A Non-Invasive Method for Estimating Cardiopulmonary Variables Using Breath-by-Breath Injection of Two Tracer Gases.
Conventional methods for estimating cardiopulmonary variables usually require complex gas analyzers and the active co-operation of the patient. Therefore, they are not compatible with the crowded environment of the intensive care unit (ICU) or operating theatre, where patient co-operation is typically impossible. However, it is these patients that would benefit the most from accurate estimation of cardiopulmonary variables, because of their critical condition. This paper describes the results of a collaborative development between an anesthesiologists and biomedical engineers to create a compact and non-invasive system for the measurement of cardiopulmonary variables such as lung volume, airway dead space volume, and pulmonary blood flow. In contrast with conventional methods, the compact apparatus and non-invasive nature of the proposed method allow it to be used in the ICU, as well as in general clinical settings. We propose the use of a non-invasive method, in which tracer gases are injected into the patient's inspired breath, and the concentration of the tracer gases is subsequently measured. A novel breath-by-breath tidal ventilation model is then used to estimate the value of a patient's cardiopulmonary variables. Experimental results from an artificial lung demonstrate minimal error in the estimation of known parameters using the proposed method. Results from analysis of a cohort of 20 healthy volunteers (within the Oxford University Hospitals NHS Trust) show that the values of estimated cardiopulmonary variables from these subjects lies within the expected ranges. Advantages of this method are that it is non-invasive, compact, portable, and can perform analysis in real time with less than 1 min of acquired respiratory data.


Molecular dating when rates vary.
Molecular-dating techniques potentially enable us to estimate the time of origin of any biological lineage. Such techniques were originally premised on the assumption of a 'molecular clock'; that is, the assumption that genetic change accumulated steadily over time. However, it is becoming increasingly clear that constant rates of molecular evolution might be the exception rather than the rule. Recently, new methods have appeared that enable the incorporation of variable rates into molecular dating. Direct comparisons between these methods are difficult, because they differ in so many respects. However, the assumptions about rate change on which they rely fall into a few broad categories. Improving our understanding of molecular evolution will be an important next step towards evaluating and improving these methods.


Trophinin and tastin, a novel cell adhesion molecule complex with potential involvement in embryo implantation.
Two human epithelial cell lines, trophoblastic teratocarcinoma HT-H and endometrial adenocarcinoma SNG-M cells, adhere to each other at their respective apical cell surfaces in a divalent cation-independent manner. Two novel molecules responsible for the adhesion between these two cell types were identified by expression cDNA cloning. One, named trophinin, is an intrinsic membrane protein and mediates homophilic self-binding. Another, named tastin, is a cytoplasmic protein and is necessary for trophinin to function as a cell adhesion molecule. Trophinin and tastin appear to be associated with the cytoskeleton in HT-H and SNG-M cells. These molecules are normally not expressed in various types of human cells in tissues, with the exception of macrophages. Strong expression of these molecules was detected in the trophectoderm surface of monkey blastocyst. These molecules are also expressed in human endometrial surface epithelium on day 16/17 at the early secretory phase of human endometrium, the time consistent with that expected for the "implantation window."


N-alkylated dipeptide amides and related structures as imitations of the melanocortins' active core.
Thirty-three low molecular mass structures combining both peptide and peptoid features were prepared and tested on human melanocortin receptors MC1,3-5R. Most of them displayed low micromolar activity with preference for diamines, guanidino and 2-naphthyl derivatives compared to monoacetylated, amino and 3-indolyl counterparts. Some contained L- or D-histidine residues, but the change did not influence affinity. QSAR modelling yielded excellent models for the MC3-5 receptors explaining R2Y=0.89-0.91 and predicting Q2=0.77-0.80 of the affinity variations. One compound displayed MC1R selectivity (13-fold and more). An NMR study of showed that it exists as a mixture of four rotamers at its tertiary amide bonds. Comparisons with earlier data for melanocortin core tetrapeptide analogues indicate that the novel peptide-peptoids interact with the melanocortin receptors in a different way.


Genotyping for Cx26 and Cx30 mutations in cases with congenital hearing loss.
Hearing loss is the most frequent sensory defect in human being. The 13q11-q12 region contains the GJB2 and GJB6 genes, which code connexin 26 (CX26) and connexin 30 (CX30) proteins, respectively. The 35delG, 167delT, and 235delC mutations in the Cx26 gene are the main cause for sporadic nonsyndromic hearing loss (NSHL) in many populations. The 342-kb deletion [del(GJB6-D13S1830)] of the Cx30 gene is the second most common connexin mutation after the 35delG mutation in some NSHL populations. In our study 47 hearing-impaired students were included. The Cx26 gene and the Cx30 gene were analyzed for presence of the 35delG, 167delT, and 342-kb deletion [del(GJB6-D13S1830)]. Genotyping were performed for detecting 35delG, 167delT, and del(GJB6-D13S1830) mutations using the PCR-ELISA techniques. According to the results obtained from 47 cases, the 35delG mutation was detected in 7 cases ( approximately 14.9%). Four of these mutations were determined as homozygote mutant ( approximately 8.5%), and three were determined as heterozygote mutant ( approximately 6.4%). However, 167delT and del(GJB6-D13S1830) mutations were not detected in the study group. These results support the overwhelming majority of 35delG in our study group from deafness school in our study. In conclusion, the 35delG mutation was determined as the most frequently shown mutation that leads to congenital hearing loss as in previous studies from Turkey.


Magnetic resonance imaging planning in children with complex congenital heart disease - A new approach.
To compare a standard sequential 2D Planning Method (2D-PM) with a 3D offline Planning Method (3D-PM) based on 3D contrast-enhanced magnetic resonance angiography (CE-MRA) in children with congenital heart disease (CHD). In 14 children with complex CHD (mean: 2.6 years, range: 3 months to 7.6 years), axial and coronal cuts were obtained with single slice spin echo sequences to get the final double oblique longitudinal cut of the targeted anatomical structure (2D-PM, n = 31). On a separate workstation, similar maximal intensity projection (MIP) images were generated offline from a 3D CE-MRA. MIP images were localizers for repeated targeted imaging using the previous spin echo sequence (3D-PM). Finally, image coverage, spatial orientation and acquisition time were compared for 2D-PM and 3D-PM. 2D-PM and 3D-PM images were similar: both perfectly covered the selected anatomic regions and no spatial differences were found (p>0.05). The mean time for creation of the final imaging plane was 241 ± 31 s (2D-PM) compared to 71 ± 18 s (3D-PM) (p<0.05). 3D-PM shows similar results compared to 2D-PM, but allows faster and offline planning thereby reducing the scan time significantly. As newly developed high-resolution 3D datasets can also be used further improvement of this technology is expected.


Accumulation of neurotoxic organochlorines and trace elements in brain of female European eel (Anguilla anguilla).
Xenobiotics such as organochlorine compounds (OCs) and metals have been suggested to play a significant role in the collapse of European eel stocks in the last decades. Several of these pollutants could affect functioning of the nervous system. Still, no information is so far available on levels of potentially neurotoxic pollutants in eel brain. In present study, carried out on female eels caught in Belgian rivers and canals, we analyzed brain levels of potentially-neurotoxic trace elements (Ag, Al, As, Cd, Co, Cr, Cu, Fe, Hg, MeHg, Mn, Ni, Pb, Sn, Sb, Zn) and OCs (Polychlorinated biphenyls, PCBs; Hexachlorocyclohexanes, HCHs; Dichlorodiphenyltrichloroethane and its metabolites, DDTs). Data were compared to levels in liver and muscle tissues. Eel brain contained very high amounts of OCs, superior to those found in the two other tissues. Interestingly, the relative abundance of PCB congeners markedly differed between tissues. In brain, a predominance of low chlorinated PCBs was noted, whereas highly chlorinated congeners prevailed in muscle and liver. HCHs were particularly abundant in brain, which contains the highest amounts of β-HCH and ϒ-HCH. p,p'-DDTs concentration was similar between brain and muscle (i.e., about twice that of liver). A higher proportion of p,p'-DDT was noticed in brain. Except for Cr and inorganic Hg, all potentially neurotoxic metals accumulated in brain to levels equal to or lower than hepatic levels. Altogether, results indicate that eel brain is an important target for organic and, to a lesser extent, for inorganic neurotoxic pollutants.


Effect of exercise on serum concentration of cartilage oligomeric matrix protein in Thoroughbreds.
To evaluate changes in serum cartilage oligomeric matrix protein (COMP) concentrations in response to exercise in horses. 15 horses in experiment 1 and 27 horses in experiment 2. In experiment 1, 15 Thoroughbreds free of orthopedic disease underwent a standardized exercise protocol. Running velocity and heart rate (HR) were recorded, and blood samples were collected immediately before (baseline) and 1, 5, and 24 hours after a single episode of exercise. In experiment 2, 27 horses underwent 9 stages of a training program in which each stage consisted of 4 to 8 consecutive daily workouts followed by a rest day. Blood samples were collected immediately before the first and final daily workouts in each stage. Serum COMP concentrations were measured via inhibition ELISA with a monoclonal antibody (14G4) against equine COMP. In experiment 1, mean serum COMP concentration was significantly higher than baseline 1 and 5 hours after exercise and returned to baseline concentrations 24 hours after exercise. Mean serum baseline COMP concentration increased as the velocity of running at maximum HR and at an HR of 200 beats/min increased, being significantly higher during the third and fourth exercise tests than during the first. In experiment 2, mean baseline COMP concentration at the final workout of each stage was significantly higher than that at the first workout, beginning with stage 3. Serum COMP concentrations changed significantly in response to exercise. Exercise may enhance movement of COMP into the circulation as well as change the basal turnover rate of COMP.


The growth of mammals following a period of nutritional limitation.
A theory of predicting the growth of an animal following a period of nutritional limitation is proposed here. The theory consists of the following three propositions. Proposition 1: there is a scale of size, such that the growth rate following a period of nutritional limitation is that of normal animals at the same size. Proposition 2: any abnormalities in body composition at the end of the period of limitation will be corrected over time. Proposition 3: the rates at which abnormalities in body composition at the end of the period of limiting conditions are corrected, are always set by the conditions of rehabilitation. The propositions are connected with three other problems viz normal growth, growth under some limitation and the conditions needed by an animal to perform at its potential. These problems are dealt here by: (i) using a simple description of normal growth, (ii) considering the limitation by its effects rather by attempting to predict the effects of a given limitation, and (iii) using simple assumptions about protein and energy scales and requirements. The predictions of the theory are compared with experimental data, and a good qualitative agreement is found. Important conclusions from the theory are: (a) that it is essential to measure as fully as may be the state of the animal at the end of the period of limitation and to compare it with the state of normal animals and (b) that careful attention needs to be given to the choice and the description of the treatment, or preferably treatments, during the rehabilitation period.


