A clinical investigation of obstructive sleep apnea syndrome (OSAS) and upper airway resistance syndrome (UARS) patients.
Objective: (i) Evaluation of the clinical differences and similarities presented by patients diagnosed as OSAS and UARS subjects. (ii) Evaluation of the ability of a sleep disorders specialist to dissociate the two syndromes based upon clinical evaluation.Population: 314 subjects were included. They were referred to a sleep disorders clinic with complaints of loud snoring during a 3 month period.Method: The evaluation consisted of: (i) Clinical interview and evaluation. (ii) Administration of validated questionnaires (Sleep Disorders Questionnaire and Epworth Sleepiness Scale). (iii) Establishment of clinical diagnostic and results of polygraphic recording.Results: After clinical evaluation and polygraphic recordings (performed within 3 weeks of initial evaluation) patients were subdivided into two groups: 176 OSAS and 128 UARS. The misclassification of patients by specialists correlated with body mass index (BMI) measurement, with an over classification of patient as OSAS when a high BMI was noted and vice-versa for UARS. The only significant difference between OSAS and UARS patients was an older age and a wider neck circumference in the OSAS group than in UARS patients.Conclusion: Clinical presentation including daytime sleepiness complaint and ESS score is similar for patients with and without drop of oxygen saturation below 90% during sleep. There was always a male predominance within both syndromes, but more women were diagnosed with UARS than with OSAS.


Mosquitoes on a plane: Disinsection will not stop the spread of vector-borne pathogens, a simulation study.
Mosquito-borne diseases are increasingly being recognized as global threats, with increased air travel accelerating their occurrence in travelers and their spread to new locations. Since the early days of aviation, concern over the possible transportation of infected mosquitoes has led to recommendations to disinsect aircraft. Despite rare reports of mosquitoes, most likely transported on aircraft, infecting people far from endemics areas, it is unclear how important the role of incidentally transported mosquitoes is compared to the role of traveling humans. We used data for Plasmodium falciparum and dengue viruses to estimate the probability of introduction of these pathogens by mosquitoes and by humans via aircraft under ideal conditions. The probability of introduction of either pathogen by mosquitoes is low due to few mosquitoes being found on aircraft, low infection prevalence among mosquitoes, and high mortality. Even without disinsection, introduction via infected human travelers was far more likely than introduction by infected mosquitoes; more than 1000 times more likely for P. falciparum and more than 200 times more likely for dengue viruses. Even in the absence of disinsection and under the most favorable conditions, introduction of mosquito-borne pathogens via air travel is far more likely to occur as a result of an infected human travelling rather than the incidental transportation of infected mosquitoes. Thus, while disinsection may serve a role in preventing the spread of vector species and other invasive insects, it is unlikely to impact the spread of mosquito-borne pathogens.


Enterococcus faecium secreted antigen A generates muropeptides to enhance host immunity and limit bacterial pathogenesis.
We discovered that Enterococcus faecium (E. faecium), a ubiquitous commensal bacterium, and its secreted peptidoglycan hydrolase (SagA) were sufficient to enhance intestinal barrier function and pathogen tolerance, but the precise biochemical mechanism was unknown. Here we show E. faecium has unique peptidoglycan composition and remodeling activity through SagA, which generates smaller muropeptides that more effectively activates nucleotide-binding oligomerization domain-containing protein 2 (NOD2) in mammalian cells. Our structural and biochemical studies show that SagA is a NlpC/p60-endopeptidase that preferentially hydrolyzes crosslinked Lys-type peptidoglycan fragments. SagA secretion and NlpC/p60-endopeptidase activity was required for enhancing probiotic bacteria activity against Clostridium difficile pathogenesis in vivo. Our results demonstrate that the peptidoglycan composition and hydrolase activity of specific microbiota species can activate host immune pathways and enhance tolerance to pathogens.


Choline: an essential nutrient for public health.
Choline was officially recognized as an essential nutrient by the Institute of Medicine (IOM) in 1998. There is significant variation in the dietary requirement for choline that can be explained by common genetic polymorphisms. Because of its wide-ranging roles in human metabolism, from cell structure to neurotransmitter synthesis, choline-deficiency is now thought to have an impact on diseases such as liver disease, atherosclerosis, and, possibly, neurological disorders. Choline is found in a wide variety of foods. Eggs and meats are rich sources of choline in the North American diet, providing up to 430 milligrams per 100 grams. Mean choline intakes for older children, men, women, and pregnant women are far below the adequate intake level established by the IOM. Given the importance of choline in a wide range of critical functions in the human body, coupled with less-than-optimal intakes among the population, dietary guidance should be developed to encourage the intake of choline-rich foods.


The effects of synthetic cannabinoids on the cardiovascular system: A case-control study.
Limited clinical studies have investigated the effects of synthetic cannabinoids (SCs) on the cardiovascular system (CVS). The aim of this study was to evaluate the effects of SCs on the CVS. The patient group of this single-center, prospective, case-control study consisted of adult patients presenting to the emergency department (ED) with symptoms of SC use. Vital signs and electrocardiogram (ECG) after use of SC of patients were followed. A control group with a similar number of patients and patient demographics were formed following the patient admission process. Pulse rate, arterial blood pressure (ABP), and ECG of patient and control groups were compared using Mann-Whitney U and Chi-squared tests. A total of 148 people were included in the study, 74 in the patient group and 74 in the control group. Systolic and diastolic ABPs of patient group were statistically significantly lower than those of the control group (P < 0.001). P-wave width and amplitude in the patient group were significantly higher compared to the control group (P: 0.027 and P: 0.004, respectively). QRS width on patient group ECGs was significantly higher than in the control group, while T-wave amplitude was significantly lower (P: 0.045 and P < 0.001, respectively). ST elevation was seen in 12 (16.2%) subjects in the patient group, while no ST elevation was seen in the control group (P < 0.001). SCs can reduce systemic tension and SCs may cause changes in ECG, especially P wave, ST segment, T wave, and QRS. Further large-scale studies are needed to show whether these changes are associated with fatal arrhythmias or myocardial infarction.


Viral metagenomics on animals as a tool for the detection of zoonoses prior to human infection?
Many human viral infections have a zoonotic, i.e., wild or domestic animal, origin. Several zoonotic viruses are transmitted to humans directly via contact with an animal or indirectly via exposure to the urine or feces of infected animals or the bite of a bloodsucking arthropod. If a virus is able to adapt and replicate in its new human host, human-to-human transmissions may occur, possibly resulting in an epidemic, such as the A/H1N1 flu pandemic in 2009. Thus, predicting emerging zoonotic infections is an important challenge for public health officials in the coming decades. The recent development of viral metagenomics, i.e., the characterization of the complete viral diversity isolated from an organism or an environment using high-throughput sequencing technologies, is promising for the surveillance of such diseases and can be accomplished by analyzing the viromes of selected animals and arthropods that are closely in contact with humans. In this review, we summarize our current knowledge of viral diversity within such animals (in particular blood-feeding arthropods, wildlife and domestic animals) using metagenomics and present its possible future application for the surveillance of zoonotic and arboviral diseases.


Adherence with bisphosphonate therapy in US veterans with rheumatoid arthritis.
Pharmacy Benefits Management program data for patients enrolled in the Veterans Affairs Rheumatoid Arthritis (VARA) registry were linked with clinical data to determine bisphosphonate adherence and persistence among US veterans with rheumatoid arthritis (RA) and to determine factors associated with adherence. The primary outcome measures were the duration of bisphosphonate therapy and the medication possession ratio (MPR). Patients with an MPR <0.80 were classified as nonadherent. Potential covariates considered in the analysis included patient demographics, RA disease activity and severity parameters, and factors associated with osteoporosis risk. Associations of patient factors with duration of therapy and adherence were examined using multivariable regression modeling. Bisphosphonates were prescribed to 573 (41.5%) of 1,382 VARA subjects. The mean ± SD duration of therapy for bisphosphonates was 39.2 ± 31.4 months. A longer duration of therapy correlated with older age, more years of education, and dual x-ray absorptiometry testing. The mean ± SD MPR of VARA subjects for bisphosphonate therapy was 0.69 ± 0.28; 302 (52.7%) were nonadherent. In multivariate analyses, nonadherence with bisphosphonate therapy was associated with a longer duration of RA disease (odds ratio [OR] 1.02, 95% confidence interval [95% CI] 1.00-1.04) and duration of bisphosphonate therapy >32 months (OR 1.63, 95% CI 1.04-2.57). Whites were less likely to have a low MPR compared with nonwhites (OR 0.52, 95% CI 0.30-0.88). Nonadherence with bisphosphonates was common in this cohort of RA patients and was associated with nonwhite ethnicity, a longer duration of RA disease, and a greater duration of bisphosphonate therapy.


Spatial prediction of malaria prevalence in an endemic area of Bangladesh.
Malaria is a major public health burden in Southeastern Bangladesh, particularly in the Chittagong Hill Tracts region. Malaria is endemic in 13 districts of Bangladesh and the highest prevalence occurs in Khagrachari (15.47%). A risk map was developed and geographic risk factors identified using a Bayesian approach. The Bayesian geostatistical model was developed from previously identified individual and environmental covariates (p < 0.2; age, different forest types, elevation and economic status) for malaria prevalence using WinBUGS 1.4. Spatial correlation was estimated within a Bayesian framework based on a geostatistical model. The infection status (positives and negatives) was modeled using a Bernoulli distribution. Maps of the posterior distributions of predicted prevalence were developed in geographic information system (GIS). Predicted high prevalence areas were located along the north-eastern areas, and central part of the study area. Low to moderate prevalence areas were predicted in the southwestern, southeastern and central regions. Individual age and nearness to fragmented forest were associated with malaria prevalence after adjusting the spatial auto-correlation. A Bayesian analytical approach using multiple enabling technologies (geographic information systems, global positioning systems, and remote sensing) provide a strategy to characterize spatial heterogeneity in malaria risk at a fine scale. Even in the most hyper endemic region of Bangladesh there is substantial spatial heterogeneity in risk. Areas that are predicted to be at high risk, based on the environment but that have not been reached by surveys are identified.


