Hydraulic traits are coupled with plant anatomical traits under drought-rewatering cycles in Ginkgo biloba L.
Investigating the responses of plant anatomical traits of trees to drought-rewatering cycles helps us to understand their responses to climate change; however, such work has not been adequately reported. In this study, Ginkgo biloba L. saplings were subjected to moderate, severe, extreme and lethal drought conditions by withholding water according to the percentage loss of hydraulic conductivity (PLC) and rewatering on a regular basis. Samples of phloem, cambium and xylem were collected to quantify their cellular properties including cambium and phloem cell vitality, xylem growth ring width, pit aspiration rates and pit membrane thickness using light microscopy and transmission microscopy. The results showed that the mortality rate of G. biloba saplings reached 90% at approximately P88 (xylem water potential inducing 88% loss of hydraulic conductivity). The onset of cambium and phloem cell mortality might be in accordance with that of xylem embolism. Close negative correlations between xylem water potential and PLC and between xylem water potential and cambium and phloem mortality suggested that xylem hydraulic traits are coupled with anatomical traits under declining xylem water potential. Cambium and phloem cell vitality as well as xylem growth ring width decreased significantly with increasing drought conditions. However, xylem pit membrane thickness, cambial zone width and cambial cell geometry were not affected by the drought-rewatering cycles. The tracheid radial diameter, intertracheid cell wall thickness and tracheid density decreased significantly during both drought conditions and rewatering conditions. In addition to hydraulic traits, cambium and phloem cell vitality can be used as anatomical traits to evaluate the mortality of G. biloba under drought. Future work is proposed to observe the dynamics of pit aspiration rates under drought-rewatering cycles in situ to deepen our understanding of the essential role of bordered pits in the 'air-seeding' mechanism.


Antitumor and anti-angiogenic potentials of isolated crude saponins and various fractions of Rumex hastatus D. Don.
Cancer, being the foremost challenge of the modern era and the focus of world-class investigators, gargantuan research is in progress worldwide to explore novel therapeutic for its management. The exploitation of natural sources has been proven to be an excellent approach to treat or minify the excessive angiogenesis and proliferation of cells. Similarly, based the ethnomedicinal uses and literature survey, the current study is designed to explore the anti-tumor and anti-angiogenic potentials of Rumex hastatus. Anti-tumor and anti-angiogenic activities were carried out using potato-disc model and chorioallantoic membrane (CAM) assay respectively. Moreover, R. hastatus was also assessed for antibacterial activity against Agrobacterium tumefaciens (tumor causing bacterial strain). The positive controls used in anti-tumor, anti-angiogenic and antibacterial activities were vincristine sulphate, dexamethasone and cefotaxime respectively. The crude saponins (Rh.Sp), methanolic extract (Rh.Cr) and other solvent extracts like n-hexane (Rh.Hex), chloroform (Rh.Chf), ethylacetate (Rh.EtAc) and aqueous fraction (Rh.Aq) exhibited notable anti-tumor and anti-angiogenic activities. In potato tumor assay, the chloroform and saponin fractions were observed to be the most effective showing 86.7 and 93.3 % tumor inhibition at 1000 µg/ml with IC50 values 31.6 and 18.1 µg/ml respectively. Similarly, these two samples i.e., chloroform and saponins also excelled among the entire test samples in anti-angiogenic evaluation exhibiting 81.6 % (IC50 = 17.9 µg/ml) and 78.9 % (IC50 = 64.9 µg/ml) at 1000 µg/ml respectively. In contrast, the antibacterial investigations revealed a negligible potential against A. tumefaciens. Based on our results we can claim that R. hastatus possesses both anti-tumor and anti-angiogenic potentials. In all of the solvent fractions, Rh.Chf and Rh.Sp were most effective against tumor and angiogenesis while having negligible activity against A. tumefaciens. It can be concluded that Rh.Chf and Rh.Sp might be potential targets in the isolation of natural product having anti-neoplastic action.