Should excessive worry be required for a diagnosis of generalized anxiety disorder? Results from the US National Comorbidity Survey Replication.
Excessive worry is required by DSM-IV, but not ICD-10, for a diagnosis of generalized anxiety disorder (GAD). No large-scale epidemiological study has ever examined the implications of this requirement for estimates of prevalence, severity, or correlates of GAD. Data were analyzed from the US National Comorbidity Survey Replication, a nationally representative, face-to-face survey of adults in the USA household population that was fielded in 2001-2003. DSM-IV GAD was assessed with Version 3.0 of the WHO Composite International Diagnostic Interview. Non-excessive worriers meeting all other DSM-IV criteria for GAD were compared with respondents who met full GAD criteria as well as with other survey respondents to consider the implications of removing the excessiveness requirement. The estimated lifetime prevalence of GAD increases by approximately 40% when the excessiveness requirement is removed. Excessive GAD begins earlier in life, has a more chronic course, and is associated with greater symptom severity and psychiatric co-morbidity than non-excessive GAD. However, non-excessive cases nonetheless evidence substantial persistence and impairment of GAD, high rates of treatment-seeking, and significantly elevated co-morbidity compared with respondents without GAD. Non-excessive cases also have sociodemographic characteristics and familial aggregation of GAD comparable to excessive cases. Individuals who meet all criteria for GAD other than excessiveness have a somewhat milder presentation than those with excessive worry, yet resemble excessive worriers in a number of important ways. These findings challenge the validity of the excessiveness requirement and highlight the need for further research into the optimal definition of GAD.


Innate immune activation in Alzheimer's disease.
Alzheimer's disease (AD) is known as the most predominant cause of dementia among the aged people. Previously, two hallmarks of AD pathology including extracellular amyloid-β (Aβ) deposition and neurofibrillary tangles (NFTs) inside neurons have been identified. With a better understanding of this disease, neuroinflammation has been a focus, and as its initial event, innate immune activation plays an indispensable role. In brain, as an endogenous stimulator, extracellular Aβ deposition activates innate immunity through binding to the pattern recognition receptors (PRR), thus leading to the production and release of substantial inflammatory mediators (NO and ROS) and cytokines (IL-1β, IL-10, IL-33 and TNF-α) contributing to the development of AD. Epidemiologic evidence has suggested an affirmative influence of non-steroidal anti-inflammatory drugs (NSAIDs) on delaying the progression of AD. Therefore, blocking the inflammatory process may be an effective way to delay or even cure AD. In this review, we mainly elucidate the mechanism underlying these immune responses in AD pathogenesis and attempt to seek the therapeutic methods targeting neuroinflammation.


Efficacy and Safety of Ombitasvir, Paritaprevir, and Ritonavir in an Open-Label Study of Patients With Genotype 1b Chronic Hepatitis C Virus Infection With and Without Cirrhosis.
Interferon-free treatment options are rapidly evolving for patients with chronic hepatitis C virus (HCV) genotype 1b (GT1b) infection with cirrhosis and for nonresponders to prior pegylated interferon and ribavirin therapy. We performed a phase 2b, open-label trial of the combination of ombitasvir (a NS5A replication complex inhibitor), paritaprevir, and ritonavir (an NS3/4A protease inhibitor)-an interferon- and ribavirin-free regimen-in difficult-to-treat patients, including prior null responders and patients with cirrhosis. In an international study, 82 patients without cirrhosis (42 treatment-naive and 40 prior null responders) and 99 with cirrhosis (47 treatment-naive and 52 treatment-experienced with prior relapse or a null or partial response) with chronic HCV GT1b infection received ombitasvir (25 mg), paritaprevir (150 mg), and ritonavir (100 mg) once daily for 12 weeks (without cirrhosis) or 24 weeks (with cirrhosis). The primary efficacy endpoint was sustained virologic response 12 weeks after the end of treatment (SVR12). In treatment-naive and null responder patients without cirrhosis, rates of SVR12 were 95.2% and 90.0%, respectively. In treatment-naive and treatment-experienced patients with cirrhosis, rates of SVR12 were 97.9% and 96.2%, respectively. No clinically meaningful differences in rates of SVR12 were observed between patients with or without cirrhosis. Virologic relapse occurred in 3 null responders without cirrhosis and 1 with cirrhosis; virologic breakthrough occurred in 1 null responder without cirrhosis. Common adverse events included headache, asthenia, pruritus, and diarrhea. One patient discontinued taking the drugs because of treatment-related adverse events. An interferon- and ribavirin-free regimen of ombitasvir, paritaprevir, and ritonavir, achieved high rates of SVR12 in patients with HCV GT1b infection with and without cirrhosis. This regimen was well tolerated and was associated with low rates of treatment discontinuation. ClinicalTrials.gov no: NCT01685203.


Introgression of Maize Diversity for Drought Tolerance: Subtropical Maize Landraces as Source of New Positive Variants.
Current climate change models predict an increased frequency and intensity of drought for much of the developing world within the next 30 years. These events will negatively affect maize yields, potentially leading to economic and social instability in many smallholder farming communities. Knowledge about the genetic resources available for traits related to drought tolerance has great importance in developing breeding program strategies. The aim of this research was to study a maize landrace introgression panel to identify chromosomal regions associated with a drought tolerance index. For that, we performed Genome-Wide Association Study (GWAS) on 1326 landrace progenies developed by the CIMMYT Genetic Resources Program, originating from 20 landraces populations collected in arid regions. Phenotypic data were obtained from early testcross trials conducted in three sites and two contrasting irrigation environments, full irrigation (well-watered) and reduced irrigation (drought). The populations were genotyped using the DArTSeq® platform, and a final set of 5,695 SNPs markers was used. The genotypic values were estimated using spatial adjustment in a two-stage analysis. First, we performed the individual analysis for each site/irrigation treatment combination. The best linear unbiased estimates (BLUEs) were used to calculate the Harmonic Mean of Relative Performance (HMRP) as a drought tolerance index for each testcross. The second stage was a joint analysis, which was performed using the HMRP to obtain the best linear unbiased predictions (BLUPs) of the index for each genotype. Then, GWAS was performed to determine the marker-index associations and the marker-Grain Yield (GY) associations for the two irrigation treatments. We detected two significant markers associated with the drought-tolerance index, four associated with GY in drought condition, and other four associated with GY in irrigated conditions each. Although each of these markers explained less than 0.1% of the phenotypic variation for the index and GY, we found two genes likely related to the plant response to drought stress. For these markers, alleles from landraces provide a slightly higher yield under drought conditions. Our results indicate that the positive diversity delivered by landraces are still present on the backcrosses and this is a potential breeding strategy for improving maize for drought tolerance and for trait introgression bringing new superior allelic diversity from landraces to breeding populations.


Photocatalytic applications of paper-like poly(vinylidene fluoride)-titanium dioxide hybrids fabricated using a combination of electrospinning and electrospraying.
A paper-like photocatalyst was fabricated by electrospraying an N,N'-dimethylformamide (DMF) dispersion of titanium dioxide (TiO2) nanoparticles (NPs) on a poly(vinylidene fluoride) nanofiber (PVDF NF) mat prepared by electrospinning. Morphological studies revealed that the TiO2 NPs uniformly deposited as clusters on the surface of the PVDF NF mat. The immobilized amount of TiO2 was found to be 2.08, 2.44, 3.80, and 4.73 mg per 45 cm(2) of PVDF-TiO2 hybrids for the electrospraying of 10, 20, 40, and 60 ml of TiO2-DMF, respectively. The hybrid photocatalysts were effective in degrading bisphenol A (BPA), 4-chlorophenol (4-CP), and cimetidine (CMT), which dissolved in both deionized water and secondary wastewater effluents, with activity being proportional to the quantity of TiO2 NPs immobilized. For the highest loading amount of TiO2, BPA, 4-CP, and CMT degraded completely within 100, 100, and 40 min of UV irradiation, respectively. Stable photo-oxidation of CMT was maintained through 10 repeated cycles. During these cycles, it was confirmed that there was no loss of TiO2 NPs by inductively coupled plasma optical emission spectrometry. Our results suggest that effective and stable PVDF-TiO2 hybrid photocatalysts can be fabricated on a large scale by combining electrospinning and electrospraying techniques.


A Compressed-Sensing Approach for Super-Resolution Reconstruction of Diffusion MRI.
We present an innovative framework for reconstructing high-spatial-resolution diffusion magnetic resonance imaging (dMRI) from multiple low-resolution (LR) images. Our approach combines the twin concepts of compressed sensing (CS) and classical super-resolution to reduce acquisition time while increasing spatial resolution. We use subpixel-shifted LR images with down-sampled and non-overlapping diffusion directions to reduce acquisition time. The diffusion signal in the high resolution (HR) image is represented in a sparsifying basis of spherical ridgelets to model complex fiber orientations with reduced number of measurements. The HR image is obtained as the solution of a convex optimization problem which can be solved using the proposed algorithm based on the alternating direction method of multipliers (ADMM). We qualitatively and quantitatively evaluate the performance of our method on two sets of in-vivo human brain data and show its effectiveness in accurately recovering very high resolution diffusion images.


Involvement of Matrin 3 and SFPQ/NONO in the DNA damage response.
The DNA damage response (DDR) is a complex signaling network that is induced by DNA lesions and vigorously activated by double strand breaks (DSBs). The DSB response is mobilized by the nuclear protein kinase ATM, which phosphorylates key players in its various branches. SFPQ (PSF) and NONO (p54) are nuclear proteins that interact with each other and have diverse roles in nucleic acids metabolism. The SFPQ/NONO heterodimer was previously found to enhance DNA strand break rejoining in vitro. Our attention was drawn to these two proteins as they interact with the nuclear matrix protein Matrin 3 (MATR3), which we found to be a novel ATM target. We asked whether SFPQ and NONO too are involved in the DSB response. Proteins that function at the early phase of this response are often recruited to the damaged sites. We observed rapid recruitment of SFPQ/NONO to sites of DNA damage induced by laser microbeam. In MATR3 knockdown cells SFPQ/NONO retention at DNA damage sites was prolonged. SFPQ and MATR3 depletion led to abnormal accumulation of cells at the S-phase of the cell cycle following treatment with the radiomimetic chemical neocarzinostatin. Notably, proteins involved in DSB repair via nonhomologous end-joining co-immunoprecipitated with NONO; SFPQ depletion delayed DSB repair. Collectively the data suggest that SFPQ, NONO and MATR3 are involved in the early stage of the DSB response, setting the scene for DSB repair.


