Microarray profiling of progesterone-regulated endometrial genes during the rhesus monkey secretory phase.
In the endometrium the steroid hormone progesterone (P), acting through its nuclear receptors, regulates the expression of specific target genes and gene networks required for endometrial maturation. Proper endometrial maturation is considered a requirement for embryo implantation. Endometrial receptivity is a complex process that is spatially and temporally restricted and the identity of genes that regulate receptivity has been pursued by a number of investigators. In this study we have used high density oligonucleotide microarrays to screen for changes in mRNA transcript levels between normal proliferative and adequate secretory phases in Rhesus monkey artificial menstrual cycles. Biotinylated cRNA was prepared from day 13 and days 21-23 of the reproductive cycle and transcript levels were compared by hybridization to Affymetrix HG-U95A arrays. Of approximately 12,000 genes profiled, we identified 108 genes that were significantly regulated during the shift from a proliferative to an adequate secretory endometrium. Of these genes, 39 were up-regulated at days 21-23 versus day 13, and 69 were down-regulated. Genes up-regulated in P-dominant tissue included: secretoglobin (uteroglobin), histone 2A, polo-like kinase (PLK), spermidine/spermine acetyltransferase 2 (SAT2), secretory leukocyte protease inhibitor (SLPI) and metallothionein 1G (MT1G), all of which have been previously documented as elevated in the Rhesus monkey or human endometrium during the secretory phase. Genes down-regulated included: transforming growth factor beta-induced (TGFBI or BIGH3), matrix metalloproteinase 11 (stromelysin 3), proenkephalin (PENK), cysteine/glycine-rich protein 2 (CSRP2), collagen type VII alpha 1 (COL7A1), secreted frizzled-related protein 4 (SFRP4), progesterone receptor membrane component 1 (PGRMC1), chemokine (C-X-C) ligand 12 (CXCL12) and biglycan (BGN). In addition, many novel/unknown genes were also identified. Validation of array data was performed by semi-quantitative RT-PCR of two selected up-regulated genes using temporal (cycle day specific) endometrial cDNA populations. This approach confirmed up-regulation of WAP four-disulfide core domain 2 (WFDC2) and SLPI during the expected window of receptivity. The identification of P-regulated genes and gene pathways in the primate endometrium is expected to be an important first step in elucidating the cellular processes necessary for the development of a receptive environment for implantation.


Telephone triage for management of same-day consultation requests in general practice (the ESTEEM trial): a cluster-randomised controlled trial and cost-consequence analysis.
Telephone triage is increasingly used to manage workload in primary care; however, supporting evidence for this approach is scarce. We aimed to assess the effectiveness and cost consequences of general practitioner-(GP)-led and nurse-led telephone triage compared with usual care for patients seeking same-day consultations in primary care. We did a pragmatic, cluster-randomised controlled trial and economic evaluation between March 1, 2011, and March 31, 2013, at 42 practices in four centres in the UK. Practices were randomly assigned (1:1:1), via a computer-generated randomisation sequence minimised for geographical location, practice deprivation, and practice list size, to either GP-led triage, nurse-led computer-supported triage, or usual care. We included patients who telephoned the practice seeking a same-day face-to-face consultation with a GP. Allocations were concealed from practices until after they had agreed to participate and a stochastic element was included within the minimisation algorithm to maintain concealment. Patients, clinicians, and researchers were not masked to allocation, but practice assignment was concealed from the trial statistician. The primary outcome was primary care workload (patient contacts, including those attending accident and emergency departments) in the 28 days after the first same-day request. Analyses were by intention to treat and per protocol. This trial was registered with the ISRCTN register, number ISRCTN20687662. We randomly assigned 42 practices to GP triage (n=13), nurse triage (n=15), or usual care (n=14), and 20,990 patients (n=6695 vs 7012 vs 7283) were randomly assigned, of whom 16,211 (77%) patients provided primary outcome data (n=5171 vs 5468 vs 5572). GP triage was associated with a 33% increase in the mean number of contacts per person over 28 days compared with usual care (2·65 [SD 1·74] vs 1·91 [1·43]; rate ratio [RR] 1·33, 95% CI 1·30-1·36), and nurse triage with a 48% increase (2·81 [SD 1·68]; RR 1·48, 95% CI 1·44-1·52). Eight patients died within 7 days of the index request: five in the GP-triage group, two in the nurse-triage group, and one in the usual-care group; however, these deaths were not associated with the trial group or procedures. Although triage interventions were associated with increased contacts, estimated costs over 28 days were similar between all three groups (roughly £75 per patient). Introduction of telephone triage delivered by a GP or nurse was associated with an increase in the number of primary care contacts in the 28 days after a patient's request for a same-day GP consultation, with similar costs to those of usual care. Telephone triage might be useful in aiding the delivery of primary care. The whole-system implications should be assessed when introduction of such a system is considered. Health Technology Assessment Programme UK National Institute for Health Research.


Do Non-Responders to Exercise Exist-and If So, What Should We Do About Them?
It is well established that exercise is an important component in the maintenance of good health, and yet recent studies have demonstrated that a sub-section of individuals experience no significant improvements following an exercise training intervention. Such individuals are commonly termed "non-responders". However, recently a number of researchers have taken a skeptical view as to whether exercise non-response either exists, or is clinically relevant. Here, we explore the research underpinning exercise response, to determine whether non-response to exercise actually exists. We discuss the impact of measurement error and assessment type on the identification of "non-responders", and whether such non-response is global- or modality-specific. Additionally, we discuss whether, if non-response to an exercise intervention is meaningful and relevant, certain additional interventions-in the form of increasing exercise intensity, volume, or duration-could be made in order to enhance training adaptations. Consequently, based on our interpretations of the available evidence, we suggest that it is unlikely that global non-responders to exercise exist. Furthermore, we suggest this realization effectively counters the perception that some individuals will not positively respond to exercise, and that in turn, this insight serves to encourage health professionals to create more nuanced, efficacious, and individually-focused exercise prescriptions designed to circumvent and overcome apparent non-responsiveness. Adopting a more individually-adaptive approach to exercise prescription could, subsequently, prove a powerful tool in promoting population health.


Transforming communication and safety culture in intrapartum care: a multi-organization blueprint.
Effective, patient-centered communication facilitates interception and correction of potentially harmful conditions and errors. All team members, including women, their families, physicians, midwives, nurses, and support staff, have a role in identifying the potential for harm during labor and birth. However, the results of collaborative research studies conducted by organizations that represent professionals who care for women during labor and birth indicate that health care providers may frequently witness, but may not always report, problems with safety or clinical performance. Some of these health care providers felt resigned to the continuation of such problems and fearful of retribution if they tried to address them. Speaking up to address safety and quality concerns is a dynamic social process. Every team member must feel empowered to speak up about concerns without fear of put-downs, retribution, or receiving poor-quality care. Patient safety requires mutual accountability: individuals, teams, health care facilities, and professional associations have a shared responsibility for creating and sustaining environments of mutual respect and engaging in highly reliable perinatal care. Defects in human factors, communication, and leadership have been the leading contributors to sentinel events in perinatal care for more than a decade. Organizational commitment and executive leadership are essential to creating an environment that proactively supports safety and quality. The problem is well-known; the time for action is now.


Human Capital Questionnaire: Assessment of European nurses' perceptions as indicators of human capital quality.
Healthcare accreditation models generally include indicators related to healthcare employees' perceptions (e.g. satisfaction, career development, and health safety). During the accreditation process, organizations are asked to demonstrate the methods with which assessments are made. However, none of the models provide standardized systems for the assessment of employees. In this study, we analyzed the psychometric properties of an instrument for the assessment of nurses' perceptions as indicators of human capital quality in healthcare organizations. The Human Capital Questionnaire was applied to a sample of 902 nurses in four European countries (Spain, Portugal, Poland, and the UK). Exploratory factor analysis identified six factors: satisfaction with leadership, identification and commitment, satisfaction with participation, staff well-being, career development opportunities, and motivation. The results showed the validity and reliability of the questionnaire, which when applied to healthcare organizations, provide a better understanding of nurses' perceptions, and is a parsimonious instrument for assessment and organizational accreditation. From a practical point of view, improving the quality of human capital, by analyzing nurses and other healthcare employees' perceptions, is related to workforce empowerment.


Effect of coronary artery pressure on transmural distribution of adrenergic coronary vasoconstriction in the dog.
The transmural distribution of alpha-receptor coronary vasoconstriction was studied in 14 closed-chest, morphine- and chloralose-anesthesized dogs. The alpha-receptor-blocking agent, phenoxybenzamine (0.25 mg/kg), was infused into either the anterior or circumflex coronary artery while collection of coronary sinus blood minimized recirculation. The left main coronary artery was cannulated and perfused at constant pressure. Myocardial lactate extraction and blood flow (9 micron radioactive microspheres) were measured during adrenergic activation with intracoronary norepinephrine (2 micrograms/min) at coronary artery pressures of 100, 70, 50, and 38 mm Hg. Flow was 15-25% less in the control region (alpha-receptors intact) than in the alpha-receptor-blocked region, in all layers of the left ventricular wall, at coronary pressures of 100 and 70 mm Hg. When coronary pressure was 50 mm Hg, lactate production resulted (-4% extraction) and significant alpha-receptor vasoconstriction was observed in the outer layer, but was marginal (P = 0.043) in the inner layer, of the left ventricle. Flows were not different in control and alpha-receptor-blocked regions at a coronary pressure of 38 mm Hg (lactate extraction -49%). These data indicate a uniform transmural alpha-receptor vasoconstriction at normal coronary artery pressures that diminished as the heart was progressively underperfused.


The effects of endophytes on seed production and seed predation of tall fescue and meadow fescue.
Fungal endophytes of grasses are often included in agricultural management and in ecological studies of natural grass populations. In European agriculture and ecological studies, however, grass endophytes are largely ignored. In this study, we determined endophyte infection frequencies of 13 European cultivars and 49 wild tall fescue (Schedonorus phoenix) populations in Northern Europe. We then examined seed production and seed predation of endophyte-infected (E+) and endophyte-free (E-) tall fescue (in wild grass populations and in a field experiment) and meadow fescue (Schedonorus pratensis; in a field experiment only). Endophytes were detected in only one of the 13 cultivars. In contrast, >90% of wild tall fescue plants harbored endophytes in 45 wild populations but were absent in three inland populations in Estonia. In three wild tall fescue study sites, 17%, 22%, and 56% of the seeds were preyed upon by the cocksfoot moth. Endophyte infection did not affect seed mass of tall fescue in the field experiment. However, seed predation was lower in E+ than E- grasses in the two tall fescue populations with higher predation rates. For meadow fescue, the mean number of seeds from E+ plants was higher than E- plants, but E- and E+ seeds had equal rates of predation by the moth. Our results suggest that the effects of grass endophytes on seed production and cocksfoot moth seed predation vary considerably among grass species, and the effects may depend on herbivore pressure and other environmental conditions.