Picosecond and femtosecond asymmetric switching using a semiconductor optical amplifier-based Mach-Zehnder interferometer.
In this paper, we numerically analyze nonlinear asymmetric switching using a semiconductor optical amplifier (SOA) phase-shifter-based Mach-Zehnder interferometer (MZI), for the first time, to the best of our knowledge. The self-phase modulation (SPM) effect and nonlinear phase shift in each MZI arm are investigated for different input pulse intensities and linear gains in both picosecond and femtosecond regimes. The input light signal is split unequally over the two arms, where SOAs are placed and act as nonlinear phase shifters in each arm. The finite difference beam propagation method is used to solve the modified nonlinear Schrodinger equation to analyze the wave propagation. In this work, the main nonlinear effects in SOA, such as group velocity dispersion, Kerr effect, two-photon absorption, carrier heating, and spectral hole burning, are considered. Furthermore, the effect of SPM on distortion of the pulse shape and its spectrum, which can be used for pulse shaping in a picosecond-switching scheme, is studied. We depicted red and blue shifts that each pulse experiences in the process of switching in picosecond and femtosecond regimes, respectively. Based on the results for sub-picosecond input pulses, by controlling the bias current level in the MZI arms, the pulse distortion due to nonlinear effects of SOAs can be decreased at the switch output port, and symmetric pulse can be obtained. Switching with higher speed is possible in bulk SOAs in the femtosecond regime using asymmetric MZI-switching structure.


Reliability and validity of quantifying absolute muscle hardness using ultrasound elastography.
Muscle hardness is a mechanical property that represents transverse muscle stiffness. A quantitative method that uses ultrasound elastography for quantifying absolute human muscle hardness has been previously devised; however, its reliability and validity have not been completely verified. This study aimed to verify the reliability and validity of this quantitative method. The Young's moduli of seven tissue-mimicking materials (in vitro; Young's modulus range, 20-80 kPa; increments of 10 kPa) and the human medial gastrocnemius muscle (in vivo) were quantified using ultrasound elastography. On the basis of the strain/Young's modulus ratio of two reference materials, one hard and one soft (Young's moduli of 7 and 30 kPa, respectively), the Young's moduli of the tissue-mimicking materials and medial gastrocnemius muscle were calculated. The intra- and inter-investigator reliability of the method was confirmed on the basis of acceptably low coefficient of variations (≤6.9%) and substantially high intraclass correlation coefficients (≥0.77) obtained from all measurements. The correlation coefficient between the Young's moduli of the tissue-mimicking materials obtained using a mechanical method and ultrasound elastography was 0.996, which was equivalent to values previously obtained using magnetic resonance elastography. The Young's moduli of the medial gastrocnemius muscle obtained using ultrasound elastography were within the range of values previously obtained using magnetic resonance elastography. The reliability and validity of the quantitative method for measuring absolute muscle hardness using ultrasound elastography were thus verified.


Marfan syndrome-causing mutations in fibrillin-1 result in gross morphological alterations and highlight the structural importance of the second hybrid domain.
Mutations in fibrillin-1 result in Marfan syndrome, which affects the cardiovascular, skeletal and ocular systems. The multiorgan involvement and wide spectrum of associated phenotypes highlights the complex pathogenesis underlying Marfan syndrome. To elucidate the genotype to phenotype correlations, we engineered four Marfan syndrome causing mutations into a fibrillin-1 fragment encoded by exons 18-25, a region known to interact with tropoelastin. Biophysical and biochemical approaches, including small angle x-ray scattering, analytical ultracentrifugation, and circular dichroism, were used to study the impact of these mutations upon the structure and function of the protein. Mutations G880S, C862R, and C908R, situated within the second hybrid domain, disrupted the ratio of alpha-helix to beta-sheet leading to a more compact conformation. These data clearly demonstrate the importance of the previously uncharacterized hybrid domain in fibrillin-1 structure. In contrast, mutation K1023N situated within the linker region between the third eight cysteine motif and cbEGF 11 markedly extended the length of the fragment. However, none of the mutations affected tropoelastin binding. The profound effects of all four mutations on fragment conformation suggest that they contribute to the pathogenesis of Marfan syndrome by disrupting protein folding and its assembly into fibrillin-rich microfibrils.