Poverty and non-communicable diseases in South Africa.
High levels of wealth inequality with improved health statistics in South Africa (SA) provide an important opportunity to investigate non-communicable diseases (NCDs) among the poor. This paper uses two distinct national data sets to contrast patterns of mortality in rich and poor areas and explore the associations between poverty, risk factors, health care and selected NCDs diseases in South African adults. Causes of premature mortality in 1996 experienced in the poorest magisterial districts are compared with those in the richest, using average household wealth to classify districts. Logistic and multinomial regression are used to investigate the association of a household asset index and selected chronic conditions, related risk factors and healthcare indicators using data from the 1998 South African Demographic and Health Survey. NCDs accounted for 39% and 33% of premature mortality in rich and poor districts respectively. The household survey data showed that the risk factors hypertension and obesity increased with increasing wealth, while most of the lifestyle factors, such as light smoking, domestic exposure to ;;smoky'' fuels and alcohol dependence were associated with poverty. Treatment status for hypertension and asthma was worse for poor people than for rich people. The study suggests that NCDs and lifestyle-related risk factors are prevalent among the poor in SA and treatment for chronic diseases is lacking for poor people. The observed increase in hypertension and obesity with wealth suggests that unless comprehensive health promotion strategies are implemented, there will be an unmanageable chronic disease epidemic with future socioeconomic development in SA.


Physical characterization and in vitro biological impact of highly aggregated antibodies separated into size-enriched populations by fluorescence-activated cell sorting.
An IgG2 monoclonal antibody (mAb) solution was subjected to stirring, generating high concentrations of nanometer and subvisible particles, which were then successfully size-enriched into different size bins by low-speed centrifugation or a combination of gravitational sedimentation and fluorescence-activated cell sorting (FACS). The size-fractionated mAb particles were assessed for their ability to elicit the release of cytokines from a population of donor-derived human peripheral blood mononuclear cells (PBMC) at two phases of the immune response. Fractions enriched in nanometer-sized particles showed a lower response than those enriched in micron-sized particles in this assay. Particles of 5-10 μm in size displayed elevated cytokine release profiles compared with other size ranges. Stir-stressed mAb particles had amorphous morphology, contained protein with partially altered secondary structure, elevated surface hydrophobicity (compared with controls), and trace levels of elemental fluorine. FACS size-enriched the mAb particle samples, yet did not notably alter the overall morphology or composition of particles as measured by microflow imaging, transmission electron microscopy, and scanning electron microscopy-energy dispersive X-ray spectroscopy. The utility and limitations of FACS for size separation of mAb particles and potential of in vitro PBMC studies to rank-order the immunogenic potential of various types of mAb particles are discussed.


Latent Nested Nonparametric Priors (with Discussion).
Discrete random structures are important tools in Bayesian nonparametrics and the resulting models have proven effective in density estimation, clustering, topic modeling and prediction, among others. In this paper, we consider nested processes and study the dependence structures they induce. Dependence ranges between homogeneity, corresponding to full exchangeability, and maximum heterogeneity, corresponding to (unconditional) independence across samples. The popular nested Dirichlet process is shown to degenerate to the fully exchangeable case when there are ties across samples at the observed or latent level. To overcome this drawback, inherent to nesting general discrete random measures, we introduce a novel class of latent nested processes. These are obtained by adding common and group-specific completely random measures and, then, normalizing to yield dependent random probability measures. We provide results on the partition distributions induced by latent nested processes, and develop a Markov Chain Monte Carlo sampler for Bayesian inferences. A test for distributional homogeneity across groups is obtained as a by-product. The results and their inferential implications are showcased on synthetic and real data.


Load-dependent relationships between frontal fNIRS activity and performance: A data-driven PLS approach.
Neuroimaging research frequently demonstrates load-dependent activation in prefrontal and parietal cortex during working memory tasks such as the N-back. Most of this work has been conducted in fMRI, but functional near-infrared spectroscopy (fNIRS) is gaining traction as a less invasive and more flexible alternative to measuring cortical hemodynamics. Few fNIRS studies, however, have examined how working memory load-dependent changes in brain hemodynamics relate to performance. The current study employs a newly developed and robust statistical analysis of task-based fNIRS data in a large sample, and demonstrates the utility of data-driven, multivariate analyses to link brain activation and behavior in this modality. Seventy participants completed a standard N-back task with three N-back levels (N = 1, 2, 3) while fNIRS data were collected from frontal and parietal cortex. Overall, participants showed reliably greater fronto-parietal activation for the 2-back versus the 1-back task, suggesting fronto-parietal fNIRS measurements are sensitive to differences in cognitive load. The results for 3-back were much less consistent, potentially due to poor behavioral performance in the 3-back task. To address this, a multivariate analysis (behavioral partial least squares, PLS) was conducted to examine the interaction between fNIRS activation and performance at each N-back level. Results of the PLS analysis demonstrated differences in the relationship between accuracy and change in the deoxyhemoglobin fNIRS signal as a function of N-back level in eight mid-frontal channels. Specifically, greater reductions in deoxyhemoglobin (i.e., more activation) were positively related to performance on the 3-back task, unrelated to accuracy in the 2-back task, and negatively associated with accuracy in the 1-back task. This pattern of results suggests that the metabolic demands correlated with neural activity required for high levels of accuracy vary as a consequence of task difficulty/cognitive load, whereby more automaticity during the 1-back task (less mid-frontal activity) predicted superior performance on this relatively easy task, and successful engagement of this mid-frontal region was required for high accuracy on a more difficult and cognitively demanding 3-back task. In summary, we show that fNIRS activity can track working memory load and can uncover significant associations between brain activity and performance, thus opening the door for this modality to be used in more wide-spread applications.


A novel laboratory cultivation method to examine antibiotic-resistance-related microbial risks in urban water environments.
Although microbial risk due to antibiotic-resistant microbes in water has been a serious public health concern, the current culture-dependent detecting methods using nutrient-rich conditions may not be suitable for evaluating microbial risk. In the present study, a novel cultivation method was developed to detect antibiotic-resistant microbes at various nutrient levels. A nutrient-rich medium Luria-Bertani broth (LB) was diluted at a wide range of dilution factors (10(0)-10(4)) and amended with either tetracycline or vancomycin. A standard environmental cultivation medium, R2A, was also used by amending with antibiotics. The diluted LB with antibiotics was able to detect previously known oligotrophic and antibiotic-resistant bacteria in drinking water and swine manure samples, respectively. These results validate the capability of the new method to detect antibiotic-resistant microbes in various environmental water samples. Using the developed method in assessing microbial risk due to antibiotic-resistant microbes in river and municipal wastewater plants, we found that the viable counts and antibiotic resistance fraction were significantly influenced by the type and concentration level of antibiotic exposure and the cultivation medium conditions. This suggests a further need to standardize cultivation method procedures, to assess microbial risk in water samples.


'Team capital' in quality improvement teams: findings from an ethnographic study of front-line quality improvement in the NHS.
Teamwork is important in the design and delivery of initiatives in complex healthcare systems but the specifics of quality improvement (QI) teams are not well studied. To explain the functioning of front-line healthcare teams working on patient-centred QI using Bourdieu's sociological construct of capital. One medical ward from each of six NHS Trusts in England participated in the study, purposively selected for a range of performance levels on patient experience metrics. Three ethnographers conducted focused ethnography for 1 year, using interviews and observations to explore the organisation, management and delivery of patient-centred QI projects by the six front-line teams. Data were analysed using Bourdieu's typology of the four forms of capital: economic, social, symbolic and cultural. While all teams implemented some QI activities to improve patient experience, progress was greater where teams included staff from a broad range of disciplines and levels of seniority. Teams containing both clinical and non-clinical staff, including staff on lower grades such as healthcare assistants and clerks, engaged more confidently with patient experience data than unidisciplinary teams, and implemented a more ambitious set of projects. We explain these findings in terms of 'team capital'. Teams that chose to restrict membership to particular disciplines appeared to limit their capital, whereas more varied teams were able to draw on multiple resources, skills, networks and alliances to overcome challenges. Staff of varying levels of seniority also shared and valued a broader range of insights into patient experience, including informal knowledge from daily practice. The construct of 'team capital' has the potential to enrich understanding of the mechanism of teamwork in QI work.


Multi-center, prospective, randomized, controlled investigational device exemption clinical trial comparing Mobi-C Cervical Artificial Disc to anterior discectomy and fusion in the treatment of symptomatic degenerative disc disease in the cervical spine.
Anterior cervical discectomy and fusion (ACDF) is the gold standard for treating symptomatic cervical disc degeneration. Cervical total disc replacements (TDRs) have emerged as an alternative for some patients. The purpose of this study was to evaluate the safety and effectiveness of a new TDR device compared with ACDF for treating single-level cervical disc degeneration. This was a prospective, randomized, controlled, multicenter Food and Drug Administration (FDA) regulated Investigational Device Exemption (IDE) study. A total of 245 patients were treated (164 TDR: 81 ACDF). The primary outcome measure was overall success based on improvement in Neck Disability Index (NDI), no subsequent surgical interventions, and no adverse events (AEs) classified as major complications. Secondary outcome measures included SF-12, visual analog scale (VAS) assessing neck and arm pain, patient satisfaction, radiographic range of motion, and adjacent level degeneration. Patients were evaluated preoperatively and postoperatively at 6 weeks, 3, 6, 12, 18, and 24 months. The hypothesis was that the TDR success rate was non-inferior to ACDF at 24 months. Overall success rates were 73.6% for TDR and 65.3% for ACDF, confirming non-inferiority (p < 0.0025). TDR demonstrated earlier improvements with significant differences in NDI scores at 6 weeks and 3 months, and VAS neck pain and SF-12 PCS scores at 6 weeks (p<0.05). Operative level range of motion in the TDR group was maintained throughout follow-up. Radiographic evidence of inferior adjacent segment degeneration was significantly greater with ACDF at 12 and 24 months (p < 0.05). AE rates were similar. Mobi-C TDR is a safe and effective treatment for single-level disc degeneration, producing outcomes similar to ACDF with less adjacent segment degeneration. Level I. This study adds to the literature supporting cervical TDR as a viable option to ACDF in appropriately selected patients with disc degeneration.