Empirical equation for the index of refraction of seawater.
We have determined an empirical equation for the index of refraction of water as a function of temperature, salinity, and wavelength at atmospheric pressure. The experimental data selected by Austin and Halikas ["The index of refraction of seawater," SIO Ref. 76-1 (Scripps Institution of Oceanography, La Jolla, Calif., 1976)] were fitted to power series in the variables. A ten-parameter empirical equation that reproduces the original data to within its experimental errors was obtained.


Effects of Sport-Based Exercise Interventions on Executive Function in Older Adults: A Systematic Review and Meta-Analysis.
Exercise programs of moderate-to-vigorous intensity have been shown to improve the cognitive performance of older people. However, the specific effects of sports-based exercise programs on cognitive performance, particularly executive functions, remain unclear. Therefore, the purpose of this study is to clarify the effects of sports-based exercise programs on executive functions in older adults using a systematic review and meta-analysis of the scientific literature. A systematic review was conducted between 1 March and 1 July 2022, to look for published scientific evidence that analyzed different sports programs that may have affected executive function in healthy older adults. Longitudinal studies, which assessed the effects of sports interventions on healthy older adults, were identified through a systematic search of the four principal electronic databases: Web of Science, PubMed, Scopus, and EBSCO. A total of nine studies with a total of 398 subjects met the inclusion criteria and were classified based on one or more of the following categories: working memory, inhibition, and cognitive flexibility. The DerSimonian and Laird random-effects model was performed using the Comprehensive Meta-Analysis software to facilitate the analysis of the studies. Statistical significance was set at p ≤ 0.05. In terms of working memory, a small but positive significant effect was noted for the intervention group compared to the control group (effect size (ES) = 0.35, 95% confidence interval (CI) = 0.04-0.67; p = 0.029; I2 = 36.7%); in terms of inhibition, the intervention had a small favoring but no significant effect compared to the control group (ES = 0.20, 95% CI = -0.42-0.84; p = 0.517; I2 = 78.8%); and in terms of cognitive flexibility, the intervention had a small favoring but no significant effect compared to the control group (ES = 0.39, 95% CI = -0.11-0.89; p = 0.131; I2 = 75.5%). Our findings suggest that healthy older adults should be encouraged to participate in sports to improve their working memory; however, more studies are required in this area to reach more robust conclusions. This systematic review was registered with the International Prospective Register of Systematic Reviews (registration number: CRD42022284788).


Rapid effect of inhaled ciclesonide in asthma: a randomized, placebo-controlled study.
Ciclesonide is a novel inhaled corticosteroid for the treatment of asthma, and it is important to measure the onset of effect of this therapy on airway hyperresponsiveness (AHR), exhaled nitric oxide (NO), and levels of eosinophils in induced sputum. In a randomized, double-blind, crossover study, 21 patients with mild asthma inhaled ciclesonide 320 microg (ex-actuator) qd, ciclesonide 640 microg (ex-actuator) bid, and placebo for 7 days. Exhaled NO and AHR to adenosine monophosphate (AMP), measured as the provocative concentration of AMP producing a 20% reduction in FEV1 (PC20FEV1), were assessed after inhalation on days 1, 3 and 7. Eosinophil levels in induced sputum were also measured. Ciclesonide 320 microg qd and 640 microg bid produced significantly greater improvements in PC20FEV1 compared with placebo on day 1 (within 2.5 h), and on days 3 and 7 (all p < 0.0001). On day 3, both ciclesonide doses significantly reduced exhaled NO levels by - 17.7 parts per billion (p < 0.0001) and - 15.4 parts per billion (p < 0.003) vs placebo, respectively. Significant reductions were maintained during the study with both ciclesonide doses (p < 0.01). A nonsignificant trend towards a decrease in eosinophil cell numbers was observed after 7 days of ciclesonide treatment, especially in patients receiving the higher dose. A single dose of ciclesonide decreased AHR to AMP and exhaled NO within 3 h, while FEV, improved at 3 days and 7 days.


Faecal-wood biomass co-combustion and ash composition analysis.
Fuel blending is a widely used approach in biomass combustion, particularly for feedstocks with low calorific value and high moisture content. In on-site sanitation technologies, fuel blending is proposed as a pre-treatment requirement to reduce moisture levels and improve the physiochemical properties of raw faeces prior to drying. This study investigates the co-combustion performance of wood dust: raw human faeces blends at varying air-to-fuel ratios in a bench-scale combustor test rig. It concludes with ash composition analyses and discusses their potential application and related problems. The study shows that a 50:50 wood dust (WD): raw human faeces (FC) can reduce moisture levels in raw human faeces by ∼40% prior to drying. The minimum acceptable blend for treating moist faeces without prior drying at a combustion air flow rate of 14-18 L/min is 30:70 WD: FC. For self-sustained ignition and flame propagation, the minimum combustion temperature required for conversion of the fuel to ash is ∼400 °C. The most abundant elements in faecal ash are potassium and calcium, while elements such as nickel, aluminium and iron are in trace quantities. This suggests the potential use of faecal ash as a soil conditioner, but increases the tendency for fly ash formation and sintering problems.


Impairments, disabilities and handicap pattern in konzo--a non-progressive spastic para/tetraparesis of acute onset.
To assess impairments, disabilities and handicap pattern in konzo. The study included 17 konzo subjects, of which three were males and 14 females (mean age 21, median 18 years). A detailed neurological examination was performed on all subjects. Subsequently, an assessment of impairments, disabilities and handicap was done with a constructed rating scale partially based on the ICIDH-2 framework. The overall disablement picture in all subjects consisted of motor dysfunction in lower limbs leading to limitations in walking and movement activities, and restrictions in mobility. Hip mobility was severely impaired in most cases (15/17). Although konzo subjects showed normal muscle power in upper limbs (13/17), they had impaired fine motor function (10/17). Further studies are needed to assess the effectiveness of the WHO criteria for konzo in defining its forms. The applicability of the ICIDH-2 framework in this study demonstrates the possibility of its use as a common language among researchers in the field of motor disorders. However, a revision is suggested of its taxonomy, and a definition of operational criteria to clarify the content of different qualifiers provided to assess the level of functioning or disability.


Endovascular treatment of ophthalmic artery aneurysms: ophthalmic artery patency following flow diversion versus coil embolization.
The Pipeline Embolization Device (PED) has been shown to effectively treat complex internal carotid artery aneurysms while maintaining patency of covered side branches. The purpose of this retrospective matched cohort study is to evaluate the effect of flow diversion on the patency of the ophthalmic artery when treating ophthalmic artery aneurysms. A retrospective review of our prospectively collected institutional database identified 19 ophthalmic artery aneurysms treated with a PED. These were matched according to aneurysm diameter in a 1:2 fashion to ophthalmic artery aneurysms treated via coil embolization, although it is important to note that there was a statistically significance difference in the neck diameter between the two groups (p=0.045). Clinical and angiographic outcomes were recorded and analyzed. On follow-up angiography, decreased flow through the ophthalmic artery was observed in 26% of the PED cohort and 0% of the coil embolization cohort (p=0.003). No ophthalmologic complications were noted in either cohort. Complete occlusion at 12 months was more common following PED treatment than coil embolization (74% vs 47%; p=0.089), although lower than reported in previous trials. This may be due to inflow into the ophthalmic artery keeping the aneurysm patent. Retreatments were more common following coil embolization than PED (24% vs 11%), but this was not significant (p=0.304). Permanent morbidity rates were not significantly different between the PED (11%) and coil embolization (3%) cohorts (p=0.255). Our results suggest that ophthalmic artery aneurysms may be adequately and safely treated with either the PED or coil embolization. However, treatment with the PED carries a higher risk of impeding flow to the ophthalmic artery, although this did not result in clinical sequelae in the current study.


Enhanced temporal but not attentional processing in expert tennis players.
In tennis, as in many disciplines of sport, fine spatio-temporal resolution is required to reach optimal performance. While many studies on tennis have focused on anticipatory skills or decision making, fewer have investigated the underlying visual perception abilities. In this study, we used a battery of seven visual tests that allowed us to assess which kind of visual information processing is performed better by tennis players than other athletes (triathletes) and non-athletes. We found that certain time-related skills, such as speed discrimination, are superior in tennis players compared to non-athletes and triathletes. Such tasks might be used to improve tennis performance in the future.


Impact of COVID-19 on UK stress echocardiography practice: insights from the EVAREST sites.
Healthcare delivery is being transformed by COVID-19 to reduce transmission risk but continued delivery of routine clinical tests is essential. Stress echocardiography is one of the most widely used cardiac tests in the NHS. We assessed the impact of the first (W1) and second (W2) waves of the pandemic on the ability to deliver stress echocardiography. Clinical echocardiography teams in 31 NHS hospitals participating in the EVAREST study were asked to complete a survey on the structure and delivery of stress echocardiography as well as its impact on patients and staff in July and November 2020. Results were compared to stress echocardiography activity in the same centre during January 2020. 24 completed the survey in July, and 19 NHS hospitals completed the survey in November. A 55% reduction in the number of studies performed was reported in W1, recovering to exceed pre-COVID rates in W2. The major change was in the mode of stress delivery. 70% of sites stopped their exercise stress service in W1, compared to 19% in W2. In those still using exercise during W1, 50% were wearing FFP3/N95 masks, falling to 38% in W2. There was also significant variability in patient screening practices with 7 different pre-screening questionnaires used in W1 and 6 in W2. Stress echocardiography delivery restarted effectively after COVID-19 with adaptations to reduce transmission that means activity has been able to continue, and exceed, pre-COVID-19 levels during the second wave. Further standardization of protocols for patient screening and PPE may help further improve consistency of practice within the United Kingdom.


