Application of simulated annealing to the biclustering of gene expression data.
In a gene expression data matrix, a bicluster is a submatrix of genes and conditions that exhibits a high correlation of expression activity across both rows and columns. The problem of locating the most significant bicluster has been shown to be NP-complete. Heuristic approaches such as Cheng and Church's greedy node deletion algorithm have been previously employed. It is to be expected that stochastic search techniques such as evolutionary algorithms or simulated annealing might improve upon such greedy techniques. In this paper we show that an approach based on simulated annealing is well suited to this problem, and we present a comparative evaluation of simulated annealing and node deletion on a variety of datasets. We show that simulated annealing discovers more significant biclusters in many cases. Furthermore, we also test the ability of our technique to locate biologically verifiable biclusters within an annotated set of genes.


A hybrid approach to estimate chromophoric dissolved organic matter in turbid estuaries from satellite measurements: a case study for Tampa Bay.
Remote sensing of chromophoric dissolved organic matter (CDOM) from satellite measurements for estuaries has been problematic due to optical complexity of estuarine waters and uncertainties in satellite-derived remote sensing reflectance (Rrs, sr(-1)). Here we demonstrate a hybrid approach to combine empirical and semi-analytical algorithms to derive CDOM absorption coefficient at 443 nm (a(g)(443), m(-1)) in a turbid estuary (Tampa Bay) from MODIS Aqua (MODISA) and SeaWiFS measurements. The approach first used a validated empirical algorithm and a modified quasi-analytical algorithm (QAA) to derive chlorophyll-a concentration (Chla, mg m(-3)) and particulate backscattering coefficient at 443 nm (b(bp)(443), m(-1)), respectively, from which phytoplankton pigment and non-algal particulate absorption coefficient at 443 nm (a(ph)(443) and a(d)(443), m(-1)) were derived with pre-determined bio-optical relationships. Then, the modified QAA was used to estimate the total absorption coefficient at 443 nm (a(t)(443), m(-1)). Finally, a(g)(443) was estimated as (a(t)(443) - a(ph)(443) - a(d)(443) - a(w)(443)) where a(w)(443) is the absorption coefficient of pure water (a constant). Using data collected from 71 field stations and 33 near-concurrent satellite-field matchup data pairs covering a large dynamic range (0.3 - 8 m(-1)), the approach showed ~23% RMS uncertainties in retrieving a(g)(443) when in situ Rrs data (N = 71) were used. The same approach applied to satellite Rrs yielded much higher uncertainties of a(g)(443) (~85%) due to large errors in the satellite-retrieved Rrs(443). When the Rrs(443) was derived from the satellite-retrieved Rrs(550) and then used in the hybrid approach, uncertainties in the retrieved a(g)(443) reduced to ~30% (N = 33). Application of the approach to MODISA and SeaWiFS data led to a 15-year time series of monthly mean a(g)(443) distributions in Tampa Bay between 1998 and 2012. This time series showed significant seasonal and annual variations regulated mainly by river discharge. Testing of the approach over another turbid estuary (Chesapeake Bay, the largest estuary in the U.S.) demonstrated the potential (~25% uncertainties for a limited a(g)(443) range) of using this approach to establish long-term environmental data records (EDRs) of CDOM distributions in other estuaries with similar optical complexity.


Screening seniors for risk of functional decline: results of a survey in family practice.
To measure functional status, determine risk of functional decline and assess consistency between responses and standardized instruments. A mailed survey which measured functional impairment, recent hospitalization and bereavement. A positive response on at least one of these factors indicated that the individual was "at risk" for functional decline. A random sample (n = 73) of "at risk" subjects (specifically, family practice patients aged 70 and older) were assessed by a nurse. The response rate was 89% (369/415), 59% of seniors were female and the mean age was 77.1 (SD = 5.5) years. Self-reported risk, based on activities of daily living (ADLs), was associated with impairment in at least one basic ADL (p < 0.0005) using a standardized instrument. The positive predictive value of the survey for ADL impairment was 65%. Response to a mailed survey was high and self-reported ADL risks were consistent with findings from standardized assessment tools.


Achieving across-laboratory replicability in psychophysical scaling.
It is well known that, although psychophysical scaling produces good qualitative agreement between experiments, precise quantitative agreement between experimental results, such as that routinely achieved in physics or biology, is rarely or never attained. A particularly galling example of this is the fact that power function exponents for the same psychological continuum, measured in different laboratories but ostensibly using the same scaling method, magnitude estimation, can vary by a factor of three. Constrained scaling (CS), in which observers first learn a standardized meaning for a set of numerical responses relative to a standard sensory continuum and then make magnitude judgments of other sensations using the learned response scale, has produced excellent quantitative agreement between individual observers' psychophysical functions. Theoretically it could do the same for across-laboratory comparisons, although this needs to be tested directly. We compared nine different experiments from four different laboratories as an example of the level of across experiment and across-laboratory agreement achievable using CS. In general, we found across experiment and across-laboratory agreement using CS to be significantly superior to that typically obtained with conventional magnitude estimation techniques, although some of its potential remains to be realized.


A tyrosine-phosphorylated 55-kilodalton motility-associated bovine sperm protein is regulated by cyclic adenosine 3',5'-monophosphates and calcium.
Sperm motility is regulated by protein phosphorylation. We have recently shown that a serine/threonine phosphatase system is involved in motility regulation. Two of the components of the phosphatase system, GSK-3 and PP1gamma2, are regulated by tyrosine phosphorylation. During our investigation of sperm tyrosine-phosphorylated proteins we discovered a 55-kDa protein whose tyrosine phosphorylation correlates closely to the motility state of sperm. This protein is tyrosine phosphorylated to a much higher degree in motile caudal than in immotile caput epididymal sperm. Motility inhibition of caudal epididymal sperm by protein kinase A (PKA) anchoring inhibition or by ionomycin-induced calcium overload led to the virtual disappearance of tyrosine phosphorylation of the 55-kDa protein. Conversely, treatment of sperm with motility activators, isobutylmethylxanthine or 8-bromo-cAMP, resulted in increased tyrosine phosphorylation of the protein. The protein was present in the soluble 100 000 x g supernatants of sperm extracts and was heat labile. Chromatography through diethylaminoethyl-cellulose and Western blot analysis showed that this 55-kDa protein is not a regulatory subunit of PKA or alpha-tubulin. Our results represent the identification of a soluble protein whose tyrosine phosphorylation varies directly with motility and suggest that motility regulation may involve cross talk between PKA, calcium, and tyrosine kinase pathways.


Attenuated behaviour in Cornelia de Lange and fragile X syndromes.
Catatonia-like presentations in people with autism have been increasingly recognised within research and diagnostic guidelines. The recently developed Attenuated Behaviour Questionnaire has identified that attenuated behaviour [autistic catatonia] is very prevalent in people with autism spectrum disorders (ASDs) and associated with repetitive behaviour. In the current study, we investigated attenuated behaviour within two genetic syndromes associated with ASD and examined ASD and repetitive behaviour as longitudinal predictors of attenuated behaviour. The Attenuated Behaviour Questionnaire was completed by parents/carers of 33 individuals with Cornelia de Lange syndrome (CdLS) and 69 with fragile X syndrome (FXS). Information collected from the same informants 4 years previously was utilised to examine ASD and repetitive behaviour as predictors of later attenuated behaviour, controlling for age, gender and ability. Catatonia-like attenuated behaviour was reported for individuals with CdLS (30.3%) and FXS (11.6%). Slowed movement was more prevalent in people with CdLS. No other phenotypic differences were observed. Across the two groups, repetitive behaviour predicted the presence of attenuated behaviour 4 years later, after controlling for age, gender and ability. Attenuated behaviour can be identified in individuals with CdLS and FXS and may have an effect on both adaptive behaviour and quality of life. Repetitive behaviours predicted subsequent risk within both groups and should be assessed by services as part of a pro-active strategy of support.


The prostate after castration and hormone replacement in a rat model: structural and ultrastructural analysis.
To evaluate if late hormonal replacement is able to recover the prostatic tissue modified by androgenic deprivation. 24 rats were assigned into a Sham group; an androgen deficient group, submitted to bilateral orchiectomy (Orch); and a group submitted to bilateral orchiectomy followed by testosterone replacement therapy (Orch+T). After 60 days from surgery blood was collected for determination of testosterone levels and the ventral prostate was collected for quantitative and qualitative microscopic analysis. The acinar epithelium height, the number of mast cells per field, and the densities of collagen fibers and acinar lumen were analyzed by stereological methods under light microscopy. The muscle fibers and types of collagen fibers were qualitatively assessed by scanning electron microscopy and polarization microscopy. Hormone depletion (in group Orch) and return to normal levels (in group Orch+T) were effective as verified by serum testosterone analysis. The androgen deprivation promoted several alterations in the prostate: the acinar epithelium height diminished from 16.58±0.47 to 11.48±0.29μm; the number of mast cells per field presented increased from 0.45±0.07 to 2.83±0.25; collagen fibers density increased from 5.83±0.92 to 24.70±1.56%; and acinar lumen density decreased from 36.78±2.14 to 16.47±1.31%. Smooth muscle was also increased in Orch animals, and type I collagen fibers became more predominant in these animals. With the exception of the densities of collagen fibers and acinar lumen, in animals receiving testosterone replacement therapy all parameters became statistically similar to Sham. Collagen fibers density became lower and acinar lumen density became higher in Orch+T animals, when compared to Sham. This is the first study to demonstrate a relation between mast cells and testosterone levels in the prostate. This cells have been implicated in prostatic cancer and benign hyperplasia, although its specific role is not understood. Testosterone deprivation promotes major changes in the prostate of rats. The hormonal replacement therapy was effective in reversing these alterations.