Differential amplification of gene expression in lens cell lines conditioned to survive peroxide stress.
The response of lens systems to oxidative stress is confusing. Antioxidative defense systems are not mobilized as expected, and unanticipated defenses appear important. Therefore, mouse lens cell lines conditioned to survive different peroxide stresses have been analyzed to determine their global changes in gene expression. The immortal mouse lens epithelial cell line alphaTN4-1 was conditioned to survive 125 microM H2O2 (H cells) or a combination of both 100 microM tertiary butyl hydroperoxide (TBHP) and 125 microM H2O2 (HT cells), by a methodology previously described. The total RNA was isolated from the different cell lines and analyzed with oligonucleotide mouse expression microarrays. Four microarrays were used for each cell line. Microarray results were confirmed by real-time RT-PCR. A new cell line resistant to both 125 microM H2O2 and 100 micro M TBHP was developed, because cells resistant to H2O2 were killed by TBHP. Analysis of classic antioxidative enzyme activities showed little change between cells that survive H2O2 (H) and those that survive H2O2 and TBHP (HT). Therefore, the global change in gene expression in these cell lines was determined with gene expression microarrays. The fluorescent signal changes of the genes within the three cell lines, H, HT, and control (C), were analyzed by statistical methods including Tukey analysis. It was found that from the 12,422 gene fragments and expressed sequence tags (ESTs) analyzed--based on a one-way ANOVA with a stringent cutoff of one false positive per 1000 genes and correcting for microarray background and noise--approximately 950 (7.6%) genes had a significant change in expression in comparing the C, H, and HT groups. A small group of antioxidative defense genes were found in this population, including catalase, members of the glutathione (GSH)-S-transferase family, NAD(P)H menadione oxidoreductase 1, and the ferritin light chain. The remaining genes are involved in a broad spectrum of other biological systems. In the HT versus H comparison, only a few genes were found that had increased expression in the HT line compared with expression in the H line, including GSH-S-transferase alpha 3 and hephaestin. Many genes that are frequently considered antioxidative defense genes, including most of the GSH peroxidases, unexpectedly showed little change. An unusual and generally unexpected small group of antioxidative defense genes appear to have increased expression in response to H2O2 stress. Cell lines resistant to H2O2 do not appear to survive challenge with another type of peroxide, TBHP, a lipid peroxide prototype. However, acquisition of TBHP resistance by H cells was found to be accompanied by significantly amplified expression of only a few additional antioxidative defense genes. Many of the amplified genes do not appear to be involved with antioxidative systems, reflecting the complexity of the cells' response to oxidative stress.


The plant ionome revisited by the nutrient balance concept.
Tissue analysis is commonly used in ecology and agronomy to portray plant nutrient signatures. Nutrient concentration data, or ionomes, belongs to the compositional data class, i.e., multivariate data that are proportions of some whole, hence carrying important numerical properties. Statistics computed across raw or ordinary log-transformed nutrient data are intrinsically biased, hence possibly leading to wrong inferences. Our objective was to present a sound and robust approach based on a novel nutrient balance concept to classify plant ionomes. We analyzed leaf N, P, K, Ca, and Mg of two wild and six domesticated fruit species from Canada, Brazil, and New Zealand sampled during reproductive stages. Nutrient concentrations were (1) analyzed without transformation, (2) ordinary log-transformed as commonly but incorrectly applied in practice, (3) additive log-ratio (alr) transformed as surrogate to stoichiometric rules, and (4) converted to isometric log-ratios (ilr) arranged as sound nutrient balance variables. Raw concentration and ordinary log transformation both led to biased multivariate analysis due to redundancy between interacting nutrients. The alr- and ilr-transformed data provided unbiased discriminant analyses of plant ionomes, where wild and domesticated species formed distinct groups and the ionomes of species and cultivars were differentiated without numerical bias. The ilr nutrient balance concept is preferable to alr, because the ilr technique projects the most important interactions between nutrients into a convenient Euclidean space. This novel numerical approach allows rectifying historical biases and supervising phenotypic plasticity in plant nutrition studies.