Rapid 3-D cone-beam reconstruction with the simultaneous algebraic reconstruction technique (SART) using 2-D texture mapping hardware.
Algebraic reconstruction methods, such as the algebraic reconstruction technique (ART) and the related simultaneous ART (SART). reconstruct a two-dimensional (2-D) or three-dimensional (3-D) object from its X-ray projections. The algebraic methods have, in certain scenarios, many advantages over the more popular Filtered Backprojection approaches and have also recently been shown to perform well for 3-D cone-beam reconstruction. However, so far the slow speed of these iterative methods have prohibited their routine use in clinical applications. In this paper, we address this shortcoming and investigate the utility of widely available 2-D texture mapping graphics hardware for the purpose of accelerating the 3-D algebraic reconstruction. We find that this hardware allows 3-D cone-beam reconstructions to be obtained at almost interactive speeds, with speed-ups of over 50 with respect to implementations that only use general-purpose CPUs. However, we also find that the reconstruction quality is rather sensitive to the resolution of the framebuffer, and to address this critical issue we propose a scheme that extends the precision of a given framebuffer by 4 bits, using the color channels. With this extension, a 12-bit framebuffer delivers useful reconstructions for 0.5% tissue contrast, while an 8-bit framebuffer requires 4%. Since graphics hardware generates an entire image for each volume projection, it is most appropriately used with an algebraic reconstruction method that performs volume correction at that granularity as well, such as SART or SIRT. We chose SART for its faster convergence properties.


Agonistic angiotensin II type 1 receptor autoantibodies in postpartum women with a history of preeclampsia.
Activating angiotensin II type 1 autoantibodies (AT1-AAs) develop in women with preeclampsia and may contribute to the disorder. Insulin resistance and serum concentrations of the antiangiogenic soluble fms-like tyrosine kinase 1 (sFlt-1) are also increased in women with preeclampsia compared with normal pregnancy. sFlt-1 and insulin resistance decrease substantially after delivery; however, significant group differences persist postpartum. Women who have had preeclampsia are at increased cardiovascular risk later in life. We measured AT1-AAs in groups of women with previous preeclampsia (n=29) and previous normal pregnancies (n=35) 18+/-9 months after the first completed pregnancy. These women had had sFlt-1, insulin resistance homeostasis model assessment score, and related cardiovascular risk factors measured. Activating antibodies were detected by the chronotropic response of cultured neonatal rat cardiomyocytes coupled with receptor-specific antagonists (losartan and prazosin). AT1-AAs were detected in 17.2% of women with previous preeclampsia versus 2.9% of women with previous uncomplicated pregnancies (P<0.05). In contrast, there was no difference in the prevalence of autoantibodies against the alpha1-adrenoceptor (10% of previous preeclamptic versus 14% of previous normal pregnant). Women with activating autoantibodies had significantly increased sFlt-1, reduced free vascular endothelial growth factor, and higher insulin resistance homeostasis model assessment values compared with autoantibody-negative women. These data suggest that, as with sFlt-1 and insulin resistance, the AT1-AA does not regress completely after delivery and, secondarily, that correlations exist among these variables. The impact of AT1-AA after preeclampsia, especially in the context of cardiovascular risk, remains to be determined.


Effects of a preventive intervention on adolescent substance use initiation, expectancies, and refusal intentions.
This study evaluated the effects of a school-based preventive intervention (Botvin, G. J. 1996, 2000) on growth trajectories of substance initiation (alcohol, tobacco, and marijuana), expectancies, and refusal intentions. A rural midwestern sample (N=847) provided three waves of data from middle school students. Growth curve analyses demonstrated that the intervention significantly slowed the rate of increase in substance initiation and significantly slowed the rate of decrease in refusal intentions. The intervention also slowed the rate of decrease in negative outcome expectancies, although the significance level was only marginal. A multiple group comparison showed that the impact of initial levels of substance initiation on growth trajectories of refusal intentions differed between conditions, suggesting that the intervention decreased the effect of early substance initiation on the rate of change over time for refusal intentions. Gender differences also were found, although the intervention was effective in slowing the rate of increase in initiation for both genders.


Evaluation of the effectiveness of a phonoaudiology program for the rehabilitation of dysphagia in the elderly.
The object of this study was to evaluate the effectiveness of a phonoaudiology rehabilitation program directed towards the degenerative changes of the neurophysiological mechanisms responsible for the upper digestive system compromised by the age between 80 and 90 years, in a regime of internment in a long permanence institution in the city of Rio de Janeiro, which express, bodily, their difficulty in swallowing food of liquid or pasty consistency. To achieve such an object, the current study was developed in an experimental format or design, comprised of a sample group of 23 elderly subjects, of both genders, undergoing evaluation by the phonoaudiology protocol for evaluating the risk of dysphagia (abbreviated from the Portuguese name: Protocolo de Avaliação do Risco para Disfagia=PARD), pre- and post-intervention procedure. The developed program used the indirect therapy approach based on the adaptive myotherapeutic and myofunctional stimulation, contemplating two isometric and isokinetic orofacial active myotherapeutic exercises, two passive myotherapeutic manipulations for the supra-hyoidal musculature, as well as adaptive cephalic postural maneuvres, associated to the intra-oral olfactive and gustative sensory stimulation. The evolution of the individuals was analyzed before and after the therapy by estimating the capacity of swallowing food in the liquid fluid (LF) consistency, by the paired Student t-test, giving a p=0.01, making the difference of 2.31 between the 2 evaluations statistically significant. The changes produced by the PARD program in the capacity of individuals in swallowing food of a pasty consistency were also significant when analyzing the variance of the results on the 2 instances by using the F-test (pre- and post-intervention difference=4.47, p=0.039).


Diagnostic disparity and identification of two TNNI3 gene mutations, one novel and one arising de novo, in South African patients with restrictive cardiomyopathy and focal ventricular hypertrophy.
The minimum criterion for the diagnosis of hypertrophic cardiomyopathy (HCM) is thickening of the left ventricular wall, typically in an asymmetrical or focal fashion, and it requires no functional deficit. Using this criterion, we identified a family with four affected individuals and a single unrelated individual essentially with restrictive cardiomyopathy (RCM). Mutations in genes coding for the thin filaments of cardiac muscle have been described in RCM and HCM with 'restrictive features'. One such gene encodes for cardiac troponin I (TNNI3), a sub-unit of the troponin complex involved in the regulation of striated muscle contraction. We hypothesised that mutations in TNNI3 could underlie this particular phenotype, and we therefore screened TNNI3 for mutations in 115 HCM probands. Clinical investigation involved examination, echocardiography, chest X-ray and an electrocardiogram of both the index cases and close relatives. The study cohort consisted of 113 South African HCM probands, with and without known founder HCM mutations, and 100 ethnically matched control individuals. Mutation screening of TNNI3 for diseasecausing mutations were performed using high-resolution melt (HRM) analysis. HRM analyses identified three previously described HCM-causing mutations (p.Pro82Ser, p.Arg162Gln, p.Arg170Gln) and a novel exonic variant (p.Leu144His). A previous study involving the same amino acid identified a p.Leu144Gln mutation in a patient presenting with RCM, with clinical features of HCM. We observed the novel p.Leu144His mutation in three siblings with clinical RCM and varying degrees of ventricular hypertrophy. The isolated index case with the de novo p.Arg170Gln mutation presented with a similar phenotype. Both mutations were absent in a healthy control group. We have identified a novel disease-causing p.Leu144His mutation and a de novo p.Arg170Gln mutation associated with RCM and focal ventricular hypertrophy, often below the typical diagnostic threshold for HCM. Our study provides information regarding TNNI3 mutations underlying RCM in contrast to other causes of a similar presentation, such as constrictive pericarditis or infiltration of cardiac muscle, all with marked right-sided cardiac manifestations. This study therefore highlights the need for extensive mutation screening of genes encoding for sarcomeric proteins, such as TNNI3 to identify the underlying cause of this particular phenotype.


Bladder and sexual function among women with multiple sclerosis.
Genitourinary dysfunction is common in women with multiple sclerosis (MS), yet few studies have evaluated the association between bladder and sexual dysfunction in these women. The aim of this study was to determine factors, including demographic and bladder function, associated with sexual dysfunction in a sample of women with MS. One hundred and thirty-three women with MS completed questionnaires related to overall heath status, bladder function and sexual function. Response frequencies and percentages were calculated for questionnaire responses. Multivariate logistic regression analyses were performed to determine predictors of sexual dysfunction. Sixty-one per cent of the sample indicated that they had a problem with bladder control. Forty-seven per cent of respondents indicated that their neurological problems interfered with their sex life. Over 70% of the sample reported that they enjoyed, felt aroused and experienced orgasm during sexual activity. Not having a sexual partner and the indication of bothersome neurological problems were the best predictors of sexual dysfunction. Interestingly, patients bothered by their urge incontinence had higher levels of orgasm compared to women not bothered by urge incontinence. Although over half of the women reported voiding symptoms, most still enjoyed, felt aroused and could experience orgasm. Neurological symptoms and lacking a sexual partner emerged as the best predictors of sexual dysfunction. Urge incontinence may not be a risk factor for an orgasm. Our findings elucidate the complex nature of sexual dysfunction in women with MS.


Neurokinetic and myokinetic effects of the peptide xenin on the motility of the small and large intestine of guinea pig.
The effects of xenin 25, a peptide of the neurotensin/xenopsin family, were examined on the motility of the guinea pig jejunum and colon in vitro. In the jejunum, xenin induced a biphasic response: first a small relaxation and then a large contraction. In the colon, xenin induced relaxation. The tenia coli was contracted. Deletion or amidation of the C-terminal leucine inactivated xenin. A peptide sequence of 16 C-terminal amino acids was necessary to elicit a full response in the jejunum, whereas in the colon, the potencies of all fragments of xenin 25, including the 6 C-terminal amino acid sequence (xenin 6), were not significantly different from that of xenin 25. In the jejunum, contraction was abolished or reduced by tetrodotoxin, by atropine, by met-enkephalin, by antagonists to the tachykinin receptors NK1 and NK2 and by the P2-purinoceptor antagonist suramin. L-NNA, phentolamine, methysergide, hexamethonium and apamin had no effect. In the colon, all actions of xenin were tetrodotoxin-resistant; the potency of xenin to relax was reduced by apamin and suramin. The actions of xenin on small and large bowel were attenuated by the neurotensin receptor antagonist SR 48692. The xenin analog neurotensin was significantly less potent than xenin 25 in contracting the jejunum and more potent than xenin 25 in relaxing the colon. We conclude that xenin exerts excitatory effects on a neuronal subtype receptor in the jejunum, with participation of muscarinic, purinergic and tachykinin-related mechanisms. Xenin exerts relaxing effects on the colon by interaction with a myokinetic subtype receptor involving Ca(++)-dependent K+ channels and the P2-purinoceptor.