Photochemical stroke model: flunarizine prevents sensorimotor deficits after neocortical infarcts in rats.
We produced unilateral photochemical infarcts in the hindlimb sensorimotor neocortex of 186 rats by intravenous injection of the fluorescein derivative rose bengal and focal illumination of the intact skull surface. Infarcted rats showed specific, long-lasting deficits in tactile and proprioceptive placing reactions of the contralateral limbs, mostly the hindlimb. Placing deficits were most prominent during transition to immobility and/or when independent limb movements were required. Administration of flunarizine, a Class IV calcium antagonist, 30 minutes after infarction resulted in marked sparing of sensorimotor function in 30 rats. In contrast to 20 vehicle-treated rats, which remained deficient for at least 21 days, 15 (75%) of the rats treated with 1.25 mg/kg i.v. flunarizine showed normal placing on Day 1 after infarction, whereas the remaining five (25%) recovered within 5 days. Oral treatment of 10 rats with 40 mg/kg flunarizine was also effective. Neocortical infarct volume and thalamic gliosis, assessed 21 days after infarction, did not differ between 30 flunarizine- and 30 vehicle-treated rats. However, when 4-hour-old infarcts were measured in 16 rats, posttreatment with intravenous flunarizine reduced infarct size by 31%. In combination with appropriate behavioral analyses, photochemical thrombosis may constitute a relevant stroke model, in which flunarizine preserved behavioral function during a critical period, corresponding to the spread of ischemic damage.


Structurally assisted blackness in butterfly scales.
Surfaces of low reflectance are ubiquitous in animate systems. They form essential components of the visual appearance of most living species and can explicitly influence other biological functions such as thermoregulation. The blackness associated with all opaque surfaces of low reflectivity has until now been attributed to strongly absorbing pigmentation alone. Our present study challenges this assumption, demonstrating that in addition to the requirement of absorbing pigmentation, complex nano-structures contribute to the low reflectance of certain natural surfaces. We describe preliminary findings of an investigation into the nature of the black regions observed on the dorsal wings of several Lepidoptera. Specifically, we quantify the optical absorption associated with black wing regions on the butterfly Papilio ulysses and find that the nanostructure of the wing scales of these regions contributes significantly to their black appearance.


Activation of the precursor of human stromelysin 2 and its interactions with other matrix metalloproteinases.
Matrix metalloproteinases (MMP) are synthesized as inactive zymogens (proMMP) and subsequently activated by many factors to degrade the extracellular matrix (ECM). In the present study, we have examined the intermolecular activation mechanisms of proMMP by MMP-10 (stromelysin 2). ProMMP-10 was purified from the culture media of OSC-20 human oral squamous carcinoma cells stimulated with 12-O-tetradecanoylphorbol 13-acetate. The final products are partially activated (approximately 38% of the full activity) during the purification steps and contain proMMP-10 of Mr 56,000 with minor protein bands of Mr 47,000, 24,000 and 22,000. The zymogen is activated by 4-aminophenylmercuric acetate and processed to the active forms of Mr 47,000 and 24,000. The NH2-terminal sequence of the 47,000- and 24,000-Mr species is Phe82-Ser-Ser-Phe-Pro-Gly, which is identical to that of stromelysin 2. ProMMP-9 (progelatinase B) is activated by MMP-10 to its full activity and processed to the low-Mr species of Mr 81,000, 65,000, 57,000 and 55,000, the former two of which show proteolytic activity on a gelatin zymography. The NH2-terminal sequence analysis indicates that the 81,000-, 65,000- and 57,000-M, species have the identical sequence of Phe88-Gln-Thr-Phe-Glu-Gly, suggesting the cleavage of the Arg87-Phe88 peptide bond for activation and both NH2-terminal and COOH-terminal truncation in the 65,000- and 57,000-Mr forms. MMP-10 also activates proMMP-7 (promatrilysin) up to about 60% of the full activity and generates the same active species of Mr 19,000 as that obtained by activation with 4-aminophenylmercuric acetate. Incubation of proMMP-2 (progelatinase A) or proMMP-3 with MMP-10 does not result in activation of these proMMP. These results indicate that in addition to the previously reported activation of proMMP-1 (tissue procollagenase) and proMMP-8 (neutrophil procollagenase), MMP-10 can also activate proMMP-9 and proMMP-7, and suggest the possibility that MMP-10 may replace a role of MMP-3 in the ECM degradation in concert with other MMP under various pathological conditions.


Rapid epidemiological assessment of health status in displaced populations--an evolution toward standardized minimum, essential data sets.
Rapid epidemiological assessment (REA) has evolved over the past 30 years into an essential tool of disaster management. Small area survey and sampling methods are the major application. While REA is protocol driven, needs assessment of displaced populations remains highly non-standardized. The United Nations and other international organizations continue to call for the development of standardized instruments for post-disaster needs assessment. This study examines REA protocols from leading agencies in humanitarian health assistance across an evaluation criteria of best-practice attributes. Analysis of inconsistencies and deficits leads to the derivation of a Minimum Essential Data Set (MEDS) proposed for use by relief agencies in post-disaster REA of health status in displaced populations. This data set lends itself to initial assessment, ongoing monitoring, and evaluation of relief efforts. It is expected that the task of rapid epidemiological assessment, and more generally, the professional practice of post-disaster health coordination, will be enhanced by development, acceptance, and use of standardized Minimum Essential Data Sets (MEDS).


Conversion of solid waste to activated carbon to improve landfill sustainability.
Heterogeneous composite wastes from landfills were evaluated as precursors for the generation of activated carbon (AC). A single-step chemical activation process was applied involving irradiation with microwave energy and impregnation with KOH. The average percentage yield of AC from active landfill precursor was higher than that from closed landfill for all depths sampled. Increase in impregnation ratio and irradiation power decreased the average percentage yield for both landfill precursors (active: 38.1 to 33.1%; closed: 42.1 to 33.3%). The optimum pH range for adsorption of methylene blue was pH 6-7, while adsorption increased with increase in temperature over the range 30 to 50°C. Carbonyl and hydroxyl groups were the major functional groups on the surface of AC. The properties of the AC are potentially suitable for the removal of cationic dyes and pollutants. AC generated from the landfill composite was comparable to that from other biomass being managed through AC generation. This is the first report to demonstrate the possible reuse of landfill composite as AC. The reuse option of landfill composite could provide a means of sustainable management of landfilled municipal waste.


Barriers to the early integration of palliative home care into the disease trajectory of advanced cancer patients: A focus group study with palliative home care teams.
Palliative care is still often involved late in the disease trajectory. Recently, some studies have explored the barriers to early integration of PC in the hospital setting. Because palliative care home care (PHC) is organised differently compared with PC in a hospital setting, the identification of barriers to the early integration of PHC is needed. Six focus groups were held with PHC teams in Flanders, Belgium. Discussions were transcribed verbatim and analysed using constant comparative analysis. Our findings confirm many barriers found in previous studies, such as the lack of financial resources and the perception of PC as terminal care. Oncologists' lack of knowledge about the content and role of PC is also confirmed. Furthermore, professional caregivers working in the home context are lacking information on oncology therapies necessary to provide optimal PC. A barrier for the home context is the discontinuity of care, as a result of a lack of communicational structure and a lack of central coordination. The results contribute to a better understanding of the factors hindering the provision of PHC alongside oncology care. For the home context, transmural discontinuity of care seems to be an important additional barrier.


Hyperoxia-induced LC3B interacts with the Fas apoptotic pathway in epithelial cell death.
Epithelial cell death plays a critical role in hyperoxia-induced lung injury. We investigated the involvement of the autophagic marker microtubule-associated protein-1 light chain-3B (LC3B) in epithelial cell apoptosis after hyperoxia. Prolonged hyperoxia (>95% O(2)), which causes characteristic lung injury in mice, activated morphological and biochemical markers of autophagy. Hyperoxia induced the time-dependent expression and conversion of LC3B-I to LC3B-II in mouse lung in vivo and in cultured epithelial cells (Beas-2B, human bronchial epithelial cells) in vitro. Hyperoxia increased autophagosome formation in Beas-2B cells, as evidenced by electron microscopy and increased GFP-LC3 puncta. The augmented LC3B level after hyperoxia was transcriptionally regulated and dependent in part on the c-Jun N-terminal kinase pathway. We hypothesized that LC3B plays a regulatory role in hyperoxia-induced epithelial apoptosis. LC3B siRNA promoted hyperoxia-induced cell death in epithelial cells, whereas overexpression of LC3B conferred cytoprotection after hyperoxia. The autophagic protein LC3B cross-regulated the Fas apoptotic pathway by physically interacting with the components of death-inducing signaling complex. This interaction was mediated by caveolin-1 tyrosine 14, which is a known target of phosphorylation induced by hyperoxia. Taken together, hyperoxia-induced LC3B activation regulates the Fas apoptotic pathway and thus confers cytoprotection in lung epithelial cells. The interaction of LC3B and Fas pathways requires cav-1.


[A method to evaluate visual ability in infants].
The purpose of this study is to introduce a method to evaluate visual functions in infants in the first three months of life. An adaptation of the Guide for the Assessment of Visual Ability in Infants (Gagliardo, 1997) was used. The instrument was a ring with string. It was implemented a pilot study with 33 infants, selected according to the following criteria: neonates well enough to go home within two days of birth; 1 to 3 months of chronological age; monthly evaluation with no absence; subjects living in Campinas/SP metropolitan area. In the first month we observed: visual fixation (93,9%); eye contact (90,9%); horizontal tracking (72,7%); inspects surroundings (97,0%). In the third month, we observed: inspects own hands (42,4%) and increased movements of arms (36,4%). This method allowed the evaluation of visual functions in infants, according to the chronological age. Alterations in this function will facilitate immediate referral to medical services for diagnoses.


Automatic imitation in rhythmical actions: kinematic fidelity and the effects of compatibility, delay, and visual monitoring.
We demonstrate that observation of everyday rhythmical actions biases subsequent motor execution of the same and of different actions, using a paradigm where the observed actions were irrelevant for action execution. The cycle time of the distractor actions was subtly manipulated across trials, and the cycle time of motor responses served as the main dependent measure. Although distractor frequencies reliably biased response cycle times, this imitation bias was only a small fraction of the modulations in distractor speed, as well as of the modulations produced when participants intentionally imitated the observed rhythms. Importantly, this bias was not only present for compatible actions, but was also found, though numerically reduced, when distractor and executed actions were different (e.g., tooth brushing vs. window wiping), or when the dominant plane of movement was different (horizontal vs. vertical). In addition, these effects were equally pronounced for execution at 0, 4, and 8 s after action observation, a finding that contrasts with the more short-lived effects reported in earlier studies. The imitation bias was also unaffected when vision of the hand was occluded during execution, indicating that this effect most likely resulted from visuomotor interactions during distractor observation, rather than from visual monitoring and guidance during execution. Finally, when the distractor was incompatible in both dimensions (action type and plane) the imitation bias was not reduced further, in an additive way, relative to the single-incompatible conditions. This points to a mechanism whereby the observed action's impact on motor processing is generally reduced whenever this is not useful for motor planning. We interpret these findings in the framework of biased competition, where intended and distractor actions can be represented as competing and quasi-encapsulated sensorimotor streams.


