Sleep and rest regulation in young and old oestrogen-deficient female mice.
The effect of circulating oestrogen deficiency on sleep regulation and locomotor activity was investigated in aromatase cytochrome P450 deficient mice (ArKO) and wild-type (WT) controls. Sleep was recorded in 3-month old mice during a 24-h baseline day, 6-h sleep deprivation (SD) and 18-h recovery, and activity was recorded at the age of 3, 9 and 12 months. In mice deficient of oestrogen, the total amount of sleep per 24 h was the same as in WT controls. However, in ArKO mice, sleep was enhanced in the dark period at the expense of sleep in the light phase, and was more fragmented than sleep in WT mice. This redistribution of sleep resulted in a damped amplitude of slow-wave activity (SWA; power between 0.75-4.0 Hz) in non-rapid eye movement sleep across 24 h. After SD, the rebound of sleep and SWA was similar between the genotypes, suggesting that oestrogen deficiency does not affect the mechanisms maintaining the homeostatic balance between the amount of sleep and its intensity. Motor activity decreased with age in both genotypes and was lower in ArKO mice compared to WT at all three ages. After SD, the amount of rest in 3-month old WT mice increased above baseline and was more consolidated. Both effects were less pronounced in ArKO mice, reflecting the baseline differences between the genotypes. The results indicate that despite the pronounced redistribution of sleep and motor activity in oestrogen deficient mice, the basic homeostatic mechanisms of sleep regulation in ArKO mice remain intact.


Further development of YBOCS dimensions in the OCD Collaborative Genetics study: symptoms vs. categories.
Despite progress in identifying homogeneous subphenotypes of obsessive-compulsive disorder (OCD) through factor analysis of the Yale Brown Obsessive-Compulsive Scale Symptom Checklist (YBOCS-SC), prior solutions have been limited by a reliance on presupposed symptom categories rather than discrete symptoms. Furthermore, there have been few attempts to evaluate the familiality of OCD symptom dimensions. The purpose of this study was to extend prior work by this collaborative group in category-based dimensions by conducting the first-ever exploratory dichotomous factor analysis using individual OCD symptoms, comparing these results to a refined category-level solution, and testing the familiality of derived factors. Participants were 485 adults in the six-site OCD Collaborative Genetics Study, diagnosed with lifetime OCD using semi-structured interviews. YBOCS-SC data were factor analyzed at both the individual item and symptom category levels. Factor score intraclass correlations were calculated using a subsample of 145 independent affected sib pairs. The item- and category-level factor analyses yielded nearly identical 5-factor solutions. While significant sib-sib associations were found for four of the five factors, Hoarding and Taboo Thoughts were the most robustly familial (r ICC>or=0.2). This report presents considerable converging evidence for a five-factor structural model of OCD symptoms, including separate factor analyses employing individual symptoms and symptom categories, as well as sibling concordance. The results support investigation of this multidimensional model in OCD genetic linkage studies.


Green tea extract decreases starch digestion and absorption from a test meal in humans: a randomized, placebo-controlled crossover study.
Green tea is known worldwide for its beneficial effects on human health. However, objective data evaluating this influence in humans is scarce. The aim of the study was to assess the impact of green tea extract (GTE) on starch digestion and absorption. The study comprised of 28 healthy volunteers, aged 19 to 28 years. In all subjects, a starch (13)C breath test was performed twice. Subjects randomly ingested naturally (13)C-abundant cornflakes during the GTE test (GTE 4 g) or placebo test. The cumulative percentage dose recovery (CPDR) was significantly lower for the GTE test than for the placebo test (median [interquartile range]: 11.4% [5.5-15.5] vs. 16.1% [12.7-19.5]; p = 0.003). Likewise, CPDR expressed per hour was considerably lower in each point of the measurement. In conclusion, a single dose of green tea extract taken with a test meal decreases starch digestion and absorption.


Reconstructing an ancestral mammalian immune supercomplex from a marsupial major histocompatibility complex.
The first sequenced marsupial genome promises to reveal unparalleled insights into mammalian evolution. We have used the Monodelphis domestica (gray short-tailed opossum) sequence to construct the first map of a marsupial major histocompatibility complex (MHC). The MHC is the most gene-dense region of the mammalian genome and is critical to immunity and reproductive success. The marsupial MHC bridges the phylogenetic gap between the complex MHC of eutherian mammals and the minimal essential MHC of birds. Here we show that the opossum MHC is gene dense and complex, as in humans, but shares more organizational features with non-mammals. The Class I genes have amplified within the Class II region, resulting in a unique Class I/II region. We present a model of the organization of the MHC in ancestral mammals and its elaboration during mammalian evolution. The opossum genome, together with other extant genomes, reveals the existence of an ancestral "immune supercomplex" that contained genes of both types of natural killer receptors together with antigen processing genes and MHC genes.


Patent Law's Reproducibility Paradox.
Clinical research faces a reproducibility crisis. Many recent clinical and preclinical studies appear to be irreproducible--their results cannot be verified by outside researchers. This is problematic for not only scientific reasons but also legal ones: patents grounded in irreproducible research appear to fail their constitutional bargain of property rights in exchange for working disclosures of inventions. The culprit is likely patent law's doctrine of enablement. Although the doctrine requires patents to enable others to make and use their claimed inventions, current difficulties in applying the doctrine hamper or even actively dissuade reproducible data in patents. This Article assesses the difficulties in reconciling these basic goals of scientific research and patent law. More concretely, it provides several examples of irreproducibility in patents on blockbuster drugs--Prempro, Xigris, Plavix, and Avastin--and discusses some of the social costs of the misalignment between good clinical practice and patent doctrine. Ultimately, this analysis illuminates several current debates concerning innovation policy. It strongly suggests that a proper conception of enablement should take into account after-arising evidence. It also sheds light on the true purpose--and limits--of patent disclosure. And lastly, it untangles the doctrines of enablement and utility.


Role of conserved histidine residues in the low-pH dependence of the Semliki Forest virus fusion protein.
A wide variety of enveloped viruses infects cells by taking advantage of the low pH in the endocytic pathway to trigger virus-membrane fusion. For alphaviruses such as Semliki Forest virus (SFV), acidic pH initiates a series of conformational changes in the heterodimeric virus envelope proteins E1 and E2. Low pH dissociates the E2/E1 dimer, releasing the membrane fusion protein E1. E1 inserts into the target membrane and refolds to a trimeric hairpin conformation, thus driving the fusion reaction. The means by which E1 senses and responds to low pH is unclear, and protonation of conserved E1 histidine residues has been proposed as a possible mechanism. We tested the role of four conserved histidines by mutagenesis of the wild-type (wt) SFV infectious clone to create virus mutants with E1 H3A, H125A, H331A, and H331A/H333A mutations. The H125A, H331A, and H331A/H333A mutants had growth properties similar to those of wt SFV and showed modest change or no change in the pH dependence of virus-membrane fusion. By contrast, the E1 H3A mutation produced impaired virus growth and a markedly more acidic pH requirement for virus-membrane fusion. The dissociation of the H3A heterodimer and the membrane insertion of the mutant E1 protein were comparable to those of the wt in efficiency and pH dependence. However, the formation of the H3A homotrimer required a much lower pH and showed reduced efficiency. Together, these results and the location of H3 suggest that this residue acts to regulate the low-pH-dependent refolding of E1 during membrane fusion.


Effect of nutritional supplement formula on catch-up growth in young children with nonorganic faltering growth: a prospective multicenter study.
Inadequate nutrition in infants and young children affects physical growth and neurocognitive development. Therefore, early nutritional intervention is important to promote catch-up growth in young children with faltering growth. The aim of this study was to evaluate the effect of nutritional supplementation with a pediatric concentrated and balanced nutritional supplement formula on promoting growth and improving nutritional status in children with nonorganic faltering growth. Children aged 12-36 months whose body weight-for-age was < 5th percentile on the Korean Growth Charts were enrolled. Children born premature or having organic diseases were excluded. Children were instructed to consume 400 mL of formula per day in addition to their regular diet for 6 months. Pediatricians and dietitians educated the parents and examined the subjects every 2 months. Anthropometric parameters were measured at baseline and at 2, 4, and 6 months, and laboratory tests were done at baseline and 6 months. The good consumption group included children who consumed ≥ 60% of the recommended dose of formula. Total 82 children completed the 6-month intervention. At baseline, there were no significant differences in all variables between the good consumption and poor consumption groups. Weight and weight z-scores were significantly improved in the good consumption group compared to the poor consumption group at the end of the intervention (P = 0.009, respectively). The good consumption group showed a significant trend for gaining weight (P < 0.05) and weight z-score (P < 0.05) compared to the poor consumption group during 6 months of formula intake. The concentration of blood urea nitrogen was significantly increased in the good consumption group (P = 0.001). Nutritional supplementation with a concentrated and balanced pediatric nutritional formula along with dietary education might be an effective approach to promote catch-up growth in children with nonorganic faltering growth.


Polyphenolic Composition and Antioxidant Activity (ORAC, EPR and Cellular) of Different Extracts of Argylia radiata Vitroplants and Natural Roots.
Plant biochemistry studies have increased in recent years due to their potential to improve human health. Argylia radiata is an extremophile plant with an interesting polyphenolic profile. However, its biomass is scarce and occasionally available. Argylia in vitro biomass was obtained from tissue culture and compared with in vivo roots regarding its polyphenolic and flavonoid content. Different solvents were used to prepare extracts from the in vitro tissue of callus and aerial plant organs and in vivo roots. UPLC-MS/MS was used to assess the chemical composition of each extract. ORAC-FL and scavenging of free radicals (DPPH and OH) methods were used to determine the antioxidant capacity of extracts. Furthermore, the biological activity of the extracts was established using the cellular antioxidant activity method. The vitroplants were a good source of polyphenols (25-68 mg GAE/100 g tissue FW), and methanol was the most efficient solvent. Eight polyphenolic compounds were identified, and their antioxidant properties were investigated by different chemical methods with EPR demonstrating its specific scavenging activity against free radicals. All extracts showed cellular dose-dependent antioxidant activity. The methanolic extract of vitroplants showed the highest cellular antioxidant activity (44.6% and 51%) at 1 and 10 µg/mL of extract, respectively. Vitroplants of A. radiata are proposed as a biotechnological product as a source of antioxidant compounds with multiple applications.