Sources of pharmaceutical opioids for non-medical use among young adults.
The study uses qualitative and quantitative data to describe sources of pain pills for illicit use among young adult (18- to 23-year-old) users. Respondent-driven sampling was used to recruit 383 individuals in the Columbus, Ohio, area. The sample was almost 50% Caucasian and about 55% male. Qualitative interview participants (n = 45) were selected from the larger sample. Qualitative data suggest that pharmaceutical opioid availability was so pervasive that most individuals did not have to venture outside of their immediate social networks to find people who sold or shared pills. Participants emphasized differences between those who are actively involved in obtaining pills and those who play a more passive role. Active involvement was described as going out searching for pills and paying money to obtain them. In contrast, passive role included obtaining pills when somebody offered or shared them free of charge. Multiple logistic regression analysis indicates that a more active role in obtaining pharmaceutical opioids was related to being White, more frequent use of pharmaceutical opioids, extended-release oxycodone use, and using pharmaceutical opioids to get high, as opposed to self-treating a health problem. The study results can help inform drug use epidemiology, interventions, and policy.


Transverse parton distribution functions at next-to-next-to-leading order: the quark-to-quark case.
We present a calculation of the perturbative quark-to-quark transverse parton distribution function at next-to-next-to-leading order based on a gauge invariant operator definition. We demonstrate for the first time that such a definition works beyond the first nontrivial order. We extract from our calculation the coefficient functions relevant for a next-to-next-to-next-to-leading logarithmic Q(T) resummation in a large class of processes at hadron colliders.


Anxiogenic effects of developmental bisphenol A exposure are associated with gene expression changes in the juvenile rat amygdala and mitigated by soy.
Early life exposure to Bisphenol A (BPA), a component of polycarbonate plastics and epoxy resins, alters sociosexual behavior in numerous species including humans. The present study focused on the ontogeny of these behavioral effects beginning in adolescence and assessed the underlying molecular changes in the amygdala. We also explored the mitigating potential of a soy-rich diet on these endpoints. Wistar rats were exposed to BPA via drinking water (1 mg/L) from gestation through puberty, and reared on a soy-based or soy-free diet. A group exposed to ethinyl estradiol (50 µg/L) and a soy-free diet was used as a positive estrogenic control. Animals were tested as juveniles or adults for anxiety-like and exploratory behavior. Assessment of serum BPA and genistein (GEN), a soy phytoestrogen, confirmed that internal dose was within a human-relevant range. BPA induced anxiogenic behavior in juveniles and loss of sexual dimorphisms in adult exploratory behavior, but only in the animals reared on the soy-free diet. Expression analysis revealed a suite of genes, including a subset known to mediate sociosexual behavior, associated with BPA-induced juvenile anxiety. Notably, expression of estrogen receptor beta (Esr2) and two melanocortin receptors (Mc3r, Mc4r) were downregulated. Collectively, these results show that behavioral impacts of BPA can manifest during adolescence, but wane in adulthood, and may be mitigated by diet. These data also reveal that, because ERβ and melanocortin receptors are crucial to their function, oxytocin/vasopressin signaling pathways, which have previously been linked to human affective disorders, may underlie these behavioral outcomes.


Animal seed dispersal recovery during passive restoration in a forested landscape.
Seed dispersal by animals is key for restoration of tropical forests because it maintains plant diversity and accelerates community turnover. Therefore, changes in seed dispersal during forest restoration can indicate the recovery of species interactions, and yet these changes are rarely considered in forest restoration planning. In this study, we examined shifts in the importance of different seed dispersal modes during passive restoration in a tropical chronosequence spanning more than 100 years, by modelling the proportion of trees dispersed by bats, small birds, large birds, flightless mammals and abiotic means as a function of forest age. Contrary to expectations, tree species dispersed by flightless mammals dominated after 20 years of regeneration, and tree richness and abundance dispersed by each mode mostly recovered to old growth levels between 40 and 70 years post-abandonment. Seed dispersal by small birds declined over time during regeneration, while bat dispersal played a minor role throughout all stages of succession. Results suggest that proximity to old growth forests, coupled with low hunting, explained the prevalence of seed dispersal by animals, especially by flightless mammals at this site. We suggest that aspects of seed dispersal should be monitored when restoring forest ecosystems to evaluate the reestablishment of species interactions. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.


Effects of visual feedback therapy on postural control in bilateral standing after stroke: a systematic review.
To establish whether bilateral standing with visual feedback therapy after stroke improves postural control compared with conventional therapy and to evaluate the generalization of the effects of visual feedback therapy on gait and gait-related activities. A systematic review. A computer-aided literature search was performed. Randomized controlled trials and controlled clinical trials, comparing visual feedback therapy with conventional balance treatments were included up to April 2005. The methodological quality of each study was assessed with the the Physiotherapy Evidence Database scale. Depending on existing heterogeneity, studies with a common variable of outcome were pooled by calculating the summary effect-sizes using fixed or random effects models. Eight out of 78 studies, presenting 214 subjects, were included for qualitative and quantitative analysis. The methodological quality ranged from 3 to 6 points. The meta-analysis demonstrated non-significant summary effect-sizes in favour of visual feedback therapy for weight distribution and postural sway, as well as balance and gait performance, and gait speed. The additional value of visual feedback therapy in bilateral standing compared with conventional therapy shows no statistically significant effects on symmetry of weight distribution between paretic and non-paretic leg, postural sway in bilateral standing, gait and gait-related activities. Visual feedback therapy should not be favoured over conventional therapy. The question remains as to exactly how asymmetry in weight distribution while standing is related to balance control in patients with stroke.


Prevalence, incidence, and spatial dependence of soybean mosaic virus in Iowa.
The prevalence of soybean fields with plants infected with Soybean mosaic virus (SMV) in Iowa is assumed to be random, because the primary source of the virus is SMV-infected seed. Data collected from 2,500 soybean fields sampled over a 3-year period as part of the Iowa Soybean Disease Survey (2005 to 2007) were used to evaluate this assumption. SMV was first detected in early June of each year but counties in which it was first detected varied among years. Prevalence at the county scale at end of season was 32.3, 27.3, and 89.9% in 2005, 2006, and 2007, respectively. End-of-season incidence of SMV within SMV-positive counties was 1.5 to 25.0, 1.7 to 24, and 1.8 to 58% in 2005, 2006, and 2007, respectively. The number of fields in which plants infected with SMV were detected increased at the linear rate of approximately one new field every 2 days in 2007, compared with one new field every 22 days (2005) and 21 days (2006), with coefficients of determination (R2) of 93.2 to 96.8% using the linear model. Weak spatial dependence for end-of-season SMV incidence was detected using Moran's Index, indicating that the risk for SMV incidence at the county scale within Iowa at the end of the growing season is not random.


Fructose-induced leptin resistance exacerbates weight gain in response to subsequent high-fat feeding.
It has been suggested that increased fructose intake is associated with obesity. We hypothesized that chronic fructose consumption causes leptin resistance, which subsequently may promote the development of obesity in response to a high-fat diet. Sprague-Dawley rats were fed a fructose-free control or 60% fructose diet for 6 mo and then tested for leptin resistance. Half of the rats in each group were then switched to high-fat diet for 2 wk, while the other half continued on their respective diets. Chronic fructose consumption caused leptin resistance, while serum leptin levels, weight, and adiposity were the same as in control rats that were leptin responsive. Intraperitoneal leptin injections reduced 24-h food intake in the fructose-free group (73.7 +/- 6.3 vs. 58.1 +/- 8 kcal, P = 0.02) but had no effect in fructose-fed rats (71.2 +/- 6.6 vs. 72.4 +/- 6.4 kcal, P = 0.9). Absence of anorexic response to intraperitoneal leptin injection was associated with 25.7% decrease in hypothalamic signal transducer and activator of transcription 3 phosphorylation in the high-fructose-fed rats compared with controls (P = 0.015). Subsequent exposure of the fructose-mediated, leptin-resistant rats to a high-fat diet led to exacerbated weight gain (50.2 +/- 2 g) compared with correspondingly fed leptin-responsive animals that were pretreated with the fructose-free diet (30.4 +/- 5.8 g, P = 0.012). Our data indicate that chronic fructose consumption induces leptin resistance prior to body weight, adiposity, serum leptin, insulin, or glucose increases, and this fructose-induced leptin resistance accelerates high-fat induced obesity.


Aerosol-Mediated Non-Viral Lung Gene Therapy: The Potential of Aminoglycoside-Based Cationic Liposomes.
Aerosol lung gene therapy using non-viral delivery systems represents a credible therapeutic strategy for chronic respiratory diseases, such as cystic fibrosis (CF). Progress in CF clinical setting using the lipidic formulation GL67A has demonstrated the relevance of such a strategy while emphasizing the need for more potent gene transfer agents. In recent years, many novel non-viral gene delivery vehicles were proposed as potential alternatives to GL67 cationic lipid. However, they were usually evaluated using procedures difficult or even impossible to implement in clinical practice. In this study, a clinically-relevant administration protocol via aerosol in murine lungs was used to conduct a comparative study with GL67A. Diverse lipidic compounds were used to prepare a series of formulations inspired by the composition of GL67A. While some of these formulations were ineffective at transfecting murine lungs, others demonstrated modest-to-very-efficient activities and a series of structure-activity relationships were unveiled. Lipidic aminoglycoside derivative-based formulations were found to be at least as efficient as GL67A following aerosol delivery of a luciferase-encoding plasmid DNA. A single aerosol treatment with one such formulation was found to mediate long-term lung transgene expression, exceeding half the animal's lifetime. This study clearly supports the potential of aminoglycoside-based cationic lipids as potent GL67-alternative scaffolds for further enhanced aerosol non-viral lung gene therapy for diseases such as CF.