Inheritance and characterization of strong resistance to phosphine in Sitophilus oryzae (L.).
Sitophilus oryzae (Linnaeus) is a major pest of stored grain across Southeast Asia and is of increasing concern in other regions due to the advent of strong resistance to phosphine, the fumigant used to protect stored grain from pest insects. We investigated the inheritance of genes controlling resistance to phosphine in a strongly resistant S. oryzae strain (NNSO7525) collected in Australia and find that the trait is autosomally inherited and incompletely recessive with a degree of dominance of -0.66. The strongly resistant strain has an LC50 52 times greater than a susceptible reference strain (LS2) and 9 times greater than a weakly resistant strain (QSO335). Analysis of F2 and backcross progeny indicates that two or more genes are responsible for strong resistance, and that one of these genes, designated So_rph1, not only contributes to strong resistance, but is also responsible for the weak resistance phenotype of strain QSO335. These results demonstrate that the genetic mechanism of phosphine resistance in S. oryzae is similar to that of other stored product insect pests. A unique observation is that a subset of the progeny of an F1 backcross generation are more strongly resistant to phosphine than the parental strongly resistant strain, which may be caused by multiple alleles of one of the resistance genes.


The K-SF-42.
The purpose of the present article is to propose an alternative short form for the 199-item Arizona Life History Battery (ALHB), which we are calling the K-SF-42, as it contains 42 items as compared with the 20 items of the Mini-K, the short form that has been in greatest use for the past decade. These 42 items were selected from the ALHB, unlike those of the Mini-K, making direct comparisons of the relative psychometric performance of the two alternative short forms a valid and instructive exercise. A series of secondary data analyses were performed upon a recently completed five-nation cross-cultural survey, which was originally designed to assess the role of life history strategy in the etiology of interpersonal aggression. Only data from the ALHB that were collected in all five cross-cultural replications were used for the present analyses. The single immediate objective of this secondary data analysis was producing the K-SF-42 such that it would perform optimally across all five cultures sampled, and perhaps even generalize well to other modern industrial societies not currently sampled as a result of the geographic breadth of those included in the present study. A novel method, based on the use of the Cross-Sample Geometric Mean as a criterion for item selection, was used for generating such a cross-culturally valid short form.


Enhanced cancer immunotherapy using STAT3-depleted dendritic cells with high Th1-inducing ability and resistance to cancer cell-derived inhibitory factors.
STAT3 signaling constitutes an important negative feedback mechanism for the maintenance of immune homeostasis, a suppressive signal for the Th1 immune response in murine macrophages, and a cancer immune evasion signal in various immune cells. The strategy for STAT3 signal inhibition should be considered, because these features could impede effective cancer immunotherapy. We have evaluated the effects of STAT3 inactivation in dendritic cells (DCs) on immune responses in mice and humans. DCs derived from LysMcre/STAT3(flox/flox) mice displayed higher cytokine production in response to TLR stimulation, activated T cells more efficiently, and were more resistant to the suppression of cytokine production by cancer-derived immunosuppressive factors compared with DCs from control littermates. Antitumor activities of STAT3-depleted and control DCs were compared by intratumoral administration of gp70 Ag peptide-pulsed DCs in the therapeutic MC38 tumor model. Intratumoral administration of STAT3-depleted DCs significantly inhibited MC38 tumor growth of both injected and nontreated remote tumors. The inhibition was accompanied by an increase in gp70-specific T cell response as well as in systemic Th1 immune response. STAT3-depleted human DCs with adenoviral STAT3 short hairpin RNA were also capable of producing more cytokines with TLR stimulation and more resistant to cancer-derived factors, and they induced tumor Ag-specific T cells more efficiently than control DCs. The identified role of DC STAT3 signaling in both in vivo therapeutic tumor models in mice and in vitro-specific T cell induction in humans indicates that STAT3-inactivated DCs may be a promising approach for cancer immunotherapy.


Simulations of the emission spectra of fac-tris(2-phenylpyridine) iridium and Duschinsky rotation effects using the Herman-Kluk semiclassical initial value representation method.
The phosphorescent emission spectra of fac-tris(2-phenylpyridine) iridium [fac-Ir(ppy)(3)] due to the lowest triplet T(1) and T(2) states are simulated using the harmonic oscillator approximation for the S(0), T(1), and T(2) potential energy surfaces (PESs) and taking the Duschinsky rotation into account. The simulations involve the propagation of 177-dimensional wave packets on the coupled PES according to the Herman-Kluk (HK) semiclassical (SC) initial value representation (IVR) method. The HK SC-IVR method is employed because of its accuracy for the PES with mode mixing and its efficiency in dealing with coupled degrees of freedom for large systems. The simulated emission spectrum due to T(1) reproduces the structures of the emission spectra observed experimentally, while T(2) is found very unlikely to participate in the phosphorescent emission. Although the effect of the Duschinsky mode mixing is small for the T(1) state, neglecting it blueshifts the spectrum due to the T(2) state by 800 cm(-1) and changes the relative intensities, indicating that the importance of the Duschinsky rotation is rather unpredictable and should not be overlooked. The present simulations demonstrate that the simple harmonic oscillator approximation combined with the Duschinsky rotation can adequately describe the photophysics of fac-Ir(ppy)(3) and that the HK SC-IVR method is a powerful tool in studies of this kind.


Climate change alters the trophic niche of a declining apex marine predator.
Changes in the world's oceans have altered nutrient flow, and affected the viability of predator populations when prey species become unavailable. These changes are integrated into the tissues of apex predators over space and time and can be quantified using stable isotopes in the inert feathers of historical and contemporary avian specimens. We measured δ(13) C and δ(15) N values in Flesh-footed Shearwaters (Puffinus carneipes) from Western and South Australia from 1936-2011. The Flesh-footed Shearwaters more than doubled their trophic niche (from 3.91 ± 1.37 ‰(2) to 10.00 ± 1.79 ‰(2) ), and dropped an entire trophic level in 75 years (predicted δ(15) N decreased from +16.9 ‰ to + 13.5 ‰, and δ(13) C from -16.9 ‰ to -17.9 ‰) - the largest change in δ(15) N yet reported in any marine bird, suggesting a relatively rapid shift in the composition of the Indian Ocean food web, or changes in baseline δ(13) C and δ(15) N values. A stronger El Niño-Southern Oscillation results in a weaker Leeuwin Current in Western Australia, and decreased Flesh-footed Shearwater δ(13) C and δ(15) N. Current climate forecasts predict this trend to continue, leading to increased oceanic 'tropicalization' and potentially competition between Flesh-footed Shearwaters and more tropical sympatric species with expanding ranges. Flesh-footed Shearwater populations are declining, and current conservation measures aimed primarily at bycatch mitigation are not restoring populations. Widespread shifts in foraging, as shown here, may explain some of the reported decline. An improved understanding and ability to mitigate the impacts of global climactic changes is therefore critical to the long-term sustainability of this declining species.


Time to generate ventilator-induced lung injury among mammals with healthy lungs: a unifying hypothesis.
To investigate ventilator-induced lung injury (VILI), several experimental models were designed including different mammalian species and ventilator settings, leading to a large variability in the observed time-course and injury severity. We hypothesized that the time-course of VILI may be fully explained from a single perspective when considering the insult actually applied, i.e. lung stress and strain. Studies in which healthy animals were aggressively ventilated until preterminal VILI were selected via a Medline search. Data on morphometry, ventilator settings, respiratory function and duration of ventilation were derived. For each animal group, lung stress (transpulmonary pressure) and strain (end-inspiratory lung inflation/lung resting volume ratio) were estimated. From the Medline search 20 studies including five mammalian species (sheep, pigs, rabbits, rats, mice) were selected. Time to achieve preterminal VILI varied widely (18-2,784 min), did not correlate with either tidal volume (expressed in relation to body weight) or airway pressure applied, but was weakly associated with lung stress (r (2) = 0.25, p = 0.008). In contrast, the duration of mechanical ventilation was closely correlated with both lung strain (r (2) = 0.85, p < 0.0001) and lung strain weighted for the actual time of application during each breath (r (2) = 0.83, p < 0.0001), according to exponential decay functions. When it was normalized for the lung strain applied, larger species showed a greater resistance to VILI than smaller species (medians, 25th-75th percentiles: 690, 460-2,001 min vs. 16, 4-59 min, respectively; p < 0.001). Lung strain may play a critical role as a unifying rule describing the development of VILI among mammals with healthy lungs.


Correlates of prescription opioid therapy in Veterans with chronic pain and history of substance use disorder.
Patients with a history of substance use disorder (SUD) are more likely to be prescribed opioid medications for chronic pain than patients without an SUD history; however, little is known about prescription opioid therapy in populations composed exclusively of patients with SUD. This study examined correlates of prescription opioid therapy in 214 Veterans with chronic noncancer pain and an SUD history. Participants completed psychosocial questionnaires and participated in a structured mental health diagnostic interview, and medical diagnoses and opioid pharmacy data were abstracted from their Department of Veterans Affairs electronic medical records. Participants were categorized into three groups based on opioid prescriptions in the past 90 d: no opioid therapy (n = 134), short-term (<90 d) opioid therapy (n = 31), or long-term (>/= 90 d) opioid therapy (n = 49). Relative to participants prescribed no or short-term opioid therapy, participants who were prescribed long-term opioid therapy had a greater number of pain diagnoses; reported higher levels of pain severity, interference, and catastrophizing; and endorsed lower chronic pain self-efficacy. In a multivariate model, number of pain diagnoses and pain interference were associated with a greater likelihood of being prescribed long-term opioid therapy after controlling for demographic and clinical characteristics. Findings highlight the poor pain-related functioning in patients with SUD histories who are prescribed long-term opioid therapy.


The effect of delta rays on the ionometric dosimetry of proton beams.
The interface effects arising in the measurement of absorbed dose by ionization chambers, owing to the inhomogeneity between the walls and the gas, have been evaluated by an analytical model. The geometrical situation considered here is appropriate for representing the behaviour of a plane-parallel ionization chamber exposed to a radiotherapeutic beam of protons. Two gases, dry air and tissue equivalent gas (methane based), as well as six materials commonly used in ionization chamber walls, i.e. graphite, A-150 tissue equivalent plastic, C-522 air equivalent plastic, nylon type 6, polymethyl methacrylate and polystyrene, have been examined. The analysis of the results shows that, within the limits of the detector dimensions and proton energies commonly used in the dosimetry of radiotherapeutic beams, these effects, if not taken into account in the measurement interpretation, can entail deviations of up to about 2% with respect to the correct absorbed dose in gas.