Nitrogen, phosphorus, and eutrophication in the coastal marine environment.
The distribution of inorganic nitrogen and phosphorus and bioassay experiments both show that nitrogen is the critical limiting factor to algal growth and eutrophication in coastal marine waters. About twice the amount of phosphate as can be used by the algae is normally present. This surplus results from the low nitrogen to phosphorus ratio in terrigenous contributions, including human waste, and from the fact that phosphorus regenerates more quickly than ammonia from decomposing organic matter. Removal of phosphate from detergents is therefore not likely to slow the eutrophication of coastal marine waters, and its replacement with nitrogen-containing nitrilotriacetic acid may worsen the situation.


Electro-otolithography: new insight into benign paroxysmal positioning vertigo.
Early vestibular evoked potentials were recorded with an extratympanic electrode. The recording principle was adding responses that were phase-locked to a recording frequency. The recording frequency was empirically determined to match harmonically an individual response frequency, thus allowing averaging. This new technique was evaluated in benign paroxysmal positioning vertigo. Normal data were obtained from 12 patients without vestibulocochlear symptoms and were compared with data from 18 symptomatic patients with diagnoses of benign paroxysmal positioning vertigo. All symptomatic patients were treated with a canalith repositioning maneuver, and all responded, in that they no longer had attacks of vertigo. Of the five patients who could be reassessed, all five no longer had attacks of vertigo, but three complained of persisting postural imbalance. Repeat electro-otolithography results continued to be abnormal in these patients, whereas in the remaining two patients responses were normal, consistent with the treatment outcome. The results suggest that electro-otolithography is a valuable addition to the otoneurological test battery. Successful canalith repositioning can abolish attacks of vertigo, although not necessarily a persisting imbalance, which patients frequently describe as a temporary and momentary instability. This is most likely related to a remaining otolithic deficit.


Reflection of medical error highlighted on media in Turkey: A retrospective study.
This study was performed with the aim of identifying how news on medical errors have be transmitted, and how the types, reasons, and conclusions of medical errors have been reflected to by the media in Turkey. A content analysis method was used in the study, and in this context, the data for the study was acquired by scanning five newspapers with the top editions on the national basis between the years 2012 and 2015 for the news about medical errors. Some specific selection criteria was used for the scanning of resulted news, and 116 news items acquired as a result of all the eliminations. According to the results of the study; the vast majority of medical errors (40.5%) transmitted by the news resulted from the negligence of the medical staff. The medical errors were caused by physicians in the ratio of 74.1%, they most commonly occurred in state hospitals (31.9%). Another important result of the research was that medical errors resulted in either patient death to a large extent (51.7%), or permanent damage and disability to patients (25.0%). The news concerning medical errors provided information about the types, causes, and the results of these medical errors. It also reflected the media point of view on the issue. The examination of the content of the medical errors reported by the media were important which calls for appropriate interventions to avoid and minimize the occurrence of medical errors by improving the healthcare delivery system.


FIrpic: archetypal blue phosphorescent emitter for electroluminescence.
FIrpic is the most investigated bis-cyclometallated iridium complex in particular in the context of organic light emitting diodes (OLEDs) because of its attractive sky-blue emission, high emission efficiency, and suitable energy levels. In this Perspective we review the synthesis, structural characterisations, and key properties of this emitter. We also survey the theoretical studies and summarise a series of selected monochromatic electroluminescent devices using FIrpic as the emitting dopant. Finally we highlight important shortcomings of FIrpic as an emitter for OLEDs. Despite the large body of work dedicated to this material, it is manifest that the understanding of photophysical and electrochemical processes are only broadly understood mainly because of the different environment in which these properties are measured, i.e., isolated molecules in solvent vs. device.


Anatomical basis of pedicles in breast reduction.
The mammary gland is composed of multiple tubules acinar pockets in which the secretory layer, connective tissue stroma, and fatty tissue all respond to hormonal and systemic influences. These structures are irrigated by three vascular pedicle branches, from the axillary artery, internal mammary artery, and intercostal artery. This vascular anastomotic arrangement forms the anatomical basis of the flaps used in breast reduction techniques. The veins form a strong subdermal network, latticed with the arterial network. The lymph vessels have three well-defined pedicles, skin, glandular and milk ducts that drain into internal, external, and posterior ducts. The understanding of these anatomical structures determines the selection of different pedicles in breast volume reduction and preservation of the nipple-areola complex circulation.


Validation of a Chinese version of the Quality of Well-Being Scale-Self-Administered (QWB-SA) in patients with epilepsy.
Generic preference-based health-related quality of life (HRQoL) instruments are increasingly used to estimate the quality-adjusted life years (QALYs) in cost-effectiveness/utility studies. However, no such tool has been used and validated in epilepsy patients in China. This study was conducted to validate a generic preference-based HRQoL instrument, namely the Quality of Well-Being Scale-Self-Administered (QWB-SA) in Chinese patients with epilepsy. Accepted translation procedures were followed to develop the Chinese QWB-SA. An epilepsy group (adults with established diagnosis of epilepsy) and a control group (adults without manifested cognitive problems) were recruited between July and October, 2012, from two tertiary hospitals in China. After giving informed consent, each subject completed both the QWB-SA and the EuroQol (EQ-5D) as well as provided sociodemographic data. Construct validity was examined by six (convergent) and two (discriminative) a priori hypotheses. Sensitivity was compared by ability to differentiate epilepsy-specific variable-based subgroups. Agreement between the QWB-SA and EQ-5D was assessed by intraclass correlation coefficient (ICC) and Bland-Altman plot. One hundred forty-four epilepsy patients and 323 control subjects were enrolled, respectively. The utility medians (interquartile range, IQR) for the QWB-SA and EQ-5D were 0.673 (0.172), 0.848 (0.275) for epilepsy group and 0.775 (0.258), 1.000 (0.152) for control group, respectively. The difference in utilities between the two measures were significant (p < 0.0001). Construct validity was demonstrated by six a priori hypotheses. In addition, the QWB-SA was able to discriminate across different seizure frequency and antiepileptic drug (AED) treatment subgroups. Agreement between the QWB-SA and EQ-5D was demonstrated by ICC (0.725). Finally, the multiple linear regression analysis indicated that group and the EQ-VAS had influences on the utility difference of these two measures, whereas seizure frequency and number of AEDs were predictors of HRQoL as measured by the QWB-SA. The QWB-SA is a valid and sensitive HRQoL measure in Chinese patients with epilepsy. Compared to the EQ-5D, the QWB-SA showed superiority in coverage of health dimensions, sensitivity, and ceiling effects. However, future study is still needed to ascertain its responsiveness.


A comparative study of the effects of certain halogenated benzimidazole ribosides on RNA synthesis, cell proliferation, and interferon production.
5-(or 6-)Bromo-4,5-(or 5,7-)dichloro-1-beta-D-ribofuranosylbenzimidazole, 5,6-dibromo-1-beta-D-ribofuranosylbenzimidazole, and 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole show closely similar structure-activity relationships with respect to inhibition of cellular RNA synthesis, cellular proliferation, and influenza virus multiplication, and also with respect to enhancement of interferon production. The activities ofth ese compounds are ranked 20:2.5:1. The log dose-response curves constructed for inhibiton of FS-4 cell RNA synthesis show similar slopes and a leveling off at 60-70% inhibition of RNA synthesis at the highest concentrations of each compound tested. This evidence suggests that these three derivatives act through the same mechanism. It has been shown previously that the dichloro compound selectively inhibits nuclear heterogenous RNA and messenger RNA synthesis. The concentrations of the benzimidazole ribosides at which the rate of proliferation of human fibroblasts (FS-4) is reduced by 50% are as follows: monobromodichloro: 1.7 muM (0.68 mug/ml); dibromo: 12 muM (4.9 mug/ml); dichloro: 38 muM (12 mug/ml). All compounds reduce the exponential rate of cell proliferation in a dose-dependent manner. The inhibition of cell growth is reversible upon removal of the compounds from the medium. Protocols based on any one of the three halobenzimidazole ribosides give interferon yields from poly(I)-poly(C)-induced FS-4 cells which are comparable to the high yields obtained with the conventional cycloheximide-actinomycin D protocol. The enhancement of interferon yield depends on blocking of the synthesis of RNA which is involved in the shutoff of interferon production.


Improved power by use of a weighted score test for linkage disequilibrium mapping.
Association studies offer an exciting approach to finding underlying genetic variants of complex human diseases. However, identification of genetic variants still includes difficult challenges, and it is important to develop powerful new statistical methods. Currently, association methods may depend on single-locus analysis--that is, analysis of the association of one locus, which is typically a single-nucleotide polymorphism (SNP), at a time--or on multilocus analysis, in which multiple SNPs are used to allow extraction of maximum information about linkage disequilibrium (LD). It has been shown that single-locus analysis may have low power because a single SNP often has limited LD information. Multilocus analysis, which is more informative, can be performed on the basis of either haplotypes or genotypes. It may lose power because of the often large number of degrees of freedom involved. The ideal method must make full use of important information from multiple loci but avoid increasing the degrees of freedom. Therefore, we propose a method to capture information from multiple SNPs but with the use of fewer degrees of freedom. When a set of SNPs in a block are correlated because of LD, we might expect that the genotype variation among the different phenotypic groups would extend across all the SNPs, and this information could be compressed into the low-frequency components of a Fourier transform. Therefore, we develop a test based on weighted Fourier transformation coefficients, with more weight given to the low-frequency components. Our simulation results demonstrate the validity and substantially higher power of the proposed method compared with other common methods. This method provides an additional tool to existing methods for identification of causative genetic variants underlying complex diseases.


Growth kinetics of a diesel-degrading bacterial strain from petroleum-contaminated soil.
A diesel-degrading bacterium was isolated from a diesel-contaminated site in Selangor, Malaysia. The isolate was tentatively identified as Acinetobacter sp. strain DRY12 based on partial 16S rDNA molecular phylogeny and Biolog GN microplate panels and Microlog database. Optimum growth occurred from 3 to 5% diesel and the strain was able to tolerate as high as 8% diesel. The optimal pH that supported growth of the bacterium was between pH 7.5 to 8.0. The isolate exhibited optimal growth in between 30 and 35 degrees C. The best nitrogen source was potassium nitrate (between 0.6 and 0.9% (w/v)) followed by ammonium chloride, sodium nitrite and ammonium sulphate in descending order. An almost complete removal of diesel components was seen from the reduction in hydrocarbon peaks observed using Solid Phase Microextraction Gas Chromatography analysis after 10 days of incubation. The best growth kinetic model to fit experimental data was the Haldane model of substrate inhibiting growth with a correlation coefficient value of 0.97. The maximum growth rate- micromax was 0.039 hr(-1) while the saturation constant or half velocity constant Ks and inhibition constant Ki, were 0.387% and 4.46%, respectively. MATH assays showed that 75% of the bacterium was found in the hexadecane phase indicating that the bacterium was hydrophobic. The characteristics of this bacterium make it useful for bioremediation works in the Tropics.