Glycemic index and microstructure analysis of a newly developed fiber enriched cookie.
A diet with a high glycemic index (GI) is associated with an elevated risk for obesity or type 2 diabetes. We investigated the GI of a newly-developed fiber enriched cookie and characterized the microstructure of ingredients used. In a study with 26 non-diabetic healthy volunteers it was shown that the fiber enriched cookie has a GI of 58.9 in relation to white bread as reference. Using a conversion factor of 1.4, the GI of the fiber enriched cookie in relation to a glucose-solution is 42.0 and can be classified as a low-GI food. Postprandial insulin concentration was significantly lower after consumption of fiber enriched cookies compared to white bread. Glucose release after in vitro digestion was significantly lower from fiber enriched cookies compared to other cookies tested. In addition to its high percentage of fiber, the cookies' low GI can be attributed to the limited gelatinization potential of the starch granules found in the ingredients used. Using confocal laser scanning microscopy it is shown that starch granule surface area of whole grain barley flour, spelt flour and oat flakes bears cluster-shaped protein-NSPS complexes that preferentially absorb water in conditions of water shortage and thereby prevent starch gelatinization.


An EPDM/MVQ polymer blend based magnetorheological elastomer with good thermostability and mechanical performance.
Magnetorheological elastomers (MREs) with outstanding magnetic-control properties are highly desirable for applications such as vibration attenuation, smart sensing, and soft robots. However, the low strength and thermolability of these materials still restrict their application in attenuating the vibration of large-scale devices. In this paper, we prepared an MRE based on ethylene-propylene-diene monomer (EPDM)/methylvinyl silicone rubber (MVQ) polymer blends. The resulting MRE showed good thermostability and mechanical properties. Good interfacial interaction and particle dispersion were achieved by modifying the surface of carbonyl iron powder (CIP) with silica coating by the sol-gel method. The compatibility between the EPDM and MVQ was promoted using silane coupling agents. Moreover, the resulting MRE had high mechanical strength and elongation at break. The dynamic viscoelastic properties of the MRE were tested using a rheometer. The influences of frequency, strain, matrices, temperature, and magnetic fields were discussed comprehensively, and relevant physical mechanisms were proposed. Finally, thermal aging tests were performed to evaluate the heat resistance of the MRE. Analytical results showed that the resulting MRE could be significantly applied to reduce the vibration of large devices because of its excellent mechanical properties and thermostability.


Quantitative proteomics analysis of macrophage-derived lipid rafts reveals induction of autophagy pathway at the early time of Francisella tularensis LVS infection.
Francisella tularensis is a highly infectious intracellular pathogen that has evolved an efficient strategy to subvert host defense response to survive inside the host. The molecular mechanisms regulating these host-pathogen interactions and especially those that are initiated at the time of the bacterial entry via its attachment to the host plasma membrane likely predetermine the intracellular fate of pathogen. Here, we provide the evidence that infection of macrophages with F. tularensis leads to changes in protein composition of macrophage-derived lipid rafts, isolated as detergent-resistant membranes (DRMs). Using SILAC-based quantitative proteomic approach, we observed the accumulation of autophagic adaptor protein p62 at the early stages of microbe-host cell interaction. We confirmed the colocalization of the p62 with ubiquitinated and LC3-decorated intracellular F. tularensis microbes with its maximum at 1 h postinfection. Furthermore, the infection of p62-knockdown host cells led to the transient increase in the intracellular number of microbes up to 4 h after in vitro infection. Together, these data suggest that the activation of the autophagy pathway in F. tularensis infected macrophages, which impacts the early phase of microbial proliferation, is subsequently circumvented by ongoing infection.