A review on neuroimaging-based classification studies and associated feature extraction methods for Alzheimer's disease and its prodromal stages.
Neuroimaging has made it possible to measure pathological brain changes associated with Alzheimer's disease (AD) in vivo. Over the past decade, these measures have been increasingly integrated into imaging signatures of AD by means of classification frameworks, offering promising tools for individualized diagnosis and prognosis. We reviewed neuroimaging-based studies for AD and mild cognitive impairment classification, selected after online database searches in Google Scholar and PubMed (January, 1985-June, 2016). We categorized these studies based on the following neuroimaging modalities (and sub-categorized based on features extracted as a post-processing step from these modalities): i) structural magnetic resonance imaging [MRI] (tissue density, cortical surface, and hippocampal measurements), ii) functional MRI (functional coherence of different brain regions, and the strength of the functional connectivity), iii) diffusion tensor imaging (patterns along the white matter fibers), iv) fluorodeoxyglucose positron emission tomography (FDG-PET) (metabolic rate of cerebral glucose), and v) amyloid-PET (amyloid burden). The studies reviewed indicate that the classification frameworks formulated on the basis of these features show promise for individualized diagnosis and prediction of clinical progression. Finally, we provided a detailed account of AD classification challenges and addressed some future research directions.


Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies.
The authors questioned whether an automated kinetic mode assay of combined cytomegalovirus (CMV) late viral message and immediate and early antigens might result in a more sensitive and timely CMV diagnosis relevant to speedy treatment in the transplant setting. Toward this end, two cohorts were studied using automated in situ hybridization (ISH) for CMV as well as immunohistochemistry (IHC). The first cohort of patients consisted of 19 cases that were histologically positive (CMV-associated cytopathic change). A second cohort consisted of 10 cases that were histologically negative, yet culture positive. From the first cohort of histologically positive cases, 100% were positive by both ISH and IHC run on separate slides. In the second cohort, CMV was detected overall in 70% of cases (50% by ISH alone and 30% by IHC alone). These results indicate that a combined assay of ISH and IHC can detect more cases than routine hematoxylin and eosin staining or either assay alone. In two illustrative cases, used to demonstrate the feasibility of combining ISH and IHC, the authors used a combined two-color assay (ISH and IHC) performed sequentially on the same slide. The combined assays resulted in colocalized single cell message and protein in some cells and demonstrated more positive cells overall (some positive by IHC alone, some by ISH alone, and some by both) than either assay alone. The combined dual color assay can be completed within 4 to 5 hours giving the prospect of a same day result, which is faster than shell vial technique with immunofluorescence (24 to 48 hours) or culture (7 to 14 days). This study demonstrates that combining CMV message and protein assays results in a more sensitive assay and, when carried out in the kinetic mode, allows a speedy result relevant to early anti-CMV therapy.


MicroRNAs miR-26a, miR-26b, and miR-29b accelerate osteogenic differentiation of unrestricted somatic stem cells from human cord blood.
MicroRNAs are a population of short non-coding RNAs with widespread negative regulatory impact on mRNA translation. Unrestricted somatic stem cells (USSC) are a rare population in human cord blood that can be induced into cells representative of all three germinal layers. Here we analyzed the functional impact of miRNAs on the osteogenic differentiation in USSC. Gene expression profiling identified 20 microRNAs that were consistently upregulated during osteogenic differentiation of two different USSC cell lines (SA5/73 and SA8/25). Bioinformatic target gene prediction indicated that among these microRNAs, miR-10a, -22, -26a, -26b, and -29b recognize transcripts that encode a set of proteins inhibiting osteogenesis. We subsequently verified osteo-inhibitory CDK6, CTNNBIP1, HDAC4, and TOB1 and osteo-promoting SMAD1 as targets of these microRNAs. In Western blot analyses demonstrated that endogenous levels of CDK6 and HDAC4 were downregulated during osteogenic differentiation of USSC and reduced following ectopic expression of miR-26a/b and miR-29b. In contrast, endogenous expression of SMAD1, targeted by miR-26a/b, was unaltered during osteogenic differentiation of USSC or following ectopic expression of miR-26a/b. Functional overexpression analyses using microRNA mimics revealed that miR-26a/b, as well as miR-29b strongly accelerated osteogenic differentiation of USSC as assessed by Alizarin-Red staining and calcium-release assays. miR-26a/b and miR-29b are upregulated during osteogenic differentiation of USSC and share target genes inhibiting osteogenesis. Furthermore, these microRNAs accelerate osteogenic differentiation, likely mediated by osteo-inhibitory proteins such as CDK6 and HDAC4.


Accommodation in Early Presbyopes Fit with Bilateral or Unilateral Near Add.
When fit with monovision, most early presbyopes (aged 40 to 50 years) accommodated to near objects by focusing the distance corrected eye, leaving the near corrected eye myopically defocused with reduced image quality. A few were able to switch focus to the near corrected eye retaining a consistently focused image in one eye over a wider range of distances. The aim of this study was to examine accommodation behavior, pupil responses, and resultant image quality of early presbyopes fit with either bilateral or unilateral (monovision) near adds. Accommodative response and pupil size of 19 subjects (27 to 60 years), including 13 early presbyopes (40 to 50 years), were measured using an aberrometer as a binocularly viewed 20/40 letter E was moved from 2 m to 20 cm. Each subject was fit with different refractive strategies: bilateral distance correction, bilateral +2 diopters (D) near add, and unilateral +2 D near add placed over the measured right eye or unmeasured left eye. Monochromatic image quality was quantified using the Visual Strehl ratio metric. With bilateral +2 D near add, all early presbyopes mostly refrained from accommodating (gain = 0.22 D/D) until the target approached closer than the 50-cm far point, and they then accommodated accurately until their maximum accommodative amplitude was reached. With monovision, most (10 of 13 early presbyopes) accommodated to focus the distance corrected eye, leaving the near corrected eye myopically defocused with reduced image quality. As stimulus distance became closer than their distance corrected eye's near point, they continued to exert maximum accommodation. Only two early presbyopes relaxed their accommodation to "switch" focus to the near corrected eye as target distance was reduced, and these two did not experience bilateral drop in image quality as stimulus distance became closer than the near point of the distance corrected eye. Our data suggest that many early presbyopes will not initially adopt an accommodation strategy that optimizes image quality with monovision, but consistently accommodate to focus the distance corrected eye.


Absorption and elimination of formate following oral administration of calcium formate in female human subjects.
Calcium formate is a water-soluble salt of an essential mineral nutrient with potential for use as a dietary calcium supplement. Formate ion is a product of endogenous and xenobiotic metabolism, but sustained high plasma formate concentrations (such as occur in cases of methanol poisoning) are toxic to the retina and optic nerve. Humans and primates have reduced capacity for formate oxidation compared with rodents and dogs and are thus more sensitive to methanol (and formate) intoxication. To assess the potential for accumulation of formate ion upon repeated administration of calcium formate as a potential dietary calcium supplement, we measured plasma concentrations of formate in 14 adult human subjects before and after oral administration of a single large dose of calcium formate (3900 mg; ca. 3-6 times the anticipated dose for calcium supplementation). Plasma formate concentrations increased briskly from 0.024 +/- 0.008 mM (endogenous formate) to reach C(max) (0.50 +/- 0.04 mM) at 60 min postdose and then declined with a half-life of 59 +/- 7 min. By 225 min postdose, plasma formate concentration had returned to baseline. With such a short half-life, repeated use of calcium formate as a dietary supplement, even three times daily, should not lead to progressive accumulation of formate. These findings are discussed in light of the production of formate by endogenous and xenobiotic metabolism and the kinetics of formate during methanol poisoning.


Elevated carbon dioxide concentrations indirectly affect plant fitness by altering plant tolerance to herbivory.
Global environmental changes, such as rising atmospheric CO(2) concentrations, have a wide range of direct effects on plant physiology, growth, and fecundity. These environmental changes also can affect plants indirectly by altering interactions with other species. Therefore, the effects of global changes on a particular species may depend on the presence and abundance of other community members. We experimentally manipulated atmospheric CO(2) concentration and amounts of herbivore damage (natural insect folivory and clipping to simulate browsing) to examine: (1) how herbivores mediate the effects of elevated CO(2) (eCO(2)) on the growth and fitness of Arabidopsis thaliana; and (2) how predicted changes in CO(2) concentration affect plant resistance to herbivores, which influences the amount of damage plants receive, and plant tolerance of herbivory, or the fitness consequences of damage. We found no evidence that CO(2) altered resistance, but plants grown in eCO(2) were less tolerant of herbivory-clipping reduced aboveground biomass and fruit production by 13 and 22%, respectively, when plants were reared under eCO(2), but plants fully compensated for clipping in ambient CO(2) (aCO(2)) environments. Costs of tolerance in the form of reduced fitness of undamaged plants were detected in eCO(2) but not aCO(2) environments. Increased costs could reduce selection on tolerance in eCO(2) environments, potentially resulting in even larger fitness effects of clipping in predicted future eCO(2) conditions. Thus, environmental perturbations can indirectly affect both the ecology and evolution of plant populations by altering both the intensity of species interactions as well as the fitness consequences of those interactions.


Impact of glycemic control, disease duration, and exercise on heart rate variability in children with type 1 diabetes mellitus.
Type 1 diabetes is commonly associated with autonomic neuropathy. The present study investigated the influences of glycemic control, disease duration (DD), and exercise on autonomic nervous function in children with type 1 diabetes by analysis of their heart rate variability (HRV). Seventy-nine type 1 diabetic children were recruited and categorized into four groups by HbA1c of 8% and DD of 4.5 years. HRV parameters as determined by separate frequency domain components (low frequency: LnLF, 0.04-0.15 Hz; high frequency: LnHF, 0.15-0.5 Hz; total power: LnTP, 0.04-0.5 Hz) were measured both at rest and during exercise. Pearson's correlation, one-way ANOVA, and multiple regressions with stepwise method were used for statistical analysis. While at rest, HbA1c and DD were negatively correlated with all HRV parameters. Both HbA1c and DD were significant predictors in LnTP. However, only HbA1c was a significant predictor in LnLF and LnHF. Type 1 diabetes patients with HbA1c > 8% and DD > 4.5 years had a significantly lower HRV than the other patients. During exercise, HRV reduced significantly and no significant correlation between HbA1c and HRV or between DD and HRV was observed. Also, a significant difference in HRV among the four groups was not demonstrated. The smallest decrement in HRV from resting to exercise were in subjects with HbA1c > 8% and DD > 4.5 years. HbA1c was a more dominant predictor for LnTP, LnHF and LnLF than DD in children with type 1 diabetes at rest. HRV reduced significantly from resting to exercise. However, the responses of HRV during exercise differ from the responses of HRV at rest.