Percent emphysema, airflow obstruction, and impaired left ventricular filling.
Very severe chronic obstructive pulmonary disease causes cor pulmonale with elevated pulmonary vascular resistance and secondary reductions in left ventricular filling, stroke volume, and cardiac output. We hypothesized that emphysema, as detected on computed tomography (CT), and airflow obstruction are inversely related to left ventricular end-diastolic volume, stroke volume, and cardiac output among persons without very severe lung disease. We measured left ventricular structure and function with the use of magnetic resonance imaging in 2816 persons who were 45 to 84 years of age. The extent of emphysema (expressed as percent emphysema) was defined as the percentage of voxels below -910 Hounsfield units in the lung windows on cardiac computed tomographic scans. Spirometry was performed according to American Thoracic Society guidelines. Generalized additive models were used to test for threshold effects. Of the study participants, 13% were current smokers, 38% were former smokers, and 49% had never smoked. A 10-point increase in percent emphysema was linearly related to reductions in left ventricular end-diastolic volume (-4.1 ml; 95% confidence interval [CI], -3.3 to -4.9; P<0.001), stroke volume (-2.7 ml; 95% CI, -2.2 to -3.3; P<0.001), and cardiac output (-0.19 liters per minute; 95% CI, -0.14 to -0.23; P<0.001). These associations were of greater magnitude among current smokers than among former smokers and those who had never smoked. The extent of airflow obstruction was similarly associated with left ventricular structure and function, and smoking status had similar modifying effects on these associations. Percent emphysema and airflow obstruction were not associated with the left ventricular ejection fraction. In a population-based study, a greater extent of emphysema on CT scanning and more severe airflow obstruction were linearly related to impaired left ventricular filling, reduced stroke volume, and lower cardiac output without changes in the ejection fraction.


Management factors affecting mortality, feed intake and feed conversion ratio of grow-finishing pigs.
The aim of this study was to determine the effect of animal management and farm facilities on total feed intake (TFI), feed conversion ratio (FCR) and mortality rate (MORT) of grower-finishing pigs. In total, 310 batches from 244 grower-finishing farms, consisting of 454 855 Pietrain sired pigs in six Spanish pig companies were used. Data collection consisted of a survey on management practices (season of placement, split-sex by pens, number of pig origins, water source in the farm, initial or final BW) and facilities (floor, feeder, ventilation or number of animals placed) during 2008 and 2009. Results indicated that batches of pigs placed between January and March had higher TFI (P=0.006), FCR (P=0.005) and MORT (P=0.03) than those placed between July and September. Moreover, batches of pigs placed between April and June had lower MORT (P=0.003) than those placed between January and March. Batches which had split-sex pens had lower TFI (P=0.001) and better FCR (P<0.001) than those with mixed-sex in pens; pigs fed with a single-space feeder with incorporated drinker also had the lowest TFI (P<0.001) and best FCR (P<0.001) in comparison to single and multi-space feeders without a drinker. Pigs placed in pens with <50% slatted floors presented an improvement in FCR (P<0.05) than pens with 50% or more slatted floors. Batches filled with pigs from multiple origins had higher MORT (P<0.001) than those from a single origin. Pigs housed in barns that performed manual ventilation control presented higher MORT (P<0.001) in comparison to automatic ventilation. The regression analysis also indicated that pigs which entered to grower-finisher facilities with higher initial BW had lower MORT (P<0.05) and finally pigs which were sent to slaughterhouse with a higher final BW presented higher TFI (P<0.001). The variables selected for each dependent variable explained 61.9%, 24.8% and 20.4% of the total variability for TFI, FCR and MORT, respectively. This study indicates that farms can increase growth performance and reduce mortality by improving farm facilities and/or modifying management practices.


Triheptanoin versus trioctanoin for long-chain fatty acid oxidation disorders: a double blinded, randomized controlled trial.
Observational reports suggest that supplementation that increases citric acid cycle intermediates via anaplerosis may have therapeutic advantages over traditional medium-chain triglyceride (MCT) treatment of long-chain fatty acid oxidation disorders (LC-FAODs) but controlled trials have not been reported. The goal of our study was to compare the effects of triheptanoin (C7), an anaplerotic seven-carbon fatty acid triglyceride, to trioctanoin (C8), an eight-carbon fatty acid triglyceride, in patients with LC-FAODs. A double blinded, randomized controlled trial of 32 subjects with LC-FAODs (carnitine palmitoyltransferase-2, very long-chain acylCoA dehydrogenase, trifunctional protein or long-chain 3-hydroxy acylCoA dehydrogenase deficiencies) who were randomly assigned a diet containing 20% of their total daily energy from either C7 or C8 for 4 months was conducted. Primary outcomes included changes in total energy expenditure (TEE), cardiac function by echocardiogram, exercise tolerance, and phosphocreatine recovery following acute exercise. Secondary outcomes included body composition, blood biomarkers, and adverse events, including incidence of rhabdomyolysis. Patients in the C7 group increased left ventricular (LV) ejection fraction by 7.4% (p = 0.046) while experiencing a 20% (p = 0.041) decrease in LV wall mass on their resting echocardiogram. They also required a lower heart rate for the same amount of work during a moderate-intensity exercise stress test when compared to patients taking C8. There was no difference in TEE, phosphocreatine recovery, body composition, incidence of rhabdomyolysis, or any secondary outcome measures between the groups. C7 improved LV ejection fraction and reduced LV mass at rest, as well as lowering heart rate during exercise among patients with LC-FAODs. Clinicaltrials.gov NCT01379625.


Awareness of social and emotional functioning in people with early-stage dementia and implications for carers.
This study explores the extent to which awareness of social and emotional function is reduced in early-stage dementia and whether this relates to the quality of life of the person with dementia (PwD), the quality of the relationship between the PwD and carer and carer stress. Ninety-seven participants with a diagnosis of Alzheimer's, vascular or mixed dementia rated their social functioning using the Socio-Emotional Questionnaire (SEQ). Carers provided parallel ratings, allowing calculation of discrepancy scores used to index awareness. Neuropsychiatric symptoms, PwD quality of life, the perceived quality of the relationship for both partners and carer stress were also measured. Factor analysis of the SEQ indicated three domains of social functioning: emotional recognition and empathy (ERE), social relationships (SR) and prosocial behaviour (PB). For PwD unawareness was related to cognitive dysfunction and psychiatric disturbance, but not to quality of life or quality of relationship. Lower awareness was associated with greater carer stress and poorer perceived quality of relationship. Lack of awareness of social functioning had important implications for relationship quality and levels of carer stress.


Suppressor of Hairless is required for long-term memory formation in Drosophila.
Suppressor of Hairless [Su(H)] is a DNA-binding protein of the Notch-signaling pathway, which is important for developmental processes and has been implicated in behavior plasticity. It acts as a transcriptional activator in the Notch pathway, but also as a repressor in the absence of Notch signaling. Our previous work has shown that Notch signaling contributes to long-term memory formation in the Drosophila adult brain. In the present report, we show that Su(H) null heterozygous mutants perform normally for learning, early memory, and anesthesia-resistant memory, whereas long-term memory is impaired. Interestingly, we find overexpressing wild- type Su(H) also causes long-term memory defect in Drosophila. Significantly, induction of a heat-shock inducible Su(H)(+) transgene before training can fully rescue the memory defect of Su(H) mutants, thereby demonstrating an acute role for Su(H) in behavioral plasticity. We show that Su(H) is widely expressed in the adult brain. Transgenic expression of wild-type Su(H) in the Mushroom Bodies is sufficient to rescue the memory defect of Su(H) mutants. Our data clearly demonstrate that transcriptional activity of Su(H) in Notch signaling in the mushroom bodies is critical for the formation of long-term memory.


Quantitative miRNA expression analysis using fluidigm microfluidics dynamic arrays.
MicroRNAs (miRNAs) represent a growing class of small non-coding RNAs that are important regulators of gene expression in both plants and animals. Studies have shown that miRNAs play a critical role in human cancer and they can influence the level of cell proliferation and apoptosis by modulating gene expression. Currently, methods for the detection and measurement of miRNA expression include small and moderate-throughput technologies, such as standard quantitative PCR and microarray based analysis. However, these methods have several limitations when used in large clinical studies where a high-throughput and highly quantitative technology needed for the efficient characterization of a large number of miRNA transcripts in clinical samples. Furthermore, archival formalin fixed, paraffin embedded (FFPE) samples are increasingly becoming the primary resource for gene expression studies because fresh frozen (FF) samples are often difficult to obtain and requires special storage conditions. In this study, we evaluated the miRNA expression levels in FFPE and FF samples as well as several lung cancer cell lines employing a high throughput qPCR-based microfluidic technology. The results were compared to standard qPCR and hybridization-based microarray platforms using the same samples. We demonstrated highly correlated Ct values between multiplex and singleplex RT reactions in standard qPCR assays for miRNA expression using total RNA from A549 (R = 0.98; p < 0.0001) and H1299 (R = 0.95; p < 0.0001) lung cancer cell lines. The Ct values generated by the microfluidic technology (Fluidigm 48.48 dynamic array systems) resulted in a left-shift toward lower Ct values compared to those observed by ABI 7900 HT (mean difference, 3.79), suggesting that the microfluidic technology exhibited a greater sensitivity. In addition, we show that as little as 10 ng total RNA can be used to reliably detect all 48 or 96 tested miRNAs using a 96-multiplexing RT reaction in both FFPE and FF samples. Finally, we compared miRNA expression measurements in both FFPE and FF samples by qPCR using the 96.96 dynamic array and Affymetrix microarrays. Fold change comparisons for comparable genes between the two platforms indicated that the overall correlation was R = 0.60. The maximum fold change detected by the Affymetrix microarray was 3.5 compared to 13 by the 96.96 dynamic array. The qPCR-array based microfluidic dynamic array platform can be used in conjunction with multiplexed RT reactions for miRNA gene expression profiling. We showed that this approach is highly reproducible and the results correlate closely with the existing singleplex qPCR platform at a throughput that is 5 to 20 times higher and a sample and reagent usage that was approximately 50-100 times lower than conventional assays. We established optimal conditions for using the Fluidigm microfluidic technology for rapid, cost effective, and customizable arrays for miRNA expression profiling and validation.