Biological indicators of exposure to total and respirable aluminium dust fractions in a primary aluminium smelter.
The study attempts to define biological indicators of aluminium uptake and excretion in workers exposed to airborne aluminium compounds in a primary aluminium smelter. Also, this study defines the total and respirable aluminium dust fractions in two different potrooms, and correlates their concentrations with biological indicators in this group of workers. Air was sampled at defined work sites. Non-destructive and conventional techniques were used to find total and respirable aluminium content of the dust. Blood and urine was collected from 84 volunteers employed at various work stations throughout the smelter and from two different cohorts of controls matched for sex, age, and socioeconomic status. Aluminium in serum samples and urine specimens was measured by flameless atomic absorption with a PE 4100 ZL spectrometer. The correlation of aluminium concentrations in serum and urine samples with the degree of exposure was assessed for three arbitrary exposure categories; low (0.036 mg Al/m3), medium (0.35 mg Al/m3) and high (1.47 mg Al/m3) as found in different areas of the smelter. At medium and high exposure, the ratio of respirable to total aluminium in the dust samples varied significantly. At high exposure, serum aluminium, although significantly raised, was still within the normal range of an unexposed population. The workers with low exposure excreted aluminium in urine at levels significantly higher than the controls, but still within the normal range of the population. However, potroom workers with medium and high exposure had significantly higher urinary aluminium than the normal range. It is concluded that only urinary aluminium constitutes a practical index of occupational exposure at or above 0.35 mg Al/m3, and that the respirable fraction of the dust may play a major role in the biological response to exposure to aluminium in a smelter environment.


Single image superresolution based on gradient profile sharpness.
Single image superresolution is a classic and active image processing problem, which aims to generate a high-resolution (HR) image from a low-resolution input image. Due to the severely under-determined nature of this problem, an effective image prior is necessary to make the problem solvable, and to improve the quality of generated images. In this paper, a novel image superresolution algorithm is proposed based on gradient profile sharpness (GPS). GPS is an edge sharpness metric, which is extracted from two gradient description models, i.e., a triangle model and a Gaussian mixture model for the description of different kinds of gradient profiles. Then, the transformation relationship of GPSs in different image resolutions is studied statistically, and the parameter of the relationship is estimated automatically. Based on the estimated GPS transformation relationship, two gradient profile transformation models are proposed for two profile description models, which can keep profile shape and profile gradient magnitude sum consistent during profile transformation. Finally, the target gradient field of HR image is generated from the transformed gradient profiles, which is added as the image prior in HR image reconstruction model. Extensive experiments are conducted to evaluate the proposed algorithm in subjective visual effect, objective quality, and computation time. The experimental results demonstrate that the proposed approach can generate superior HR images with better visual quality, lower reconstruction error, and acceptable computation efficiency as compared with state-of-the-art works.


Spatially resolved thickness determination of rough thin films by reflectometry with polychromatic light.
This article introduces a simple metrology set-up based on white light interference. The aim is to determine the thickness of thin transparent films by their reflection color under white illumination. Therefore, one digital color image of a large field of view is taken in nontelecentric geometry. To obtain reflected light at all angles of view without having to move the camera the substrate has to be sufficiently rough. Additionally, interference occurs at the thin transparent film covering the substrate surface from which a two-dimensional lateral thickness distribution can be calculated. This method has been used as industrial inline process control for a CdS semiconductor layer on top of a rough Cu(In,Ga)(Se,S)2 thin film solar absorber and yielded 74 nm film thickness with only a 1 nm standard deviation. Options to further reduce the standard deviation with different illuminants and other cameras are proposed.


Quantitative effect of natural killer-cell licensing on hepatocellular carcinoma recurrence after curative hepatectomy.
Natural killer (NK) cells have a potential role in immune surveillance of hepatocellular carcinoma (HCC). Self-recognition of human leukocyte antigens (HLA) through killer immunoglobulin-like receptors (KIR) confers competence to NK cells-a process termed "licensing." We investigated the effect of NK-cell licensing on the susceptibility of patients to HCC recurrence. A total of 170 Japanese patients with HCC who underwent primary curative hepatectomy between 1996 and 2010 were enrolled in this study. The median follow-up period was 5.4 years. We analyzed their KIR-HLA genotypes with sequence-specific polymorphism-based typing and estimated their susceptibility to HCC recurrence by performing propensity score-matching analyses. The presence of KIR2DL1-C2, KIR2DL2-C1, KIR3DL1-BW4, or KIR3DL2-A3/11, functional compound genotypes that intrinsically license NK cells, did not markedly affect HCC recurrence. However, the multiplicity of those compound KIR-HLA genotypes was significantly associated with the HCC recurrence rate, i.e., the cumulative risk of recurrence in patients with at least three compound genotypes was significantly lower than that in patients with one or two compound genotypes, suggesting that the effect of NK-cell licensing on HCC recurrence is quantitative. Patients at high risk of HCC recurrence after curative hepatectomy could be identified by KIR-HLA genotyping.


Whole-body exposure to 2.45 GHz electromagnetic fields does not alter radial-maze performance in rats.
Mobile communication is based on utilization of electromagnetic fields (EMFs) in the frequency range of 0.3-300 GHz. Human and animal studies suggest that EMFs, which are in the 0.1 MHz-300 GHz range, might interfere with cognitive processes. In 1994, a report by Lai et al. [Bioelectromagnetics 15 (1994) 95-104] showed that whole-body exposure of rats to pulsed 2.45 GHz microwaves (2 micros pulse width, 500 pps, and specific absorption rate [SAR] 0.6 W/kg) for 45 min resulted in altered spatial working memory assessed in a 12-arm radial-maze task. Surprisingly, there has been only one attempt to replicate this experiment so far [Bioelectromagnetics 25 (2004) 49-57]; confirmation of the Lai et al. experiment failed. In the present study, rats were tested in a 12-arm radial-maze subsequently to a daily exposure to 2.45 GHz microwaves (2 micros pulse width, 500 pps, and SAR 0.6 W/kg) for 45 min. The performance of exposed rats was comparable to that found in sham-exposed or in naive rats (no contact with the exposure system). Regarding the methodological details provided by Lai et al. on their testing protocol, our results might suggest that the microwave-induced behavioral alterations measured by these authors might have had more to do with factors liable to performance bias than with spatial working memory per se.


Glass window coatings for sunlight heat reflection and co-utilization.
Buildings that simultaneously provide natural illumination and thermal comfort for all seasons have met with increasing demand as conventional resource limitations are realized. In this context, organic and metal-dielectric coatings are tested, and a simple, coated double-glazed window with solar blinds is conceived that includes passive infrared (IR) reflection, active illumination control, and integration to the building envelope. As a result, a proper spectrally selective coating is applied to produce a low-emissivity solar window with climate-adaptive co-utilization of the reflected IR.


The murine stanniocalcin 1 gene is not essential for growth and development.
The stanniocalcin 1 (STC1) gene is expressed in a wide variety of tissues, including the kidney, prostate, thyroid, bone, and ovary. STC1 protein is considered to have roles in many physiological processes, including bone development, reproduction, wound healing, angiogenesis, and modulation of inflammatory response. In fish, STC1 is a hormone that is secreted by the corpuscles of Stannius and is involved in calcium and phosphate homeostasis. To determine the role of STC1 in mammals, we generated Stc1-null mice by gene targeting. The number of Stc1-/- mice obtained was in accordance with Mendelian ratios, and both males and females produced offspring normally. No anatomical or histological abnormalities were detected in any tissues. Our results demonstrated that Stc1 function is not essential for growth or reproduction in the mouse.


A rewiring model of intratumoral interaction networks.
Intratumoral heterogeneity (ITH) has been regarded as a key cause of the failure and resistance of cancer therapy, but how it behaves and functions remains unclear. Advances in single-cell analysis have facilitated the collection of a massive amount of data about genetic and molecular states of individual cancer cells, providing a fuel to dissect the mechanistic organization of ITH at the molecular, metabolic and positional level. Taking advantage of these data, we propose a computational model to rewire up a topological network of cell-cell interdependences and interactions that operate within a tumor mass. The model is grounded on the premise of game theory that each interactive cell (player) strives to maximize its fitness by pursuing a "rational self-interest" strategy, war or peace, in a way that senses and alters other cells to respond properly. By integrating this idea with genome-wide association studies for intratumoral cells, the model is equipped with a capacity to visualize, annotate and quantify how somatic mutations mediate ITH and the network of intratumoral interactions. Taken together, the model provides a topological flow by which cancer cells within a tumor cooperate or compete with each other to downstream pathogenesis. This topological flow can be potentially used as a blueprint for genetically intervening the pattern and strength of cell-cell interactions towards cancer control.


[Genetically modified tobacco--chance or threat for smokers?].
Tobacco (Nicotiana sp.) is of the most often used model plants for research in the field of physiology, biochemistry, molecular biology and genetic engineering. Tobacco was also the first plant subject to genetic transformation--in 1983 the plant was introduced with the kanamycin resistance gene. The internet database PubMed includes roughly 3,500 records concerning transgenic tobacco, out of which nearly 200 works were published this year. Advancement of state-of-the-art techniques of genetic engineering offers new, as yet unprecedented opportunities to take advantage of tobacco. Apparently tobacco is not only a source of carcinogenic tobacco smoke, but also a plant which due to genetic modifications may be used in a positive way in medicine and biological sciences. The aim of the article is to summarize the role of genetically modified Nicotiana genus plants in modern medicine and environmental protection. The publication describes the usage of transgenic plants in the tobacco industry and evaluates possible effects of such applications. Moreover, potential possibilities of deployment of genetically modified tobacco for production of anti-virus and antibacterial vaccinations, interferon, antibodies, etc. have been characterized.