The effect of donor content on the efficiency of P3HT:PCBM bilayers: optical and photocurrent spectral data analyses.
State-of-the-art organic solar cells mostly rely on bulk-heterojunction architectures, where the photoactive layer is cast from a solution containing both the electron donor and acceptor components and subsequently annealed. An alternative route for device preparation is the sequential deposition of the two components using "orthogonal" solvents. The morphology of sequentially deposited bilayers has been extensively studied, but the interplay between optical and electrical properties and its influence on device efficiency is still unclear. Here we present a study of poly(3-hexylthiophene) (P3HT):phenyl-C61-butyric acid methyl ester (PCBM) bilayers with variable P3HT content, including also a standard bulk-heterojunction device for comparison. Measured optical absorption, external quantum efficieny (EQE), and internal quantum efficiency (IQE) data are analysed and interpreted with the aid of numerical models. In agreement with other studies, our results suggest substantial intermixing between the PCBM and P3HT component, regardless of the P3HT content. In the bulk heterojunction and the bilayer devices with an active layer thickness of 100 nm or less, our best fits to both the optical and optoelectronic data highlight a concentration inversion, with an accumulation of PCBM on the anode side. Through the numerical analysis of device performance at short-circuit, we also find that exciton diffusion toward the P3HT:PCBM interface and geminate recombination can be the main IQE loss factors. Additional losses, attributed to bimolecular electron-hole recombination, are also observed upon increasing the P3HT content.


The MDRD equation underestimates the prevalence of CKD among blacks and overestimates the prevalence of CKD among whites compared to the CKD-EPI equation: a retrospective cohort study.
Black individuals are far more likely than white individuals to develop end stage renal disease (ESRD). However, earlier stages of chronic kidney disease (CKD) have been reported to be less prevalent among blacks. This disparity remains poorly understood. The objective of this study was to evaluate whether the lower prevalence of CKD among blacks in early stages of CKD might be due in part to an inability of the MDRD equation to accurately determine early stages of CKD in both the black and white population. We conducted a retrospective cohort study of 97, 451 patients seen in primary care clinic in Veterans Integrated Service Network 2 (VISN 2) over a 7 year period to determine the prevalence of CKD using both the Modification of Diet in Renal Disease (MDRD) Study equation and the more recently developed CKD Epidemiology Collaboration (CKD-EPI) equation. Demographic data, comorbid conditions, prescription of medications, and laboratory data were recorded. Logistic regression and quantile regression models were used to compare the prevalence of estimated glomerular filtration rate (eGFR) categories between black and white individuals. The overall prevalence of CKD was lower when the CKD-EPI equation was used. Prevalence of CKD in whites was 53.2% by MDRD and 48.4% by CKD-EPI, versus 34.1% by MDRD and 34.5% by CKD-EPI in blacks. The cumulative logistic regression and quantile regression showed that when eGFR was calculated by the EPI method, blacks were as likely to present with an eGFR value less than 60 mL/min/1.73 m2 as whites. Using the CKD-EPI equation, blacks were more likely than white individuals to have stage 3b, 4 and 5 CKD. Using the MDRD method, the prevalence in blacks was only higher than in whites for stage 4 and 5 CKD. Similar results were obtained when the analysis was confined to patients over 65 years of age. The MDRD equation overestimates the prevalence of CKD among whites and underestimates the prevalence of CKD in blacks compared to the CKD-EPI equation.


Metabolic changes in vestibular and visual cortices in acute vestibular neuritis.
Five right-handed patients with a right-sided vestibular neuritis were examined twice with fluorodeoxyglucose positron emission tomography while lying supine with eyes closed: once during the acute stage (mean, 6.6 days) and then 3 months later when central vestibular compensation had occurred. Regional cerebral glucose metabolism (rCGM) was significantly increased (p <0.001 uncorrected) during the acute stage in multisensory vestibular cortical and subcortical areas (parietoinsular vestibular cortex in the posterior insula, posterolateral thalamus, anterior cingulate gyrus [Brodmann area 32/24], pontomesencephalic brainstem, hippocampus). Simultaneously, there was a significant rCGM decrease in the visual (Brodmann area 17 to 19) and somatosensory cortex areas in the postcentral gyrus as well as in parts of the auditory cortex (transverse temporal gyrus). Fluorodeoxyglucose positron emission tomography thus allows imaging of the cortical activation pattern that is induced by unilateral peripheral vestibular loss. It was possible to demonstrate that the central vestibular system including the vestibular cortex exhibits a visual-vestibular activation-deactivation pattern during the acute stage of vestibular neuritis similar to that in healthy volunteers during unilateral labyrinthine stimulation. Contrary to experimental vestibular stimulation, the activation of the vestibular cortex was not bilateral but was unilateral and contralateral to the right-sided labyrinthine failure.


Effects of aerobic vs. resistance training on visceral and liver fat stores, liver enzymes, and insulin resistance by HOMA in overweight adults from STRRIDE AT/RT.
While the benefits of exercise are clear, many unresolved issues surround the optimal exercise prescription. Many organizations recommend aerobic training (AT) and resistance training (RT), yet few studies have compared their effects alone or in combination. The purpose of this study, part of Studies Targeting Risk Reduction Interventions Through Defined Exercise-Aerobic Training and/or Resistance Training (STRRIDE/AT/RT), was to compare the effects of AT, RT, and the full combination (AT/RT) on central ectopic fat, liver enzymes, and fasting insulin resistance [homeostatic model assessment (HOMA)]. In a randomized trial, 249 subjects [18-70 yr old, overweight, sedentary, with moderate dyslipidemia (LDL cholesterol 130-190 mg/dl or HDL cholesterol ≤ 40 mg/dl for men or ≤ 45 mg/dl for women)] performed an initial 4-mo run-in period. Of these, 196 finished the run-in and were randomized into one of the following 8-mo exercise-training groups: 1) RT, which comprised 3 days/wk, 8 exercises, 3 sets/exercise, 8-12 repetitions/set, 2) AT, which was equivalent to ∼19.2 km/wk (12 miles/wk) at 75% peak O(2) uptake, and 3) full AT + full RT (AT/RT), with 155 subjects completing the intervention. The primary outcome variables were as follows: visceral and liver fat via CT, plasma liver enzymes, and HOMA. AT led to significant reductions in liver fat, visceral fat, alanine aminotransferase, HOMA, and total and subcutaneous abdominal fat (all P < 0.05). RT resulted in a decrease in subcutaneous abdominal fat (P < 0.05) but did not significantly improve the other variables. AT was more effective than RT at improving visceral fat, liver-to-spleen ratio, and total abdominal fat (all P < 0.05) and trended toward a greater reduction in liver fat score (P < 0.10). The effects of AT/RT were statistically indistinguishable from the effects of AT. These data show that, for overweight and obese individuals who want to reduce measures of visceral fat and fatty liver infiltration and improve HOMA and alanine aminotransferase, a moderate amount of aerobic exercise is the most time-efficient and effective exercise mode.


Visual agnosia.
As was originally proposed by Lissauer, visual recognition may break down either at an apperceptive or at an associative level. At an apperceptive level, finer grain distinctions may be made; the authors distinguished here between disorders of shape recognition and perceptual integration. It is not the case, however, that all patients with visual recognition deficits have impaired perception: poor recognition and naming may also result from damage to stored perceptual knowledge (e.g., structural descriptions), from problems in accessing semantic knowledge, from perceptual knowledge, or from impairments to semantic knowledge itself. These represent different types of associative deficit. In some cases, mild damage to stored perceptual knowledge also generates problems that are more severe on naming than on recognition and more severe for some categories of objects than others.


Absence of association of a single-nucleotide polymorphism in the TERT-CLPTM1L locus with age-related phenotypes in a large multicohort study: the HALCyon programme.
Several age-related traits are associated with shorter telomeres, the structures that cap the end of linear chromosomes. A common polymorphism near the telomere maintenance gene TERT has been associated with several cancers, but relationships with other aging traits such as physical capability have not been reported. As part of the Healthy Ageing across the Life Course (HALCyon) collaborative research programme, men and women aged between 44 and 90 years from nine UK cohorts were genotyped for the single-nucleotide polymorphism (SNP) rs401681. We then investigated relationships between the SNP and 30 age-related phenotypes, including cognitive and physical capability, blood lipid levels and lung function, pooling within-study genotypic effects in meta-analyses. No significant associations were found between the SNP and any of the cognitive performance tests (e.g. pooled beta per T allele for word recall z-score = 0.02, 95% CI: -0.01 to 0.04, P-value = 0.12, n = 18,737), physical performance tests (e.g. pooled beta for grip strength = -0.02, 95% CI: -0.045 to 0.006, P-value = 0.14, n = 11,711), blood pressure, lung function or blood test measures. Similarly, no differences in observations were found when considering follow-up measures of cognitive or physical performance after adjusting for its measure at an earlier assessment. The lack of associations between SNP rs401681 and a wide range of age-related phenotypes investigated in this large multicohort study suggests that while this SNP may be associated with cancer, it is not an important contributor to other markers of aging.


Central and peripheral mechanisms of the NPY system in the regulation of bone and adipose tissue.
Skeletal research is currently undergoing a period of marked expansion. The boundaries of "bone" research are being re-evaluated and with this, a growing recognition of a more complex and interconnected biology than previously considered. One aspect that has become the focus of particular attention is the relationship between bone and fat homeostasis. Evidence from a number of avenues indicates that bone and adipose regulation are both related and interdependent. This review examines the neuropeptide Y (NPY) system, known to exert powerful control over both bone and fat tissue. The actions of this system are characterized by signaling both within specific nuclei of the hypothalamus and also the target tissues, mediated predominantly through two G-protein coupled receptors (Y1 and Y2). In bone tissue, elevated NPY levels act consistently to repress osteoblast activity. Moreover, both central Y2 receptor and osteoblastic Y1 receptor signaling act similarly to repress bone formation. Conversely, loss of NPY expression or receptor signaling induces increased osteoblast activity and bone mass in both cortical and cancellous envelopes. In fat tissue, NPY action is more complex. Energy homeostasis is powerfully altered by elevations in hypothalamic NPY, resulting in increases in fat accretion and body-wide energy conservation, through the action of locally expressed Y1 receptors, while local Y2 receptors act to inhibit NPY-ergic tone. Loss of central NPY expression has a markedly reduced effect, consistent with a physiological drive to promote fat accretion. In fat tissue, NPY and Y1 receptors act to promote lipogenesis, consistent with their roles in the brain. Y2 receptors expressed in adipocytes also act in this manner, showing an opposing action to their role in the hypothalamus. While direct investigation of these processes has yet to be completed, these responses appear to be interrelated to some degree. The starvation-based signal of elevated central NPY inducing marked inhibition of osteoblast activity, whilst promoting fat accretion, indicating skeletal tissue is a component of the energy conservation system. Moreover, when NPY expression is reduced, consistent with high calorie intake and weight gain, bone formation is stimulated, strengthening the skeleton. In conclusion, NPY acts to regulate both bone and fat tissue in a coordinated manner, and remains a strong candidate for mediating interactions between these two tissues.