Update on rehabilitation in multiple sclerosis.
Given that mobility impairment is a hallmark of multiple sclerosis, people with this disease are likely to benefit from rehabilitation therapy throughout the course of their illness. The review provides an update on rehabilitation focused on balance and walking impairment. Classical rehabilitation focusing on muscle rehabilitation, neurotherapeutic facilitation is effective and recommended. Other techniques did not prove their superiority: transcutaneal neurostimulation, repetitive magnetic stimulation, electromagnetic therapy, whole body vibration and robot-assisted gait rehabilitation and need more studies to conclude. Cooling therapy, hydrotherapy, orthoses and textured insoles could represent a complementary service to other techniques in specific conditions. Multidisciplinary rehabilitation program provides positive effects and high satisfaction for patients with multiple sclerosis but needs more evaluation. New technologies using serious game and telerehabilitation seem to be an interesting technique to promote physical activity, self-management and quality of life. Rehabilitation like other therapy needs regular clinical evaluation to adapt the program and propose appropriate techniques. Moreover, the objective of rehabilitation needs to be decided with the patient with realistic expectation.


Sparstolonin B inhibits pro-angiogenic functions and blocks cell cycle progression in endothelial cells.
Sparstolonin B (SsnB) is a novel bioactive compound isolated from Sparganium stoloniferum, an herb historically used in Traditional Chinese Medicine as an anti-tumor agent. Angiogenesis, the process of new capillary formation from existing blood vessels, is dysregulated in many pathological disorders, including diabetic retinopathy, tumor growth, and atherosclerosis. In functional assays, SsnB inhibited endothelial cell tube formation (Matrigel method) and cell migration (Transwell method) in a dose-dependent manner. Microarray experiments with human umbilical vein endothelial cells (HUVECs) and human coronary artery endothelial cells (HCAECs) demonstrated differential expression of several hundred genes in response to SsnB exposure (916 and 356 genes, respectively, with fold change ≥2, p<0.05, unpaired t-test). Microarray data from both cell types showed significant overlap, including genes associated with cell proliferation and cell cycle. Flow cytometric cell cycle analysis of HUVECs treated with SsnB showed an increase of cells in the G1 phase and a decrease of cells in the S phase. Cyclin E2 (CCNE2) and Cell division cycle 6 (CDC6) are regulatory proteins that control cell cycle progression through the G1/S checkpoint. Both CCNE2 and CDC6 were downregulated in the microarray data. Real Time quantitative PCR confirmed that gene expression of CCNE2 and CDC6 in HUVECs was downregulated after SsnB exposure, to 64% and 35% of controls, respectively. The data suggest that SsnB may exert its anti-angiogenic properties in part by downregulating CCNE2 and CDC6, halting progression through the G1/S checkpoint. In the chick chorioallantoic membrane (CAM) assay, SsnB caused significant reduction in capillary length and branching number relative to the vehicle control group. Overall, SsnB caused a significant reduction in angiogenesis (ANOVA, p<0.05), demonstrating its ex vivo efficacy.


A pair of transposons coordinately suppresses gene expression, independent of pathways mediated by siRNA in Antirrhinum.
Our knowledge is limited regarding mechanisms by which transposable elements control host gene expression. Two Antirrhinum lines, HAM2 and HAM5, show different petal colors, pale-red and white, respectively, although these lines contain the same insertion of transposon Tam3 in the promoter region of the nivea (niv) locus encoding chalcone synthase. Among 1000 progeny from HAM5 grown under the preferred conditions for the Tam3 transposition, a few showed an intermediate petal color between HAM2 and HAM5. Transposon tagging using these progeny identified a causative insertion of Tam3 for the HAM5 type (white) petal color, which was found 1.6 kb downstream of the niv gene. Insertion of Tam3 at the position 1.6 kb downstream of niv alone showed nearly wildtype petal pigmentation, and the niv expression reduced by only 50%. Severe suppression of niv observed in HAM5 required interaction of two Tam3 copies on either side of the niv coding sequence. DNA methylation and small interfering RNAs (siRNAs) were not associated with the suppression of niv expression in HAM5. Insertion of a pair of transposons in close proximity can interfere with the expression of gene located between the two copies, and also provide evidence that this interference is not directly associated with pathways mediated by siRNAs.


Does health literacy affect patients' receipt of preventative primary care? A multilevel analysis.
People with limited health literacy are more likely to be socioeconomically disadvantaged and have risk factors for preventable chronic diseases. General practice is the ideal setting to address these inequalities however these patients engage less in preventive activities and experience difficulties navigating health services. This study aimed to compare primary care patients with and without sufficient health literacy in terms of their lifestyle risk factors, and explore factors associated with receiving advice and referral for these risk factors from their GPs. A mailed survey of 739 patients from 30 general practices across four Australian states was conducted in 2012. Health literacy was measured using the Health Literacy Management Scale. Patients with a mean score of <4 within any domain were defined as having insufficient health literacy. Multilevel logistic regression was used to adjust for clustering of patients within practices. Patients with insufficient health literacy (n = 351; 48%) were more likely to report being overweight or obese, and less likely to exercise adequately. Having insufficient health literacy increased a patient's chance of receiving advice on diet, physical activity or weight management, and referral to and attendance at lifestyle modification programs. Not speaking English at home; being overweight or obese; and attending a small sized practice also increased patients' chances of receiving advice on these lifestyle risks. Few (5%, n = 37) of all patients reported being referred to lifestyle modification program and of those around three-quarters had insufficient health literacy. Overweight or obese patients were more likely to be referred to lifestyle modification programs and patients not in paid employment were more likely to be referred to and attend lifestyle programs. Patients with insufficient health literacy were more likely to report receiving advice and being referred by GPs to attend lifestyle modification. Although the number of patients referred from this sample was very low, these findings are positive in that they indicate that GPs are identifying patients with low health literacy and appropriately referring them for assistance with lifestyle modification. Future research should measure the effectiveness of these lifestyle programs for patients with low health literacy.


Diet and risk of renal cell cancer: a population-based case-control study.
In a few previous studies on diet and renal cell cancer, an inconsistent positive association with meat, milk, and protein and a negative association with vegetable and fruit consumption have been found. Whereas earlier studies have dealt with recent diet only, our study explored the effect of foods consumed both during the usual adult lifetime and 20 years prior to interview. The study included 379 individuals with incident histologically verified renal cell cancer and 350 control subjects residing in eight counties in Sweden between June 1989 and December 1991. Usual adult dietary intake and dietary habits 20 years prior to interview were assessed by a structured face-to-face interview and a self-administered questionnaire, respectively. Odds ratios were estimated through unconditional logistic regression. We have not observed an association of renal cell cancer with milk or total meat consumption per se; however, frequent intake of fried/sautéed meat increased the risk of renal cell cancer by about 60%; frequent consumption of poultry was also associated with an increased risk (P for trend, 0.05). A significantly protective effect on risk of renal cell cancer was observed with increasing consumption of fruit (P for trend, 0.05). When analyzed by smoking status, total fruit and especially citrus fruit consumption among nonsmokers showed an even stronger protective effect; the highest quartiles of total fruit, apple, and citrus fruit consumption entailed a 50-60% reduction in risk of renal cell cancer compared with the lowest quartiles. There was a suggestion of a protective effect of high total vegetable consumption. A protective effect of vitamin C and alpha-tocopherol was also more pronounced in nonsmokers (P for trend, 0.004 and 0.007, respectively). Our study adds to the evidence that diet may have an important role in the etiology of renal cell cancer.


The importance of placebo effects in pain treatment and research.
To estimate the importance and implications of placebo effects in pain treatment and research from the existing literature, with emphasis on their magnitude and duration, the conditions influencing them, and proposed explanations. English-language articles and books identified through MEDLINE (1980 through 1993) and PsycLIT (1967 through 1993) database searching, bibliography review, and expert consultation. Articles were included if they pertained to the review objectives. Placebo response rates vary greatly and are frequently much higher than the often-cited one third. Placebos have time-effect curves, and peak, cumulative, and carryover effects similar to those of active medications. As with medication, surgery can produce substantial placebo effects, and this possibility is commonly overlooked in case series reports on back surgery. Individuals are not consistent in their placebo responses, and a placebo-responder personality has not been identified. Models advanced to explain placebo effects emphasize the role of anxiety, expectations, and learning. Placebo effects influence patient outcomes after any treatment, including surgery, that the clinician and patient believe is effective. Placebo effects plus disease natural history and regression to the mean can result in high rates of good outcomes, which may be misattributed to specific treatment effects. The true causes of improvements in pain after treatment remain unknown in the absence of independently evaluated randomized controlled trials.


Phylogenomic Analysis of Extraintestinal Pathogenic Escherichia coli Sequence Type 1193, an Emerging Multidrug-Resistant Clonal Group.
The fluoroquinolone-resistant sequence type 1193 (ST1193) of Escherichia coli, from the ST14 clonal complex (STc14) within phylogenetic group B2, has appeared recently as an important cause of extraintestinal disease in humans. Although this emerging lineage has been characterized to some extent using conventional methods, it has not been studied extensively at the genomic level. Here, we used whole-genome sequence analysis to compare 355 ST1193 isolates with 72 isolates from other STs within STc14. Using core genome phylogeny, the ST1193 isolates formed a tightly clustered clade with many genotypic similarities, unlike ST14 isolates. All ST1193 isolates possessed the same set of three chromosomal mutations conferring fluoroquinolone resistance, carried the fimH64 allele, and were lactose non-fermenting. Analysis revealed an evolutionary progression from K1 to K5 capsular types and acquisition of an F-type virulence plasmid, followed by changes in plasmid structure congruent with genome phylogeny. In contrast, the numerous identified antimicrobial resistance genes were distributed incongruently with the underlying phylogeny, suggesting frequent gain or loss of the corresponding resistance gene cassettes despite retention of the presumed carrier plasmids. Pangenome analysis revealed gains and losses of genetic loci occurring during the transition from ST14 to ST1193 and from the K1 to K5 capsular types. Using time-scaled phylogenetic analysis, we estimated that current ST1193 clades first emerged approximately 25 years ago. Overall, ST1193 appears to be a recently emerged clone in which both stepwise and mosaic evolution have contributed to epidemiologic success.


