dosage of vitamin C was significantly correlated with both plasma (R=0.662; p=0.0015) and CSF level (R=0.639, p=0.0024). Plasma and CSF vitamin C levels were also highly correlated R=0.793, p<0.0001). Similarly, daily dosage of Vitamin E was significantly correlated with plasma vitamin E (R=0.681; p=0.0009) and showed a trend toward correlation with CSF vitamin E (R=0.422, p=0.06). There were no significant differences between groups in absolute CSF or plasma levels of any analyte. However, the CSF: plasma ratio of vitamin C was significantly greater in the AD patients compared to the controls (p=0.048). In a subset of AD patients, hippocampal volume was significantly correlated with plasma (R2=0.833; p=0.004) and CSF (R2 =0.603; p=0.04) vitamin C levels, and inversely correlated with CSF:plasma vitamin C ratio (R2 =0.717; p=0.016). We conclude that oral vitamin C supplements are delivered to the brain, and speculate that the increased CSF: plasma ratio of vitamin C in AD reflects increased antioxidant consumption by the AD brain.


The PACE study: a randomised clinical trial of cognitive activity (CA) for older adults with mild cognitive impairment (MCI).
Research evidence from observational studies suggests that cognitive activity reduces the risk of cognitive impairment in later life as well as the rate of cognitive decline of people with dementia. The Promoting Healthy Ageing with Cognitive Exercise (PACE) study has been designed to determine whether a cognitive activity intervention decreases the rate of cognitive decline amongst older adults with mild cognitive impairment (MCI). The study will recruit 160 community-dwelling men and women aged 65 years of age or over with mild cognitive impairment (MCI). Participants will be randomly allocated to two treatment groups: non-specific education and cognitive activity. The intervention will consist of ten 90-minute sessions delivered twice per week over a period of five weeks. The primary outcome measure of the study is the change from baseline in the total score on the Cambridge Cognitive Score (CAMCOG). Secondary outcomes of interest include changes in memory, attention, executive functions, mood and quality of life. Primary endpoints will be collected 12, 52 and 104 weeks after the baseline assessment. The proposed project will produce the best available evidence on the merits of increased cognitive activity as a strategy to prevent cognitive decline among older adults with MCI. We anticipate that the results of this study will have implications for the development of evidence-based preventive strategies to reduce the rate of cognitive decline amongst older people at risk of dementia. ACTRN12608000556347.


Small misfolded Tau species are internalized via bulk endocytosis and anterogradely and retrogradely transported in neurons.
The accumulation of Tau into aggregates is associated with key pathological events in frontotemporal lobe degeneration (FTD-Tau) and Alzheimer disease (AD). Recent data have shown that misfolded Tau can be internalized by cells in vitro (Frost, B., Jacks, R. L., and Diamond, M. I. (2009) J. Biol. Chem. 284, 12845-12852) and propagate pathology in vivo (Clavaguera, F., Bolmont, T., Crowther, R. A., Abramowski, D., Frank, S., Probst, A., Fraser, G., Stalder, A. K., Beibel, M., Staufenbiel, M., Jucker, M., Goedert, M., and Tolnay, M. (2009) Nat. Cell Biol. 11, 909-913; Lasagna-Reeves, C. A., Castillo-Carranza, D. L., Sengupta, U., Guerrero-Munoz, M. J., Kiritoshi, T., Neugebauer, V., Jackson, G. R., and Kayed, R. (2012) Sci. Rep. 2, 700). Here we show that recombinant Tau misfolds into low molecular weight (LMW) aggregates prior to assembly into fibrils, and both extracellular LMW Tau aggregates and short fibrils, but not monomers, long fibrils, nor long filaments purified from brain extract are taken up by neurons. Remarkably, misfolded Tau can be internalized at the somatodendritic compartment, or the axon terminals and it can be transported anterogradely, retrogradely, and can enhance tauopathy in vivo. The internalized Tau aggregates co-localize with dextran, a bulk-endocytosis marker, and with the endolysosomal compartments. Our findings demonstrate that exogenous Tau can be taken up by cells, uptake depends on both the conformation and size of the Tau aggregates and once inside cells, Tau can be transported. These data provide support for observations that tauopathy can spread trans-synaptically in vivo, via cell-to-cell transfer.


Evaluation of linear array human papillomavirus genotyping using automatic optical imaging software.
Variations in biological behavior suggest that each carcinogenic human papillomavirus (HPV) type should be considered individually in etiologic studies. HPV genotyping assays might have clinical applications if they are approved for use by the FDA. A widely used genotyping assay is the Roche Linear Array HPV genotyping test (LA). We used LA to genotype the HPV isolates from cervical specimens from women with the full spectrum of cervical disease: cervical cancer, cervical intraepithelial neoplasia (CIN), and HPV infections. To explore the feasibility and value of the automated reading of the LA results, we custom-designed novel, optical imaging software that provides optical density measurements of LA bands. We compared unmagnified visual examination with the automated measurements. The two measurements were highly associated. By either method, the threshold between a negative and a positive result was fairly sharp, with a clear bimodal distribution. Visually, most positive results were judged to be strong or medium, with fewer equivocal results categorized as weak (9.5% of positive samples), very weak (6.5% of positive samples), or extremely weak (7.7% of positive samples). The automated measurements of the intensities were significantly associated with the strength of the visual categories (P < 0.001). At the extremes of the automated signal intensities (< or = 20 units or > or = 120 units), the bands were almost always categorized visually as negative and positive, respectively. In the equivocal zone (20 to 119 units), specimens were more increasingly likely to be judged to be visually positive as the number of other, definite infections on the same strip increased (P for trend < 0.001). Multiple, concurrent infections comprise > or = 25% of HPV infections; thus, any systematic visual tendency that influences their evaluation when the result is equivocal should be minimized. Therefore, automated reading is probably worth development if easy-to-calibrate hardware and software can be optimized.