Folate intake and risk of oral and pharyngeal cancer.
Diet has been recognised as having a role in the aetiology of oral and pharyngeal cancer, and dietary factors may account for 10-15% of cases in Europe. Folate deficiency has been linked to risk of several cancers, but has not been studied adequately with respect to oral cancer. This case-control study, conducted in Italy and French-speaking Switzerland, included 749 patients with incident cancer of the oral cavity and pharynx, and 1772 hospital controls with acute, non-neoplastic conditions. The interviews used a validated food frequency questionnaire. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using multiple logistic regression. The ORs were 0.68 (95% CI 0.52-0.88) for the intermediate tertile and 0.53 (95% CI 0.40-0.69) for the highest tertile of dietary folate intake, compared with the lowest tertile. No heterogeneity was found in strata of gender, age, methionine intake or alcohol consumption. The combined OR for low-folate and high-alcohol intake versus high-folate and low-alcohol intake was 22.3 (95% CI 13.1-38). Our study supports a protective role of folate against oral and pharyngeal carcinogenesis. Compared with low folate intake, a consistent reduction in risk was already observed from intermediate levels of intake, suggesting that cancer risk may be related to relative folate deficiency.


Molecular architecture of the Jumonji C family histone demethylase KDM5B.
The full length human histone 3 lysine 4 demethylase KDM5B (PLU-1/Jarid1B) has been studied using Hydrogen/Deuterium exchange mass spectrometry, homology modelling, sequence analysis, small angle X-ray scattering and electron microscopy. This first structure on an intact multi-domain Jumonji histone demethylase reveal that the so-called PLU region, in the central region of KDM5B, has a curved α-helical three-dimensional structure, that acts as a rigid linker between the catalytic core and a region comprising four α-helices, a loop comprising the PHD2 domain, two large intrinsically disordered loops and the PHD3 domain in close proximity. The dumbbell shaped and curved KDM5B architecture observed by electron microscopy is complementary to the nucleosome surface and has a striking overall similarity to that of the functionally related KDM1A/CoREST complex. This could suggest that there are similarities between the demethylation mechanisms employed by the two histone 3 lysine 4 demethylases at the molecular level.


Identifying Mendelian disease genes with the variant effect scoring tool.
Whole exome sequencing studies identify hundreds to thousands of rare protein coding variants of ambiguous significance for human health. Computational tools are needed to accelerate the identification of specific variants and genes that contribute to human disease. We have developed the Variant Effect Scoring Tool (VEST), a supervised machine learning-based classifier, to prioritize rare missense variants with likely involvement in human disease. The VEST classifier training set comprised ~ 45,000 disease mutations from the latest Human Gene Mutation Database release and another ~45,000 high frequency (allele frequency >1%) putatively neutral missense variants from the Exome Sequencing Project. VEST outperforms some of the most popular methods for prioritizing missense variants in carefully designed holdout benchmarking experiments (VEST ROC AUC = 0.91, PolyPhen2 ROC AUC = 0.86, SIFT4.0 ROC AUC = 0.84). VEST estimates variant score p-values against a null distribution of VEST scores for neutral variants not included in the VEST training set. These p-values can be aggregated at the gene level across multiple disease exomes to rank genes for probable disease involvement. We tested the ability of an aggregate VEST gene score to identify candidate Mendelian disease genes, based on whole-exome sequencing of a small number of disease cases. We used whole-exome data for two Mendelian disorders for which the causal gene is known. Considering only genes that contained variants in all cases, the VEST gene score ranked dihydroorotate dehydrogenase (DHODH) number 2 of 2253 genes in four cases of Miller syndrome, and myosin-3 (MYH3) number 2 of 2313 genes in three cases of Freeman Sheldon syndrome. Our results demonstrate the potential power gain of aggregating bioinformatics variant scores into gene-level scores and the general utility of bioinformatics in assisting the search for disease genes in large-scale exome sequencing studies. VEST is available as a stand-alone software package at http://wiki.chasmsoftware.org and is hosted by the CRAVAT web server at http://www.cravat.us.


Community knowledge, attitude and practice about malaria in a low endemic setting of Shewa Robit Town, northeastern Ethiopia.
Since malaria is one of the foremost public health problems in Ethiopia, assessment of situation of the disease, and communities' knowledge and perceptions about malaria is necesary to institute appropriate preventive and control measures. Thus, the aim of this study was to assess malaria prevalence and knowledge, attitude and practice (KAP) about the disease among ShewaRobit Town community, northeastern Ethiopia. A community-based cross-sectional study was conducted in Shewa Robit Town from October to November 2011. A multi-stage random sampling technique was used to select the study participants. A total of 425 individuals were examined for malaria using thin and thick Giemsa stained blood film, and 284 of the participants were interviewed to assess their KAP about malaria. Logistic regression analysis was used to assess predictor factors for malaria prevalence. All respondents had ever heard of malaria. Most of the respondents (85.2%) attributed the cause of malaria to mosquito bite. However, some of the respondents (>20%) mentioned lack of personal hygiene, exposure to cold weather, hunger, chewing maize stalk, body contact with malaria patient and flies as the causes of malaria. Sleeping under mosquito nets, draining stagnant water and indoor residual spraying were the most frequently mentioned malaria preventive measures perceived and practiced by the respondents. Among 425 individuals examined for malaria, only 2.8% were positive for Plasmodium parasites. Living in houses made of wall without hole, sprayed with insecticide within the last 12 hours and located at a distance of greater than 500 meters from potential mosquito breeding sites as well as knowing and using of mosquito net were significant predictors of low malaria prevalence among the study participants. A high level of knowledge about the cause, transmission and preventive methods of malaria was detected among the community in Shewa Robit Town. However, a considerable proportion had misconception about the cause and transmission of malaria suggesting the necessity of health education to raise the community's awareness about the disease.


A quantitative 3D motility analysis of Trypanosoma brucei by use of digital in-line holographic microscopy.
We present a quantitative 3D analysis of the motility of the blood parasite Trypanosoma brucei. Digital in-line holographic microscopy has been used to track single cells with high temporal and spatial accuracy to obtain quantitative data on their behavior. Comparing bloodstream form and insect form trypanosomes as well as mutant and wildtype cells under varying external conditions we were able to derive a general two-state-run-and-tumble-model for trypanosome motility. Differences in the motility of distinct strains indicate that adaption of the trypanosomes to their natural environments involves a change in their mode of swimming.


Dorsal and ventral stream sensitivity in normal development and hemiplegia.
Form and motion coherence thresholds can provide comparable measures of global visual processing in the ventral and dorsal streams respectively. Normal development of thresholds was tested in 360 normally developing children aged 4-11 and in normal adults. The two tasks showed similar developmental trends, with some greater variability and a slight delay in motion coherence compared to form coherence performance, in reaching adult levels. To examine the proposal of dorsal stream vulnerability related to specific developmental disorders, we compared 24 children with hemiplegic cerebral palsy with the normally developing group. Hemiplegic children performed significantly worse than controls on the motion coherence task for their age, but not on the form coherence task; however, within this group no specific brain area was significantly associated with poor motion compared to form coherence performance. These results suggest that extrastriate mechanisms mediating these thresholds normally develop in parallel, but that the dorsal stream has a greater, general vulnerability to early neurological impairment.


Economic evaluation of human papillomavirus vaccination in developed countries.
With promising efficacy results from randomized control trials of human papillomavirus (HPV) vaccines and the availability of new screening paradigms, policymakers are being asked to make recommendations and decisions regarding the optimal strategies to reduce HPV infection and disease. Such decisions are increasingly being made with significant input from mathematical and economic models. The demand for modeling has resulted in the publication of numerous mathematical models looking at the cost-effectiveness of HPV vaccination. To review published models that have been used to evaluate the cost-effectiveness of HPV vaccination in developed countries and highlight points of consensus and disagreement in methods and findings. This review consists of cost-effectiveness studies published in the peer-reviewed literature before August 2008. Despite variations in methods, modeling studies are producing consistent conclusions: (1) vaccinating young girls against HPV is likely to be cost- effective; (2) vaccinating boys will most likely not be cost- effective in countries that can reach high coverage rates in girls, and (3) results are most sensitive to the duration of vaccine protection. However, results from analyses examining the effectiveness and cost-effectiveness of vaccinating boys when coverage rates are low (< or = 80%) and catch-up strategies have reached conflicting conclusions.


Feasibility of assessing the community composition of prasinophytes at the Helgoland Roads sampling site with a DNA microarray.
The microalgal class Prasinophyceae (Chlorophyta) contains several picoeukaryotic species, which are known to be common in temperate and cold waters and have been observed to constitute major fractions of marine picoplankton. However, reliable detection and classification of prasinophytes are mainly hampered by their small size and few morphological markers. Consequently, very little is known about the abundance and ecology of the members of this class. In order to facilitate the assessment of the abundance of the Prasinophyceae, we have designed and evaluated an 18S rRNA gene-targeted oligonucleotide microarray consisting of 21 probes targeting different taxonomic levels of prasinophytes. The microarray contains both previously published probes from other hybridization methods and new probes, which were designed for novel prasinophyte groups. The evaluation of the probe set was done under stringent conditions with 18S PCR fragments from 20 unialgal reference cultures used as positive targets. This microarray has been applied to assess the community composition of prasinophytes at Helgoland, an island in the North Sea where time series data are collected and analyzed daily but only for the nano- and microplankton-size fractions. There is no identification of prasinophytes other than to record them numerically in the flagellate fraction. The samples were collected every 2 weeks between February 2004 and December 2006. The study here demonstrates the potential of DNA microarrays to be applied as a tool for quick general monitoring of this important picoplanktonic algal group.