Brain structural abnormalities in behavior therapy-resistant obsessive-compulsive disorder revealed by voxel-based morphometry.
Although several functional imaging studies have demonstrated that behavior therapy (BT) modifies the neural circuits involved in the pathogenesis of obsessive-compulsive disorder (OCD), the structural abnormalities underlying BT-resistant OCD remain unknown. In this study, we examined the existence of regional structural abnormalities in both the gray matter and the white matter of patients with OCD at baseline using voxel-based morphometry in responders (n=24) and nonresponders (n=15) to subsequent BT. Three-dimensional T1-weighted magnetic resonance imaging was performed before the completion of 12 weeks of BT. Relative to the responders, the nonresponders exhibited significantly smaller gray matter volumes in the right ventromedial prefrontal cortex, the right orbitofrontal cortex, the right precentral gyrus, and the left anterior cingulate cortex. In addition, relative to the responders, the nonresponders exhibited significantly smaller white matter volumes in the left cingulate bundle and the left superior frontal white matter. These results suggest that the brain structures in several areas, including the orbitofrontal cortex, anterior cingulate cortex, and cingulate bundles, are related to the lack of a response to BT in patients with OCD. The use of a voxel-based morphometry approach may be advantageous to understanding differences in brain abnormalities between responders and nonresponders to BT.


Qualitative and Quantitative Evaluation of Blob-Based Time-of-Flight PET Image Reconstruction in Hybrid Brain PET/MR Imaging.
Many neurological diseases affect small structures in the brain and, as such, reliable visual evaluation and accurate quantification are required. Recent technological developments made the clinical use of hybrid positron emission tomography/magnetic resonance (PET/MR) systems possible, providing both functional and anatomical information in a single imaging session. Nevertheless, there is a trade-off between spatial resolution and image quality (contrast and noise), which is dictated mainly by the chosen acquisition and reconstruction protocols. Image reconstruction algorithms using spherical symmetric basis functions (blobs) for image representation have a number of additional parameters that impact both the qualitative and quantitative image characteristics. Hence, a detailed investigation of the blob-based reconstruction characteristics using different parameters is needed to achieve optimal reconstruction results. This work evaluated the impact of a range of blob parameters on image quality and quantitative accuracy of brain PET images acquired on the Ingenuity Time-of-Flight (TOF) PET/MR system. Two different phantoms were used to simulate brain imaging applications. Image contrast and noise characteristics were assessed using an image quality phantom. Quantitative performance in a clinical setting was investigated using the Hoffman 3D brain phantom at various count levels. Furthermore, the visual quality of four clinical studies was scored blindly by two experienced physicians to qualitatively evaluate the influence of different reconstruction protocols, hereby providing indications on parameters producing the best image quality. Quantitative evaluation using the image quality phantom showed that larger basis function radii result in lower contrast recovery (∼2%) and lower variance levels (∼15%). The brain phantom and clinical studies confirmed these observations since lower contrast was seen between anatomical structures. High and low count statistics gave comparable values. The qualitative evaluation of the clinical studies, based on the assessment performed by the physicians, showed a preference towards lower image variance levels with a slightly lower contrast, favoring higher radii and four iterations. This study systematically evaluated a number of basis function parameters and their quantitative and qualitative effect within PET image reconstruction in the context of brain imaging. A range of blob parameters can minimize error and provided optimal image quality, where the anatomical structures could be recognized but the exact delineation of these structures is simplified in scans with lower image variance levels and thus, higher radii should be preferred. With the optimization of blob parameters, the reconstructed images were found to be qualitatively improved (optimum parameters {d = 2.0375, alpha = 10.4101, radius = 3.9451}) as assessed by the physicians compared to the current clinical protocol. However, this qualitative improvement is task specific, depending on the desired image characteristics to be extracted. Finally, this work could be used as a guide for application-specific optimal parameter selection.


Application of high-throughput 16S rRNA sequencing to identify fecal contamination sources and to complement the detection of fecal indicator bacteria in rural groundwater.
Residents in rural communities across Canada collect potable water from aquifers. Fecal contaminants from sewage and agricultural runoffs can penetrate aquifers, posing a public health risk. Standard methods for detecting fecal contamination test for fecal indicator bacteria (FIB), but the presence of these do not identify sources of contamination. In contrast, DNA-based diagnostic tools can achieve this important objective. We employed quantitative polymerase chain reaction (qPCR) and high-throughput DNA sequencing to trace fecal contamination sources in Wainfleet, a rural Ontario township that has been under the longest active boil water advisory in Canada due to FIB contamination in groundwater wells. Using traditional methods, we identified FIBs indicating persistent fecal pollution in well waters. We used 16S rRNA sequencing to profile groundwater microbial communities and identified Campylobacteraceae as a fecal contamination DNA marker in septic tank effluents (STEs). We also identified Turicibacter and Gallicola as a potential cow and chicken fecal contamination marker, respectively. Using human specific Bacteroidales markers, we identified leaking septic tanks as the likely primary fecal contamination source in some of Wainfleet's groundwater. Overall, the results support the use of sequencing-based methods to augment traditional water quality testing methods and help end-users assess fecal contamination levels and identify point and non-point pollution sources.


The Y Chromosome Modulates Splicing and Sex-Biased Intron Retention Rates in Drosophila.
The Drosophila Y chromosome is a 40-Mb segment of mostly repetitive DNA; it harbors a handful of protein-coding genes and a disproportionate amount of satellite repeats, transposable elements, and multicopy DNA arrays. Intron retention (IR) is a type of alternative splicing (AS) event by which one or more introns remain within the mature transcript. IR recently emerged as a deliberate cellular mechanism to modulate gene expression levels and has been implicated in multiple biological processes. However, the extent of sex differences in IR and the contribution of the Y chromosome to the modulation of AS and IR rates has not been addressed. Here we showed pervasive IR in the fruit fly Drosophila melanogaster with thousands of novel IR events, hundreds of which displayed extensive sex bias. The data also revealed an unsuspected role for the Y chromosome in the modulation of AS and IR. The majority of sex-biased IR events introduced premature termination codons and the magnitude of sex bias was associated with gene expression differences between the sexes. Surprisingly, an extra Y chromosome in males (X^YY genotype) or the presence of a Y chromosome in females (X^XY genotype) significantly modulated IR and recapitulated natural differences in IR between the sexes. Our results highlight the significance of sex-biased IR in tuning sex differences and the role of the Y chromosome as a source of variable IR rates between the sexes. Modulation of splicing and IR rates across the genome represent new and unexpected outcomes of the Drosophila Y chromosome.


Spin-dependent exciton formation in pi-conjugated compounds.
The efficiency of light-emitting diodes (LEDs) made from organic semiconductors is determined by the fraction of injected electrons and holes that recombine to form emissive spin-singlet states rather than non-emissive spin-triplet states. If the process by which these states form is spin-independent, the maximum efficiency of organic LEDs will be limited to 25 per cent. But recent reports have indicated fractions of emissive singlet states ranging from 22 to 63 per cent, and the reason for this variation remains unclear. Here we determine the absolute fraction of singlet states generated in a platinum-containing conjugated polymer and its corresponding monomer. The spin-orbit coupling introduced by the platinum atom allows triplet-state emission, so optically and electrically generated luminescence from both singlet and triplet states can be compared directly. We find an average singlet generation fraction of 22 +/- 1 per cent for the monomer, but 57 +/- 4 per cent for the polymer. This suggests that recombination is spin-independent for the monomer, but that a spin-dependent process, favouring singlet formation, is effective in the polymer. We suggest that this process is a consequence of the exchange interaction, which will operate on overlapping electron and hole wavefunctions on the same polymer chain at their capture radius.


Identification of ubiquinol binding motifs at the Qo-site of the cytochrome bc1 complex.
Enzymes of the bc1 complex family power the biosphere through their central role in respiration and photosynthesis. These enzymes couple the oxidation of quinol molecules by cytochrome c to the transfer of protons across the membrane, to generate a proton-motive force that drives ATP synthesis. Key for the function of the bc1 complex is the initial redox process that involves a bifurcated electron transfer in which the two electrons from a quinol substrate are passed to different electron acceptors in the bc1 complex. The electron transfer is coupled to proton transfer. The overall mechanism of quinol oxidation by the bc1 complex is well enough characterized to allow exploration at the atomistic level, but details are still highly controversial. The controversy stems from the uncertain binding motifs of quinol at the so-called Qo active site of the bc1 complex. Here we employ a combination of classical all atom molecular dynamics and quantum chemical calculations to reveal the binding modes of quinol at the Qo-site of the bc1 complex from Rhodobacter capsulatus. The calculations suggest a novel configuration of amino acid residues responsible for quinol binding and support a mechanism for proton-coupled electron transfer from quinol to iron-sulfur cluster through a bridging hydrogen bond from histidine that stabilizes the reaction complex.


Association between the intact foreskin and inferior standards of male genital hygiene behaviour: a cross-sectional study.
This study was undertaken to determine whether non-circumcised men have inferior standards of genital hygiene behaviour, as measured by reported washing of the whole penis, compared with circumcised men. Male attenders at a sexually transmitted infections (STI) clinic at Ealing Hospital, London had routine STI tests and examinations performed and were asked about the frequency and thoroughness of genital washing. One hundred and fifty non-circumcised and 75 circumcised men were enrolled. Not always washing the whole penis, including retracting the foreskin in non-circumcised men every time they washed (defined as inferior genital hygiene behaviour) was more common in non-circumcised (26%) than circumcised men (4%) (crude odds ratio = 8.43, 95% confidence interval: 2.51-28.3, P<0.001) and those with balanitis (42% and 5%, P=0.036). Circumcised men were more likely than non-circumcised men to wash the genital area more than once a day (37% and 19%, P=0.011). Studies investigating the relationship between male circumcision status and other outcomes, for example HIV infection, should include assessment of genital hygiene.