Gut microbes and adverse food reactions: Focus on gluten related disorders.
Immediately following birth, the gastrointestinal tract is colonized with a complex community of bacteria, which helps shape the immune system. Under conditions of health, the immune system is able to differentiate between innocuous antigens, including food protein and commensals, and harmful antigens such as pathogens. However, patients with celiac disease (CD) develop an intolerance to gluten proteins which results in a pro-inflammatory T-cell mediated immune response with production of anti-gluten and anti-tissue transglutaminase antibodies. This adaptive immune response, in conjunction with activation of innate inflammatory cells, lead to destruction of the small intestinal mucosa. Overall 30% of the global population has genetic risk to develop CD. However, only a small proportion develop CD, suggesting that additional environmental factors must play a role in disease pathogenesis. Alterations in small intestinal microbial composition have recently been associated with active CD, indicating a possible role for the microbiota in CD. However, studies demonstrating causality are lacking. This review will highlight the recent data on the potential role of the microbiota in CD pathogenesis, the potential mechanisms, and discuss future research directions.


Patterns of colorectal cancer test use, including CT colonography, in the 2010 National Health Interview Survey.
Recommended colorectal cancer (CRC) screening tests for adults ages 50 to 75 years include home fecal occult blood tests (FOBT), sigmoidoscopy with FOBT, and colonoscopy. A newer test, computed tomographic (CT) colonography, has been recommended by some, but not all, national organizations. We analyzed 2010 National Health Interview Survey data, including new CT colonography questions, from respondents ages 50 to 75 years (N = 8,952). We (i) assessed prevalence of CRC test use overall, by test type, and by sociodemographic and health care access factors and (ii) assessed reported reasons for not having a CRC test. The age-standardized percentage of respondents reporting FOBT, sigmoidoscopy, or colonoscopy within recommended time intervals was 58.3% [95% confidence interval (CI), 57.0-59.6]. Colonoscopy was the most commonly reported test [within past 10 years: 54.6% (95% CI, 53.2-55.9)]. Home FOBT and sigmoidoscopy with FOBT were less frequently used [FOBT within past year: 8.8% (95% CI, 8.1-9.6); sigmoidoscopy within past 5 years with FOBT within past 3 years: 1.3% (95% CI, 1.0-1.6)]. CT colonography was rare: 1.3% (95% CI, 1.0-1.7). Increasing age, education, income, having health care insurance, and having a usual source of health care were associated with higher CRC test use. Test use within recommended time intervals was particularly low among individuals ages 50 to 64 years without health care insurance [21.2% (95% CI, 18.3-24.4)]. The most common reason for nonuse was "no reason or never thought about it." About 40% of Americans ages 50 to 75 years do not meet the recommendations for having CRC screening tests. Expanded health care coverage and greater awareness of CRC screening are needed to further decrease CRC mortality.


Estrous cycle stage-dependent manner of type I interferon-stimulated genes induction in the bovine endometrium.
Interferon tau (IFN-τ) is a ruminant-specific type I IFN secreted by a conceptus before its attachment to the uterus. IFN-τ induces the expression of IFN-stimulated genes (ISGs) via the type I IFN receptor (IFNAR), which is composed of IFNAR1 and IFNAR2 subunits in the endometrium. However, expression patterns of IFNARs during the estrous cycle have not been reported. We hypothesized that the response to a type I IFN changes along with IFNARs and the IFN-regulatory factors (IRFs) driving transcription of IFN signal-related genes and modulating a type I IFN signal during the estrous cycle. We investigated the estrous cycle stage-dependent type I IFN induction of ISGs and expression patterns of IFN signal-related genes in bovine endometrial tissues. Endometrial tissue pieces collected from bovine uteri at each estrous stage (early, mid, and late) were cultured with or without recombinant bovine IFN-α or concentrated pregnant uterine flushing (PUF) on day 18 after confirming the presence of a conceptus. IFN-α and PUF each significantly increased the expression of ISGs in endometrial tissues. The induction levels of the typical ISGs (MX1-a and ISG15) were significantly higher at the mid stage and correlated with high expression of IRFs at the mid stage. The immunostaining of IFNARs showed strong fluorescence intensities in luminal and glandular epithelia at the early and mid stages. Collectively, these results suggest that the endometrium exhibits estrous cycle stage-dependent responsiveness to type I IFN that may be associated with the expression of IFNARs and IRFs for pregnancy recognition.