Detection of Escherichia coli O157:H7 by multiplex PCR.
In order to develop a PCR assay for Escherichia coli O157:H7, a portion of the 60-MDa plasmid harbored by enterohemorrhagic E. coli (EHEC) was sequenced and PCR primers were designed. A multiplex PCR method was then designed by employing primers specific for the EHEC eaeA gene, conserved sequences of Shiga-like toxins I (SLT-I) and II (SLT-II), and the 60-MDa plasmid. PCR products of 1,087 bp (eaeA), 227 and/or 224 bp (SLT-I and/or SLT-II), and 166 bp (plasmid) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC of serogroup O157.


Viral Cultures for Coronavirus Disease 2019 Infectivity Assessment: A Systematic Review.
We aimed to review the evidence from studies relating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) culture with the results of reverse-transcription polymerase chain reaction (RT-PCR) and other variables that may influence the interpretation of the test, such as time from symptom onset. We searched LitCovid, medRxiv, Google Scholar, and the World Health Organization coronavirus disease 2019 (COVID-19) database for COVID-19 up to 10 September 2020. We included studies attempting to culture or observe SARS-CoV-2 in specimens with RT-PCR positivity. Studies were dual-extracted and the data summarized narratively by specimen type. Where necessary, we contacted corresponding authors of included papers for additional information. We assessed quality using a modified Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS 2) risk-of-bias tool. We included 29 studies reporting attempts at culturing, or observing tissue infection by, SARS-CoV-2 in sputum, nasopharyngeal or oropharyngeal, urine, stool, blood, and environmental specimens. The quality of the studies was moderate with lack of standardized reporting. The data suggest a relationship between the time from onset of symptom to the timing of the specimen test, cycle threshold (Ct), and symptom severity. Twelve studies reported that Ct values were significantly lower and log copies higher in specimens producing live virus culture. Two studies reported that the odds of live virus culture were reduced by approximately 33% for every 1-unit increase in Ct. Six of 8 studies reported detectable RNA for >14 days, but infectious potential declined after day 8 even among cases with ongoing high viral loads. Four studies reported viral culture from stool specimens. Complete live viruses are necessary for transmission, not the fragments identified by PCR. Prospective routine testing of reference and culture specimens and their relationship to symptoms, signs, and patient co-factors should be used to define the reliability of PCR for assessing infectious potential. Those with high Ct are unlikely to have infectious potential.


Accurate quantum-chemical description of gold complexes with pyridine and its derivatives.
Interaction of gold with pyridine and its derivatives was studied by means of different wavefunction-based correlation methods and standar DFT functionals as well as accounting for dispersion correction. Comparison of the calculated binding energies with benchmark CCSD(T)results allows us to find an appropriate computational method, when considering the two structures reflecting the interaction of gold with the lone pair at nitrogen, on the one hand, and with the π-system of pyridine, on the other hand. Additional binding sites were evaluated, when performing potential energy surface calculations and structure optimizations. The enhancement of the interaction energy due to donor substituents in the 4-position of the pyridine molecule has been investigated.


Long-chain polyunsaturated fatty acids: selected mechanisms of action on bone.
Evidence presented over the past 20 years has shown that long-chain polyunsaturated fatty acids (LCPUFAs), especially the n-3 fatty acids such as eicospentaenoic acid (EPA) and docosahexaenoic acid (DHA) are beneficial for bone health. Some studies in humans indicate that LCPUFAs can increase bone formation, affect peak bone mass in adolescents and reduce bone loss as measured using bone mineral densitometry. The cellular mechanisms of action of the LCPUFAs, however, are complex and involve modulation of fatty acid metabolites such as prostaglandins, resolvins and protectins, several signalling pathways, cytokines and growth factors. LCPUFAs affect receptor activator of nuclear factor κβ (RANK), a receptor found on the osteoclast, the cell causing bone resorption, which controls osteoclast formation. Lipoxygenase (LOX) generated lipid mediators (resolvins, lipoxins, protectins and docosanoids) have both anti-inflammatory and pro-resolving activities. Both resolvins and lipoxins inhibit inflammation-induced bone resorption. Arachidonic acid significantly upregulates inducible NO synthase (iNOS) mRNA expression in human osteoblast-like cells, thereby possibly enhancing osteoclastic activity. The protective effect of EPA on osteoblastogenesis could be mediated by the biphasic cross-talk between PGE(2) and NO production involving COX-2 and iNOS pathways. Other mediators of osteoblast maturation include PPARα ligands such as linoleic acid and possibly DHA in association with bone morphogenic proteins. Since DHA is a weaker ligand for PPARγ, more uncommitted mesenchymal stem cells are thought to differentiate into osteoblasts rather than adipocytes. This review addresses selected cellular mechanisms that may explain the beneficial effects of the LCPUFAs on bone.


An unattended device for sleep-related breathing disorders: validation study in suspected obstructive sleep apnoea syndrome.
Portable devices for the diagnosis of obstructive sleep apnoea (OSA) are considered to be an acceptable alternative to polysomnography (PSG), but their validation is essential. The aim of our study was to validate a device specifically designed for OSA diagnosis. Twenty nine suspected OSA patients were studied with simultaneous nocturnal PSG and an unattended recording device (MicroDigitrapper-S) (M-S). The device measured body position, snoring sound, oronasal flow, thoracic and abdominal effort, heart rate and percentage arterial oxygen saturation (Sa,O2%). We compared the apnoea plus hypopnoea index (AHI) and Sa,O2% results of PSG with that of the system's automatic analysis (M-SA). We also performed a semiautomatic analysis (M-SS) with visual editing of the raw data. Results at different AHI cut-off levels were analysed to obtain an indication of accuracy in diagnosis and severity. Both M-SA and M-SS showed a sensitivity and specificity of 100% at the cut-off level of AHI > 10. When increasing the cut-off levels, M-SA sensitivity decreased (55% for AHI > 40), while specificity remained high (95%). This was improved to a clinically acceptable level of agreement by M-SS analysis (sensitivity 91% and specificity 94%). In conclusion, the MicroDigitrapper-S device showed a good sensitivity and specificity for the diagnosis of OSA. However, the device could not predict the severity of OSA precisely enough. In severe cases (apnoea plus hypopnoea index > 40), semi-automatic scoring was necessary to obtain a more accurate detection of the severity of the disease.


Interactions in sheep between tall fescue ergot alkaloids and hepatotoxic carbon tetrachloride and Senecio pyrrolizidine alkaloids.
The interaction between ergot alkaloids in endophyte-infected (E+) tall fescue (Festuca arundinacea) seed and pyrrolizidine alkaloids (PA) in tansy ragwort (TR; Senecio jacobaea) when simultaneously fed to sheep was investigated. Because of the hepatogenic effects of prolactin (PRL), it was hypothesized that low serum PRL induced by ergot alkaloids would increase the susceptibility of sheep to hepatotoxicity induced by PA. Sheep are normally resistant to PA-induced hepatotoxicity. Twenty-four wether lambs weighing 34.1 +/- 2.3 kg were used after being randomly assigned to 1 of 6 treatments of 4 wethers each. The diets offered were a control basal diet containing endophyte-free (E-) tall fescue seed, control + carbon tetrachloride (CCL4), E+ tall fescue basal diet, E+2CCl4, E(-)+TR, or E+2TR. The diets were composed of 50% alfalfa meal, 34.5% rolled barley, 5% soybean meal, 4% cane molasses, 0.5% trace mineralized salt and 6% tall fescue (E- or E+) seed. In the 2 TR treatments 25% of the alfalfa was replaced by TR. Interaction between ergot alkaloids in E+ tall fescue and PA in TR was assessed by gamma glutamyl transferase (GGTP) activity and/or sulfobromophthalein (BSP) clearance rate, both of which measure liver function. Serum GGTP activity was measured on days 14 and 28; plasma BSP clearance was monitored on days 28 and 70 by collection of blood at 4, 8 and 16 min after i.v. BSP injection. Serum PRL assays were performed on days 14, 70 and 84. Serum GGTP activity was elevated by CCl4 drenching on days 14 and 28. On day 28 TR feeding reduced GGTP activity, but there was no difference between the 2 TR treatments. There was no difference in the mean BSP half-times (t1/2) and % BSP retentions on day 28 among any of the 6 treatments. On day 70, there was no difference in the t1/2 or % BSP retention 4 min after BSP injection among any of the treatments. At 8 min after BSP injection, however, % BSP retention was significantly higher for the control +CCl4 than for the E+2CCl4 treatment. At 16 min after BSP injection, the E+ treatment had a higher mean % BSP retention value than the E+2CCl4 treatment; there were no differences among the other treatments. Serum PRL levels were reduced by E+ or CCl4 on all 3 dates of PRL evaluation. The lack of interaction between ergot alkaloids in E+ tall fescue seed and PA in TR as assessed by GGTP activity and/or BSP clearance in sheep may imply that the target sites o the 2 alkaloids are metabolically different. The low serum PRL with E+ did not increase susceptibility of sheep to CCl4 or PA.


Molecular characterization of the beta-lactamases from clinical isolates of Moraxella (Branhamella) catarrhalis obtained from 24 U.S. medical centers during 1994-1995 and 1997-1998.
The beta-lactamases from 403 Moraxella (Branhamella) catarrhalis clinical isolates obtained during 1994-1995 and 1997-1998 U.S. multicenter surveillance studies were characterized by isoelectric focusing. The overall prevalences of the BRO-1 and BRO-2 enzymes among beta-lactamase-positive isolates were estimated to be 97.5 and 2.5%, respectively. The minimum inhibitory concentrations (MICs) of ampicillin for all BRO-2-producing isolates were </=1 microg/ml; however, numerous beta-lactamase-positive isolates for which the ampicillin MICs were </=1 microg/ml produced the BRO-1 enzyme (88. 1%).