Sublingual immunotherapy: recent advances.
The practice of administering sublingual immunotherapy for respiratory allergy is gaining more and more diffusion worldwide as a consequence of the robust demonstration of clinical efficacy and safety provided by recent high-powered and well-designed studies, confirming for individual seasonal allergens the results of previous metanalyses in adult and pediatric populations. Preliminary evidence derives from recent rigorous trials on perennial allergens, like house dust mites, and specifically designed studies addressed the benefits on asthma. Emerging research suggests that SLIT may have a future role in other allergic conditions such as atopic dermatitis, food, latex and venom allergy. Efforts to develop a safer and more effective SLIT for inhalant allergens have led to the development of allergoids, recombinant allergens and formulations with adjuvants and substances targeting antigens to dendritic cells that possess a crucial role in initiating immune responses. The high degree of variation in the evaluation of clinical effects and immunological changes requires further studies to identify the candidate patients to SLIT and biomarkers of short and long term efficacy. Appropriate management strategies are urgently needed to overcome the barriers to SLIT compliance.


Experimental and theoretical studies of the structural and electronic properties of vanadium-benzene sandwich clusters and their anions: V(n)Bz(n)(0/-) (n = 1-5) and V(n)Bz(n-1)(0/-) (n = 2-5).
One end open V(n)Bz(n)(-) (n = 1-5; Bz = benzene) and both ends open V(n)Bz(n-1)(-) (n = 2-5) vanadium-benzene cluster anions were studied using anion photoelectron spectroscopy and density functional calculations. The smaller (n ≤ 3) V(n)Bz(n) and V(n)Bz(n-1) clusters and corresponding anions were found to have structural isomers, whereas full-sandwiched V(n)Bz(n+1) clusters preferred to form multiple-decker sandwich structures. Several isomeric V2Bz2 structures were identified theoretically and the anion photoelectron spectra of V2Bz2(0/-) were explained well by the coexistence of two isomeric structures: (1) a V2-core structure sandwiched between benzene molecules and (2) an alternating sandwich structure with the spin state strongly dependent on the structure. The adiabatic electron affinity of both V(n)Bz(n) and V(n)Bz(n-1) was found to increase with the cluster size at larger sizes (n = 4 or 5) and approaches to that of V(n)Bz(n+1). The evolution of the structural and electronic properties of V(n)Bz(m) and V(n)Bz(m)(-) (m = n and n - 1) with size is discussed in comparison with V(n)Bz(n+1) and V(n)Bz(n+1)(-).


Identification and quantification of phytochemicals in nutraceutical products from green tea by UHPLC-Orbitrap-MS.
A method has been developed and validated for the simultaneous detection and quantification of phytochemicals in nutraceutical products obtained from green tea. For that purpose, ultra-high performance liquid chromatography coupled to single-stage Orbitrap high resolution mass spectrometry (UHPLC-Orbitrap-MS) has been used. A database containing 37 compounds has been used for the detection and identification of the target compounds. The developed methodology was based on solid-liquid extraction, using a mixture of methanol:H2O (80:20, v/v, pH 4), followed by dilution (10 times) with a mixture of ammonium acetate:methanol (50:50, v/v). Chromatographic conditions were optimised and full scan accurate mass data acquisition using electrospray ionisation in positive and negative ion mode was used. Moreover, all-ion fragmentation mode was used to get information of fragment ions, and they were used for identification purposes. The developed method was validated, obtaining repeatability (intra-day) and inter-day precision values (expressed as relative standard deviation, RSD) lower than 16% and 20%, respectively. Lower limits were also evaluated and limits of detection (LODs) ranged from 1 to 50 μg L(-1), while limits of quantification (LOQs) ranged from 2 to 150 μg L(-1). Recovery was performed at five levels and it ranged from 70% to 109%. Finally, this method was used to evaluate the phytochemical content in 10 samples (tablets or capsules), showing concentrations of (+)-catechin, (-)-epicatechin, gallic acid, (-)-gallocatechin and quercetin-3-O-rutinoside, ranging from 258 (C6) to 10,729 (C6) mg kg(-1).


Revision anterior cruciate ligament reconstruction surgery.
In 1995, it was reported that 60,000 to 75,000 anterior cruciate ligament (ACL) reconstructions were being performed annually in the United States. Successful long-term results are achieved in 75% to 95% of these patients, but 8% have unsatisfactory results due to recurrent instability and graft failure. With the increasing popularity of this procedure, ACL revision surgery has also become increasingly common. While the techniques described for ACL revision have been varied, the overall results in the literature do not compare favorably with primary ACL reconstruction. The proper execution of revision ACL reconstruction requires precise preoperative planning to assess the cause of initial failure and avoid repeating the same mistakes with revision reconstruction. Graft choice, hardware removal, revision notchplasty, tunnel placement, and method of fixation are key points for a successful result. The causes of ACL failure, the technical aspects of revision ACL surgery, and the reported results of revision ACL surgery are reviewed.


Ethnoracial differences in PTSD symptoms and trauma-related cognitions in treatment-seeking active duty military personnel for PTSD.
It is uncertain whether ethnoracial factors should be considered by clinicians assessing and treating posttraumatic stress disorder (PTSD) among service members. The purpose of this study was to shed light on ethnoracial variation in the presentation of PTSD symptoms, trauma-related cognitions, and emotions among treatment-seeking active duty military personnel. Participants were 303 male active duty military members with PTSD participating in a clinical trial (60% were self-identified as White, 19% as African American, and 21% as Hispanic/Latino). In the parent study, participants completed a baseline assessment that included clinician-administered and self-report measures of PTSD, trauma-related cognitions, and emotions. Multivariate hierarchical regression models were used to examine ethnoracial differences in these variables, covarying age, education, military grade, combat exposure, and exposure to other potentially traumatic events. Hispanic/Latino and African American participants reported more reexperiencing symptoms, more fear, and more guilt and numbing than White participants. All effect sizes were in the small to medium range. These findings suggest ethnoracial variation in PTSD symptom burden and posttraumatic cognitions among treatment-seeking service members with PTSD. Attending to cultural factors related to differences in PTSD presentation and cognitive coping strategies during the assessment and treatment process could increase rapport and lead to more comprehensive trauma processing. (PsycINFO Database Record


Genetic Analysis of Pyricularia grisea Population by rep-PCR During Development of Resistance to Scytalone Dehydratase Inhibitors of Melanin Biosynthesis.
In 2001, field isolates of Pyricularia grisea resistant to scytalone dehydratase inhibitors of melanin biosynthesis (MBI-D) were reported in Saga prefecture, Kyushu. Among 1,175 isolates collected from six prefectures of Kyushu in 2002 and 2003, 647 were resistant to MBI-D fungicides, each due to a single point mutation of the scytalone dehydratase (SDH) gene. On the basis of repetitive element-based polymerase chain reaction (rep-PCR) fingerprint data, the haplotypes of the resistant isolates showed high genetic diversity, indicating that the resistance existed in a multigenetic background. Three predominant haplotypes mainly contributed to the widespread resistance in Kyushu; haplotype Sa4 was observed frequently in Saga, Sa18 was predominant in Oita and Miyazaki, and Sa5 was widely distributed among all four prefectures. Also, phylogenetic analysis showed that both the resistant and sensitive isolates were clustered together in a closely related group. These results suggest that isolates possessing the SDH mutation would have been selected and then multiplied rapidly in each region of Kyushu as a result of the widespread introduction of MBI-D fungicides in a short period.


The effect of perceived person-job fit on employee attitudes toward change in trauma centers.
Employee attitudes toward change are critical for health care organizations implementing new procedures and practices. When employees are more positive about the change, they are likely to behave in ways that support the change, whereas when employees are negative about the change, they will resist the changes. This study examined how perceived person-job (demands-abilities) fit influences attitudes toward change after an externally mandated change. Specifically, we propose that perceived person-job fit moderates the negative relationship between individual job impact and attitudes toward change. We examined this issue in a sample of Level 1 trauma centers facing a regulatory mandate to develop an alcohol screening and brief intervention program. A survey of 200 providers within 20 trauma centers assessed perceived person-job fit, individual job impact, and attitudes toward change approximately 1 year after the mandate was enacted. Providers who perceived a better fit between their abilities and the new job demands were more positive about the change. Further, the impact of the alcohol screening and brief intervention program on attitudes toward change was mitigated by perceived fit, where the relationship between job impact and change attitudes was more negative for providers who perceived a worse fit as compared with those who perceived a better fit. Successful implementation of changes to work processes and procedures requires provider support of the change. Management can enhance this support by improving perceived person-job fit through ongoing training sessions that enhance providers' abilities to implement the new procedures.


Sweet orosensation induces Arc expression in dorsal hippocampal CA1 neurons in an experience-dependent manner.
There is limited knowledge regarding how the brain controls the timing of meals. Similarly, there is a large gap in our understanding of how top-down cognitive processes, such as memory influence energy intake. We hypothesize that dorsal hippocampal (dHC) neurons, which are critical for episodic memory, form a memory of a meal and inhibit meal onset during the postprandial period. In support, we showed previously that reversible inactivation of these neurons during the period following a sucrose meal accelerates the onset of the next meal. If dHC neurons form a memory of a meal, then consumption should induce synaptic plasticity in dHC neurons. To test this, we determined (1) whether a sucrose meal increases the expression of the synaptic plasticity marker activity-regulated cytoskeleton-associated protein (Arc) in dHC CA1 neurons, (2) whether previous experience with sucrose influences sucrose-induced Arc expression, and (3) whether the orosensory stimulation produced by the noncaloric sweetener saccharin is sufficient to induce Arc expression. Male Sprague-Dawley rats were trained to consume a sweetened solution at a scheduled time daily. On the experimental day, they were given a solution for 7 min, euthanized, and then fluorescence in situ hybridization procedures were used to measure meal-induced Arc mRNA. Compared to caged control rats, Arc expression was significantly higher in rats that consumed sucrose or saccharin. Interestingly, rats given additional experience with sucrose had less Arc expression than rats with less sucrose experience, even though both groups consumed similar amounts on the experimental day. Thus, this study is the first to suggest that orosensory stimulation produced by consuming a sweetened solution and possibly the hedonic value of that sweet stimulation induces synaptic plasticity in dHC CA1 neurons in an experience-dependent manner. Collectively, these findings are consistent with our hypothesis that dHC neurons form a memory of a meal.