Characterizing a rat Brca2 knockout model.
Evidence exists that BRCA2 carriers may have an elevated risk of breast, ovarian, colon, prostate, and pancreatic cancer. In general, carriers are defined as individuals with protein truncating mutations within the BRCA2 gene. Many Brca2 knockout lines have been produced and characterized in the mouse. We previously produced a rat Brca2 knockout strain in which there is a nonsense mutation in exon 11 between BRC repeats 2 and 3, and a truncated protein is produced. Interestingly, while such a mutation in homozygous mice would lead to limited survival of approximately 3 months, the Brca2-/- rats are 100% viable and the vast majority live to over 1 year of age. Brca2-/- rats show a phenotype of growth inhibition and sterility in both sexes. Aspermatogenesis in the Brca2-/- rats is due to a failure of homologous chromosome synapsis. Long-term phenotypes include underdeveloped mammary glands, cataract formation and lifespan shortening due to the development of tumors and cancers in multiple organs. The establishment of the rat Brca2 knockout model provides a means to study the role of Brca2 in increasing cancer susceptibility and inducing a novel ocular phenotype not previously associated with this gene.


Simulated microgravity combined with polyglycolic acid scaffold culture conditions improves the function of pancreatic islets.
The in vitro culture of pancreatic islets reduces their immunogenicity and prolongs their availability for transplantation. Both simulated microgravity (sMG) and a polyglycolic acid scaffold (PGA) are believed to confer advantages to cell culture. Here, we evaluated the effects of sMG combined with a PGA on the viability, insulin-producing activity and morphological alterations of pancreatic islets. Under PGA-sMG conditions, the purity of the islets was ≥85%, and the islets had a higher survival rate and an increased ability to secrete insulin compared with islets cultured alone in the static, sMG, or PGA conditions. In addition, morphological analysis under scanning electron microscopy (SEM) revealed that the PGA-sMG treatment preserved the integral structure of the islets and facilitated islet adhesion to the scaffolds. These results suggest that PGA-sMG coculture has the potential to improve the viability and function of islets in vitro and provides a promising method for islet transplantation.


Feeling Is Believing: Evaluative Conditioning and the Ethics of Pharmaceutical Advertising.
A central goal in regulating direct-to-consumer advertising of prescription pharmaceuticals (DTCA) is to ensure that explicit drug claims are truthful. Yet imagery can also alter viewer attitudes, and the degree to which this occurs in DTCA is uncertain. Addressing this data gap, we provide evidence that positive feelings produced by images can promote favourable beliefs about pharmaceuticals. We had participants view a fictitious anti-influenza drug paired with unrelated images that elicited either positive, neutral or negative feelings. Participants who viewed positive images rated the influenza drug as significantly more effective, safe, and beneficial than did participants who viewed negative images. This effect, known as evaluative conditioning, is well described in experimental social psychology but has not previously been shown with pharmaceuticals. We discuss how evaluative conditioning in DTCA may compromise viewer autonomy, and canvass possible regulatory responses.