Victim voice in reenvisioning responses to sexual and physical violence nationally and internationally.
Internationally and in the United States many victims of sexual assault and domestic violence are unserved, underserved, or ill-served, especially those from the most vulnerable populations. Programs developed in the United States are routinely exported to developing countries but often without success. Notably, the failures seen internationally resemble those in the United States and are related to structural and attitudinal-cultural factors. Many victims do not disclose, and if they do seek services, they often report that available options mismatch their objectives, present accessibility challenges, disempower their pursuit of justice, and fail to augment needed resources. A deeper understanding of obstacles to effective service provision is needed if the United States is to continue to be an international partner in victim response and violence prevention. This article builds on what is known about service delivery challenges in U.S. programs to envision a path forward that concomitantly accommodates anticipation of shrinking resources, by (a) reviewing illustrative services and feedback from victims about utilizing them; (b) examining structural inequalities and the intersections of personal and contextual features that both increase vulnerability to victimization and decrease accessibility and acceptability of services; (c) advocating for reintroduction of direct victim voice into response planning to enhance reach and relevance; and (d) reorienting delivery systems, community partnerships, and Coordinated Community Response teams. The authors suggest as the way forward pairing direct victim voice with open-minded listening to expressed priorities, especially in vulnerable populations, and designing services accordingly. Through a process that prioritizes adaptation to diverse needs and cultures, U.S models can increase desirability, equity, and thrift at home as well as enhance international relevance. (PsycINFO Database Record


Adenosine-mediated inhibition of cytotoxic activity and cytokine production by IL-2/NKp46-activated NK cells: involvement of protein kinase A isozyme I (PKA I).
Adenosine suppresses the production of various cytokines/ chemokines and inhibits the cytotoxic activity of murine and human NK cells activated with IL-2 or Ly49D, NKp46-receptor crosslinking, respectively. These effects are mediated by the type A2A adenosine receptor via stimulation of adenylyl cyclase, increased production of cAMP, and activation of PKA. PKA I, but not PKA II, participates in the inhibitory effects of adenosine. Blocking regulatory, but not catalytic, subunits of PKA I abrogates the inhibitory effects of adenosine. These findings suggest that tumor-produced adenosine inhibits the activity of NK and other effector cells and thereby protects tumors from immune-mediated destruction.


Analysis of the transgene insertion pattern in a transgenic mouse strain using long-read sequencing.
Transgene insertion patterns are critical for the analysis of transgenic animals because the influence of transgenes may change depending on the insertion pattern (such as copy numbers and orientations of concatenations) and the insertion position in the genome. We previously reported a genomic walking strategy to locate transgenes in the genomes of transgenic mice (Exp. Anim. 53: 103-111, 2004) and to analyze transgene insertion patterns (Exp. Anim. 55: 65-69, 2006). With such strategies, however, we could not determine the copy number of transgenes or global genome modification induced by transgene insertion due to read-length limitation. In this study, we used a long-read sequencer (MinION, Oxford Nanopore Technologies) to overcome this limitation. We obtained 922,210 reads using MinION with genomic DNA from a transgenic mouse strain (4C30, Proc. Jpn. Acad. Ser. B. Phys. Biol. Sci. 87: 550-562, 2011). Among the reads, we found one 21,457-bp read containing the transgene using a local BLAST search. Nucleotide dot plot analysis revealed that the transgene was inserted in the genome as a tandem concatemer with an almost entire construct (15-3,508 of 3,508 bp) and a partial fragment (4-660, 657 bp). Ensembl's BLAST search against the C57BL/6N genome revealed a 9,388-bp deletion at the insertion position in the intron of the Sgcd gene, confirming that mutations such as a large genomic deletion could occur at the time of transgene insertion. Thus, long-read sequencers are useful tools for the analysis of transgene insertion patterns.


Global sea level trend in the past century.
Data derived from tide-gauge stations throughout the world indicate that the mean sea level rose by about 12 centimeters in the past century. The sea level change has a high correlation with the trend of global surface air temperature. A large part of the sea level rise can be accounted for in terms of the thermal expansion of the upper layers of the ocean. The results also represent weak indirect evidence for a net melting of the continental ice sheets.


Automatic detection of abnormalities in chest radiographs using local texture analysis.
A fully automatic method is presented to detect abnormalities in frontal chest radiographs which are aggregated into an overall abnormality score. The method is aimed at finding abnormal signs of a diffuse textural nature, such as they are encountered in mass chest screening against tuberculosis (TB). The scheme starts with automatic segmentation of the lung fields, using active shape models. The segmentation is used to subdivide the lung fields into overlapping regions of various sizes. Texture features are extracted from each region, using the moments of responses to a multiscale filter bank. Additional "difference features" are obtained by subtracting feature vectors from corresponding regions in the left and right lung fields. A separate training set is constructed for each region. All regions are classified by voting among the k nearest neighbors, with leave-one-out. Next, the classification results of each region are combined, using a weighted multiplier in which regions with higher classification reliability weigh more heavily. This produces an abnormality score for each image. The method is evaluated on two databases. The first database was collected from a TB mass chest screening program, from which 147 images with textural abnormalities and 241 normal images were selected. Although this database contains many subtle abnormalities, the classification has a sensitivity of 0.86 at a specificity of 0.50 and an area under the receiver operating characteristic (ROC) curve of 0.820. The second database consist of 100 normal images and 100 abnormal images with interstitial disease. For this database, the results were a sensitivity of 0.97 at a specificity of 0.90 and an area under the ROC curve of 0.986.


Virus Adaptation and Selection Following Challenge of Animals Vaccinated against Classical Swine Fever Virus.
Vaccines against classical swine fever have proven very effective in protecting pigs from this deadly disease. However, little is known about how vaccination impacts the selective pressures acting on the classical swine fever virus (CSFV). Here we use high-throughput sequencing of viral genomes to investigate evolutionary changes in virus populations following the challenge of naïve and vaccinated pigs with the highly virulent CSFV strain "Koslov". The challenge inoculum contained an ensemble of closely related viral sequences, with three major haplotypes being present, termed A, B, and C. After the challenge, the viral haplotype A was preferentially located within the tonsils of naïve animals but was highly prevalent in the sera of all vaccinated animals. We find that the viral population structure in naïve pigs after infection is very similar to that in the original inoculum. In contrast, the viral population in vaccinated pigs, which only underwent transient low-level viremia, displayed several distinct changes including the emergence of 16 unique non-synonymous single nucleotide polymorphisms (SNPs) that were not detectable in the challenge inoculum. Further analysis showed a significant loss of heterogeneity and an increasing positive selection acting on the virus populations in the vaccinated pigs. We conclude that vaccination imposes a strong selective pressure on viruses that subsequently replicate within the vaccinated animal.


Why do older women have poor implantation rates? A possible role of the mitochondria.
Mitochondria are organelles responsible for oxidative phosphorylation, the main energy source for all eukaryotic cells. In oocytes and embryos, it seems that mitochondria provide sufficient energy for fecundation by supporting spindle formation during meiosis II, and for implantation. Since mitochondria are inherited from mother to child, it is important that oocyte mitochondria should be intact. Older women seem to have more mitochondrial DNA mutations, which can be responsible for poor implantation and aneuploidy, two conditions that occur more often in this group. In the present report we propose a new model to explain why older women have poor implantation rates.


Aperture phase modulation with adaptive optics: a novel approach for speckle reduction and structure extraction in optical coherence tomography.
Speckle is an inevitable consequence of the use of coherent light in imaging and acts as noise that corrupts image formation in most applications. Optical coherence tomographic imaging, as a technique employing coherence time gating, suffers from speckle. We present here a novel method of suppressing speckle noise intrinsically compatible with adaptive optics (AO) for confocal coherent imaging: modulation of the phase in the system pupil aperture with a segmented deformable mirror (DM) to introduce minor perturbations in the point spread function. This approach creates uncorrelated speckle patterns in a series of images, enabling averaging to suppress speckle noise while maintaining structural detail. A method is presented that efficiently determines the optimal range of modulation of DM segments relative to their AO-optimized position so that speckle noise is reduced while image resolution and signal strength are preserved. The method is active and independent of sample properties. Its effectiveness and efficiency are quantified and demonstrated by both ex vivo non-biological and in vivo biological applications.


[Atherosclerosis-related aortic dissection].
Penetrating atherosclerotic aortic ulcers (PAU) can cause aortic dissection. Of 38 autopsy cases with aortic dissection, 6 (15.8%) had severe atherosclerotic changes, resembling those of PAU, at the site of entry (SE). Clinicopathological data on these patients were compared with those on 32 cases with nonatheromatous dissection (5 with Marfan syndrome or its forme fruste and 27 without Marfan syndrome) and 13 with atherosclerotic saccular aneurysms. For control study, the aorta of a 44-year-old woman who died of pulmonary cancer was used. Compare to nonatheromatous dissection, atherosclerosis-related aortic dissections were found in older women. Four cases were complicated by saccular aneurysms of the aorta. The SE was located in the ascending aorta in 1 and the descending aorta in 5. These sites usually were ulcerated atheromatous plaques or longitudinal fissures rather than transverse tears. Immunohistochemical examination of the SE revealed that MMP-1, 2, 9 and TIMP-2 were expressed in macrophages and/or interstitium, similar to the findings in atheromatous plaque or PAU. We propose that atherosclerosis-related aortic dissection differs from the usual classical aortic dissection. Patients with this lesion have a high risk of re-dissection from the new SE in the same lesion.


Evaluation of a new generation of oral contraceptives. The Advisory Board for the New Progestins.
To assess the differences and similarities in efficacy, safety, and metabolic effects of oral contraceptives (OCs) containing the new progestins desogestrel, gestodene, and norgestimate. All formulations reviewed contained no more than 35 micrograms ethinyl estradiol. Data were reported from approximately 100 published reports, dating from 1980, of comparative and noncomparative clinical studies on phasic and fixed-dose preparations culled from computer searches of several sources, including MEDLINE and Excerpta Medica. An attempt was made to select the most meaningful studies in terms of length, size, methodology, and quality of description. All clinical studies were considered; in general, review articles were not. Some preclinical studies were also included. No abstracts were used. The indices chosen for examination were contraceptive efficacy, cycle control, coagulation, carbohydrate and lipid metabolism, and androgenicity. The new formulations were found to be comparable in efficacy to each other and to established agents. They also appeared to be less androgenic than current OCs and to have less impact on carbohydrate and lipoprotein metabolism. Cycle control was similar to that of older products. Changes in coagulation-promoting and antithrombotic factors were minor. Clinical relevance of the results could not be determined because of small sample size and methodologic differences between studies.