Strategies for switching from conventional antipsychotic drugs or risperidone to olanzapine.
This study compared the efficacy and safety of 4 therapeutically relevant strategies for switching clinically stable patients from a conventional antipsychotic drug or risperidone to olanzapine. Two hundred nine outpatients with a DSM-IV diagnosis of schizophrenia or schizo-affective disorder who were clinically stable while being treated with a conventional antipsychotic drug or risperidone were openly randomly assigned to either abrupt or gradual discontinuation of their prior antipsychotic drug. Patients were further randomly assigned in a double-blind fashion to immediate olanzapine initiation (olanzapine, 10 mg q.d. for 3 weeks) or stepwise initiation (a sequence of 1 week each on placebo; olanzapine, 5 mg q.d.; and olanzapine, 10 mg q.d.). The efficacy of these 4 switching paradigms was assessed using the Clinical Global Impressions (CGI)-Improvement scale, Patient's Global Impressions (PGI)-Improvement scale, and Positive and Negative Syndrome Scale (PANSS). Safety assessments included ratings for extrapyramidal symptoms, cognitive impairment, adverse events, laboratory parameters, weight change, and vital signs. The paradigm of gradual antipsychotic drug discontinuation combined with an initial full dose of olanzapine, 10 mg/day, had the most favorable efficacy and tolerability profile overall. By week 3, the majority of completing patients on all 4 switching paradigms were either improved or clinically unchanged (> 90%). No clinically significant differences between switching paradigms were seen in laboratory values or vital signs. In this study, switching clinically stable outpatients with a diagnosis of schizophrenia or schizoaffective disorder to olanzapine was most successful when a full therapeutic dose of olanzapine was immediately initiated while gradually discontinuing prior conventional antipsychotic drug or risperidone treatment. Overall, switching was achieved without increased vulnerability to relapse or to occurrence of clinically burdensome antipsychotic drug withdrawal symptoms in the majority of patients.


Initiation of hypertension in utero and its amplification throughout life.
To determine whether the relation between high blood pressure and low birth weight is initiated in utero or during infancy, and whether it changes with age. A longitudinal study of children and three follow up studies of adults. Farnborough, Preston, and Hertfordshire, England, and a national sample in Britain. 1895 children aged 0-10 years, 3240 men and women aged 36 years, 459 men and women aged 46-54 years, and 1231 men and women aged 59-71 years. The birth weight of all subjects had been recorded. Systolic blood pressure. At all ages beyond infancy people who had lower birth weight had higher systolic blood pressure. Systolic blood pressure was not related to growth during infancy independently of birth weight. The relation between systolic pressure and birth weight became larger with increasing age so that, after current body mass was allowed for, systolic pressure at ages 64-71 years decreased by 5.2 mm Hg (95% confidence interval 1.8 to 8.6) for every kg increase in birth weight. Essential hypertension is initiated in fetal life. A raised blood pressure is then amplified from infancy to old age, perhaps by a positive feedback mechanism.


Do physicians induce demand for medical services?
In this paper we provide a critique of an article by Roger Feldman and Frank Sloan that appeared in the Summer 1988 issue of this journal. In that article, the authors contend that there is little evidence to support the notion of supplier-induced demand or the contention that physicians generate demand to avoid the impact on their incomes of government price controls. In this critique, we argue that the evidence on supplier-induced demand and physician responses to price controls does not support the conclusions drawn by Feldman and Sloan. We conclude with a discussion of the implications of the debate for policy formulation and future research.


The Arabidopsis aleurone layer responds to nitric oxide, gibberellin, and abscisic acid and is sufficient and necessary for seed dormancy.
Seed dormancy is a common phase of the plant life cycle, and several parts of the seed can contribute to dormancy. Whole seeds, seeds lacking the testa, embryos, and isolated aleurone layers of Arabidopsis (Arabidopsis thaliana) were used in experiments designed to identify components of the Arabidopsis seed that contribute to seed dormancy and to learn more about how dormancy and germination are regulated in this species. The aleurone layer was found to be the primary determinant of seed dormancy. Embryos from dormant seeds, however, had a lesser growth potential than those from nondormant seeds. Arabidopsis aleurone cells were examined by light and electron microscopy, and cell ultrastructure was similar to that of cereal aleurone cells. Arabidopsis aleurone cells responded to nitric oxide (NO), gibberellin (GA), and abscisic acid, with NO being upstream of GA in a signaling pathway that leads to vacuolation of protein storage vacuoles and abscisic acid inhibiting vacuolation. Molecular changes that occurred in embryos and aleurone layers prior to germination were measured, and these data show that both the aleurone layer and the embryo expressed the NO-associated gene AtNOS1, but only the embryo expressed genes for the GA biosynthetic enzyme GA3 oxidase.


Moral experience and ethical challenges in an emergency department in Pakistan: emergency physicians' perspectives.
Emergency departments (ED) are often stressful environments posing unique ethical challenges-issues that primarily raise moral rather than clinical concerns-in patient care. Despite this, there are very few reports of what emergency physicians find ethically challenging in their everyday work. Emergency medicine (EM) is a relatively young but rapidly growing specialty that is gaining acceptance worldwide. The aim of this study was to explore the perspectives of EM residents and physicians regarding the common ethical challenges they face during patient care in one of only two academic EM departments in Pakistan. These challenges could then be addressed in residents' training and departmental practice guidelines. A qualitative research design was employed and in-depth interviews were conducted with ED physicians. Participants were encouraged to think of specific examples from their work, to highlight the particular ethical concerns raised and to describe in detail the process by which those concerns were addressed or left unresolved. Transcripts were analysed using grounded theory methods. Thirteen participants were interviewed and they described four key challenges: how to provide highest quality care with limited resources; how to be truthful to patients; what to do when it is not possible to provide or continue treatment to patients; and when (and when not) to offer life-sustaining treatments. Participants' accounts provided important insights into how physicians tried to resolve these challenges in the 'local moral world' of an ED in Pakistan. The study highlights the need for developing systematic and contextually appropriate mechanisms for resolving common ethical challenges in the EDs and for training residents in moral problem solving.


Infantile hemangioma in a v2 distribution: treatment with nadolol.
Infantile hemangiomas are common benign tumours of infancy affecting up to 10% of children. They are typically not present at birth but undergo a rapid proliferation stage and then plateau in growth before resolving spontaneously. Recently, beta-blockers have been favoured over systemic corticosteroids for treatment of disfiguring or life-threatening infantile hemangiomas. We present a case of an 11-week-old female with a 7 week history of an evolving hemangioma along a facial V2 distribution. Physical exam revealed a well-defined bright red plaque over the right zygoma and lower eyelid. MRI, echocardiograph, and liver ultrasound were normal. Patient was treated with nadolol and had a rapid and substantial regression of the hemangioma. Nadolol is an effective treatment option for disfiguring facial infantile hemangioma. The use of beta-blockers as treatment offers clues into the pathogenesis of infantile hemangioma, which is not yet completely understood.


Antilope--a Lagrangian relaxation approach to the de novo peptide sequencing problem.
Peptide sequencing from mass spectrometry data is a key step in proteome research. Especially de novo sequencing, the identification of a peptide from its spectrum alone, is still a challenge even for state-of-the-art algorithmic approaches. In this paper, we present ANTILOPE, a new fast and flexible approach based on mathematical programming. It builds on the spectrum graph model and works with a variety of scoring schemes. ANTILOPE combines Lagrangian relaxation for solving an integer linear programming formulation with an adaptation of Yen's k shortest paths algorithm. It shows a significant improvement in running time compared to mixed integer optimization and performs at the same speed like other state-of-the-art tools. We also implemented a generic probabilistic scoring scheme that can be trained automatically for a data set of annotated spectra and is independent of the mass spectrometer type. Evaluations on benchmark data show that ANTILOPE is competitive to the popular state-of-the-art programs PepNovo and NovoHMM both in terms of runtime and accuracy. Furthermore, it offers increased flexibility in the number of considered ion types. ANTILOPE will be freely available as part of the open source proteomics library OpenMS.


Pregnancy dating by fetal crown-rump length: a systematic review of charts.
Fetal crown-rump length (CRL) measurement by ultrasound in the first trimester is the standard method for pregnancy dating; however, a multitude of CRL equations to estimate gestational age (GA) are reported in the literature. To evaluate the methodological quality used in studies reporting CRL equations to estimate GA using a set of predefined criteria. Searches of MEDLINE, EMBASE, and CINAHL databases, from 1948 to 31 January 2011, and secondary reference sources, were performed. Observational ultrasound studies, where the primary aim was to create equations for GA estimation using a CRL measurement. Included studies were scored against predefined independently agreed methodological criteria: an overall quality score was calculated for each study. The searches yielded 1142 citations. Two reviewers screened the papers and independently assessed the full-text versions of 29 eligible studies. The highest potential for bias was noted in inclusion and exclusion criteria, and in maternal demographic characteristics. No studies had systematic ultrasound quality-control measures. The four studies with the highest scores (lowest risk of bias) satisfied 18 or more of the 29 criteria; these showed lower variation in GA estimation than the remaining, lower-scoring studies. This was particularly evident at the extremes of GA. Considerable methodological heterogeneity and limitations exist in studies reporting CRL equations for estimating GA, and these result in a wide range of estimated GAs for any given CRL; however, when studies with the highest methodological quality are used, this range is reduced.