Breathing exercises for asthma: a randomised controlled trial.
The effect of breathing modification techniques on asthma symptoms and objective disease control is uncertain. A prospective, parallel group, single-blind, randomised controlled trial comparing breathing training with asthma education (to control for non-specific effects of clinician attention) was performed. Subjects with asthma with impaired health status managed in primary care were randomised to receive three sessions of either physiotherapist-supervised breathing training (n = 94) or asthma nurse-delivered asthma education (n = 89). The main outcome was Asthma Quality of Life Questionnaire (AQLQ) score, with secondary outcomes including spirometry, bronchial hyper-responsiveness, exhaled nitric oxide, induced sputum eosinophil count and Asthma Control Questionnaire (ACQ), Hospital Anxiety and Depression (HAD) and hyperventilation (Nijmegen) questionnaire scores. One month after the intervention there were similar improvements in AQLQ scores from baseline in both groups but at 6 months there was a significant between-group difference favouring breathing training (0.38 units, 95% CI 0.08 to 0.68). At the 6-month assessment there were significant between-group differences favouring breathing training in HAD anxiety (1.1, 95% CI 0.2 to 1.9), HAD depression (0.8, 95% CI 0.1 to 1.4) and Nijmegen (3.2, 95% CI 1.0 to 5.4) scores, with trends to improved ACQ (0.2, 95% CI 0.0 to 0.4). No significant between-group differences were seen at 1 month. Breathing training was not associated with significant changes in airways physiology, inflammation or hyper-responsiveness. Breathing training resulted in improvements in asthma-specific health status and other patient-centred measures but not in asthma pathophysiology. Such exercises may help patients whose quality of life is impaired by asthma, but they are unlikely to reduce the need for anti-inflammatory medication.


Berry polyphenols metabolism and impact on human gut microbiota and health.
Berries are rich in phenolic compounds such as phenolic acids, flavonols and anthocyanins. These molecules are often reported as being responsible for the health effects attributed to berries. However, their poor bioavailability, mostly influenced by their complex chemical structures, raises the question of their actual direct impact on health. The products of their metabolization, however, may be the most bioactive compounds due to their ability to enter the blood circulation and reach the organs. The main site of metabolization of the complex polyphenols to smaller phenolic compounds is the gut through the action of microorganisms, and reciprocally polyphenols and their metabolites can also modulate the microbial populations. In healthy subjects, these modulations generally lead to an increase in Bifidobacterium, Lactobacillus and Akkermansia, therefore suggesting a prebiotic-like effect of the berries or their compounds. Finally, berries have been demonstrated to alleviate symptoms of gut inflammation through the modulation of pro-inflammatory cytokines and have chemopreventive effects towards colon cancer through the regulation of apoptosis, cell proliferation and angiogenesis. This review recapitulates the knowledge available on the interactions between berries polyphenols, gut microbiota and gut health and identifies knowledge gaps for future research.


Divergent Nodes of Non-autonomous UPRER Signaling through Serotonergic and Dopaminergic Neurons.
In multicellular organisms, neurons integrate a diverse array of external cues to affect downstream changes in organismal health. Specifically, activation of the endoplasmic reticulum (ER) unfolded protein response (UPRER) in neurons increases lifespan by preventing age-onset loss of ER proteostasis and driving lipid depletion in a cell non-autonomous manner. The mechanism of this communication is dependent on the release of small clear vesicles from neurons. We find dopaminergic neurons are necessary and sufficient for activation of cell non-autonomous UPRER to drive lipid depletion in peripheral tissues, whereas serotonergic neurons are sufficient to drive protein homeostasis in peripheral tissues. These signaling modalities are unique and independent and together coordinate the beneficial effects of neuronal cell non-autonomous ER stress signaling upon health and longevity.


Sequence determination and comparison of the exfoliative toxin A and toxin B genes from Staphylococcus aureus.
The DNA encoding the exfoliative toxin A gene (eta) of Staphylococcus aureus was cloned into bacteriophage lambda gt11 and subsequently into plasmid pLI50 on a 1,391-base-pair DNA fragment of the chromosome. Exfoliative toxin A is expressed in the Escherichia coli genetic background, is similar in length to the toxin purified from culture medium, and is biologically active in an animal assay. The nucleotide sequence of the DNA fragment containing the gene was determined. The protein deduced from the nucleotide sequence is a polypeptide of 280 amino acids. The mature protein is 242 amino acids. The DNA sequence of the exfoliative toxin B gene was also determined. Corrections indicate that the amino acid sequence of exfoliative toxin B is in accord with chemical sequence data.