Role of 14-3-3 proteins in eukaryotic signaling and development.
14-3-3 genes encode a ubiquitous family of highly conserved eukaryotic proteins from fungi to humans and plants with several molecular and cellular functions. Most notably, 14-3-3 proteins bind to phosphoserine/phosphothreonine motifs in a sequence-specific manner. More than 100 14-3-3 binding partners involved in signal transduction, cell cycle regulation, apoptosis, stress responses, and malignant transformation have been identified. The 14-3-3 proteins form homodimers and heterodimers, and there is redundancy of the binding specificity and function of different 14-3-3 proteins because of their highly similar amino acid sequence and tertiary structure. 14-3-3 proteins can regulate target protein function by several mechanisms. Although the molecular and cellular functions of 14-3-3 proteins have been well studied, there have been fewer studies addressing the in vivo role of 14-3-3s. Here we review what is known about 14-3-3 proteins during eukaryotic development.


The role of proteoglycans in aging, degeneration and repair of the intervertebral disc.
The ability of the nucleus pulposus of the intervertebral disc to resist compressive loads is due to its high content of the proteoglycan aggrecan. Degeneration of the intervertebral disc is preceded and accompanied by a loss of aggrecan due to proteolysis. Biological repair of intervertebral disc degeneration should strive to restore aggrecan content to its optimal functional level. One approach to such repair is to supplement the degenerate nucleus with cells that are capable of aggrecan synthesis. Such cells can be supported in a biomolecular scaffold, but it is essential that the scaffold is compatible with high aggrecan retention if a functional tissue is to be attained.


Making sense of the past: hyperstability of ancestral thioredoxins explained by free energy simulations.
Thioredoxin (Trx), a small and globular protein, is present in all kinds of organisms, from Archea to higher mammals. Throughout evolution, the Trx sequence has undergone subtle modifications to adapt to varying environmental conditions. The high degree of sequence conservation makes Trx very amenable to ancestral protein reconstruction techniques. In this work, we address the study of the structural and energetic determinants of thermostability in E. coli Trx using a dataset of mutations inspired by ancestral reconstruction. We compute, from first principles, the expected contribution of 19 different amino acid substitutions to the stability (ΔΔG) and the melting temperature (ΔTm) of the protein. We also describe the specific changes in structure and protein dynamics responsible for the stabilizing or destabilizing effects of these mutations. Our results point to local and independent changes for most of the variants. Our predictions are accurate enough to substantiate the proposal of new hypotheses regarding evolutionary relationships between mutations, as in the case of T89R, P68A and G74S or K90L and F102A, and reach beyond the initial set to suggest improved variants, such as K90I or K90Y.


AML1-ETO reprograms hematopoietic cell fate by downregulating scl expression.
AML1-ETO is one of the most common chromosomal translocation products associated with acute myelogenous leukemia (AML). Patients carrying the AML1-ETO fusion gene exhibit an accumulation of granulocyte precursors in the bone marrow and the blood. Here, we describe a transgenic zebrafish line that enables inducible expression of the human AML1-ETO oncogene. Induced AML1-ETO expression in embryonic zebrafish causes a phenotype that recapitulates some aspects of human AML. Using this highly tractable model, we show that AML1-ETO redirects myeloerythroid progenitor cells that are developmentally programmed to adopt the erythroid cell fate into the granulocytic cell fate. This fate change is characterized by a loss of gata1 expression and an increase in pu.1 expression in myeloerythroid progenitor cells. Moreover, we identify scl as an early and essential mediator of the effect of AML1-ETO on hematopoietic cell fate. AML1-ETO quickly shuts off scl expression, and restoration of scl expression rescues the effects of AML1-ETO on myeloerythroid progenitor cell fate. These results demonstrate that scl is an important mediator of the ability of AML1-ETO to reprogram hematopoietic cell fate decisions, suggesting that scl may be an important contributor to AML1-ETO-associated leukemia. In addition, treatment of AML1-ETO transgenic zebrafish embryos with a histone deacetylase inhibitor, Trichostatin A, restores scl and gata1 expression, and ameliorates the accumulation of granulocytic cells caused by AML1-ETO. Thus, this zebrafish model facilitates in vivo dissection of AML1-ETO-mediated signaling, and will enable large-scale chemical screens to identify suppressors of the in vivo effects of AML1-ETO.


Winning hearts and minds: using psychology to promote voluntary organ donation.
Recent psychological research concerning determinants of and barriers to organ donation is reviewed with the intention of ascertaining acceptable and potentially effective ways of improving organ retrieval. On the basis of this review, five recommendations are made. (1) Individuals' donation wishes, where explicit, should be decisive. (2) Next of kin should witness donor decisions. (3) Mandated choice should replace voluntary 'opting-in'. (4) Initial donation choices should be repeatedly re-evaluated. (5) Those involved in organ procurement should distance themselves from model of bodies as machines or gardens and embrace models where bodies are viewed as sacred extensions of self. In combination, these recommendations are argued to be preferable in several ways to opt-out/presumed consent procurement options.


RF coil design for accurate parallel imaging on 13 C MRSI using 23 Na sensitivity profiles.
To develop a coil-based method to obtain accurate sensitivity profiles in 13 C MRI at 3T from the endogenous 23 Na. An eight-channel array is designed for 13 C MR acquisitions. As application examples, the array is used for two-fold accelerated acquisitions of both hyperpolarized 13 C metabolic imaging of pig kidneys and the human brain. A flexible coil array was tuned optimally for 13 C at 3T (32.1 MHz), with the coil coupling coefficients matched to be nearly identical at the resonance frequency of 23 Na (33.8 MHz). This is done by enforcing a high decoupling (obtained through highly mismatched preamplifiers) and adjusting the coupling frequency response. The SNR performance is compared to reference coils. The measured sensitivity profiles on a phantom showed high spatial similarity for 13 C and 23 Na resonances, with average noise correlation of 9 and 11%, respectively. For acceleration factors 2, 3, and 4, the obtained maximum g-factors were 1.0, 1.1, and 2.6, respectively. The 23 Na profiles obtained in vivo could be used successfully to perform two-fold acceleration of hyperpolarized 13 C 3D acquisitions of both pig kidneys and a healthy human brain. A receive array has been developed in such a way that the 13 C sensitivity profiles could be accurately obtained from measurements at the 23 Na frequency. This technique facilitates accelerated acquisitions for hyperpolarized 13 C imaging. The SNR performance obtained at the 13 C frequency, compares well to other state-of-the-art coils for the same purpose, showing slightly better superficial and central SNR.


2019 Focused Update of the Guidelines of the Taiwan Society of Cardiology for the Diagnosis and Treatment of Heart Failure.
Heart failure is a growing epidemic, especially in Taiwan because of the aging population. The 2016 Taiwan Society of Cardiology - Heart Failure with reduced Ejection Fraction (TSOC-HFrEF) registry showed that the guideline-recommended therapies were prescribed suboptimally both at the time of hospital discharge and during follow-up. We, therefore, conducted this 2019 focused update of the guidelines of the Taiwan Society of Cardiology for the diagnosis and treatment of heart failure to reinforce the importance of new diagnostic and therapeutic modalities of heart failure. The 2019 focused update discusses new diagnostic criteria, pharmacotherapy, non-pharmacological management, and certain co-morbidities of heart failure. Angiotensin receptor neprilysin inhibitor and If channel inhibitor is introduced as new and recommended medical therapies. Latest criteria of cardiac resynchronization therapy, implantable cardioverter-defibrillator, heart transplantation, and ventricular assist device therapy are reviewed in the non-pharmacological management chapter. Co-morbidities in heart failure are discussed including chronic kidney disease, diabetes, chronic obstructive pulmonary disease, and sleep-disordered breathing. We also explain the adequate use of oxygen therapy and non-invasive ventilation in heart failure management. A particular chapter for chemotherapy-induced cardiac toxicity is incorporated in the focused update to emphasize the importance of its recognition and management. Lastly, implications from the TSOC-HFrEF registry and post-acute care of heart failure are discussed to highlight the importance of guideline-directed medical therapy and the benefits of multidisciplinary disease management programs. With guideline recommendations, we hope that the management of heart failure can be improved in our society.


Controlling the processing of collagen-hydroxyapatite scaffolds for bone tissue engineering.
Scaffolds are an important aspect of the tissue engineering approach to tissue regeneration. This study shows that it is possible to manufacture scaffolds from type I collagen with or without hydroxyapatite (HA) by critical point drying. The mean pore sizes of the scaffolds can be altered from 44 to 135 microm depending on the precise processing conditions. Such pore sizes span the range that is likely to be required for specific cells. The mechanical properties of the scaffolds have been measured and behave as expected of foam structures. The degradation rate of the scaffolds by collagenase is independent of pore size. Dehydrothermal treatment (DHT), a common method of physically crosslinking collagen, was found to denature the collagen at a temperature of 120 degrees C resulting in a decrease in the scaffold's resistance to collagenase. Hybrid scaffold structures have also been manufactured, which have the potential to be used in the generation of multi-tissue interfaces. Microchannels are neatly incorporated via an indirect solid freeform fabrication (SFF) process, which could aid in reducing the different constraints commonly observed with other scaffolds.


MicroRNA epigenetics: a new avenue for wound healing research.
A group of small non-coding RNA molecules, termed microRNAs (miRNAs), have generated considerable interest in recent years due to their central role in a growing number of biologic processes. Serving as post-transcriptional regulators of gene expression, miRNAs have also emerged as critical factors in the pathogenesis of many diseases. As a result, they show great potential as accurate diagnostic and prognostic markers, as well as viable therapeutic targets for treating disease. It has been proposed that miRNAs play a significant role in cutaneous wound repair and that aberrant miRNA expression may result in disorganized or poor healing. Specific patterns of miRNA expression have been identified in wound healing models. miRNAs are important regulators of leucocyte function and the cytokine network, and are necessary for endothelial cell migration and capillary formation. These molecules also control proliferation and differentiation of wound-specific cells and can determine extracellular matrix composition. This article reviews the evidence for miRNA regulation of inflammation, angiogenesis, fibroblast function, keratinocyte function, and apoptosis, which are essential components for effective wound repair. The future potential for improving wound healing outcomes using miRNA-based therapies is also discussed.