The glucocorticoid receptor and AP-1 are involved in a positive regulation of the muscle regulatory gene myf5 in cultured myoblasts.
The muscle regulatory factor, myf5, is involved in the establishment of skeletal muscle precursor cells. Little is known, however, about the control of the expression of the gene encoding this basic helix-loop-helix (bHLH) factor. We have addressed this question in the mouse myogenic cell line, C2, and in a derivative of this cell line where the myf5 gene is the only muscle-specific bHLH factor to be expressed at the myoblast stage. We present evidence that the synthetic glucocorticoid dexamethasone, and the pharmacological agent anisomycin, act synergistically to rapidly up-regulate the levels of myf5 transcript and protein. The glucocorticoid antagonist RU 486 abolishes this synergy, demonstrating the involvement of the glucocorticoid receptor. The expression of a dominant negative mutant of c-jun which interferes with the transactivating properties of all AP-1 family members also blocks the induction of myf5 by anisomycin and dexamethasone. An activator of protein kinase C (PKCs), 12-O-tetradecanoyl phorbol 13-acetate (TPA), abolishes the up-regulation of myf5 gene expression by dexamethasone and anisomycin, and its effect is counteracted by an inhibitor of PKCs, GF 109203X. These results point to the possible involvement of PKCs in the negative control of myf5. Evidence that both positive and negative regulation of myf5 transcripts, described here, does not require the fresh synthesis of transcription factors suggests that myf5 may behave like an immediate early gene.


Social evolution in multispecies biofilms.
Microbial ecology is revealing the vast diversity of strains and species that coexist in many environments, ranging from free-living communities to the symbionts that compose the human microbiome. In parallel, there is growing evidence of the importance of cooperative phenotypes for the growth and behavior of microbial groups. Here we ask: How does the presence of multiple species affect the evolution of cooperative secretions? We use a computer simulation of spatially structured cellular groups that captures key features of their biology and physical environment. When nutrient competition is strong, we find that the addition of new species can inhibit cooperation by eradicating secreting strains before they can become established. When nutrients are abundant and many species mix in one environment, however, our model predicts that secretor strains of any one species will be surrounded by other species. This "social insulation" protects secretors from competition with nonsecretors of the same species and can improve the prospects of within-species cooperation. We also observe constraints on the evolution of mutualistic interactions among species, because it is difficult to find conditions that simultaneously favor both within- and among-species cooperation. Although relatively simple, our model reveals the richness of interactions between the ecology and social evolution of multispecies microbial groups, which can be critical for the evolution of cooperation.


The Use of Structural Priming and Focused Recasts to Facilitate the Production of Subject- and Object-Focused Relative Clauses by School-Age Children With and Without Developmental Language Disorder.
Purpose The purpose of this study was to examine the use of structural priming combined with a focused recasting procedure to elicit subject- and object-focused, center-embedded relative clauses from students with developmental language disorders (DLDs) and typically developing (TD) students. Method A total of 26 children (13 DLD, 13 TD), ranging in age from 6;10 to 10;11 (years;months), participated in this study. All children completed a priming and recasting task that targeted subject- and object-focused relatives. The stimuli were presented in two blocks, which each contained 40 trials for each sentence type. Results Children with DLD and their TD peers were significantly more accurate producing subject-focused than object-focused relatives. An analysis of rate of learning indicated that significantly more trials were required for subjects in the DLD group to demonstrate consistent performance on both subject- and object-focused relatives. Conclusions The study supports the feasibility of combining an implicit priming task with an explicit recasting task for teaching subject-focused relative clauses to children with DLD. However, it is likely that additional instruction and/or more trials will be necessary for children with DLD to attain consistent performance levels. Supplemental Material https://doi.org/10.23641/asha.12670847.


Optimal ratio of transcription factors for somatic cell reprogramming.
Somatic cell reprogramming is achieved by four reprogramming transcription factors (RTFs), Oct3/4, Sox2, Klf4, and c-Myc. However, in addition to the induction of pluripotent cells, these RTFs also generate pseudo-pluripotent cells, which do not show Nanog promoter activity. Therefore, it should be possible to fine-tune the RTFs to produce only fully pluripotent cells. For this study, a tagging system was developed to sort induced pluripotent stem (iPS) cells according to the expression levels of each of the four RTFs. Using this system, the most effective ratio (Oct3/4-high, Sox2-low, Klf4-high, c-Myc-high) of the RTFs was 88 times more efficient at producing iPS cells than the worst effective ratio (Oct3/4-low, Sox2-high, Klf4-low, c-Myc-low). Among the various RTF combinations, Oct3/4-high and Sox2-low produced the most efficient results. To investigate the molecular basis, microarray analysis was performed on iPS cells generated under high (Oct3/4-high and Sox2-low) and low (Oct3/4-low and Sox2-high) efficiency reprogramming conditions. Pathway analysis revealed that the G protein-coupled receptor (GPCR) pathway was up-regulated significantly under the high efficiency condition and treatment with the chemokine, C-C motif ligand 2, a member of the GPCR family, enhanced somatic cell reprogramming 12.3 times. Furthermore, data from the analysis of the signature gene expression profiles of mouse embryonic fibroblasts at 2 days after RTF infection revealed that the genetic modifier, Whsc1l1 (variant 1), also improved the efficiency of somatic cell reprogramming. Finally, comparison of the overall gene expression profiles between the high and low efficiency conditions will provide novel insights into mechanisms underlying somatic cell reprogramming.


Effects of a large-scale distribution of water filters and natural draft rocket-style cookstoves on diarrhea and acute respiratory infection: A cluster-randomized controlled trial in Western Province, Rwanda.
Unsafe drinking water and household air pollution (HAP) are major causes of morbidity and mortality among children under 5 in low and middle-income countries. Household water filters and higher-efficiency biomass-burning cookstoves have been widely promoted to improve water quality and reduce fuel use, but there is limited evidence of their health effects when delivered programmatically at scale. In a large-scale program in Western Province, Rwanda, water filters and portable biomass-burning natural draft rocket-style cookstoves were distributed between September and December 2014 and promoted to over 101,000 households in the poorest economic quartile in 72 (of 96) randomly selected sectors in Western Province. To assess the effects of the intervention, between August and December, 2014, we enrolled 1,582 households that included a child under 4 years from 174 randomly selected village-sized clusters, half from intervention sectors and half from nonintervention sectors. At baseline, 76% of households relied primarily on an improved source for drinking water (piped, borehole, protected spring/well, or rainwater) and over 99% cooked primarily on traditional biomass-burning stoves. We conducted follow-up at 3 time-points between February 2015 and March 2016 to assess reported diarrhea and acute respiratory infections (ARIs) among children <5 years in the preceding 7 days (primary outcomes) and patterns of intervention use, drinking water quality, and air quality. The intervention reduced the prevalence of reported child diarrhea by 29% (prevalence ratio [PR] 0.71, 95% confidence interval [CI] 0.59-0.87, p = 0.001) and reported child ARI by 25% (PR 0.75, 95% CI 0.60-0.93, p = 0.009). Overall, more than 62% of households were observed to have water in their filters at follow-up, while 65% reported using the intervention stove every day, and 55% reported using it primarily outdoors. Use of both the intervention filter and intervention stove decreased throughout follow-up, while reported traditional stove use increased. The intervention reduced the prevalence of households with detectable fecal contamination in drinking water samples by 38% (PR 0.62, 95% CI 0.57-0.68, p < 0.0001) but had no significant impact on 48-hour personal exposure to log-transformed fine particulate matter (PM2.5) concentrations among cooks (β = -0.089, p = 0.486) or children (β = -0.228, p = 0.127). The main limitations of this trial include the unblinded nature of the intervention, limited PM2.5 exposure measurement, and a reliance on reported intervention use and reported health outcomes. Our findings indicate that the intervention improved household drinking water quality and reduced caregiver-reported diarrhea among children <5 years. It also reduced caregiver-reported ARI despite no evidence of improved air quality. Further research is necessary to ascertain longer-term intervention use and benefits and to explore the potential synergistic effects between diarrhea and ARI. Clinical Trials.gov NCT02239250.


Identifying the most productive breeding sites for malaria mosquitoes in The Gambia.
Ideally larval control activities should be targeted at sites that generate the most adult vectors, thereby reducing operational costs. Despite the plethora of potential mosquito breeding sites found in the floodplains of the Gambia River, about 150 km from its mouth, during the rainy season, only a small proportion are colonized by anophelines on any day. This study aimed to determine the characteristics of larval habitats most frequently and most densely populated by anopheline larvae and to estimate the numbers of adults produced in different habitats. A case-control design was used to identify characteristics of sites with or without mosquitoes. Sites were surveyed for their physical water properties and invertebrate fauna. The characteristics of 83 sites with anopheline larvae (cases) and 75 sites without (controls) were collected between June and November 2005. Weekly adult productivity was estimated with emergence traps in water-bodies commonly containing larvae. The presence of anopheline larvae was associated with high invertebrate diversity (Odds Ratio, OR 11.69, 95% CI 5.61-24.34, p < 0.001), the presence of emergent vegetation (OR 2.83, 95% CI 1.35-5.95, p = 0.006), and algae (at borderline significance; OR 1.87, 95% CI 0.96-3.618, p = 0.065). The density of larvae was reduced in sites that were larger than 100 m in perimeter (OR 0.151; 95% CI 0.060-0.381, p < 0.001), where water was tidal (OR 0.232; 95% CI 0.101-0.533, p = 0.001), vegetation shaded over 25% of the habitat (OR 0.352; 95% CI 0.136-0.911, p = 0.031) and water conductivity was above 2,000 muS/cm (OR 0.458; 95% CI 0.220-0.990, p = 0.048). Pools produced the highest numbers of Anopheles gambiae adults compared with rice fields, floodwater areas close to the edge of the floodplain or close to the river, and stream fringes. Pools were characterized by high water temperature and turbidity, low conductivity, increased presence of algae, and absence of tidal water. There are few breeding sites that produce a high number of adult vectors in the middle reaches of the river in The Gambia, whereas those with low productivity are larger in area and can be found throughout the rainy season. Even though risk factors could be identified for the presence and density of larvae and productivity of habitats, the results indicate that anti-larval interventions in this area of The Gambia cannot be targeted in space or time during the rainy season.