Distinct Functions for Mammalian CLASP1 and -2 During Neurite and Axon Elongation.
Mammalian cytoplasmic linker associated protein 1 and -2 (CLASP1 and -2) are microtubule (MT) plus-end tracking proteins that selectively stabilize MTs at the edge of cells and that promote MT nucleation and growth at the Golgi, thereby sustaining cell polarity. In vitro analysis has shown that CLASPs are MT growth promoting factors. To date, a single CLASP1 isoform (called CLASP1α) has been described, whereas three CLASP2 isoforms are known (CLASP2α, -β, and -γ). Although CLASP2β/γ are enriched in neurons, suggesting isoform-specific functions, it has been proposed that during neurite outgrowth CLASP1 and -2 act in a redundant fashion by modulating MT dynamics downstream of glycogen synthase kinase 3 (GSK3). Here, we show that in differentiating N1E-115 neuroblastoma cells CLASP1 and CLASP2 differ in their accumulation at MT plus-ends and display different sensitivity to GSK3-mediated phosphorylation, and hence regulation. More specifically, western blot (WB) analysis suggests that pharmacological inhibition of GSK3 affects CLASP2 but not CLASP1 phosphorylation and fluorescence-based microscopy data show that GSK3 inhibition leads to an increase in the number of CLASP2-decorated MT ends, as well as to increased CLASP2 staining of individual MT ends, whereas a reduction in the number of CLASP1-decorated ends is observed. Thus, in N1E-115 cells CLASP2 appears to be a prominent target of GSK3 while CLASP1 is less sensitive. Surprisingly, knockdown of either CLASP causes phosphorylation of GSK3, pointing to the existence of feedback loops between CLASPs and GSK3. In addition, CLASP2 depletion also leads to the activation of protein kinase C (PKC). We found that these differences correlate with opposite functions of CLASP1 and CLASP2 during neuronal differentiation, i.e., CLASP1 stimulates neurite extension, whereas CLASP2 inhibits it. Consistent with knockdown results in N1E-115 cells, primary Clasp2 knockout (KO) neurons exhibit early accelerated neurite and axon outgrowth, showing longer axons than control neurons. We propose a model in which neurite outgrowth is fine-tuned by differentially posttranslationally modified isoforms of CLASPs acting at distinct intracellular locations, thereby targeting MT stabilizing activities of the CLASPs and controlling feedback signaling towards upstream kinases. In summary, our findings provide new insight into the roles of neuronal CLASPs, which emerge as regulators acting in different signaling pathways and locally modulating MT behavior during neurite/axon outgrowth.


Profile: the Chakaria Health and Demographic Surveillance System.
Chakaria Health and Demographic Surveillance System (CHDSS), located on the south-eastern coast of the Bay of Bengal, was established in 1999 and is one of the field sites of International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDRB). The surveillance covers 118 315 residents living in 19 847 households. Data on socio-demographic and health indicators including birth, death, migration, marriage, maternal health, education and employment are recorded through quarterly household visits. The primary objective of CHDSS is to monitor the changes in socio-demographic indicators, inequalities in health and impact of public health interventions. A demographic change was accompanied by a shift from traditional to modern society during the past decade, but inequality in health still persists. The findings from the surveillance are shared regularly among the local and global communities. Data are also available upon request to ICDDRB and INDEPTH for use by researchers and policy makers.


Curcumin is a biologically active copper chelator with antitumor activity.
Curcumin is a natural product with antitumor activity. The compound targets multiple cell signaling pathways, including cell survival and proliferation, caspase activation and oncogene expression. As a β-diketone, curcumin also exists as a keto-enol tautomer that chelates transition metal ions with high affinity. Copper has an integral role in promoting tumor growth and angiogenesis. This study aims to investigate whether curcumin exerts its antitumor activity through copper chelation. Copper chelation ability of curcumin was validated by measuring US/VIS spectrum. The antitumor activity and in vivo copper removal ability of curcumin was determined in a murine xenograft model. The effect of curcumin on copper-induced MAPK activation and cell proliferation was determined in cell culture system. Administration of curcumin to tumor-bearing animals resulted in suppression of A549 xenograft growth, an effect that was also observed in animals treated with ammonium tetrathiomolybdate (TM), a metal chelator used for copper storage disorders clinically. The inhibition on tumor growth was associated with reduction of copper concentrations in the serum of treated groups. In cell culture studies, we showed that copper promoted cell proliferation through Erk/MAPK activation. Treatment with curcumin or U0126, a specific MAPK inhibitor, or suppression of cellular uptake of copper by siRNA knockdown of copper transporter protein 1 (CTR1) blocked copper-induced cell proliferation. This study therefore demonstrates curcumin antitumor effect to its copper chelation capability. These results also implicate copper chelation as a general mechanism for their action of some biologically active polyphenols like flavonoids.


Resistance of gammaA/gamma' fibrin clots to fibrinolysis.
Elevated plasma fibrinogen levels are a major risk factor for thrombosis. This report shows two mechanisms by which fibrinogen can affect the fibrinolysis rate in vitro and thus may lead to thrombosis. First, the lysis rate of fibrin decreases as the initial concentration of fibrinogen increases. Second, a minor variant form of fibrinogen decreases the rate of fibrinolysis. This variant, gammaA/gamma' fibrinogen, has one altered gamma chain and is known to bind to factor XIII zymogen. In a fibrinolysis assay containing purified thrombin, fibrinogen, tissue-type plasminogen activator, and plasminogen, clots from gammaA/gammaA and gammaA/gamma' fibrinogen lysed at similar rates. However, when factor XIII was added, slower lysis was seen in gammaA/gamma' fibrin clots when compared with gammaA/gammaA fibrin clots. A D-dimer agglutination assay showed that the gammaA/gamma' clots were more highly cross-linked than the gammaA/gammaA clots. The lysis rates of gammaA/gamma' clots were similar to gammaA/gammaA clots in the presence of N-ethylmaleimide, a specific inhibitor of factor XIIIa. The gammaA/gamma' fibrin clots made in the presence of factor XIII showed increased proteolytic resistance to both plasmin and trypsin. Clots made from afibrinogenemic plasma reconstituted with gammaA/gamma' fibrinogen also showed significant resistance to lysis compared with gammaA/gammaA fibrinogen. These data demonstrate gammaA/gamma' fibrin is resistant to fibrinolysis, possibly as a result of concentrating factor XIII on the clot. The total fibrinogen concentration and the amount of gammaA/gamma' fibrinogen increase clot stability in vitro and thus may contribute independently to the risk of thrombosis in humans.


Procion orange tracer dye technique vs. identification of intrafibrillar fibronectin in the assessment of sarcolemmal damage.
The use of Procion orange dye (POD) is one of the most widely accepted techniques to assess sarcolemmal damage. This phenomenon has been related to functional adaptation in skeletal muscles. The POD method includes intravenous injection of this colorant in vivo, enabling its identification inside those fibres with membrane leaks (fluorescence). However, the safety of the use of POD has not been proven. This study was designed to compare POD with a safer alternative, involving the identification of intracellular fibronectin using specific antibodies. Eight Swiss mice were submitted to electrical stimulation of the lower limbs at different frequencies (10-80 Hz). Subsequently, the POD solution was infused, and samples from the vastus medialis muscle were obtained 24 h later. Samples were processed and serial sections were analysed using immunohistochemistry (monoclonal antibodies against fibronectin) and epifluorescence microscopy. Ninety-eight per cent of the fibres were equally classified by both techniques, which in addition showed good correlation (percentages of damaged fibres, r = 0.998, P < 0.001) and concordance (R1 = 0.82) in quantitative terms. Although the two techniques compared here are based on different principles, both are comparable in assessing sarcolemmal damage. This would facilitate comparisons between human and experimental studies. In addition, the fibronectin technique appears to be a suitable alternative for long-term studies including repeated biopsies.


White-light imaging by use of a multiple passband acousto-optic tunable filter.
White-light imaging was accomplished by operation of a TeO(2) acousto-optic tunable filter (AOTF) with 40 simultaneous overlapping passbands from 400 to 700 nm. The AOTF was chromatically compensated by a wedge applied to the output surface of the AOTF, and the measured spatial resolution correlated well with predictions. Switching off specific rf's applied to the AOTF produced optical rejection corresponding to the inactive passbands. A rejection ratio of 30 dB was demonstrated, and the rejection level was found to be controlled by leakage through the sidelobes of adjacent passbands.


Parental intention to support video game play by children with autism spectrum disorder: an application of the theory of planned behavior.
The purpose of this study was to determine parental attitudes regarding engagement with video games by their children with autism spectrum disorder (ASD) and whether attitudes vary based on ASD symptom severity. Online survey methodology was used to gather information from parents of children with ASD between the ages of 8 and 12 years. The finalized data set included 152 cases. Descriptive statistics and frequency analyses were used to examine participant demographics and video game play. Descriptive and inferential statistics were used to evaluate questions on the theory of planned behavior. Regression analyses determined the predictive ability of the theory of planned behavior constructs, and t tests provided additional descriptive information about between-group differences. Children with ASD play video games. There are no significant differences in the time, intensity, or types of games played based on severity of ASD symptoms (mild vs. moderate). Parents of children with ASD had positive attitudes about video game play. Parents of children with ASD appear to support video game play. On average, parents indicated video game play was positive for their children with ASD, particularly if they believed the games were having a positive impact on their child's development.


Serum Vitamin D and Age-Related Muscle Loss in Afro-Caribbean Men: The Importance of Age and Diabetic Status.
Prospective studies examining the potential association of vitamin D with age-related muscle loss have shown inconsistent results. To examine the association between baseline serum 25-hydroxyvitamin D (25(OH)D), 1,25-dihydroxyvitamin D (1,25(OH)2D), and prospective change in lean mass with aging in African ancestry population. We also determined if associations were modulated by age and diabetes mellitus (DM). Prospective observational cohort study. Data were collected from a random sub-sample of 574 men, participants of the Tobago Bone Health Study (TBHS). 574 Afro-Caribbean men, aged 43+ years (mean age: 59.1 ± 10.5), who were randomly selected as the participants in both the baseline and the follow-up visits. Baseline fasting serum 25(OH)D was measured using liquid chromatography mass spectrometry (LC-MS/MS), and and 1,25(OH)2D was measured using radioimmunosassay (RIA). Changes in dual-energy X-ray absorptiometry (DXA)-measured appendicular lean mass (ALM), and total body lean mass (TBLM) were measured over an average of 6.0 ± 0.5 years. The associations of 25(OH)D and 1,25(OH)2D with ALM and TBLM were assessed by multiple linear regression model after adjusting for potential confounders. When stratifying all men into two groups by age, greater baseline 25(OH)D and 1,25(OH)2D levels were associated with smaller losses of ALM and TBLM in older (age 60+ years) but not in younger (age 43 - 59 years) men. When stratifying by DM status, the associations of 25(OH)D and 1,25(OH)2D with declines in ALM and TBLM were statistically significant only in prediabetic, but not among normal glycemic or diabetic men. Higher endogenous vitamin D concentrations are associated with less lean mass loss with aging among older and prediabetic Afro-Caribbean men independent of potential confounders. Our findings raise a possibility that maintaining high serum vitamin D level might be important for musculoskeletal health in elderly and prediabetic African ancestry men.