Audio-visual onset differences are used to determine syllable identity for ambiguous audio-visual stimulus pairs.
Content and temporal cues have been shown to interact during audio-visual (AV) speech identification. Typically, the most reliable unimodal cue is used more strongly to identify specific speech features; however, visual cues are only used if the AV stimuli are presented within a certain temporal window of integration (TWI). This suggests that temporal cues denote whether unimodal stimuli belong together, that is, whether they should be integrated. It is not known whether temporal cues also provide information about the identity of a syllable. Since spoken syllables have naturally varying AV onset asynchronies, we hypothesize that for suboptimal AV cues presented within the TWI, information about the natural AV onset differences can aid in speech identification. To test this, we presented low-intensity auditory syllables concurrently with visual speech signals, and varied the stimulus onset asynchronies (SOA) of the AV pair, while participants were instructed to identify the auditory syllables. We revealed that specific speech features (e.g., voicing) were identified by relying primarily on one modality (e.g., auditory). Additionally, we showed a wide window in which visual information influenced auditory perception, that seemed even wider for congruent stimulus pairs. Finally, we found a specific response pattern across the SOA range for syllables that were not reliably identified by the unimodal cues, which we explained as the result of the use of natural onset differences between AV speech signals. This indicates that temporal cues not only provide information about the temporal integration of AV stimuli, but additionally convey information about the identity of AV pairs. These results provide a detailed behavioral basis for further neuro-imaging and stimulation studies to unravel the neurofunctional mechanisms of the audio-visual-temporal interplay within speech perception.


Case Studies in Physiology: Physiological and clinical effects of temporary diaphragm pacing in two patients with ventilator-induced diaphragm dysfunction.
Ventilator-induced diaphragm dysfunction (VIDD) is increasingly recognized as an important side-effect of invasive ventilation in critically ill patients and is associated with poor outcomes. Whether patients with VIDD benefit from temporary diaphragm pacing is uncertain. Intramuscular diaphragmatic electrodes were implanted for temporary stimulation with a pacing device (TransAeris System) in two patients with VIDD. The electrodes were implanted via laparoscopy (first patient) or via bilateral thoracoscopy (second patient). Stimulation parameters were titrated according to tolerance. Diaphragm thickening fraction by ultrasound, maximum inspiratory pressure (Pimax) and diaphragm electromyography (EMGdi) signal analysis were used to monitor the response to diaphragm pacing. Both patients tolerated diaphragm pacing. In the first patient, improvements in diaphragm excursions were noted once pacing was initiated and diaphragm thickening fraction did not further deteriorate over time. The diaphragm thickening fraction improved in the second patient, and Pimax as well as EMGdi analysis suggested improved muscle function. This patient could be fully weaned from the ventilator. These case reports present the first experience with temporary diaphragm pacing in critically ill patients with VIDD. Our results should be taken cautiously given the reduced sample size, but provide the proof of concept to put forward the hypothesis that a course of diaphragm pacing may be associated with improved diaphragmatic function. Our findings of the tolerance to the procedure and the beneficial physiological effects are not prove of safety and efficacy, but may set the ground to design and conduct larger studies.NEW & NOTEWORTHY Diaphragmatic electrode implantation and temporary diaphragm pacing have not been previously used in ICU patients with VIDD. Patients were monitored using a multimodal monitoring approach including ultrasound of the diaphragm, measurement of maximum inspiratory pressure and EMG signal analysis. Our results suggest that diaphragm pacing may improve diaphragmatic function, with the potential to prevent and treat VIDD in critically ill patients. Safety and efficacy of this intervention is yet to be proven in larger studies.


Physician perspectives on the burden and management of asthma in six countries: The Global Asthma Physician Survey (GAPS).
Despite recognition of asthma as a growing global issue and development of global guidelines, asthma treatment practices vary between countries. Several studies have reported patients' perspectives on asthma control. This study presents physicians' perspectives and strategies for asthma management. Physicians seeing ≥4 adult patients with asthma per month in Australia, Canada, China, France, Germany, and Japan were surveyed (N=1809; ≈300 per country). A standardised questionnaire was developed for this study and administered by telephone, online or face-to-face. Statistics were weighted to account for the sampling scheme. Physicians estimated that 71% of their adult patients received maintenance medication, with adherence monitored by 76-97% of physicians. Perceived major barriers to patient adherence included: patients taking treatment as needed; acceptance of symptoms; and patients not perceiving treatment benefits. Written action plans (37%) and technology (15%) were seldom employed by physicians to aid patients' asthma management. Physicians rarely (10%) used validated patient-reported questionnaires to monitor asthma control, instead monitoring selected symptoms, exacerbations, and/or lung function measurements. Awareness of single maintenance and reliever therapy (SMART/MART) varied among countries (56-100%); although most physicians (72%) had prescribed SMART/MART, the majority (91%) co-prescribed a short-acting bronchodilator at least some of the time. These results show that physicians generally do not employ standardised tools to monitor asthma control or to manage its treatment and that despite high awareness of SMART/MART, the strategy appears to be commonly misapplied. Better education for patients and physicians is required to improve asthma management and resulting patient outcomes.


Understanding the prebiotic potential of different dietary fibers using an in vitro continuous adult fermentation model (PolyFermS).
Consumption of fermentable dietary fibers (DFs), which can induce growth and/or activity of specific beneficial populations, is suggested a promising strategy to modulate the gut microbiota and restore health in microbiota-linked diseases. Until today, inulin and fructo-oligosaccharides (FOS) are the best studied DFs, while little is known about the gut microbiota-modulating effects of β-glucan, α-galactooligosaccharide (α-GOS) and xylo-oligosaccharide (XOS). Here, we used three continuous in vitro fermentation PolyFermS model to study the modulating effect of these DFs on two distinct human adult proximal colon microbiota, independently from the host. Supplementation of DFs, equivalent to a 9 g daily intake, induced a consistent metabolic response depending on the donor microbiota. Irrespective to the DF supplemented, the Bacteroidaceae-Ruminococcaceae dominated microbiota produced more butyrate (up to 96%), while the Prevotellaceae-Ruminococcaceae dominated microbiota produced more propionate (up to 40%). Changes in abundance of specific bacterial taxa upon DF supplementation explained the observed changes in short-chain fatty acid profiles. Our data suggest that the metabolic profile of SCFA profile may be the most suitable and robust read-out to characterize microbiota-modulating effects of a DF and highlights importance to understand the inter-individual response to a prebiotic treatment for mechanistic understanding and human application.


Retinol, alpha-tocopherol, lycopene, and alpha- and beta-carotene simultaneously determined in plasma by isocratic liquid chromatography.
Retinol, alpha-tocopherol, lycopene, and alpha- and beta-carotene can be simultaneously determined in human plasma by reversed-phase liquid chromatography. Plasma--0.5 mL plus added internal standard, retinyl acetate--is deproteinized with 0.5 mL of ethanol, then extracted with 1.0 mL of petroleum ether. The organic layer is removed and evaporated, the residue is redissolved in 0.25 mL of ethanol, and 8-microL samples are injected into a 60 X 4.6 mm column of Hypersil ODS 3-microns particles at 35 degrees C. An isocratic methanol mobile phase, flow rate 0.9 mL/min, is used for the 9-min run. Retinol and retinyl acetate are monitored at 305 nm, the tocopherols at 292 nm, and the carotenoids at 460 nm. Between-run CVs were 3.1, 6.9, 6.1, and 6.5% for retinol, alpha-tocopherol, lycopene, and beta-carotene, respectively. Small sample requirement, simplicity of extraction, short run time, and good reproducibility make this procedure ideal for clinical or research use.


Robot-assisted versus conventional laparoscopic Nissen fundoplication: a comparative retrospective study on costs and time consumption.
Laparoscopic Nissen fundoplication offers clear benefits for our patients, but requires advanced laparoscopic skills. Robotic assistance in laparoscopic antireflux surgery improves dexterity skills and shortens learning curve, possibly leading to faster, more precise, and safer laparoscopic surgery. We review our first 11 cases of robot-assisted laparoscopic Nissen fundoplication using the 4-armed daVinci surgical system, comparing them with patients who underwent conventional laparoscopic Nissen fundoplication for gastroesophageal reflux disease in the same period. Robot-assisted laparoscopic Nissen fundoplication did not result in more complications. However, the use of robotic assistance took an extra-47 minutes to complete the operation and costs were raised with an accessory euro 987.47. The use of robotic assistance in laparoscopic antireflux surgery is safe and feasible, but results in longer operating time and higher costs compared with conventional laparoscopic surgery without proven benefit at this moment.


The t(5;17) variant of acute promyelocytic leukemia expresses a nucleophosmin-retinoic acid receptor fusion.
We have studied an acute promyelocytic leukemia (APL) patient with a variant t(5;17)(q32;q12). This translocation fuses the gene for the nucleolar phosphoprotein nucleophosmin (NPM) to the retinoic acid receptor alpha (RARA). Two alternatively spliced transcripts are expressed, which differ in 129 bases immediately upstream of the RARA sequence. The NPM sequences contained in the shorter NPM-RAR cDNA are identical to the NPM sequences contained in the NPM-ALK fusion gene expressed in t(2;5) lymphomas. The RARA sequences are the same as the RARA sequences found in the PML-RAR and PLZF-RAR fusion seen in t(15;17) and t(11;17) APL, respectively. Both NPM-RAR transcripts fuse NPM and RARA sequence in the same reading frame, to generate translation products of 57 kD and 62 kD. Both NPM-RAR proteins are expressed in the patient's leukemic cells, along with wild-type RARA derived from the uninvolved allele. In transcriptional assays using a retinoic acid response element reporter construct, both NPM-RAR fusion proteins act as retinoic acid-dependent transcriptional activators. This case defines a third class of APL rearrangements, all of which generate fusion proteins of RARA.


Treatment of ankylosing spondylitis.
Ankylosing spondylitis is a chronic, inflammatory, rheumatic disease that can reduce the quality of life and increase the risk of disability and mortality. It also causes direct and indirect economic losses due to health expenses and as a result of workforce loss. Management of this disease consists of pharmacological and nonpharmacological modalities. Until recently, pharmacological treatment options have been very limited. However, development of novel biological drugs revolutionized the management of this disease. The aim of this review article is to present an updated overview of the pharmacologic treatment of ankylosing spondylitis. Nonpharmacological treatment modalities including physiotherapy and exercise are only briefly mentioned and surgical treatment is not discussed.