Immune-Inflammatory and Metabolic Effects of High Dose Furosemide plus Hypertonic Saline Solution (HSS) Treatment in Cirrhotic Subjects with Refractory Ascites.
Patients with chronic liver diseases are usually thin as a result of hypermetabolism and malnutrition expressed by reduced levels of leptin and impairment of other adyponectins such as visfatin. We evaluated the metabolic and inflammatory effects of intravenous high-dose furosemide plus hypertonic saline solutions (HSS) compared with repeated paracentesis and a standard oral diuretic schedule, in patients with cirrhosis and refractory ascites. 59 consecutive cirrhotic patients with refractory ascites unresponsive to outpatient treatment. Enrolled subjects were randomized to treatment with intravenous infusion of furosemide (125-250mg⁄bid) plus small volumes of HSS from the first day after admission until 3 days before discharge (Group A, n:38), or repeated paracentesis from the first day after admission until 3 days before discharge (Group B, n: 21). Plasma levels of ANP, BNP, Leptin, visfatin, IL-1β, TNF-a, IL-6 were measured before and after the two type of treatment. Subjects in group A were observed to have a significant reduction of serum levels of TNF-α, IL-1β, IL-6, ANP, BNP, and visfatin, thus regarding primary efficacy endpoints, in Group A vs. Group B we observed higher Δ-TNF-α, Δ-IL-1β, Δ-IL-6, Δ-ANP, Δ-BNP, Δ-visfatin, Δ-Leptin at discharge. Our findings underline the possible inflammatory and metabolic effect of saline overload correction in treatment of cirrhosis complications such as refractory ascites, suggesting a possible role of inflammatory and metabolic-nutritional variables as severity markers in these patients.


Large increase of rotavirus diarrhoea in the hospital setting associated with emergence of G12 genotype in a highly vaccinated population in Nicaragua.
Rotaviruses (RVs) are a major cause of severe diarrhoea in young children. Nicaragua introduced routine immunization with the pentavalent RV vaccine (RV5) in 2006, which greatly reduced the incidence of diarrhoea. A remaining concern has been the possible emergence of new RV strains to which the vaccination has less effect. In this study, 837 children with diarrhoea in hospital settings were investigated for RV between May 2011 and July 2013. RVs were subsequently typed by multiplex PCR and/or sequencing. Fecal anti-RV IgA titres for a subset of RV-infected (n = 137) and noninfected children (n = 52) were determined with an in-house enzyme-linked immunosorbent assay. The RV detection rate was 8% in 2011, followed by a sharp increase to 29% in 2012 and 19% in 2013. This was associated with emergence and predominance of genotype G12 RV, from 0% in 2011 to 66% in 2012 and 82% in 2013, infecting children from 1 month to 10 years of age. Two sequenced G12 strains showed a Wa-like genome with genotype G12-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1, similar to the globally emerging G12 strains. Fecal anti-RV IgA analysis showed that most G12-infected and noninfected children had been in contact with either vaccine or wild RV strains, but such antibodies did not prevent symptomatic G12 infection. A marked increase of RV was evident in the hospital setting associated with a nationwide emergence and predominance of RV G12 genotype in a population with high RV5 vaccine coverage.


Analysis of saponins from black bean by electrospray ionization and fast atom bombardment tandem mass spectrometry.
Saponins from black bean (Vigna mungo L. Hepper) were analyzed using positive and negative ion fast atom bombardment mass spectrometry (FAB-MS) and liquid chromatography/mass spectrometry. Methanol was used to extract the saponins from defatted black bean, which was partially purified by extraction with n-butanol, and the extract was dialyzed with 3000 M(r) cut-off tubing. The dialyzate was analyzed using mass spectrometry. According to FAB-MS/MS, mixtures from black bean contain soyasaponin I as the predominant saponin. In addition, MS/MS analysis was performed in which the structures of saponins of black bean cotyledon were determined to be soyasaponin I, soyasaponin II, soyasaponin V, 3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-galactopyranosyl-(1 --> 2)-beta-D-glucuronopyranosyl]complogenin (saponin A) and 3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl-(1 --> 2)-beta-D-glucopyranosyl]oleanolic acid (saponin B). For the black bean shell and the root of black bean sprout, analysis confirmed the saponins of soyasaponin I, soyasaponin II, soyasaponin V, saponin A, saponin B, acetylsoyasaponin A(4) and soyasaponin beta(g). Moreover, all the studied saponins were found in the stem and leaves of the black bean sprouts, except soyasaponin beta(g) and acetylsoyasaponin A(4), respectively.


Exercise-induced angiogenesis-related growth and transcription factors in skeletal muscle, and their modification in muscle pathology.
Angiogenesis is the process of formation of new blood vessels; it is generally a rare occurrence in the adult, although it is a common adaptive response to exercise training in skeletal muscle. Current thinking is that angiogenesis is mediated by diffusible angiogenic factors and that the angiogenic activity is regulated through the balance between stimulatory and inhibitory factors. Recent studies have shown that up-regulation of angiogenic factors occurs in response to increased muscle activity in skeletal muscle. The major putative angiogenic factor, vascular endothelial growth factor (VEGF), seems to increase to a greater extent and more consistently than other measured angiogenic factors, such as fibroblast growth factor-2 (FGF-2) and transforming growth factor-beta1 (TGF-beta1). While the regulating mechanisms in this response are not clear, present data indicate reduced oxygen tension and/or related metabolic alterations in the skeletal muscle as possible stimuli. Data on other angiogenic growth factors are limited, but an increase in endothelial cell-stimulating angiogenic growth factor (ESAF) has been observed in response to increased blood flow and muscle stretching. Therefore, different exercise associated stimuli may all contribute to exercise-induced angiogenesis in skeletal muscle, but possibly through differing angiogenic factors and mechanisms. Understanding these processes is important for the elucidation of mechanisms mediating exercise responsiveness in skeletal muscle, but also for the potential that such understanding might bring to the treatment and prevention of human diseases such as intermittent claudication.


Diffusion of oxygen through activated sludge flocs: experimental measurement, modeling, and implications for simultaneous nitrification and denitrification.
Diffusion of dissolved oxygen through activated sludge flocs was studied, as it represents a potential mechanism for simultaneous nitrification and denitrification in activated sludge systems. Dissolved oxygen profiles through six floc particles collected at different times from a full-scale activated sludge plant demonstrated that that the dissolved oxygen concentration declines through all floc particles. For larger floc particles (2-mm diameter and greater), the dissolved oxygen concentration reached near-zero values at depths depending on process operating conditions. A mathematical model based on diffusion of dissolved oxygen, organic substrate (methanol), ammonia, nitrite, and nitrate through a spherical floc and consumption of dissolved oxygen by heterotrophs and autotrophs accurately predicted the dissolved oxygen profile and required adjustment of only one model parameter--the concentration of heterotrophs. A different dissolved oxygen decline pattern was exhibited for the smaller floc particles characterized, with the dissolved oxygen reaching a non-zero plateau toward the center of the floc. This pattern was not reproduced with the mathematical model developed and suggests that additional mechanisms are responsible for the transport of dissolved oxygen into the center of these flocs. Implications of these results regarding the occurrence of simultaneous nitrification and denitrification include consideration of the factors that affect floc size and distribution (simultaneous nitrification and denitrification is maximized with larger floc particles), coupling of the International Water Association (London) activated models to predict activated sludge composition with diffusion models to consider intrafloc effects, and the effects of substrate diffusion on the apparent half-saturation constant for various substrates in activated sludge systems.


Model-based error diffusion for high fidelity lenticular screening.
Digital halftoning is the process of converting a continuous-tone image into an arrangement of black and white dots for binary display devices such as digital ink-jet and electrophotographic printers. As printers are achieving print resolutions exceeding 1,200 dots per inch, it is becoming increasingly important for halftoning algorithms to consider the variations and interactions in the size and shape of printed dots between neighboring pixels. In the case of lenticular screening where statistically independent images are spatially multiplexed together, ignoring these variations and interactions, such as dot overlap, will result in poor lenticular image quality. To this end, we describe our use of model-based error-diffusion for the lenticular screening problem where statistical independence between component images is achieved by restricting the diffusion of error to only those pixels of the same component image where, in order to avoid instabilities, the proposed approach involves a novel error-clipping procedure.


Contemporary management of severe influenza disease in the intensive care unit.
Despite continued efforts to optimize vaccination composition, severe influenza disease requiring intensive care unit (ICU) admission remains a clinical issue. Influenza epidemics and pandemics worldwide continue to challenge clinicians with managing infected patients requiring ICU care. While routine use of antiviral therapy is deployed in ambulatory outpatients, their use in the ICU in patients with hypoxemic respiratory failure is less well established. Additionally, these therapies primarily target the neuraminidase protein, while contemporary research is increasingly demonstrating potential therapeutic benefits of targeting the hemagglutinin protein. These data have given rise to a growing interest in the use of immune modulating therapies for treatment of severe influenza. Additionally, pandemic outbreaks have revealed the growing need for salvage management, wherein lies the potential role for venovenous extracorporeal membrane oxygenation therapy in refractory respiratory failure. In this report, we review the contemporary ICU care of the severe influenza patient.


Post-September 11, 2001, Incidence of Systemic Autoimmune Diseases in World Trade Center-Exposed Firefighters and Emergency Medical Service Workers.
To estimate the incidence of selected systemic autoimmune diseases (SAIDs) in approximately 14,000 male rescue/recovery workers enrolled in the Fire Department of the City of New York (FDNY) World Trade Center (WTC) Health Program and to compare FDNY incidence to rates from demographically similar men in the Rochester Epidemiology Project (REP), a population-based database in Olmsted County, Minnesota. We calculated incidence for specific SAIDs (rheumatoid arthritis, psoriatic arthritis, systemic lupus erythematosus, and others) and combined SAIDs diagnosed from September 12, 2001, through September 11, 2014, and generated expected sex- and age-specific rates based on REP rates. Rates were stratified by level of WTC exposure (higher vs lower). Standardized incidence ratios (SIRs), which are the ratios of the observed number of cases in the FDNY group to the expected number of cases based on REP rates, and 95% CIs were calculated. We identified 97 SAID cases. Overall, FDNY rates were not significantly different from expected rates (SIR, 0.97; 95% CI, 0.77-1.21). However, the lower WTC exposure group had 9.9 fewer cases than expected, whereas the higher WTC exposure group had 7.7 excess cases. Most studies indicate that the healthy worker effect reduces the association between exposure and outcome by about 20%, which we observed in the lower WTC exposure group. Overall rates masked differences in incidence by level of WTC exposure, especially because the higher WTC exposure group was relatively small. Continued surveillance for early detection of SAIDs in high WTC exposure populations is required to identify and treat exposure-related adverse effects.