Inhibitory Interplay between Orexin Neurons and Eating.
In humans and rodents, loss of brain orexin/hypocretin (OH) neurons causes pathological sleepiness [1-4], whereas OH hyperactivity is associated with stress and anxiety [5-10]. OH cell control is thus of considerable interest. OH cells are activated by fasting [11, 12] and proposed to stimulate eating [13]. However, OH cells are also activated by diverse feeding-unrelated stressors [14-17] and stimulate locomotion and "fight-or-flight" responses [18-20]. Such OH-mediated behaviors presumably preclude concurrent eating, and loss of OH cells produces obesity, suggesting that OH cells facilitate net energy expenditure rather than energy intake [2, 21-23]. The relationship between OH cells and eating, therefore, remains unclear. Here we investigated this issue at the level of natural physiological activity of OH cells. First, we monitored eating-associated dynamics of OH cells using fiber photometry in free-feeding mice. OH cell activity decreased within milliseconds after eating onset, and remained in a down state during eating. This OH inactivation occurred with foods of diverse tastes and textures, as well as with calorie-free "food," in both fed and fasted mice, suggesting that it is driven by the act of eating itself. Second, we probed the implications of natural OH cell signals for eating and weight in a new conditional OH cell-knockout model. Complete OH cell inactivation in adult brain induced a hitherto unrecognized overeating phenotype and caused overweight that was preventable by mild dieting. These results support an inhibitory interplay between OH signals and eating, and demonstrate that OH cell activity is rapidly controllable, across nutritional states, by voluntary action.


[Body image, nutritional status and practices for weight control among Brazilian adolescents].
The objective of this work is to describe the agreement between body image (BI) and nutritional status (NS) as well as to verify the association of BI and NS with behaviors regarding body weight (BRBW) among Brazilian adolescents. In 2009, a random sample of students in 9th year at public and private schools from 26 Brazilian state capitals and Federal District was studied. It was used a self-administered questionnaire with information about BI, BRBW and socioeconomic factors. Weight and height were measured. Logistic regression analyses were performed to evaluate associations between variables and weighted kappa test was used to verify the agreement between NS and BI. Twenty-four percent of the adolescents had excessive weight (EW) and 17.7% considered themselves fat. Almost 2/3 performed some action to weight control and 7% showed extreme behavior for weight control (EBWC). Low agreement between the NS and BI was observed. Nearly half and 27% of those with EW considered themselves as having normal weight and being thin, respectively. Greater proportion of EW adolescents had EBWC when compared with those with normal weight and underweight. The same was observed in relation to BI. Low agreement between EN and BI was observed. Practices for weight control were observed even among individuals with adequate NS.


Gamma knife stereotactic radiosurgery for intracranial hemangiopericytoma.
Intracranial hemangiopericytoma (HPC) is a rare tumors that comprises under 1% of all primary brain tumors. This tumor is notable for its aggressive clinical behavior, such as a high rate of recurrence after surgical resection and proclivity for extracanial metastases. For the management of recurrent intracranial tumors, we applied Gamma Knife radiosurgery (GKRS) in cases in which the tumors were well-circumscribed and measured less than 3 centimeters in diameter. In this study, we evaluated the efficacy and role of GKRS in controlling recurrent HPC.Materials and Methods: Between April 2004 and July 2010, we treated seven patients with intracranial HPC using GKRS. All patients underwent surgical resection prior to GKRS. The mean age of the patients at the first GKRS was 43.4 (range, 29 to 68) years. At the first GKRS, six patients of seven patients had one tumor, and the remaining patient had four tumors. Therefore, a total of 10 tumors were detected at the first GKRS, all of which were treated. The median follow-up time was 52.1 months (range, 13 to 71 months). During follow-up, three of the seven patients underwent a total of 22 repeat GKRS procedures for newly developed tumors or 4 lesions with regrowth. Ultimately, a total of 32 GKRS procedures were performed in the seven patients. The mean radiosurgery target volume was 4.1 ml (range, 0.3-23.9ml), and the mean marginal dose was 16.5 Gy (range, 10-20). Five patients were alive at the end of the follow-up visit, without any serious neurological deficits. One patient presented with extracranial metastasis and died from cerebrospinal dissemination of the tumor 71 months after the first GKRS. Another patient died from colon cancer during follow-up. The 1-, 3-, and 5-year local tumor control rates were 100%, 92% and 69.7%, respectively. GKRS is an effective management option for patients with recurrent hemangiopericytoma.


[Choice of initial regimen for antiretroviral-naïve HIV patients: Analysis of motivation].
Several therapeutic combination antiretroviral therapy regimen are available for initial treatment in naïve HIV infected patients. The choice of a particular regimen remains often subjective. The aim of this study was to determine factors associated with the choice of molecules in initial ARV prescriptions. From 01/01 to 30/10/2014, every initial cART prescription was analyzed regarding patients and physicians characteristics. Then, prescriptions were evaluated by an independent committee of ART prescribers. One hundred and thirty two consecutive initial prescriptions by 34 physicians of 11 medical centers were included: 71 M, migrants: 57 %, MSM: 21 %, CD4<200/mm3: 26 %, HIV RNA>100 000 cp/mL (33 %). cART regimen were: NRTI/PI (43 %), NRTI/NNRTI (29.5 %), NRTI/integrase inhibitor (23 %). 75 % of initial cART regimen were consistent with expert guidelines recommendations. The choice of initial cART was not influenced by the type of HIV contamination risk group, patient's geographic origin, CD4 levels. In contrast, working or not (P=0.007), pregnancy wish (P=0.07), pregnancy (P=0.001), HIV RNA levels (P=0.02) and HIV primary infection (P=0.049) influenced the initial choice. Neither physician's age, nor physician's experience influenced this choice. The prescription's non accordance to 2013 French guidelines was mainly related to integrase inhibitor utilisation (P= 0.0001). Overall, cART initial choice is mostly consistent with guidelines. Primary HIV infection, procreation features and high viral load are the main factors influencing this choice. New regimen with better tolerability is prescribed even if it is not yet included in the guidelines.


Influence of anesthesia on ocular effects and temperature in rabbit eyes exposed to microwaves.
To investigate the effect of systemic anesthesia on ocular effects and temperature in rabbit eyes exposed to microwaves, one eye each of 43 male pigmented rabbits (Dutch, 1.8-2.2 kg) was exposed at 2.45 GHz for 60-20 min (300 mW/cm2; 108 W/kg), either under anesthesia (ketamine hydrochloride (5 mg/kg) + xylazine (0.23 mg/kg)) or without anesthesia. Changes in the anterior segment were evaluated by image analysis utilizing a Scheimpflug camera, specular microscopy, and a laser flare cell meter. Temperatures within the eye were measured during microwave exposure by a Fluoroptic thermometer. The exposed eyes showed miosis, conjunctival congestion, corneal edema, and an increase in the light scattering of the anterior shallow cortex in the pupillary area of the lens. The group under systemic anesthesia showed much stronger symptoms than those treated without anesthesia. All of the anterior ocular changes disappeared within a week. The highest temperature during exposure was in the vitreous, followed by the anterior chamber, and the retrobulbar cavity of the orbit. The ocular temperatures of the rabbits under systemic anesthesia were 2-9 degrees C higher than those without anesthesia. Body temperature showed an increase of 1 degrees C during the exposure. Acute high intensity microwave exposure temporarily induced anterior segments inflammation and lens changes. The more pronounced ocular effects in the anesthetized rabbits were associated with the significantly higher ocular temperatures in the anesthetized animals. The influence of systemic anesthesia on ocular changes should be considered.


Ultrafast Photoinduced Electron Transfer in a π-Conjugated Oligomer/Porphyrin Complex.
Controlling charge transfer (CT), charge separation (CS), and charge recombination (CR) at the donor-acceptor interface is extremely important to optimize the conversion efficiency in solar cell devices. In general, ultrafast CT and slow CR are desirable for optimal device performance. In this Letter, the ultrafast excited-state CT between platinum oligomer (DPP-Pt(acac)) as a new electron donor and porphyrin as an electron acceptor is monitored for the first time using femtosecond (fs) transient absorption (TA) spectroscopy with broad-band capability and 120 fs temporal resolution. Turning the CT on/off has been shown to be possible either by switching from an organometallic oligomer to a metal-free oligomer or by controlling the charge density on the nitrogen atom of the porphyrin meso unit. Our time-resolved data show that the CT and CS between DPP-Pt(acac) and cationic porphyrin are ultrafast (approximately 1.5 ps), and the CR is slow (ns time scale), as inferred from the formation and the decay of the cationic and anionic species. We also found that the metallic center in the DPP-Pt(acac) oligomer and the positive charge on the porphyrin are the keys to switching on/off the ultrafast CT process.


Bile salt secretion in cirrhosis of the liver.
Secretion of bile salts into the duodenum was studied in eight normal subjects, in 10 patients with cirrhosis, and in two cholecystectomized subjects. Duodenal juice was aspirated continuously through a double-lumen tube during an unstimulated period, after an intravenous injection of pancreozymin/cholecystokinin, and during a continuous intravenous infusion of secretin given at a rate of 3 units per kilogram body weight per hour. Precautions were taken to try to ensure quantitative recovery during the studies, and recovery of an infused nonabsorbable marker was greater than 80% in all subjects. Secretin induced a flow of a greater volume of juice in the cirrhotic patients than in the normal group (49 to 57 ml per 10 minutes compared with 28 to 49 ml per 10 minutes). This change may have resulted from a higher effective dose of secretin if it is assumed that the cirrhotic liver fails to catabolize secretin. The bile acid response to pancreozymin/cholecystokinin followed by secretin in the cirrhotic subjects resembled that seen in patients after cholecystectomy in whom pancreozymin/cholecystokinin induces only a slight increase in bile salt output but in whom the output of bile salts during rest and secretin stimulation is markedly greater than normal. This response in cirrhosis is probably best interpreted as due to impaired function of the gallbladder. The total amount of bile salt liberated over the two hours of the test in the cirrhotic patients was similar to normal The concentration of bile salt after pancreozymin/cholecystokinin was less than in normal subjects, but similar to that in cholecystectomized patients. It is unlikely therefore that deficient output or concentration of bile salt can be held responsible for steatorrhea in cirrhosis. THERE WAS A MARKED DECREASE IN THE DEOXYCHOLATE CONJUGATES AND A REDUCTION IN THE GLYCINE: taurine ratio in the bile of cirrhotic patients. The former change may reflect a change in bacterial flora and the latter a defect in hepatic conjugating mechanisms.


Humoral immune system in inflammatory bowel disease: I. Complement levels.
Serum levels of complement components Clq, C4, C3, and Properdin factor B, from the classical and alternative pathways of complement activation, have been estimated in patients with ulcerative colitis and Crohn's disease. C3, factor B, and to some extent C4 concentrations all increased when the disease was active. In remission the levels of these components did not differ from hospital control patients. There was no evidence for the preferential consumption of the proteins of either pathway of activation, even in those patients with evidence of circulating immune complexes.