Comparison of corneal epitheliotrophic capacities among human platelet lysates and other blood derivatives.
To evaluate the corneal epitheliotropic abilities of two commercialized human platelet lysates (HPLs) and to compare the results with other blood derivatives, including human peripheral serum (HPS) and bovine fetal serum (FBS). In vitro, human corneal epithelial cells were incubated in various concentrations (0%, 3%, 5% and 10%) of blood derivatives. Two commercialized HPLs, including UltraGRO TM (Helios, Atlanta, GA) and PLTMax (Mill Creek, Rochester, MI), were tested and compared with HPS and FBS. Scratch-induced directional wounding assay was performed to evaluate cellular migration. MTS assay was used to evaluate cellular proliferation. Cellular differentiation was examined by scanning electron microscopy, inverted microscopy and transepithelial electrical resistance. Sprague-Dawley rats were used to evaluate the effects of the blood derivatives on corneal epithelial wound healing in vivo. Different blood derivatives were applied topically every 2 hours for 2 days after corneal epithelial debridement. The concentrations of epidermal growth factor (EGF), transforming growth factor -β1 (TGF-β1), fibronectin, platelet-derived growth factor-AB (PDGF-AB), PDGF-BB, and hyaluronic acid in different blood derivatives were evaluated by enzyme-linked immunosorbent assay (ELISA). In vitro experiments demonstrated statistically comparable epitheliotropic characteristics in cellular proliferation, migration, and differentiation for the two commercialized HPLs compared to FBS and HPS. Cells cultured without any serum were used as control group. The epitheliotropic capacities were statistically higher in the two commercialized HPLs compared to the control group (p<0.05). Among the different concentrations of blood derivatives, the preparations with 3% yielded better outcomes compared to 5% and 10%. In rats, HPLs also caused improved but not statistically significant wound healing compared to HPS. All the blood derivatives had better wound healing ratios than the control group (p<0.05). In the quantification of epitheliotropic factors, UltraGRO and PLTMax had significantly higher levels of EGF, TGF- β1, fibronectin than human peripheral serum (p<0.05). Both commercialized HPLs showed comparable corneal epitheliotropic abilities and wound healing rates compared to HPS and FBS in the in vivo and in vitro studies. Our results suggest that HPLs may have the potential to replace HPS in the treatment of corneal epithelial problems.


Graphical methods for reducing, visualizing and analyzing large data sets using hierarchical terminologies.
To explore new graphical methods for reducing and analyzing large data sets in which the data are coded with a hierarchical terminology. We use a hierarchical terminology to organize a data set and display it in a graph. We reduce the size and complexity of the data set by considering the terminological structure and the data set itself (using a variety of thresholds) as well as contributions of child level nodes to parent level nodes. We found that our methods can reduce large data sets to manageable size and highlight the differences among graphs. The thresholds used as filters to reduce the data set can be used alone or in combination. We applied our methods to two data sets containing information about how nurses and physicians query online knowledge resources. The reduced graphs make the differences between the two groups readily apparent. This is a new approach to reduce size and complexity of large data sets and to simplify visualization. This approach can be applied to any data sets that are coded with hierarchical terminologies.


Biomedical laboratory science education: standardising teaching content in resource-limited countries.
There is a worldwide shortage of qualified laboratory personnel to provide adequate testing for the detection and monitoring of diseases. In an effort to increase laboratory capacity in developing countries, new skills have been introduced into laboratory services. Curriculum revision with a focus on good laboratory practice is an important aspect of supplying entry-level graduates with the competencies needed to meet the current needs. Gaps in application and problem-solving competencies of newly graduated laboratory personnel were discovered in Ethiopia, Tanzania and Kenya. New medical laboratory teaching content was developed in Ethiopia, Tanzania and Kenya using national instructors, tutors, and experts and consulting medical laboratory educators from the United States of America (USA). Workshops were held in Ethiopia to create standardised biomedical laboratory science (BMLS) lessons based on recently-revised course objectives with an emphasis on application of skills. In Tanzania, course-module teaching guides with objectives were developed based on established competency outcomes and tasks. In Kenya, example interactive presentations and lesson plans were developed by the USA medical laboratory educators prior to the workshop to serve as resources and templates for the development of lessons within the country itself. The new teaching materials were implemented and faculty, students and other stakeholders reported successful outcomes. These approaches to updating curricula may be helpful as biomedical laboratory schools in other countries address gaps in the competencies of entry-level graduates.


Modification in the frequency of Hb C and Hb S in Burkina Faso: an influence of migratory fluxes and improvement of patient health care.
The incidence of hemoglobinopathies (Hb C and Hb S) is relatively high in West Africa. In order to calculate the gene frequency of these hemoglobinopathies, 6619 students from 23 local schools in Ouagadougou, Burkina Faso, West Africa, and 2582 individuals living in five villages near Ouagadougou, all situated in Savanna, were studied. As expected, the gene frequency in the city schools was 0.111 for the betaC gene and 0.051 for the betaS gene; in the five villages it was 0.122 for the betaC gene and 0.047 for the betaS gene. This data is somewhat different from that published in a previous study by Labie et al. [2] in the humid Savanna region, that showed a higher prevalence of betaC (0.14) than betaS (0.03), and is in contrast to data from the arid Sahel region that showed a higher prevalence of betaS (0.1) compared to betaC (0.05). The higher rate of betaS and lower rate of betaC in students in Ouagadougou, and in the individuals living in the five villages near Ouagadougou, suggest the possible influence of migratory fluxes of the betaS gene from the country region of Sahel. The dramatic increase in the prevalence of Hb SS patients, not reported in the study of Labie et al., [2] may be the result of reduced mortality due to environmental change. In addition, the improved health conditions of Hb SC and the increased life expectancy of Hb SS, may also have facilitated the increase of the betaS gene and the focus on secondary prevention for the control of correlated diseases.


High Throughput UPLC®-MSMS Method for the Analysis of Phosphatidylethanol (PEth) 16:0/18:1, a Specific Biomarker for Alcohol Consumption, in Whole Blood.
Phosphatidylethanol (PEth) is an alcohol biomarker formed in the presence of ethanol in the body. Both due to its specificity and because it has a detection window of up to several weeks after alcohol intake, its application potential is broader than for other ethanol biomarkers. The aim of this study was to develop and validate a robust method for PEth in whole blood with fast and efficient sample extraction and a short analytical runtime, suitable for high throughput routine purposes. A validated ultra-performance liquid chromatography tandem mass spectrometry (UPLC®-MSMS) method for quantification of PEth 16:0/18:1 in the range 0.05-4.00 μM (R2 ≥ 0.999) is presented. PEth 16:0/18:1 and the internal standard (IS) PEth-d5 (0.55 μM), were extracted from whole blood (150 μL) by simple protein precipitation with 2-propanol (450 μL). Chromatography was achieved using a BEH-phenyl (2.1 × 30 mm, 1.7 μm) column and a gradient elution combining ammonium formate (5 mM, pH 10.1) and acetonitrile at a flow rate of 0.5 mL/min. Runtime was 2.3 min. The mass spectrometer was monitored in negative mode with multiple reaction monitoring (MRM). The m/z 701.7 > 255.2 and 701.7 > 281.3 transitions were monitored for PEth 16:0/18:1 and the m/z 706.7 > 255.3 for PEth-d5. Limit of quantification was 0.03 μM (coefficient of variation, CV = 6.7%, accuracy = 99.3%). Within-assay and between-assay imprecision were 0.4-3.3% (CV ≤ 7.1%). Recoveries were 95-102% (CV ≤ 4.9%). Matrix effects after IS correction ranged from 107% to 112%. PEth 16:0/18:1 in patient samples were stable for several days at 30°C. Repeated freezing (-80°C) and thawing did not affect the concentration. After thawing and analysis patient samples were stable at 4-8°C for at least 4 weeks. Results from a proficiency test program, showing |Z| values ≤1.2, confirm the validity of the method. Analysis of the first 3,169 samples sent to our laboratory for routine use has demonstrated its properties as a robust method suitable for high throughput purposes.


Glucosylceramide modulates membrane traffic along the endocytic pathway.
Glycosphingolipids are endocytosed and targeted to the Golgi apparatus, but are mistargeted to lysosomes in numerous sphingolipidoses. Substrate reduction therapy utilizes imino sugars to inhibit glucosylceramide synthase and potentially abrogate the effects of storage. Gaucher disease is a hereditary deficiency in glucocerebrosidase leading to glucosylceramide accumulation; however, Gaucher fibroblasts exhibited normal Golgi transport of lactosylceramide. To better understand the effects of glycosphingolipid accumulation on intracellular trafficking and the use of imino sugar inhibitors, we studied sphingolipid endocytosis in fibroblast and macrophage models for Gaucher disease. Treatment of fibroblasts or RAW macrophages with conduritol B epoxide, an inhibitor of lysosomal glucocerebrosidase, resulted in a change in the endocytic targeting of lactosylceramide from the Golgi to the lysosomes. Co-treatment of macrophages with conduritol B-epoxide and 12-25 microM N-butyldeoxygalactonojirimycin, an inhibitor of glycosphingolipid biosynthesis, prevented the mistargeting of lactosylceramide to the lysosomes and restored trafficking to the Golgi. Surprisingly, higher doses (>25 microM) of NB-DGJ induced targeting of lactosylceramide to the lysosomes, even in the absence of conduritol B-epoxide. These data demonstrate that both increases and decreases in glucosylceramide levels can dramatically alter the endocytic targeting of lactosylceramide and suggest a role for glucosylceramide in regulation of membrane transport.


Preoperative assessment and perioperative management of cardiovascular risk.
Preoperative cardiovascular evaluation of patients scheduled to undergo surgery requires a multidisciplinary approach involving anesthetists, surgeons, and cardiologists. Within the last 5 to 10 years, there have been considerable scientific advances in this field, and the European Society of Cardiology (ESC) and the American College of Cardiology/American Heart Association(ACC/AHA) have updated their guidelines.(1,11) Greater emphasis now lies on preoperative clinical risk stratification and less on routine cardiac testing. Prophylactic coronary revascularization is now also seen as rarely indicated simply to lower the risk of surgery.(1) This review will give a brief summary of the guidelines and suggests a practical stepwise approach to evaluate the patient preoperatively.