Blood pressure screening practices of a group of dental hygienists: a pilot study.
Recent research suggests that one in five Americans has hypertension, according to the American Heart Association. When hypertension is undiagnosed or uncontrolled, it places patients at risk for other cardiovascular diseases, contributing to an increase in mortality. Dental hygienists are in an ideal setting to screen for this silent disease. This study was designed to determine how frequently a group of practicing dental hygienists performs screenings for hypertension on their patients, and to determine the barriers that prevent this screening from occurring. One hundred one dental hygienists were questioned with a written survey about their blood pressure screening practices and their reasons for not taking blood pressure readings, if applicable. Sixty-seven dental hygienists completed the survey. Survey results revealed that the majority of dental hygienists were not recording blood pressure readings, even though their dental hygiene school curriculum had emphasized doing so for all patients. The most frequently cited reasons for not performing a routine blood pressure screening were insufficient time in the appointment and the minimal value given to the procedure by their employers. To work against obstacles that prevent the provision of this service, dental and dental hygiene faculty must increase their efforts to inculcate in their students the value of blood pressure screening. Further studies are needed to determine if the findings of this study are indicative of only one segment of dental hygiene practitioners, or if they represent the norm in the profession.


Supplementation of soy protein with branched-chain amino acids alters protein metabolism in healthy elderly and even more in patients with chronic obstructive pulmonary disease.
It is often suggested that chronic wasting diseases [eg, chronic obstructive pulmonary disease (COPD)] may benefit from branched-chain amino acid (BCAA) administration via improved protein metabolism. The aim was to examine whether adding BCAAs to a soy protein meal would enhance protein anabolism in COPD patients and in healthy elderly persons. Eight normal-weight COPD patients and 8 healthy control subjects were examined on 2 test days. Simultaneous continuous intravenous infusion of l-[ring-(2)H(5)]phenylalanine (Phe) and l-[ring-(2)H(2)]tyrosine tracers was done postabsorptively and at 2 h of ingestion of a maltodextrin soy or maltodextrin soy + BCAA protein meal (rate of ingestion: 0.02 g protein.kg body weight(-1).20 min(-1)) in a crossover design. Together with the meal, oral ingestion of 1-[(13)C]Phe was performed to measure first-pass Phe splanchnic extraction (SPE(Phe)). The endogenous rate of Phe appearance [reflecting whole-body protein breakdown (WbPB)], whole-body protein synthesis (WbPS), and net WbPS (WbPS - WbPB) were calculated. Arterialized venous blood was sampled for amino acid enrichment and concentration analyses. Soy feeding induced a reduction in WbPB and an increase in WbPS. BCAA supplementation of soy protein resulted in a significantly higher (P < 0.05) increase in WbPS than did soy protein alone in COPD patients but not in the healthy elderly. BCAA supplementation did not significantly alter the change in WbPB or net WbPS. Furthermore, BCAA supplementation decreased (absolute) SPE(Phe) (P < 0.05) but did not change the percentage Phe hydroxylation in the splanchnic area, which indicates a BCAA-related reduction in splanchnic protein synthesis. BCAA supplementation to soy protein enhances WbPS in patients with COPD and alters interorgan protein metabolism in favor of the peripheral (muscle) compartment in healthy elderly and even more in COPD patients.


An optimized in vitro blood-brain barrier model reveals bidirectional transmigration of African trypanosome strains.
The transmigration of African trypanosomes across the human blood-brain barrier (BBB) is the critical step during the course of human African trypanosomiasis. The parasites Trypanosoma brucei gambiense and T. b. rhodesiense are transmitted to humans during the bite of tsetse flies. Trypanosomes multiply within the bloodstream and finally invade the central nervous system (CNS), which leads to the death of untreated patients. This project focused on the mechanisms of trypanosomal traversal across the BBB. In order to establish a suitable in vitro BBB model for parasite transmigration, different human cell lines were used, including ECV304, HBMEC and HUVEC, as well as C6 rat astrocytes. Validation of the BBB models with Escherichia coli HB101 and E. coli K1 revealed that a combination of ECV304 cells seeded on Matrigel as a semi-synthetic basement membrane and C6 astrocytes resulted in an optimal BBB model system. The BBB model showed selective permeability for the pathogenic E. coli K1 strain, and African trypanosomes were able to traverse the optimized ECV304-C6 BBB efficiently. Furthermore, coincubation indicated that paracellular macrophage transmigration does not facilitate trypanosomal BBB traversal. An inverse assembly of the BBB model demonstrated that trypanosomes were also able to transmigrate the optimized ECV304-C6 BBB backwards, indicating the relevance of the CNS as a possible reservoir of a relapsing parasitaemia.


Impact of functional foods and nutraceuticals on high blood pressure with a special focus on meta-analysis: review from a public health perspective.
In recent times many researchers are expressing immense interest in nutraceuticals and functional foods for combating various diseases or abnormal conditions, especially against hypertension (HT). Persistent HT is medically referred to as chronic high blood pressure (BP) and considered to be one of the major risk factors for the deadliest diseases including cardiovascular disease (CVD) and cerebrovascular diseases. Hence HT poses a serious socio-economic burden worldwide, particularly to developing countries. The current treatment strategy for HT includes standard anti-hypertensive drugs, which are associated with many adverse effects and lower drug adherence rates. Therefore, an alternative or complementary natural therapy (functional foods or nutraceuticals or dietary supplements) would be the alternate choice along with a modified lifestyle pattern that might help to manage or combat HT and its related complications. During this review, the author would like to shed light on the basic science behind HT including pathophysiology and the impact of dietary salt on HT and the impact of various functional foods or nutraceuticals against HT in humans (meta-analysis and systemic review). This contribution gives a better idea (public health perspective) for choosing the best functional foods/nutraceuticals for the prevention, management or delaying the onset of HT and its associated conditions along with modified lifestyle patterns and standard anti-hypertensive drugs.


Combined inhibition of Notch and JAK/STAT is superior to monotherapies and impairs pancreatic cancer progression.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a high rate of metastasis. Recent studies have indicated that Notch and janus kinase 2 (JAK2)/signal transducers and activators of transcription 3 (STAT3) signaling pathways are both important for the initiation and progression of PDAC. The purpose of this study was to determine the outcome of targeting these two tumor signaling pathways simultaneously both in vitro and in vivo. We assessed the combinational effects of the γ-secretase inhibitor IX (GSI IX) and JAK2 inhibitor (AG-490) on growth and epithelial plasticity of human pancreatic cancer cell lines, and in a genetically engineered mouse model (Pdx1-Cre, LSL-KrasG12D, p53(lox/+)) of PDAC. Dual treatment with GSI IX and AG-490 significantly impaired cell proliferation, migration, invasion, soft agar growth and apoptosis when compared with monotherapies. Most importantly, combinational treatment significantly attenuates tumor progression in vivo and suppresses conversion from acinar-ductal-metaplasia to PDAC. Our results suggest that targeting Notch and JAK2/STAT3 signaling pathways simultaneously is superior to single inhibitions, supporting combined treatment by GSI IX and AG-490 as a potential therapeutic approach for PDAC. However, the study design limits the direct transfer into the clinic and the impact of dual treatment in patients with PDAC remains still to be determined.


Peripheral T-Cell Lymphoma, Not Otherwise Specified and Concurrent Seminoma in Testis.
Concurrent seminoma and malignant lymphoma of the testis is rare. We present a case of concurrent seminoma and peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) in a 54-year-old man who complained of painless left testicular enlargement. Radical left orchiectomy was performed. Macroscopically, the tumor (4.0 × 3.0 cm) was creamy, soft, and homogeneous, and microscopic evaluation revealed an alveolar structure of large cells that formed sheets, as well as colonization by other abnormal cells in a 1.0 × 1.0 cm area. The portion of the tumor comprising large abnormal cells was diagnosed as a seminoma, which was positive for c-kit by immunohistochemistry; the other portion was diagnosed as CD3/CD8, TIA, and granzyme B-positive PTCL-NOS. These two portions were clearly differentiated from one another. The patient received CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone) therapy and achieved complete response for 50 months. To our knowledge, this is the first reported case of synchronous advanced seminoma and PTCL.


Virus-specific immune memory at peripheral sites of herpes simplex virus type 2 (HSV-2) infection in guinea pigs.
Despite its importance in modulating HSV-2 pathogenesis, the nature of tissue-resident immune memory to HSV-2 is not completely understood. We used genital HSV-2 infection of guinea pigs to assess the type and location of HSV-specific memory cells at peripheral sites of HSV-2 infection. HSV-specific antibody-secreting cells were readily detected in the spleen, bone marrow, vagina/cervix, lumbosacral sensory ganglia, and spinal cord of previously-infected animals. Memory B cells were detected primarily in the spleen and to a lesser extent in bone marrow but not in the genital tract or neural tissues suggesting that the HSV-specific antibody-secreting cells present at peripheral sites of HSV-2 infection represented persisting populations of plasma cells. The antibody produced by these cells isolated from neural tissues of infected animals was functionally relevant and included antibodies specific for HSV-2 glycoproteins and HSV-2 neutralizing antibodies. A vigorous IFN-γ-secreting T cell response developed in the spleen as well as the sites of HSV-2 infection in the genital tract, lumbosacral ganglia and spinal cord following acute HSV-2 infection. Additionally, populations of HSV-specific tissue-resident memory T cells were maintained at these sites and were readily detected up to 150 days post HSV-2 infection. Unlike the persisting plasma cells, HSV-specific memory T cells were also detected in uterine tissue and cervicothoracic region of the spinal cord and at low levels in the cervicothoracic ganglia. Both HSV-specific CD4+ and CD8+ resident memory cell subsets were maintained long-term in the genital tract and sensory ganglia/spinal cord following HSV-2 infection. Together these data demonstrate the long-term maintenance of both humoral and cellular arms of the adaptive immune response at the sites of HSV-2 latency and virus shedding and highlight the utility of the guinea pig infection model to investigate tissue-resident memory in the setting of HSV-2 latency and spontaneous reactivation.