Mechanism of histamine actions in human coronary arteries.
Helical strips of human coronary arteries contracted in response to histamine concentration dependently, they relaxed with low concentrations and contracted with high concentrations. Treatment with cimetidine potentiated contraction in the strips with intact and damaged endothelium to a similar extent and attenuated relaxation. Removal of endothelium abolished relaxation and potentiated contraction in the cimetidine-treated strips. Methylene blue increased the contractile response to histamine in the strips with endothelium but did not alter the response in the damaged-endothelium strips. Histamine-induced relaxations in the intact strips were suppressed or abolished by treatment with ETYA, AA861, a lipoxygenase inhibitor, and by chlorpheniramine but were unaffected by indomethacin. Chlorpheniramine also abolished amine-induced contraction. It may be concluded that histamine-induced contraction in human coronary arteries is mediated by H1 receptors in smooth muscle, and relaxation is mediated by H2 receptors in smooth muscle and H1 receptors in endothelium. Also, stimulation of the endothelial H1 receptor liberates vasodilator substance and possibly activates smooth muscle guanylate cyclase to accumulate cellular cyclic guanosine monophosphate.


Racial differences in fibroid prevalence and ultrasound findings in asymptomatic young women (18-30 years old): a pilot study.
1) To determine the prevalence of fibroids in asymptomatic young black and white women (ages 18-30 y); 2) to determine other differences in uterine and adnexal anatomy; and 3) to obtain preliminary data for sample size calculations. Pilot cross-sectional study. Academic medical center. One hundred one nonparous black and white women, ages 18-30 years, with no known diagnosis of fibroids or clinically suggestive symptoms. A transvaginal ultrasound was performed in the follicular phase in all subjects. 1) Presence of fibroids; 2) endometrial thickness; 3) ovarian findings. Of the 101 participants (mean age 24.5 ± 3.5 y), 43% self-identified as black and 57% as white. The prevalence of ultrasound-diagnosed fibroids was 15% overall (26% in black women and 7% in white women). The mean fibroid size was 2.3 ± 2.1 cm. There was a significant difference in endometrial thickness between races, even after adjusting for contraception use and fibroid presence. Racial differences in fibroid prevalence exist even before women become symptomatic. Findings of thicker endometrium in black women could have clinical implications and warrants further investigation.


Evaluation of the effect of home bleaching gel on microleakage of different glass ionomers reinforced with micro-hydroxyapatite.
This study aimed to evaluate the effect of home bleaching gel on microleakage of glass-ionomer cements reinforced with micro-hydroxyapatite (HAP). Class V cavities prepared on the forty extracted third molars were restored in four groups (n = 10): Group 1, Zirconomer; Group 2, resin-modified glass ionomer (RMGI); Group 3, Zirconomer + micro-HAP (20% WT); and Group 4, RMGI + micro-HAP (20% WT). After thermocycling (1000 cycles at 5 ± 2/55 ± 2°C, a dwell time of 30 s), each group was randomly divided into two groups. The first half was kept in distilled water and the restorations of the second half were bleached with carbamide peroxide 15% (14 days, each time 6 h/day). A uniform thickness of bleaching agent (0.5-1 mm) was applied on the surfaces of the restorations, extending 1 mm beyond the margins, and the bleaching agent was exchanged every 6 h. Microleakage was evaluated using dye penetration technique that is based on the amount of dye penetration (0.5% basic fuchsine solution) from the occlusal/gingival margins up to the axial wall. In distilled water, no significant difference was found between the occlusal microleakage scores (P > 0.05). The lowest and highest gingival scores in distilled water were observed in Group 4 and Group 3, respectively. In bleaching environment, there was no significant difference between four groups (P > 0.05). Comparing each glass ionomer in two environments revealed statistically significant differences in gingival and occlusal microleakage of Group 4 and in occlusal microleakage of Group 1 (P < 0.05). Micro-HAP incorporation did not affect the microleakage of the RMGI and Zirconomer in bleaching environment and their occlusal microleakage in distilled water. The lowest and highest gingival scores in distilled water were observed for the RMGI + micro-HAP and Zirconomer + micro-HAP, respectively. Bleaching procedure negatively affects the microleakage score of RMGI + micro-HAP and occlusal microleakage of Zirconomer.


Are protected areas required to maintain functional diversity in human-modified landscapes?
The conversion of forest to agriculture across the world's tropics, and the limited space for protected areas, has increased the need to identify effective conservation strategies in human-modified landscapes. Isolated trees are believed to conserve elements of ecological structure, providing micro-sites for conservation in matrix landscapes, and facilitating seed dispersal and forest restoration. Here we investigate the role of isolated Ficus trees, which are of critical importance to tropical forest ecosystems, in conserving frugivore composition and function in a human-modified landscape in Assam, India. We surveyed the frugivorous birds feeding at 122 isolated Ficus trees, 33 fruit trees, and 31 other large trees across a range of 32 km from the nearest intact forest. We found that Ficus trees attracted richer and more abundant assemblages of frugivores than the other tree categories. However, incidence function estimates revealed that forest specialist species decreased dramatically within the first kilometre of the forest edge. Despite this, species richness and functional diversity remained consistent across the human-modified landscape, as habitat generalists replaced forest-dependent frugivores, and accounted for most of the ecological function found in Ficus trees near the forest edge. We recommend that isolated Ficus trees are awarded greater conservation status, and suggest that their conservation can support ecologically functional networks of frugivorous bird communities.


Exercise differentially affects metabolic functions and white adipose tissue in female letrozole- and dihydrotestosterone-induced mouse models of polycystic ovary syndrome.
Here we hypothesized that exercise in dihydrotestosterone (DHT) or letrozole (LET)-induced polycystic ovary syndrome mouse models improves impaired insulin and glucose metabolism, adipose tissue morphology, and expression of genes related to adipogenesis, lipid metabolism, Notch pathway and browning in inguinal and mesenteric fat. DHT-exposed mice had increased body weight, increased number of large mesenteric adipocytes. LET-exposed mice displayed increased body weight and fat mass, decreased insulin sensitivity, increased frequency of small adipocytes and increased expression of genes related to lipolysis in mesenteric fat. In both models, exercise decreased fat mass and inguinal and mesenteric adipose tissue expression of Notch pathway genes, and restored altered mesenteric adipocytes morphology. In conclusion, exercise restored mesenteric adipocytes morphology in DHT- and LET-exposed mice, and insulin sensitivity and mesenteric expression of lipolysis-related genes in LET-exposed mice. Benefits could be explained by downregulation of Notch, and modulation of browning and lipolysis pathways in the adipose tissue.


Resistance to the isocitrate dehydrogenase 1 mutant inhibitor ivosidenib can be overcome by alternative dimer-interface binding inhibitors.
Ivosidenib, an inhibitor of isocitrate dehydrogenase 1 (IDH1) R132C and R132H variants, is approved for the treatment of acute myeloid leukaemia (AML). Resistance to ivosidenib due to a second site mutation of IDH1 R132C, leading to IDH1 R132C/S280F, has emerged. We describe biochemical, crystallographic, and cellular studies on the IDH1 R132C/S280F and R132H/S280F variants that inform on the mechanism of second-site resistance, which involves both modulation of inhibitor binding at the IDH1 dimer-interface and alteration of kinetic properties, which enable more efficient 2-HG production relative to IDH1 R132C and IDH1 R132H. Importantly, the biochemical and cellular results demonstrate that it should be possible to overcome S280F mediated resistance in AML patients by using alternative inhibitors, including some presently in phase 2 clinical trials.


A clinical and histopathologic comparison of cardiac sarcoidosis and idiopathic giant cell myocarditis.
The goal of this study was to determine the prognostic value of clinical data available at presentation and histology in cardiac sarcoidosis (CS) and idiopathic giant cell myocarditis (IGCM). The prognosis of patients with nonischemic cardiomyopathy is partly dependent on the histologic diagnosis. Survival in IGCM is poor. The prognosis of a histologically related entity, cardiac sarcoidosis (CS), is less well established, and the prognostic value of the distinction between CS and IGCM on endomyocardial biopsy (EMB) is unknown. We identified 115 patients from the Multicenter IGCM Registry with CS (n = 42) and IGCM (n = 73). We compared the clinical data for these two groups using Cox proportional-hazards models to assess the association between histologic diagnosis and survival. In order to determine whether histologic features could reliably differentiate these two entities, two cardiac pathologists semiquantitatively graded the inflammatory infiltrate components and compared the results between groups. Black race was more frequent in the CS group (31% vs. 4%, p < 0.0001). Syncope and atrioventricular block were also more frequently observed in CS than IGCM (31% vs. 5%, p = 0.0002 and 50% vs. 15%, p < 0.0001, respectively). Left-sided heart failure was more common in IGCM (40% vs. 64%, p = 0.013). In CS patients diagnosed by EMB, the five-year transplant-free survival after diagnosis was 69.8% versus 21.9% for IGCM (p < 0.0001, log-rank test). In multivariate models, presentation with heart failure predicted IGCM, and presentation with heart block or more than nine weeks of symptoms predicted CS. Eosinophils, myocyte damage, and foci of lymphocytic myocarditis were more frequent in IGCM, while granulomas and fibrosis were more frequent in CS. Transplant-free survival is better for patients with CS than for IGCM diagnosed by EMB. Presentation with heart failure predicted IGCM, and presentation with heart block or more than nine weeks of symptoms predicted CS.


Iron phosphate nanoparticles for food fortification: Biological effects in rats and human cell lines.
Nanotechnology offers new opportunities for providing health benefits in foods. Food fortification with iron phosphate nanoparticles (FePO4 NPs) is a promising new approach to reducing iron deficiency because FePO4 NPs combine high bioavailability with superior sensory performance in difficult to fortify foods. However, their safety remains largely untested. We fed rats for 90 days diets containing FePO4 NPs at doses at which iron sulfate (FeSO4), a commonly used food fortificant, has been shown to induce adverse effects. Feeding did not result in signs of toxicity, including oxidative stress, organ damage, excess iron accumulation in organs or histological changes. These safety data were corroborated by evidence that NPs were taken up by human gastrointestinal cell lines without reducing cell viability or inducing oxidative stress. Our findings suggest FePO4 NPs appear to be as safe for ingestion as FeSO4.