Lipopolysaccharide (LPS)-binding protein and soluble CD14 function as accessory molecules for LPS-induced changes in endothelial barrier function, in vitro.
Bacterial LPS induces endothelial cell (EC) injury both in vivo and in vitro. We studied the effect of Escherichia coli 0111:B4 LPS on movement of 14C-BSA across bovine pulmonary artery EC monolayers. In the presence of serum, a 6-h LPS exposure augmented (P < 0.001) transendothelial 14C-BSA flux compared with the media control at concentrations > or = 0.5 ng/ml, and LPS (10 ng/ml) exposures of > or = 2-h increased (P < 0.005) the flux. In the absence of serum, LPS concentrations of up to 10 micrograms/ml failed to increase 14C-BSA flux at 6 h. The addition of 10% serum increased EC sensitivity to the LPS stimulus by > 10,000-fold. LPS (10 ng/ml, 6 h) failed to increase 14C-BSA flux at serum concentrations < 0.5%, and maximum LPS-induced increments could be generated in the presence of > or = 2.5%. LPS-binding protein (LBP) and soluble CD14 (sCD14) could each satisfy this serum requirement; either anti-LBP or anti-CD14 antibody each totally blocked (P < 0.00005) the LPS-induced changes in endothelial barrier function. LPS-LBP had a more rapid onset than did LPS-sCD14. The LPS effect in the presence of both LBP and sCD14 exceeded the effect in the presence of either protein alone. These data suggest that LBP and sCD14 each independently functions as an accessory molecule for LPS presentation to the non-CD14-bearing endothelial surface. However, in the presence of serum both molecules are required.


LoRTE: Detecting transposon-induced genomic variants using low coverage PacBio long read sequences.
Population genomic analysis of transposable elements has greatly benefited from recent advances of sequencing technologies. However, the short size of the reads and the propensity of transposable elements to nest in highly repeated regions of genomes limits the efficiency of bioinformatic tools when Illumina or 454 technologies are used. Fortunately, long read sequencing technologies generating read length that may span the entire length of full transposons are now available. However, existing TE population genomic softwares were not designed to handle long reads and the development of new dedicated tools is needed. LoRTE is the first tool able to use PacBio long read sequences to identify transposon deletions and insertions between a reference genome and genomes of different strains or populations. Tested against simulated and genuine Drosophila melanogaster PacBio datasets, LoRTE appears to be a reliable and broadly applicable tool to study the dynamic and evolutionary impact of transposable elements using low coverage, long read sequences. LoRTE is an efficient and accurate tool to identify structural genomic variants caused by TE insertion or deletion. LoRTE is available for download at http://www.egce.cnrs-gif.fr/?p=6422.


[Hodgkin's disease in a patient with sarcoidosis].
We present the case of a 37 years old woman with mediastinal lymphadenopathy since the age of 13. At de age of 14, the patient presented a left cervical lymphadenopathy and the onset of episodic nodose erythema. One year later, histological examinations of an enlarged lymph node revealed sarcoidal granulomata. Corticotherapy was prescribed and the patient became asymptomatic. At the age of 31, the patient complained of asthenia and fever with left parotid tumefaction. Corticotherapy was therefore resumed with good response. Three years later, the patient presented left parotid tumefaction, left cervical lymphadenopathy and painful eye congestion. The patient was submitted to parotidectomy and lymphadenectomy. One month later, an ulceration with progressive enlargement developed in the surgical scar, and the patient suffered from asthenia, anorexia, weight loss, fever, nocturnal sweating and cervical and axillary lymphadenopathy. At Santa Maria Hospital, the patient's condition was diagnosed as Hodgkin's disease, mixed cellularity subtype, stage II Bb with local cutaneous involvement. The prescribed treatment was MOPP/ABV-8 cycles- and complete remission was achieved. The patient is still asymptomatic after a follow-up of 6 years. Cutaneous involvement in Hodgkin's disease is rare and generally associated with advanced disease and poor prognosis, facts that did not occur in this case. The differential diagnosis between sarcoidosis and sarcoid reaction in Hodgkin's disease, the accidental coexistence of both entities and the evolution of sarcoidosis into lymphoma are also discussed. The sarcoidosis immune defect may be the result of the same immune disorder. No relationships between sarcoidosis and cutaneous involvement in Hodgkin's disease were found in the literature.