Influence of sports flooring and shoes on impact forces and performance during jump tasks.
We aim to determine the influence of sports floorings and sports shoes on impact mechanics and performance during standardised jump tasks. Twenty-one male volunteers performed ankle jumps (four consecutive maximal bounds with very dynamic ankle movements) and multi-jumps (two consecutive maximal counter-movement jumps) on force plates using minimalist and cushioned shoes under 5 sports flooring (SF) conditions. The shock absorption properties of the SF, defined as the proportion of peak impact force absorbed by the tested flooring when compared with a concrete hard surface, were: SF0 = 0% (no flooring), SF1 = 19%, SF2 = 26%, SF3 = 37% and SF4 = 45%. Shoe and flooring effects were compared using 2x5 repeated-measures ANOVA with post-hoc Bonferroni-corrected comparisons. A significant interaction between SF and shoe conditions was found for VILR only (p = 0.003). In minimalist shoes, SF influenced Vertical Instantaneous Loading Rate (VILR) during ankle jumps (p = 0.006) and multi-jumps (p<0.001), in accordance with shock absorption properties. However, in cushioned shoes, SF influenced VILR during ankle jumps only (p<0.001). Contact Time was the only additional variable affected by SF, but only during multi-jumps in minimalist shoes (p = 0.037). Cushioned shoes induced lower VILR (p<0.001) and lower Contact Time (p≤0.002) during ankle jumps and multi-jumps compared to minimalist shoes. During ankle jumps, cushioned shoes induced greater Peak Vertical Ground Reaction Force (PVGRF, p = 0.002), greater Vertical Average Loading Rate (p<0.001), and lower eccentric (p = 0.008) and concentric (p = 0.004) work. During multi-jumps, PVGRF was lower (p<0.001) and jump height was higher (p<0.001) in cushioned compared to minimalist shoes. In conclusion, cushioning influenced impact forces during standardised jump tasks, whether it was provided by the shoes or the sports flooring. VILR is the variable that was the most affected.


Alteration of lymphocyte trafficking by sphingosine-1-phosphate receptor agonists.
Blood lymphocyte numbers, essential for the development of efficient immune responses, are maintained by recirculation through secondary lymphoid organs. We show that lymphocyte trafficking is altered by the lysophospholipid sphingosine-1-phosphate (S1P) and by a phosphoryl metabolite of the immunosuppressive agent FTY720. Both species were high-affinity agonists of at least four of the five S1P receptors. These agonists produce lymphopenia in blood and thoracic duct lymph by sequestration of lymphocytes in lymph nodes, but not spleen. S1P receptor agonists induced emptying of lymphoid sinuses by retention of lymphocytes on the abluminal side of sinus-lining endothelium and inhibition of egress into lymph. Inhibition of lymphocyte recirculation by activation of S1P receptors may result in therapeutically useful immunosuppression.


A sequence-activated AND logic dual-channel fluorescent probe for tracking programmable drug release.
The translation of biomarker sensing into programmable diagnostics or therapeutic applications in vivo is greatly challenging, especially for eliminating the 'false positive' signals from OR logic gates. Herein we present a sense-of-logic dual-channel nanoprobe, operating through a sequence-activated AND logic gate by responding ultra-sensitively to pH changes and being subsequently triggered with biothiol for the controllable release of anti-cancer drugs. Specifically, programmable drug release is conducted in a multistage tumor microenvironment (acidic endocytic organelles followed by abnormal glutathione-overexpressing cell cytosol), which is synchronous with dual-channel near-infrared (NIR) fluorescence output. This approach represents the merging of sensing and release, including logically enabled molecular design, biomarker sensing, and controllable drug release. Impressively, the sequential AND logic feature within an unprecedented framework provides feedback on the diversity and complexity of biological milieu, along with remarkably enhancing the tumor therapeutic efficiency via its precise targeting ability and programmable drug release.


Physical interaction of RECQ5 helicase with RAD51 facilitates its anti-recombinase activity.
Homologous recombination (HR) provides an efficient mechanism for error-free repair of DNA double-strand breaks (DSBs). However, HR can be also harmful as inappropriate or untimely HR events can give rise to lethal recombination intermediates and chromosome rearrangements. A critical step of HR is the formation of a RAD51 filament on single-stranded (ss)DNA, which mediates the invasion of a homologous DNA molecule. In mammalian cells, several DNA helicases have been implicated in the regulation of this process. RECQ5, a member of the RecQ family of DNA helicases, interacts physically with the RAD51 recombinase and disrupts RAD51 presynaptic filaments in a reaction dependent on ATP hydrolysis. Here, we have precisely mapped the RAD51-interacting domain of RECQ5 and generated mutants that fail to interact with RAD51. We show that although these mutants retain normal ATPase activity, they are impaired in their ability to displace RAD51 from ssDNA. Moreover, we show that ablation of RECQ5-RAD51 complex formation by a point mutation alleviates the inhibitory effect of RECQ5 on HR-mediated DSB repair. These findings provide support for the proposal that interaction with RAD51 is critical for the anti-recombinase attribute of RECQ5.


Gene dosage of the amyloid beta precursor protein in Alzheimer's disease.
The progressive deposition in the human brain of amyloid filaments composed of the amyloid beta protein is a principal feature of Alzheimer's disease (AD). Densitometric analysis of Southern blots probed with a complementary DNA for the amyloid protein has been carried out to determine the relative dosage of this gene in genomic DNA of 14 patients with AD, 12 aged normal subjects, and 10 patients with trisomy 21 (Down syndrome). Whereas patients in the last group showed the expected 1.5-fold increase in dosage of this gene, none of the patients with AD had a gene dosage higher than that of the normal controls. These results do not support the hypothesis that the genetic defect in AD involves duplication of a segment of chromosome 21 containing the amyloid gene. Alternative mechanisms for the brain-specific increase in amyloid protein deposition in AD should be considered.


Distinct Contributions of Tryptophan Residues within the Dimerization Domain to Nanog Function.
The level of the transcription factor Nanog directly determines the efficiency of mouse embryonic stem cell self-renewal. Nanog protein exists as a dimer with the dimerization domain composed of a simple repeat region in which every fifth residue is a tryptophan, the tryptophan repeat (WR). Although WR is necessary to enable Nanog to confer LIF-independent self-renewal, the mechanism of dimerization and the effect of modulating dimerization strength have been unclear. Here we couple mutagenesis with functional and dimerization assays to show that the number of tryptophans within the WR is linked to the strength of homodimerization, Sox2 heterodimerization and self-renewal activity. A reduction in the number of tryptophan residues leads initially to a gradual reduction in activity before a precipitous reduction in activity occurs upon reduction in tryptophan number below eight. Further functional attrition follows subsequent tryptophan number reduction with substitution of all tryptophan residues ablating dimerization and self-renewal function completely. A strong positional influence of tryptophans exists, with residues at the WR termini contributing more to Nanog function, particularly at the N-terminal end. Limited proteolysis demonstrates that a structural core of Nanog encompassing the homeodomain and the tryptophan repeat can support LIF-independent colony formation. These results increase understanding of the molecular interactions occurring between transcription factor subunits at the core of the pluripotency gene regulatory network and will enhance our ability to control pluripotent cell self-renewal and differentiation.


Optical design of a light-emitting diode lamp for a maritime lighthouse.
Traffic signaling is an emerging field for light-emitting diode (LED) applications. This sustainable power-saving illumination technology can be used in maritime signaling thanks to the recently updated norms, where the possibility to utilize LED sources is explicitly cited, and to the availability of high-power white LEDs that, combined with suitable lenses, permit us to obtain well-collimated beams. This paper describes the optical design of a LED-based lamp that can replace a traditional lamp in an authentic marine lighthouse. This source recombines multiple separated LEDs realizing a quasi-punctual localized source. Advantages can be lower energy consumption, higher efficiency, longer life, fewer faults, slower aging, and minor maintenance costs. The proposed LED source allows us to keep and to utilize the old Fresnel lenses of the lighthouse, which very often have historical value.


Human temporal lobe activation by speech and nonspeech sounds.
Functional organization of the lateral temporal cortex in humans is not well understood. We recorded blood oxygenation signals from the temporal lobes of normal volunteers using functional magnetic resonance imaging during stimulation with unstructured noise, frequency-modulated (FM) tones, reversed speech, pseudowords and words. For all conditions, subjects performed a material-nonspecific detection response when a train of stimuli began or ceased. Dorsal areas surrounding Heschl's gyrus bilaterally, particularly the planum temporale and dorsolateral superior temporal gyrus, were more strongly activated by FM tones than by noise, suggesting a role in processing simple temporally encoded auditory information. Distinct from these dorsolateral areas, regions centered in the superior temporal sulcus bilaterally were more activated by speech stimuli than by FM tones. Identical results were obtained in this region using words, pseudowords and reversed speech, suggesting that the speech-tones activation difference is due to acoustic rather than linguistic factors. In contrast, previous comparisons between word and nonword speech sounds showed left-lateralized activation differences in more ventral temporal and temporoparietal regions that are likely involved in processing lexical-semantic or syntactic information associated with words. The results indicate functional subdivision of the human lateral temporal cortex and provide a preliminary framework for understanding the cortical processing of speech sounds.


Spinal deformity and instability after multilevel cervical laminectomy.
Sixty-four patients who had undergone multilevel cervical laminectomy were studied for postoperative spinal deformity and instability. Special attention was given to patients with cervical spondylosis (CS), ossification of the posterior longitudinal ligament (OPLL), and spinal cord tumors. Twenty-three (36%) of 64 patients showed postoperative changes in curvature type and 9 (14%) had developed spinal deformity (kyphotic or meandering-type curvature). In two juvenile patients, the deformity developed soon after operation and spinal fusion was required to prevent neurologic complications. In the adult cases, contrary to the hitherto accepted concept, long-term follow-up revealed the tendency of the deformity to develop more frequently in OPLL cases than in CS cases. Mobility of the cervical spine was reduced considerably after laminectomy, both in CS and OPLL cases. There was no adult patient who required further operation for severe deformity or instability after laminectomy. Extensive laminectomy, even including the C2 lamina, seemed to have no adverse effect on the stability of the cervical spine.