Anaesthesia for evacuation of incomplete miscarriage.
An incomplete miscarriage occurs when all the products of conception are not expelled through the cervix. Curettage or vacuum aspiration have been used to remove retained tissues. The anaesthetic techniques used to facilitate this procedure have not been systematically evaluated in order to determine which provide better outcomes to the patients. To assess the effects of general anaesthesia, sedation or analgesia, regional or paracervical block anaesthetic techniques, or differing regimens of these, for surgical evacuation of incomplete miscarriage. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (23 January 2012), CENTRAL (The Cochrane Library 2012, Issue 1), PubMed (1966 to 23 January 2012), EMBASE (1974 to 23 January 2012), CINAHL (1982 to 23 January 2012), LILACS (1982 to 23 January 2012) and reference lists of retrieved studies. All published and unpublished randomised controlled trials (RCTs) or cluster-RCTs comparing the use of any anaesthetic technique (defined by authors as general anaesthesia, sedation/analgesia, regional or paracervical local block (PCB) procedures) to perform surgical evacuation of an incomplete miscarriage. We excluded quasi-randomised trials and studies that were only available as abstracts. Two review authors independently assessed studies for inclusion and assessed risk of bias. Data were independently extracted and checked for accuracy. We included seven trials involving 800 women. The comparisons revealed a very high clinical heterogeneity. As a result of the heterogeneity in the randomisation unit, we did not combine trials but reported the individual trial results in the 'Data and analysis' section and in the text. Half of trials have unclear or high risk of bias in several domains.We did not find any trial reporting data about maternal mortality. In terms of postoperative pain, PCB does not improve the control of postoperative pain when it is compared against sedation/analgesia or versus no anaesthesia/no analgesia. In the comparison of PCB with lidocaine versus PCB with saline solution, significant differences favouring the group with lidocaine were found in one trial (moderate or severe postoperative pain) (risk ratio (RR) 0.32; 95% confidence interval (CI) 0.18 to 0.59).When opioids were used, postoperative nausea and vomiting was more frequent in two trials comparing those versus PCB. In terms of requirement of blood transfusion, two trials showed conflicting results. Particular considerations that influence the choice of anaesthesia for this procedure such as availability, effectiveness, safety, side effects, practitioner's choice, costs and woman's preferences of each technique should continue to be used until more evidence supporting the use of one technique or another.


Disagreement Between Randomized and Observational Evidence on the Use of Bilateral Internal Thoracic Artery Grafting: A Meta-Analytic Approach.
Background The ART (Arterial Revascularization Trial) showed no difference in survival at 10 years between patients assigned to the single versus bilateral internal thoracic artery grafting strategies. This finding is in contrast with the results of most observational studies, where the use of 2 internal thoracic arteries has been associated with improved survival. Methods and Results We selected propensity-matched studies from the most comprehensive observational meta-analysis on the long-term outcomes of patients receiving 1 versus 2 internal thoracic arteries. Individual participant survival data from each study and the ART were reconstructed using an iterative algorithm that was applied to solve the Kaplan-Meier equations. The reconstructed individual participant survival data were aggregated to obtain combined survival curves and Cox regression hazard ratios with 95% CIs. Individual participant survival data were obtained from 14 matched observational studies (24 123 patients) and the ART. The 10-year survival of the control group of ART was significantly higher than that of the matched observational studies (hazard ratio, 0.86; 95% CI, 0.80-0.93). The 10-year survival of the experimental group of ART was significantly lower than that of the bilateral internal thoracic artery group of the observational studies (hazard ratio, 1.11; 95% CI, 1.03-1.20). Conclusions Both the improved outcome of the control arm and the lower beneficial effect of the intervention had played a role in the difference between observational evidence and ART.


A Small-Molecule Antagonist of the β-Catenin/TCF4 Interaction Blocks the Self-Renewal of Cancer Stem Cells and Suppresses Tumorigenesis.
Wnt/β-catenin signaling is a highly conserved pathway essential for embryogenesis and tissue homeostasis. However, deregulation of this pathway can initiate and promote human malignancies, especially of the colon and head and neck. Therefore, Wnt/β-catenin signaling represents an attractive target for cancer therapy. We performed high-throughput screening using AlphaScreen and ELISA techniques to identify small molecules that disrupt the critical interaction between β-catenin and the transcription factor TCF4 required for signal transduction. We found that compound LF3, a 4-thioureido-benzenesulfonamide derivative, robustly inhibited this interaction. Biochemical assays revealed clues that the core structure of LF3 was essential for inhibition. LF3 inhibited Wnt/β-catenin signals in cells with exogenous reporters and in colon cancer cells with endogenously high Wnt activity. LF3 also suppressed features of cancer cells related to Wnt signaling, including high cell motility, cell-cycle progression, and the overexpression of Wnt target genes. However, LF3 did not cause cell death or interfere with cadherin-mediated cell-cell adhesion. Remarkably, the self-renewal capacity of cancer stem cells was blocked by LF3 in concentration-dependent manners, as examined by sphere formation of colon and head and neck cancer stem cells under nonadherent conditions. Finally, LF3 reduced tumor growth and induced differentiation in a mouse xenograft model of colon cancer. Collectively, our results strongly suggest that LF3 is a specific inhibitor of canonical Wnt signaling with anticancer activity that warrants further development for preclinical and clinical studies as a novel cancer therapy.


Patients' perception of health services for sexually transmitted infections in Singapore.
The Department of Sexually Transmitted Infections (STI) Control (DSC) Clinic is the only public STI clinic in Singapore. This study aimed to determine patients' perception of the clinic as well as the factors that may influence their choice of DSC Clinic over other medical facilities. The concerns of certain risk groups were also assessed. Self-administered anonymous questionnaires were offered to Singaporeans and Permanent Residents over 18 years of age who were seeking treatment at the DSC Clinic. 1,000 responses were collected over a period of four weeks in May 2009. More than two-thirds of the patients had a positive experience at the DSC Clinic and would recommend the clinic to their family and friends. Positive attributes included competence of staff and the convenience of a one-stop treatment facility. The patients' visits to the DSC Clinic also prompted them to engage in safer sexual practices. Stigma was surprisingly not an issue among more than three-quarters of the patients. Confidentiality of medical records was a major concern, with more than half of the patients unwilling to share their records with other healthcare providers, employers or insurance companies. The majority of patients would like to see the provision of clinics catering to special groups such as young people, women as well as men-who-have-sex-with-men. The majority of patients were satisfied with the health services provided at the DSC Clinic. Confidentiality in consultation was of prime importance to patients.


fMRI responses to words repeated in a congruous semantic context are abnormal in mild Alzheimer's disease.
We adapted an event-related brain potential word repetition paradigm, sensitive to early Alzheimer's disease (AD), for functional MRI (fMRI). We hypothesized that AD would be associated with reduced differential response to New/Old congruous words. Fifteen mild AD patients (mean age=72.9) and 15 normal elderly underwent 1.5T fMRI during a semantic category decision task. We found robust between-groups differences in BOLD response to congruous words. In controls, the New>Old contrast demonstrated larger responses in much of the left-hemisphere (including putative P600 generators: parahippocampal, cingulate, fusiform, perirhinal, middle temporal (MTG) and inferior frontal gyri (IFG)); the Old>New contrast showed modest activation, mainly in right parietal and prefrontal cortex. By contrast, there were relatively few regions of significant New>Old responses in AD patients, mainly in the right-hemisphere, and their Old>New contrast did not demonstrate a right-hemisphere predominance. Across subjects, the spatial extent of New>Old responses in left medial temporal lobe (MTL) correlated with subsequent recall and recognition (r's>or=0.60). In controls, the magnitude of New-Old response in left MTL, fusiform, IFG, MTG, superior temporal and cingulate gyrus correlated with subsequent cued recall and/or recognition (0.51<or=r's<or=0.78). A distributed network of mostly left-hemisphere structures, which are putative P600 generators, appears important for successful verbal encoding (with New>Old responses to congruous words in normal elderly). This network appears dysfunctional in mild AD patients, as reflected in decreased word repetition effects particularly in left association cortex, paralimbic and MTL structures.


No effect of ablation of surfactant protein-D on acute cerebral infarction in mice.
Crosstalk between the immune system in the brain and the periphery may contribute to the long-term outcome both in experimental and clinical stroke. Although, the immune defense collectin surfactant protein-D (SP-D) is best known for its role in pulmonary innate immunity, SP-D is also known to be involved in extrapulmonary modulation of inflammation in mice. We investigated whether SP-D affected cerebral ischemic infarction and ischemia-induced inflammatory responses in mice. The effect of SP-D was studied by comparing the size of ischemic infarction and the inflammatory and astroglial responses in SP-D knock out (KO) and wild type (WT) mice subjected to permanent middle cerebral artery occlusion. SP-D mRNA production was assessed in isolated cerebral arteries and in the whole brain by PCR, and SP-D protein in normal appearing and ischemic human brain by immunohistochemistry. Changes in plasma SP-D and TNF were assessed by ELISA and proximity ligation assay, respectively. Infarct volumetric analysis showed that ablation of SP-D had no effect on ischemic infarction one and five days after induction of ischemia. Further, ablation of SP-D had no effect on the ischemia-induced increase in TNF mRNA production one day after induction of ischemia; however the TNF response to the ischemic insult was affected at five days. SP-D mRNA was not detected in parenchymal brain cells in either naïve mice or in mice subjected to focal cerebral ischemia. However, SP-D mRNA was detected in middle cerebral artery cells in WT mice and SP-D protein in vascular cells both in normal appearing and ischemic human brain tissue. Measurements of the levels of SP-D and TNF in plasma in mice suggested that levels were unaffected by the ischemic insult. Microglial-leukocyte and astroglial responses were comparable in SP-D KO and WT mice. SP-D synthesis in middle cerebral artery cells is consistent with SP-D conceivably leaking into the infarcted area and affecting local cytokine production. However, there was no SP-D synthesis in parenchymal brain cells and ablation of SP-D had no effect on ischemic cerebral infarction.