Economic impact of heart failure with preserved ejection fraction: insights from the ALDO-DHF trial.
Although heart failure (HF) with preserved ejection fraction (HFpEF) is a leading cause for hospitalization, its overall costs remain unclear. Therefore, we assessed the health care-related costs of ambulatory HFpEF patients and the effect of spironolactone. The aldosterone receptor blockade in diastolic HF trial is a multicentre, prospective, randomized, double-blind, placebo-controlled trial conducted between March 2007 and April 2011 at 10 sites in Germany and Austria that included 422 ambulatory patients [mean age: 67 years (standard deviation: 8); 52% women]. All subjects suffered from chronic New York Heart Association (NYHA) class II or III HF and preserved left ventricular ejection fraction of 50% or greater. They also showed evidence of diastolic dysfunction. Patients were randomly assigned to receive 25 mg of spironolactone once daily (n = 213) or matching placebo (n = 209) with 12 months of follow-up. We used a single-patient approach to explore the resulting general cost structure and included medication, number of general practitioner and cardiologist visits, and hospitalization in both acute and rehabilitative care facilities. The average annual costs per patient in this cohort came up to €1, 118 (±2,475), and the median costs were €332. We confirmed that the main cost factor was hospitalization and spironolactone did not affect the overall costs. We identified higher HF functional class (NYHA), male patients with low haemoglobin level, with high oxygen uptake (VO2 max) and coronary artery disease, hyperlipidaemia, and atrial fibrillation as independent predictors for higher costs. In this relatively young, oligosymptomatic, and with regard to the protocol without major comorbidities patient cohort, the overall costs are lower than expected compared with the HFrEF population. Further investigation is needed to investigate the impact of, for example, comorbidities and their effect over a longer period of time. Simultaneously, this analysis suggests that prevention of comorbidities are necessary to reduce costs in the health care system.


Hippocampal-prefrontal engagement and dynamic causal interactions in the maturation of children's fact retrieval.
Children's gains in problem-solving skills during the elementary school years are characterized by shifts in the mix of problem-solving approaches, with inefficient procedural strategies being gradually replaced with direct retrieval of domain-relevant facts. We used a well-established procedure for strategy assessment during arithmetic problem solving to investigate the neural basis of this critical transition. We indexed behavioral strategy use by focusing on the retrieval frequency and examined changes in brain activity and connectivity associated with retrieval fluency during arithmetic problem solving in second- and third-grade (7- to 9-year-old) children. Children with higher retrieval fluency showed elevated signal in the right hippocampus, parahippocampal gyrus (PHG), lingual gyrus (LG), fusiform gyrus (FG), left ventrolateral PFC (VLPFC), bilateral dorsolateral PFC (DLPFC), and posterior angular gyrus. Critically, these effects were not confounded by individual differences in problem-solving speed or accuracy. Psychophysiological interaction analysis revealed significant effective connectivity of the right hippocampus with bilateral VLPFC and DLPFC during arithmetic problem solving. Dynamic causal modeling analysis revealed strong bidirectional interactions between the hippocampus and the left VLPFC and DLPFC. Furthermore, causal influences from the left VLPFC to the hippocampus served as the main top-down component, whereas causal influences from the hippocampus to the left DLPFC served as the main bottom-up component of this retrieval network. Our study highlights the contribution of hippocampal-prefrontal circuits to the early development of retrieval fluency in arithmetic problem solving and provides a novel framework for studying dynamic developmental processes that accompany children's development of problem-solving skills.


Pigment Composition of Nine Brown Algae from the Iberian Northwestern Coastline: Influence of the Extraction Solvent.
Brown algae are ubiquitously distributed in the NW coastline of the Iberian Peninsula, where they stand as an underexploited resource. In this study, five solvents were applied to the extraction of pigments from nine brown algae, followed by their determination and quantification by HPLC-DAD. A total of 13 compounds were detected: Six were identified as chlorophylls, six were classified as xanthophylls, and one compound was reported as a carotene. Fucoxanthin was reported in all extracts, which is the most prominent pigment of these algae. Among them, L. saccharina and U. pinnatifida present the highest concentration of fucoxanthin (4.5-4.7 mg∙g-1 dry weight). Ethanol and acetone were revealed as the most efficient solvents for the extraction of pigments, showing a maximal value of 11.9 mg of total pigments per gram of dry alga obtained from the ethanolic extracts of H. elongata, followed by the acetonic extracts of L. ochroleuca. Indeed, ethanol was also revealed as the most efficient solvent according to its high extraction yield along all species evaluated. Our results supply insights into the pigment composition of brown algae, opening new perspectives on their commercial exploitation by food, pharmaceutical, and cosmeceutical industries.


Visual pain score during transrectal ultrasound-guided prostate biopsy using no anaesthesia or three different types of local anaesthetic application.
To evaluate the pain score during transrectal ultrasound (TRUS)-guided prostate biopsy using three different anaesthetic applications and no anaesthesia. One-hundred men undergoing TRUS-guided prostate biopsy were prospectively enrolled in this study. Patients were randomized to four groups. Group 1 (n = 25) received no anaesthesia, group 2 (n = 25) was administered a perianal lidocaine injection, group 3 (n = 25) was administered a periprostatic lidocaine injection, and group 4 (n = 25) was administered a combination of perianal-intrarectal lidocaine-prilocaine cream. The anaesthetic application was given 5 min before the procedure. All patients were asked to indicate the level of pain experienced on a visual analogue scale (VAS) during three situations, including when the TRUS probe was inserted and 15 min and 2 weeks after biopsy. Mean pain scores were similar in all groups at the time of probe insertion. Mean pain scores showed statistically significant differences between group 1 and the other groups, except for group 4, 15 min after the procedure. Group 3 showed better pain control 15 min after biopsy and this difference was statistically significant (p < 0.043).The VAS scores were similar 2 weeks after the procedure in all groups. Anaesthetic application before TRUS-guided prostate biopsy may be advocated. The application of periprostatic lidocaine seems to be the most advantageous method for lowering the perception of pain.


Extended benefit from sequential administration of docetaxel after standard fluorouracil, epirubicin, and cyclophosphamide regimen for node-positive breast cancer: the 8-year follow-up results of the UNICANCER-PACS01 trial.
The initial report from the Programme Action Concertée Sein (PACS) PACS01 trial demonstrated a benefit at 5 years for disease-free survival (DFS) and overall survival (OS) rates with the sequential administration of docetaxel after FEC100 (fluorouracil 500 mg/m(2), epirubicin 100 mg/m(2), and cyclophosphamide 500 mg/m(2)) for patients with node-positive, operable breast cancer. We evaluate here the impact of this regimen at 8 years. Between June 1997 and March 2000, a total of 1,999 patients (age <65) with localized, resectable, non-pretreated, unilateral breast cancer were randomly assigned to receive either standard FEC100 for 6 cycles or 3 cycles of FEC100 followed by 3 cycles of 100 mg/m(2) docetaxel (FEC-D), both given every 21 days. Radiotherapy was mandatory after conservative surgery and tamoxifen was given for 5 years to hormone receptor (HR)-positive patients. Five-year DFS was the trial's main endpoint. Updated 8-year survival data are presented. With a median follow-up of 92.8 months, 639 patients experienced at least one event. A total number of 383 deaths were registered. Eight-year DFS rates were 65.8% with FEC alone and 70.2% with FEC-D. OS rates at 8 years were 78% with FEC alone and 83.2% with FEC-D. Cox regression analysis adjusted for age and number of positive nodes showed a 15% reduction in the relative risk of relapse and a 25% reduction in the relative risk of death in favor of FEC-D. Significant relative risk reductions were observed in the HR-positive, HER2-positive, and Ki67 ≥20% subpopulations. Benefits for DFS and OS rates with the sequential FEC-D regimen are fully confirmed at 8 years.


Stretchable/flexible silver nanowire Electrodes for energy device applications.
Research on sustainable and high-efficiency energy devices has recently emerged as an important global issue. These devices are now moving beyond the form of a bulk, rigid platform to a portable, flexible/stretchable format that is easily available in our daily lives. Similar to the development of an active layer for the production of next-generation energy devices, the fabrication of flexible/stretchable electrodes for the easy flow of electrons is also very important. Silver nanowire electrodes have high electronic conductivity even in a flexible/stretchable state due to their high aspect ratio and percolation network structures compared to conventional electrodes. Herein, we summarize the research in the field of flexible/stretchable electronics on energy devices fabricated using silver nanowires as the electrodes. Additionally, for a systematic presentation of the current research trends, this review classifies the surveyed research efforts into the categories of energy production, storage, and consumption.


Copper loading of hearts increases postischemic reperfusion injury.
We studied the role of copper as a potential mediator of postischemic reperfusion injury in the isolated, perfused rat heart. Hearts were equilibrated with Krebs-Henseleit buffer for 10 minutes and then loaded with copper by way of perfusion with buffer containing 20 microM copper(II)-bis-histidial for 30 minutes. Control hearts were perfused with Krebs-Henseleit buffer alone during the loading period. Hearts than were washed with buffer for 10 minutes and subjected to 20 minutes of normothermic global ischemia followed by 30 minutes of reperfusion. Atomic absorption spectroscopy revealed a 67% increase in total copper content in loaded hearts by the end of the wash. By the end of the 30-minute period of reperfusion, control hearts demonstrated a 50-60% recovery of myocardial function as determined by peak systolic pressure, contractility, and heart rate. In contrast, copper-loaded hearts exhibited virtually no functional recovery within the 30-minute time period. Using salicylate as a probe, we determined that peak and duration of .OH formation appears to be increased in copper-loaded hearts during reperfusion. Furthermore, efflux of lactic dehydrogenase was significantly increased in copper-loaded hearts. Our results clearly demonstrate that increasing cardiac content of copper results in enhanced postischemic reperfusion injury associated with increased formation of .OH, thus suggesting an important catalytic role for this transition metal.