Dissimilar effects of organohalogenated compounds on thyroid hormones in glaucous gulls.
The glaucous gull (Larus hyperboreus) is an arctic top predator and scavenger exposed to high levels of mixtures of organohalogenated contaminants (OHCs) of which many interfere with the thyroid hormone (TH) system. In the present study, we applied statistical modeling to investigate the potential combined influence of the mixture of chlorinated, brominated and perfluorinated organic compounds in plasma of glaucous gulls on their plasma TH concentrations. In females, there were significant negative associations between several organochlorinated compounds (OCs) and free thyroxin (FT4) and triiodothyronine (FT3), indicating additive negative effects on FT4 and FT3. However, in these females there was also a significant positive association between perfluorooctane sulfonate (PFOS) and FT3. The inverse associations between several OCs and FT3 and the contrasting positive association between PFOS and FT3, indicate that these two groups of OHCs may have dissimilar and antagonistic effects on FT3 in female glaucous gulls. In males, there were no associations between any of the OHCs and the THs. That OHCs affect THs in a complex manner involving both additive and antagonistic effects add to the challenge of interpreting the overall functional effect of thyroid disruptive chemicals in wildlife. However, experimental studies are needed to confirm or disprove such effects.


Ventricular access devices are safe and effective in the treatment of posthemorrhagic ventricular dilatation prior to shunt placement.
Intraventricular hemorrhage of prematurity (IVH) is a diagnosis that has become more frequent in recent years. Advances in medical care have led to survival of increasingly premature infants, as well as infants with more complex medical conditions. Treatment with a ventricular access device (VAD) was reported almost 3 decades ago; however, it is unclear how effective this treatment is in the current population of premature infants. At our institution (from 2004 to present), we treat posthemorrhagic hydrocephalus (PHH) with a VAD. In order to look at safety and efficacy, we retrospectively combed the medical records of premature children, admitted to Lucile Packard Children's Hospital from January 2005 to December 2009, and identified 310 premature children with IVH. Of these, 28 children required treatment for PHH with a VAD. There were no infections associated with placement of these devices and a very low rate of other complications, such as need for repositioning (7.41%) or replacement (3.75%). Our data show that treatment with a VAD is very safe, with few complications and can be used to treat PHH in this very complex infant population.


Insights into DNA substrate selection by APOBEC3G from structural, biochemical, and functional studies.
Human apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3 (A3) proteins are a family of cytidine deaminases that catalyze the conversion of deoxycytidine (dC) to deoxyuridine (dU) in single-stranded DNA (ssDNA). A3 proteins act in the innate immune response to viral infection by mutating the viral ssDNA. One of the most well-studied human A3 family members is A3G, which is a potent inhibitor of HIV-1. Each A3 protein prefers a specific substrate sequence for catalysis-for example, A3G deaminates the third dC in the CCCA sequence motif. However, the interaction between A3G and ssDNA is difficult to characterize due to poor solution behavior of the full-length protein and loss of DNA affinity of the truncated protein. Here, we present a novel DNA-anchoring fusion strategy using the protection of telomeres protein 1 (Pot1) which has nanomolar affinity for ssDNA, with which we captured an A3G-ssDNA interaction. We crystallized a non-preferred adenine in the -1 nucleotide-binding pocket of A3G. The structure reveals a unique conformation of the catalytic site loops that sheds light onto how the enzyme scans substrate in the -1 pocket. Furthermore, our biochemistry and virology studies provide evidence that the nucleotide-binding pockets on A3G influence each other in selecting the preferred DNA substrate. Together, the results provide insights into the mechanism by which A3G selects and deaminates its preferred substrates and help define how A3 proteins are tailored to recognize specific DNA sequences. This knowledge contributes to a better understanding of the mechanism of DNA substrate selection by A3G, as well as A3G antiviral activity against HIV-1.


Knowing where the nose is.
Improvements in imaging technology and the development of powerful machine learning algorithms are revolutionizing the study of animal behavior in the laboratory. These innovations promise to reveal both global and local features of action relevant to understanding how the brain functions. A study in BMC Biology describes one such tool called OptiMouse, which is an open source platform that uses video to capture key features of mouse behavior, including information relevant to olfactory investigation.See research article: 10.1186/s12915-017-0377-3.


Glucose and NAADP trigger elementary intracellular β-cell Ca2+ signals.
Pancreatic β-cells release insulin upon a rise in blood glucose. The precise mechanisms of stimulus-secretion coupling, and its failure in Diabetes Mellitus Type 2, remain to be elucidated. The consensus model, as well as a class of currently prescribed anti-diabetic drugs, are based around the observation that glucose-evoked ATP production in β-cells leads to closure of cell membrane ATP-gated potassium (KATP) channels, plasma membrane depolarisation, Ca2+ influx, and finally the exocytosis of insulin granules. However, it has been demonstrated by the inactivation of this pathway using genetic and pharmacological means that closure of the KATP channel alone may not be sufficient to explain all β-cell responses to glucose elevation. We have previously proposed that NAADP-evoked Ca2+ release is an important step in stimulus-secretion coupling in pancreatic β-cells. Here we show using total internal reflection fluorescence (TIRF) microscopy that glucose as well as the Ca2+ mobilising messenger nicotinic acid adenine dinucleotide phosphate (NAADP), known to operate in β-cells, lead to highly localised elementary intracellular Ca2+ signals. These were found to be obscured by measurements of global Ca2+ signals and the action of powerful SERCA-based sequestration mechanisms at the endoplasmic reticulum (ER). Building on our previous work demonstrating that NAADP-evoked Ca2+ release is an important step in stimulus-secretion coupling in pancreatic β-cells, we provide here the first demonstration of elementary Ca2+ signals in response to NAADP, whose occurrence was previously suspected. Optical quantal analysis of these events reveals a unitary event amplitude equivalent to that of known elementary Ca2+ signalling events, inositol trisphosphate (IP3) receptor mediated blips, and ryanodine receptor mediated quarks. We propose that a mechanism based on these highly localised intracellular Ca2+ signalling events mediated by NAADP may initially operate in β-cells when they respond to elevations in blood glucose.


Molecular determinants of pathogenicity in the polymerase complex.
Viral pathogenesis involves numerous interactions between viral and cellular factors. In recent years, the influenza virus polymerase complex has emerged as a major determinant of interspecies transmission and pathogenicity. The viral RNA-dependent RNA polymerase, in concert with the nucleoprotein, mediates transcription and replication of the viral RNA genome in the nucleus of the infected cell. The activity by which the viral polymerase complex performs these processes in mammalian cells is considered to be a major contributor to viral pathogenicity in mammals. In this chapter, we summarise our current understanding on the pathogenicity determinants in the viral polymerase complex and highlight some of its cellular interaction partners. We particularly discuss the role of importin-α isoforms in host adaptation and pathogenesis as well as the role of the viral polymerase in regulating cellular responses to viral infection.


Somatic Mutations and Neoepitope Homology in Melanomas Treated with CTLA-4 Blockade.
Immune checkpoint inhibitors are promising treatments for patients with a variety of malignancies. Toward understanding the determinants of response to immune checkpoint inhibitors, it was previously demonstrated that the presence of somatic mutations is associated with benefit from checkpoint inhibition. A hypothesis was posited that neoantigen homology to pathogens may in part explain the link between somatic mutations and response. To further examine this hypothesis, we reanalyzed cancer exome data obtained from our previously published study of 64 melanoma patients treated with CTLA-4 blockade and a new dataset of RNA-Seq data from 24 of these patients. We found that the ability to accurately predict patient benefit did not increase as the analysis narrowed from somatic mutation burden, to inclusion of only those mutations predicted to be MHC class I neoantigens, to only including those neoantigens that were expressed or that had homology to pathogens. The only association between somatic mutation burden and response was found when examining samples obtained prior to treatment. Neoantigen and expressed neoantigen burden were also associated with response, but neither was more predictive than somatic mutation burden. Neither the previously described tetrapeptide signature nor an updated method to evaluate neoepitope homology to pathogens was more predictive than mutation burden. Cancer Immunol Res; 5(1); 84-91. ©2016 AACR.


Faecal phytic acid and its relation to other putative markers of risk for colorectal cancer.
Phytic acid, a major constituent of cereals, pulses, and seeds has been advocated as an important antioxidant component of dietary fibre that affords possible protection against colorectal cancer. This is supported by experimental studies showing it has antineoplastic activity in animal models of both colon and breast cancer. To date the concentration of faecal phytic acid in human clinical groups has not been evaluated. Therefore the faecal phytic acid content of adenoma patients drawn from a placebo controlled calcium intervention trial was evaluated. Phytic acid was measured in faecal extracts by an improved ion-pair high performance liquid chromatography method. Phytic acid was detected in the range 0.68-4.00 mumol/g wet faeces and 55-2038 mumol/day. Linear regression analyses showed no association between stool phytic acid and lipid content. Strong correlations were seen, however, between phytic acid and iron content, both on a concentration (r = 0.52; p = 0.00004) and daily excretion (r = 0.76; p = 5.5 x 10(-12) basis. Phytic acid was also strongly correlated with the daily excretion of calcium (r = 0.59; p = 1.36 x 10(-6) and magnesium (r = 0.42; p = 0.001). Cell proliferation in the sigmoid colon, an intermediate biomarker of colorectal cancer was not significantly associated with faecal phytic acid, minerals or lipid content in this compromised clinical group. This improved method, developed for the determination of phytic acid in faeces should allow further studies on the role of phytic acid in the aetiology of colorectal cancer to be conducted on a population or case control basis.


Detection of genetically modified maize and soybean in feed samples.
Despite the controversy about genetically modified (GM) plants, they are still incrementally cultivated. In recent years, many food and feed products produced by genetic engineering technology have appeared on store shelves. Controlling the production and legal presentation of GM crops are very important for the environment and human health, especially in terms of long-term consumption. In this study, 11 kinds of feed obtained from different regions of Turkey were used for genetic analysis based on foreign gene determination. All samples were screened by conventional polymerase chain reaction (PCR) technique for widely used genetic elements; cauliflower mosaic virus 35S promoter (CaMV35S promoter), and nopaline synthase terminator (T-NOS) sequences for GM plants. After determination of GM plant-containing samples, nested PCR and conventional PCR analysis were performed to find out whether the samples contained Bt176 or GTS-40-3-2 for maize and soy, respectively. As a result of PCR-based GM plant analysis, all samples were found to be transgenic. Both 35S- and NOS-containing feed samples or potentially Bt176-containing samples, in other words, were analyzed with Bt176 insect resistant cryIAb gene-specific primers via nested PCR. Eventually, none of them were found Bt176-positive. On the other hand, when we applied conventional PCR to the same samples with the herbicide resistance CTP4-EPSPS construct-specific primers for transgenic soy variety GTS-40-3-2, we found that all samples were positive for GTS-40-3-2.