The MAP kinase p38 is part of Drosophila melanogaster's circadian clock.
All organisms have to adapt to acute as well as to regularly occurring changes in the environment. To deal with these major challenges organisms evolved two fundamental mechanisms: the p38 mitogen-activated protein kinase (MAPK) pathway, a major stress pathway for signaling stressful events, and circadian clocks to prepare for the daily environmental changes. Both systems respond sensitively to light. Recent studies in vertebrates and fungi indicate that p38 is involved in light-signaling to the circadian clock providing an interesting link between stress-induced and regularly rhythmic adaptations of animals to the environment, but the molecular and cellular mechanisms remained largely unknown. Here, we demonstrate by immunocytochemical means that p38 is expressed in Drosophila melanogaster's clock neurons and that it is activated in a clock-dependent manner. Surprisingly, we found that p38 is most active under darkness and, besides its circadian activation, additionally gets inactivated by light. Moreover, locomotor activity recordings revealed that p38 is essential for a wild-type timing of evening activity and for maintaining ∼ 24 h behavioral rhythms under constant darkness: flies with reduced p38 activity in clock neurons, delayed evening activity and lengthened the period of their free-running rhythms. Furthermore, nuclear translocation of the clock protein Period was significantly delayed on the expression of a dominant-negative form of p38b in Drosophila's most important clock neurons. Western Blots revealed that p38 affects the phosphorylation degree of Period, what is likely the reason for its effects on nuclear entry of Period. In vitro kinase assays confirmed our Western Blot results and point to p38 as a potential "clock kinase" phosphorylating Period. Taken together, our findings indicate that the p38 MAP Kinase is an integral component of the core circadian clock of Drosophila in addition to playing a role in stress-input pathways.


Geologic Evolution of North America: Geologic features suggest that the continent has grown and differentiated through geologic time.
The oldest decipherable rock complexes within continents (more than 2.5 billion years old) are largely basaltic volcanics and graywacke. Recent and modern analogs are the island arcs formed along and adjacent to the unstable interface of continental and oceanic crusts. The major interfacial reactions (orogenies) incorporate pre-existing sial, oceanic crust, and mantle into crust of a more continental type. Incipient stages of continental evolution, more than 3 billion years ago, remain obscure. They may involve either a cataclysmic granite-forming event or a succession of volcanic-sedimentary and granite-forming cycles. Intermediate and recent stages of continental evolution, as indicated by data for North America, involve accretion of numerous crustal interfaces with fragments of adjacent continental crust and their partial melting, reinjection, elevation, unroofing, and stabilization. Areas of relict provinces defined by ages of granites suggest that continental growth is approximately linear. But the advanced differentiation found in many provinces and the known overlaps permit wide deviation from linearity in the direction of a more explosive early or intermediate growth.


The Environmental Exposures and Inner- and Intercity Traffic Flows of the Metro System May Contribute to the Skin Microbiome and Resistome.
The skin functions as the primary interface between the human body and the external environment. To understand how the microbiome varies within urban mass transit and influences the skin microbiota, we profiled the human palm microbiome after contact with handrails within the Hong Kong Mass Transit Railway (MTR) system. Intraday sampling time was identified as the primary determinant of the variation and recurrence of the community composition, whereas human-associated species and clinically important antibiotic resistance genes (ARGs) were captured as p.m. signatures. Line-specific signatures were notably correlated with line-specific environmental exposures and city characteristics. The sole cross-border line appeared as an outlier in most analyses and showed high relative abundance and a significant intraday increment of clinically important ARGs (24.1%), suggesting potential cross-border ARG transmission, especially for tetracycline and vancomycin resistance. Our study provides an important reference for future public health strategies to mitigate intracity and cross-border pathogen and ARG transmission.


Fast and global authenticity screening of honey using ¹H-NMR profiling.
An innovative analytical approach was developed to tackle the most common adulterations and quality deviations in honey. Using proton-NMR profiling coupled to suitable quantification procedures and statistical models, analytical criteria were defined to check the authenticity of both mono- and multi-floral honey. The reference data set used was a worldwide collection of more than 800 honeys, covering most of the economically significant botanical and geographical origins. Typical plant nectar markers can be used to check monofloral honey labeling. Spectral patterns and natural variability were established for multifloral honeys, and marker signals for sugar syrups were identified by statistical comparison with a commercial dataset of ca. 200 honeys. Although the results are qualitative, spiking experiments have confirmed the ability of the method to detect sugar addition down to 10% levels in favorable cases. Within the same NMR experiments, quantification of glucose, fructose, sucrose and 5-HMF (regulated parameters) was performed. Finally markers showing the onset of fermentation are described.


Waterpipe Tobacco Smoke Inhalation Triggers Thrombogenicity, Cardiac Inflammation and Oxidative Stress in Mice: Effects of Flavouring.
The consumption of water-pipe smoking (WPS) has been promoted by the use of flavoured tobacco. However, little is known about the impact of flavouring on the cardiovascular toxicity induced by WPS inhalation. Here, we compared the cardiovascular effects and underlying mechanism of actions of plain (P) (unflavoured) versus apple-flavoured (AF) WPS (30 minutes/day, 5 days/week for 1 month) in mice. Control mice were exposed to air. Both P- and AF-WPS inhalation induced an increase in systolic blood pressure, thrombogenicity and plasma concentration of fibrinogen and von Willebrand factor. In heart homogenates, AF-WPS inhalation caused an increase of 8-isoprostane and a decrease in the levels of reduced glutathione (GSH) and superoxide dismutase (SOD). Nevertheless, P-WPS decreased only the activity of SOD. The concentrations of tumour necrosis factor α and interleukin 1β were increased only in heart homogenates of mice exposed to AF-WPS. Although both P- and AF-WPS increased the concentration of troponin I in heart homogenates and induced DNA damage, the concentration of cleaved caspase 3 was only increased in mice exposed to AF-WPS. Immunohistochemical analysis of the hearts showed that both P- and AF- WPS inhalation decreased the expression of SOD. Moreover, the expression of nuclear factor erythroid-derived 2-like 2 at nuclear level in the heart was higher in both AF-WPS and P-WPS compared with control group, and the effect observed in AF-WPS group was more significant than that seen in P-WPS group. Likewise, the concentration of heme oxygenase-1 was significantly increased in both P-WPS and AF-WPS groups compared with control group, and the effect seen in AF-group was higher than that observed in P-WPS group. In conclusion, our findings showed that both P- and AF-WPS induce thrombogenicity and cardiac injury, and that this toxicity is potentiated by the presence of flavouring.


Southern rice black-streaked dwarf virus alters insect vectors' host orientation preferences to enhance spread and increase rice ragged stunt virus co-infection.
In recent years, Southern rice black-streaked dwarf virus (SRBSDV), a tentative species in the genus Fijivirus (family Reoviridae), has spread rapidly and caused serious rice losses in eastern and southeastern Asia. With this virus spread, Rice ragged stunt virus (RRSV, genus Oryzavirus, family Reoviridae) became more common in southern China, usually in co-infection with the former. SRBSDV and RRSV are transmitted by two different species of planthoppers, white-backed planthopper (WBPH, Sogatella furcifera) and brown planthopper (BPH, Nilaparvata lugens), respectively, in a persistent, circulative, propagative manner. In this study, using a Y-shape olfactometer-based device, we tested the host preference of three types of macropterous WBPH adults for healthy or SRBSDV-infected rice plants. The results showed that virus-free WBPHs significantly preferred infected rice plants to healthy plants, whereas both the viruliferous and nonviruliferous WBPHs preferred healthy plants to infected plants. In additional tests, we found that the BPHs significantly preferred healthy plants when they were virus free, whereas RRSV-carrying BPHs preferred SRBSDV-infected rice plants. From these findings, we propose that plant viruses may alter host selection preference of vectors to enhance their spread and that of insects vectoring another virus to result in co-infection with more than one virus.


The alphavbeta5 integrin functions as an endocytic receptor for vitronectin.
Endocytosis and degradation of vitronectin by human skin fibroblasts are regulated by the beta5 integrin. To determine whether the beta5 integrin is directly mediating the internalization of vitronectin, both vitronectin and the beta5 integrin were localized by indirect immunofluorescence during the endocytic process. This analysis showed that both vitronectin and beta5 were found in intracellular vesicles within 5 minutes of the addition of exogenous vitronectin to fibroblast cell layers. By 15 minutes, approximately 20% of the vitronectin-containing vesicles stained positively for beta5. In contrast, the beta3 integrin was not found in any intracellular vesicles. Within 30 minutes, more than 50% of vitronectin-containing vesicles also stained for lamp-1, indicating that internalized vitronectin traveled to lysosomes. Inhibition of clathrin assembly by either potassium depletion or hypertonic buffer inhibited vitronectin internalization, suggesting that vitronectin internalization occurred through coated pits. Confocal analysis confirmed the colocalization of vitronectin and alphavbeta5 in intracellular compartments and further demonstrated that the highest colocalization of the two proteins occurred within 1.8 microm from the ventral surface of the cell, suggesting endocytosis occurred at the substrate level. Pretreatment of cells with the PI-3 kinase inhibitor, wortmannin, resulted in a marked increase in the coincidence of vitronectin and beta5 staining within vesicles and prevented the accumulation of vitronectin within lysosomes. This suggests that following internalization, vitronectin and the alphavbeta5 integrin are segregated to different cellular compartments. This study provides the first evidence that the alphavbeta5 vitronectin receptor directly mediates the internalization of vitronectin.


Neuregulin-1 enhances differentiation of cardiomyocytes from embryonic stem cells.
Neuregulin-1 (NRG-1) is a multifunctional regulator that acts through receptor tyrosine kinases of the epidermal growth factor (EGF/ErbB) receptor family in diverse tissue. ErbB receptors are expressed in developing embryoid bodies (EBs), and the importance of the NRG-1/ErbB signaling axis in heart development has been investigated, but the underlying molecular mechanism is poorly studied. NRG-1 treatment at 100 ng/ml significantly increased the number of beating EBs of differentiated murine embryonic stem cells (ESCs). Furthermore, NRG-1 up-regulated the expression of the cardiac-restricted transcription factors Nkx2.5 and GATA-4 and factors involved in differentiated cardiac cells (alpha-MHC, beta-MHC and alpha-actinin); NRG-1-induced increase of Nkx2.5 transcription was inhibited by treatment with the PI3 K inhibitor or ErbB receptor inhibitor. Western blot analysis confirmed that the expression of phospho-Akt in the beating foci was increased in the presence of NRG-1. Our results suggest that NRG-1 promotes cardiomyocyte differentiation of ESCs and the ErbB/PI3 K/Akt signaling pathway is one of the underlying molecular mechanisms.