Multifactorial analysis of the learning curve for robot-assisted laparoscopic biliopancreatic diversion with duodenal switch.
To assess the impact of surgeon, patient, and case-specific factors on the learning curve of robot-assisted laparoscopic biliopancreatic diversion with duodenal switch (RA-LBPD/DS). The BPD/DS has better resolution of diabetes and hypercholesterolemia, and the best long-term weight loss compared to the laparoscopic gastric band or the Roux-en-Y gastric bypass. Despite excellent results, the BPD/DS is least commonly performed because of greater malabsorption, longer operative duration, and higher technical complication rates. A reduction in technical complications and operative duration will enable the BPD/DS to be offered more frequently. Consecutive patients (N = 120) undergoing RA-LBPD/DS between October 2000 and August 2008 were analyzed using univariate and multivariate logistic regression to determine the influence of surgeon and patient factors on complications and operative duration. Independent variables were case number, age, gender, body mass index, American Society of Anesthesiologists (ASA) score, difficult anatomy, and need for extensive adhesiolysis. Dependent variables were complications (leaks, bleeding, and conversion) and operative duration. The best-fit model predicted the risk factors for complications, and a risk-adjusted cumulative sum analysis estimated the learning curve. : Operative duration decreased an average of 3 minutes with each successive case (P < 0.001, R² = 0.63) and with patient's female gender. Adhesiolysis, difficult anatomy, liver biopsy, and higher ASA score increased operative duration. The incidence of high blood loss (13.3%), conversion (2.2%), and leaks (5.8%) were experienced by a total of 22 patients (18.3%). There was no mortality. Complications declined after 50 cases and were strongly predicted by increasing surgeon case number. The learning curve for the RA-LBPD/DS is 50 cases. Risk factors influencing outcomes were identified.


White matter microstructural abnormalities in blast-exposed combat veterans: accounting for potential pre-injury factors using consanguineous controls.
Assess the prevalence of white matter microstructural changes in combat veterans, within the context of a highly matched control group comprising unexposed close relatives. This prospective study had institutional review board approval, included written informed consent, and is HIPAA-compliant. Diffusion tensor imaging was analyzed in 16 male blast-exposed combat veterans of Operation Iraqi Freedom/Operation Enduring Freedom (mean age 31.0 years) and 18 unexposed males (mean age 30.4 years) chosen on the basis of a consanguineous relationship to a member of the subject group. Whole-brain voxel-based comparison of fractional anisotropy (FA) was performed using both group and individual analyses. Areas where effects on FA were detected were subsequently characterized by extracting radial diffusivity (RD), axial diffusivity (AD), and mean diffusivity (MD) from the regions of abnormal FA. Controls did not differ from veterans on any background demographic factor. In voxel-based group comparison, we identify high fractional anisotropy (FA) in veterans compared to controls (p < 0.01). Within individual veterans, we find multiple areas of both abnormally high and low FA (p < 0.01) in a heterogeneous distribution, consistent with multifocal traumatic axonal injury. In individualized analyses, low FA areas demonstrate high radial diffusivity, whereas high FA areas demonstrate low RD in both group and individual analyses. Combat-related blast exposure is associated with microstructural white matter abnormalities, and the nature of the control group decreases the likelihood that the findings reflect underlying background differences. Abnormalities are heterogeneously distributed across patients, consistent with TAI, and include areas of low and high FA.


Comparison of Campylobacter populations isolated from a free-range broiler flock before and after slaughter.
Relatively little is known about the Campylobacter genotypes colonizing extensively reared broiler flocks and their survival through the slaughter process, despite the increasing demand for free-range and organic products by the consumer. Campylobacter isolates from a free-range boiler flock, sampled before and after slaughter, were genotyped by MLST (multilocus sequence typing) and sequence analysis of the flaA short variable region (SVR). The Campylobacter genotypes isolated before and after slaughter were diverse, with up to five sequence types (STs) (seven-locus allelic profiles resulting from MLST) identified per live bird, up to eight STs identified per carcass and 31 STs identified in all. The majority (72.0%) of isolates sampled from carcasses post-slaughter were indistinguishable from those isolated from the live flock before slaughter by ST and flaA SVR type, however, sampling 'on-farm' failed to capture all of the diversity seen post-slaughter. There were statistically significant increases in the genetic diversity of Campylobacter (p=0.005) and the proportion of C. coli (p=0.002), with some evidence for differential survival of genotypes contaminating the end product. C. coli genotypes isolated after slaughter were more similar to those from free-range and organic meat products sampled nationally, than from the live flock sampled previously. This study demonstrated the utility of MLST in detecting genetic diversity before and after the slaughter process.


Leukocyte Cell-Derived Chemotaxin-2 Amyloidosis (ALECT2) in a Patient With Lung Adenocarcinoma: An Autopsy Report and Literature Review.
Amyloidosis caused by deposition of leukocyte cell-derived chemotaxin-2 amyloidosis (ALECT2) is the most recently described form of systemic amyloidosis and has quickly emerged as a common and possibly underdiagnosed cause of slowly declining renal function, particularly in patients of Hispanic ancestry. We describe the autopsy findings in a 70-year-old Hispanic woman who died of metastatic lung adenocarcinoma and was incidentally found to have extensive amyloid deposition in the kidneys, liver, spleen, adrenal glands, small intestine, gallbladder, lungs, bilateral ovaries, and uterus. The type of amyloid was confirmed to be ALECT2 by mass spectrometry. The pattern of amyloid deposition in the kidneys and the liver was typical for what has been described for ALECT2. In addition, a unique pattern of amyloid deposition was observed in spleen, adrenal glands, small intestine, gallbladder, lungs, ovaries, and uterus. The pattern of amyloid deposition in ALECT2 is distinct from amyloid A and amyloid light-chain and needs to be recognized to avoid misdiagnosis as amyloid light-chain or amyloid A amyloidosis. After recognition, an accurate diagnosis by mass spectrometry and/or immunohistochemical staining is essential to guide treatment and avoid toxic therapies.


A critical role for thymic stromal lymphopoietin in nickel-induced allergy in mice.
Ni is the most frequent cause of contact allergy induced by metals. However, the underlying mechanism of this induction is unknown. Our previous research demonstrates that activation of dendritic cells (DCs) through p38MAPK/MKK6 is required for Ni-induced allergy in mice. In the current study, we investigated the cellular and molecular mechanisms underlying Ni-induced allergy using a mouse model that involves injecting Ni into the ear, with or without Freund's incomplete or complete adjuvants. Nickel had greater potential to cause allergic reactions compared with palladium and gold. Among the proteins expressed at higher levels in mice with Ni-induced allergy, we focused on thymic stromal lymphopoietin (TSLP), which is produced in abundance by keratinocytes. We detected increased expression of the TSLP receptor (TSLPR) in DCs from cervical lymph nodes of mice with Ni-induced allergy, suggesting that DCs in ear tissues were activated through TSLPR signaling induced by keratinocyte-derived TSLP. Furthermore, delayed-type hypersensitivity reactions in mice with Ni-induced allergy were decreased significantly by injection of a Tslp-short interfering RNA along with atelocollagen in the ear skin. These results suggest that Ni allergy may be triggered by a TSLP/TSLPR-mediated interaction between epithelial and immune cells.


Human Purkinje in silico model enables mechanistic investigations into automaticity and pro-arrhythmic abnormalities.
Cardiac Purkinje cells (PCs) are implicated in lethal arrhythmias caused by cardiac diseases, mutations, and drug action. However, the pro-arrhythmic mechanisms in PCs are not entirely understood, particularly in humans, as most investigations are conducted in animals. The aims of this study are to present a novel human PCs electrophysiology biophysically-detailed computational model, and to disentangle ionic mechanisms of human Purkinje-related electrophysiology, pacemaker activity and arrhythmogenicity. The new Trovato2020 model incorporates detailed Purkinje-specific ionic currents and Ca2+ handling, and was developed, calibrated and validated using human experimental data acquired at multiple frequencies, both in control conditions and following drug application. Multiscale investigations were performed in a Purkinje cell, in fibre and using an experimentally-calibrated population of PCs to evaluate biological variability. Simulations demonstrate the human Purkinje Trovato2020 model is the first one to yield: (i) all key AP features consistent with human Purkinje recordings; (ii) Automaticity with funny current up-regulation (iii) EADs at slow pacing and with 85% hERG block; (iv) DADs following fast pacing; (v) conduction velocity of 160 cm/s in a Purkinje fibre, as reported in human. The human in silico PCs population highlights that: (1) EADs are caused by ICaL reactivation in PCs with large inward currents; (2) DADs and triggered APs occur in PCs experiencing Ca2+ accumulation, at fast pacing, caused by large L-type calcium current and small Na+/Ca2+ exchanger. The novel human Purkinje model unlocks further investigations into the role of cardiac Purkinje in ventricular arrhythmias through computer modeling and multiscale simulations.


Prediction and Validation of Native and Engineered Cas9 Guide Sequences.
Cas9-based technologies rely on native elements of Type II CRISPR-Cas bacterial immune systems, including the trans-activating CRISPR RNA (tracrRNA), CRISPR RNA (crRNA), Cas9 protein, and protospacer-adjacent motif (PAM). The tracrRNA and crRNA form an RNA duplex that guides the Cas9 endonuclease to complementary nucleic acid sequences. Mechanistically, Cas9 initiates interactions by binding to the target PAM sequence and interrogating the target DNA in a 3'-to-5' manner. Complementarity between the guide RNA and the target DNA is key. In natural systems, precise cleavage occurs when the target DNA sequence contains a PAM flanking a sequence homologous to the crRNA spacer sequence. Currently, the majority of commercial Cas9-based genome-editing tools are derived from the Type II CRISPR-Cas system of Streptococcus pyogenes However, a diverse set of Type II CRISPR-Cas systems exist in nature that are potentially valuable for genome engineering applications. Exploitation of these systems requires prediction and validation of both native and engineered dual and single guide RNAs to drive Cas9 functionality. Here, we discuss how to identify the elements of these immune systems to develop next-generation Cas9-based genome-editing tools. We first discuss how to predict tracrRNA sequences and suggest a method for designing single guide RNAs containing only critical structural modules. We then outline how to predict the PAM sequence, which is crucial for determining potential targets for Cas9. Finally, validation of the system elements through transcriptome analysis and interference assays is essential for developing next-generation Cas9-based genome-editing tools.