Effects of testosterone treatment on bone mineral density in hypogonadal men receiving intrathecal opioids.
Opioid induced depression of sex hormones is a common finding in chronic pain patients receiving long-term opioids by oral, parenteral and even intrathecal routes of administration. The hypothalamic suppression by opioids leads to a hypogonadal state with low testosterone levels in males and subsequent low bone mineral density (BMD). We have studied the effects of intrathecally administered opioids on BMD in a group of male chronic pain patients. In addition, we have studied the effects of supplementary testosterone on bone metabolism to see if the adverse effects of intrathecal opioids can be reversed. Eleven of the 27 patients were on supplementary testosterone having previously been diagnosed as hypogonadal with low serum testosterone. Duration of testosterone supplementation was greater than 2 years in all 11 patients. Both serum total and free testosterone levels were higher in patients on supplementary testosterone than in patients who did not receive this treatment. Of the 16 patients not on testosterone supplement, 14 (87%) had low serum testosterone levels (<10 nmol/L) and 11 (69%) had low or osteoporotic T scores. Within this group, low free testosterone was associated with low BMD scores, and this persisted after correcting for age. Eight of the patients on testosterone supplement had normal BMDT scores and three (27%) had low or osteoporotic T scores. T and age-corrected BMDZ scores were significantly greater in the 11 patients on testosterone supplements than BMD scores in the other 16 patients. Testosterone supplementation was found to largely correct the effects of intrathecal opioids on testosterone levels and BMD.


Individualized causal discovery with latent trajectory embedded Bayesian networks.
Bayesian networks have been widely used to generate causal hypotheses from multivariate data. Despite their popularity, the vast majority of existing causal discovery approaches make the strong assumption of a (partially) homogeneous sampling scheme. However, such assumption can be seriously violated, causing significant biases when the underlying population is inherently heterogeneous. To this end, we propose a novel causal Bayesian network model, termed BN-LTE, that embeds heterogeneous samples onto a low-dimensional manifold and builds Bayesian networks conditional on the embedding. This new framework allows for more precise network inference by improving the estimation resolution from the population level to the observation level. Moreover, while causal Bayesian networks are in general not identifiable with purely observational, cross-sectional data due to Markov equivalence, with the blessing of causal effect heterogeneity, we prove that the proposed BN-LTE is uniquely identifiable under relatively mild assumptions. Through extensive experiments, we demonstrate the superior performance of BN-LTE in causal structure learning as well as inferring observation-specific gene regulatory networks from observational data.


"I Would Just Feel Really Relaxed and at Peace": Findings From a Pilot Prison Yoga Program in Australia.
International research provides support for yoga as a well-being intervention in prison. No systematic research has been undertaken in Australia to assess the effectiveness of prison yoga programs. In 2017, the authors, in partnership with Australian Capital Territory (ACT) Corrective Services and the Yoga Foundation, introduced a weekly pilot yoga program at the ACT prison. This article presents quantitative and qualitative findings from the program. Although the small sample size (n = 8) is acknowledged, our findings indicate that participants attained statistically and clinically significant benefit from the program, demonstrated by improvements in their levels of depression, anxiety, self-esteem, goal-direction, negative affect, and non-acceptance. They also reported improved flexibility, sleep and relaxation, pain reduction, and identified improvements in their mental well-being, commenting that the program made them feel "calm" and "at peace." The article concludes by advocating for the expansion of such programs in Australian prisons and further research on such programs.


Trans-ethnic fine mapping of a quantitative trait locus for circulating angiotensin I-converting enzyme (ACE).
Circulating angiotensin I-converting enzyme (ACE) levels are influenced by a major quantitative trait locus (QTL) that maps to the ACE gene. Phylogenetic and measured haplotype analyses have suggested that the ACE-linked QTL lies downstream of a putative ancestral breakpoint located near to position 6435. However, strong linkage disequilibrium between markers in the 3' portion of the gene has prevented further resolution of the QTL in Caucasian subjects. We have examined 10 ACE gene polymorphisms in Afro-Caribbean families recruited in JAMAICA: Variance components analyses showed strong evidence of linkage and association to circulating ACE levels. When the linkage results were contrasted with those from a set of British Caucasian families, there was no evidence for heterogeneity between the samples. However, patterns of allelic association between the markers and circulating ACE levels differed significantly in the two data sets. In the British families, three markers [G2215A, Alu insertion/deletion and G2350A] were in complete disequilibrium with the ACE-linked QTL. In the Jamaican families, only marker G2350A showed strong but incomplete disequilibrium with the ACE-linked QTL. These results suggest that additional unobserved polymorphisms have an effect on circulating ACE levels in Jamaican families. Furthermore, our results show that a variance components approach combined with structured, quantitative comparisons between families from different ethnic groups may be a useful strategy for helping to determine which, if any, variants in a small genomic region directly influence a quantitative trait.


Caffeine Mitigates Lung Inflammation Induced by Ischemia-Reperfusion of Lower Limbs in Rats.
Reperfusion of ischemic limbs can induce inflammation and subsequently cause acute lung injury. Caffeine, a widely used psychostimulant, possesses potent anti-inflammatory capacity. We elucidated whether caffeine can mitigate lung inflammation caused by ischemia-reperfusion (IR) of the lower limbs. Adult male Sprague-Dawley rats were randomly allocated to receive IR, IR plus caffeine (IR + Caf group), sham-operation (Sham), or sham plus caffeine (n = 12 in each group). To induce IR, lower limbs were bilaterally tied by rubber bands high around each thigh for 3 hours followed by reperfusion for 3 hours. Caffeine (50 mg/kg, intraperitoneal injection) was administered immediately after reperfusion. Our histological assay data revealed characteristics of severe lung inflammation in the IR group and mild to moderate characteristic of lung inflammation in the IR + Caf group. Total cells number and protein concentration in bronchoalveolar lavage fluid of the IR group were significantly higher than those of the IR + Caf group (P < 0.001 and P = 0.008, resp.). Similarly, pulmonary concentrations of inflammatory mediators (tumor necrosis factor-α, interleukin-1β, and macrophage inflammatory protein-2) and pulmonary myeloperoxidase activity of the IR group were significantly higher than those of the IR + Caf group (all P < 0.05). These data clearly demonstrate that caffeine could mitigate lung inflammation induced by ischemia-reperfusion of the lower limbs.


Clustered p53 immunostaining: a novel pattern associated with prostate cancer progression.
Abnormal p53 protein accumulation is typically defined as present when greater than 5 or 10% of cancer cells stain positively. We present a novel approach whereby immunopositivity is defined when 15 or more cells within a 300 x 400-micrometer(2) field exhibit p53 protein accumulation; a feature that we have called "clustered" staining. We assessed p53 immunostaining of moderately differentiated, clinically localized prostate cancers derived from two patient groups: those without cancer recurrence 5 years after radical prostatectomy, and those in whom cancer had recurred following radical prostatectomy. Clustered p53 immunopositivity was present in 10 (63%) of 16 patients in the recurrent group and in only 7 (21%) of 33 in the nonrecurrent group. Clustered p53 staining was clearly associated with cancer recurrence (P < 0.01). This refinement of a commonly used assay may help define the biological aggressiveness of a cancer.


Genome variation and population structure among 1142 mosquitoes of the African malaria vector species Anopheles gambiae and Anopheles coluzzii.
Mosquito control remains a central pillar of efforts to reduce malaria burden in sub-Saharan Africa. However, insecticide resistance is entrenched in malaria vector populations, and countries with a high malaria burden face a daunting challenge to sustain malaria control with a limited set of surveillance and intervention tools. Here we report on the second phase of a project to build an open resource of high-quality data on genome variation among natural populations of the major African malaria vector species Anopheles gambiae and Anopheles coluzzii We analyzed whole genomes of 1142 individual mosquitoes sampled from the wild in 13 African countries, as well as a further 234 individuals comprising parents and progeny of 11 laboratory crosses. The data resource includes high-confidence single-nucleotide polymorphism (SNP) calls at 57 million variable sites, genome-wide copy number variation (CNV) calls, and haplotypes phased at biallelic SNPs. We use these data to analyze genetic population structure and characterize genetic diversity within and between populations. We illustrate the utility of these data by investigating species differences in isolation by distance, genetic variation within proposed gene drive target sequences, and patterns of resistance to pyrethroid insecticides. This data resource provides a foundation for developing new operational systems for molecular surveillance and for accelerating research and development of new vector control tools. It also provides a unique resource for the study of population genomics and evolutionary biology in eukaryotic species with high levels of genetic diversity under strong anthropogenic evolutionary pressures.


Plasma ghrelin levels after diet-induced weight loss or gastric bypass surgery.
Weight loss causes changes in appetite and energy expenditure that promote weight regain. Ghrelin is a hormone that increases food intake in rodents and humans. If circulating ghrelin participates in the adaptive response to weight loss, its levels should rise with dieting. Because ghrelin is produced primarily by the stomach, weight loss after gastric bypass surgery may be accompanied by impaired ghrelin secretion. We determined the 24-hour plasma ghrelin profiles, body composition, insulin levels, leptin levels, and insulin sensitivity in 13 obese subjects before and after a six-month dietary program for weight loss. The 24-hour ghrelin profiles were also determined in 5 subjects who had lost weight after gastric bypass and 10 normal-weight controls; 5 of the 13 obese subjects who participated in the dietary program were matched to the subjects in the gastric-bypass group and served as obese controls. Plasma ghrelin levels rose sharply shortly before and fell shortly after every meal. A diet-induced weight loss of 17 percent of initial body weight was associated with a 24 percent increase in the area under the curve for the 24-hour ghrelin profile (P=0.006). In contrast, despite a 36 percent weight loss after gastric bypass, the area under the curve for the ghrelin profile in the gastric-bypass group was 77 percent lower than in normal-weight controls (P<0.001) and 72 percent lower than in matched obese controls (P=0.01). The normal, meal-related fluctuations and diurnal rhythm of the ghrelin level were absent after gastric bypass. The increase in the plasma ghrelin level with diet-induced weight loss is consistent with the hypothesis that ghrelin has a role in the long-term regulation of body weight. Gastric bypass is associated with markedly suppressed ghrelin levels, possibly contributing to the weight-reducing effect of the procedure.