Residual association at C9orf72 suggests an alternative amyotrophic lateral sclerosis-causing hexanucleotide repeat.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of motor neurons. Single-nucleotide polymorphism rs3849942 is associated with ALS, tagging a hexanucleotide repeat mutation in the C9orf72 gene. It is possible that there is more than 1 disease-causing genetic variation at this locus, in which case association might remain after removal of cases carrying the mutation. DNA from patients with ALS was therefore tested for the mutation. Genome-wide association testing was performed first using all samples, and then restricting the analysis to samples not carrying the mutation. rs3849942 and rs903603 were strongly associated with ALS when all samples were included (rs3849942, p = [3 × 2] × 10(-6), rank 7/442,057; rs903603, p = [7 × 6] × 10(-8), rank 2/442,057). Removal of the mutation-carrying cases resulted in loss of association for rs3849942 (p = [2 × 6] × 10(-3), rank 1225/442,068), but had little effect on rs903603 (p = [1 × 9] × 10(-5), rank 8/442,068). Those with a risk allele of rs903603 had an excess of apparent homozygosity for wild type repeat alleles, consistent with polymerase chain reaction failure of 1 allele because of massive repeat expansion. These results indicate residual association at the C9orf72 locus suggesting a second disease-causing repeat mutation.


Melatonin: reducing the toxicity and increasing the efficacy of drugs.
Melatonin (N-acetyl-5-methoxytryptamine) is a molecule with a very wide phylogenetic distribution from plants to man. In vertebrates, melatonin was initially thought to be exclusively of pineal origin recent studies have shown, however, that melatonin synthesis may occur in a variety of cells and organs. The concentration of melatonin within body fluids and subcellular compartments varies widely, with blood levels of the indole being lower than those at many other sites. Thus, when defining what constitutes a physiological level of melatonin, it must be defined relative to a specific compartment. Melatonin has been shown to have a variety of functions, and research in the last decade has proven the indole to be both a direct free radical scavenger and indirect antioxidant. Because of these actions, and possibly others that remain to be defined, melatonin has been shown to reduce the toxicity and increase the efficacy of a large number of drugs whose side effects are well documented. Herein, we summarize the beneficial effects of melatonin when combined with the following drugs: doxorubicin, cisplatin, epirubicin, cytarabine, bleomycin, gentamicin, ciclosporin, indometacin, acetylsalicylic acid, ranitidine, omeprazole, isoniazid, iron and erythropoietin, phenobarbital, carbamazepine, haloperidol, caposide-50, morphine, cyclophosphamide and L-cysteine. While the majority of these studies were conducted using animals, a number of the investigations also used man. Considering the low toxicity of melatonin and its ability to reduce the side effects and increase the efficacy of these drugs, its use as a combination therapy with these agents seems important and worthy of pursuit.


A 15 Year Review (2006-2020) of Patient-Reported Outcome (PRO) in United States Oncology Product Labeling and Trends in Sponsor Size and Oncology Experience.
Despite wide use of PRO tools in clinical development, resulting data is rarely incorporated into the US label. This study reviewed oncology product labels approved by the Food and Drug Administration (FDA) between 2006 and 2020 to determine if the number of PRO included in labeling has meaningfully changed. Sponsors were assessed to identify demographic trends in achieving PRO label success. FDA-approved drugs were searched utilizing the Drugs@FDA database by month from January 2015 to December 2020 for novel drug and biologic approvals. Products approved between 2006-2014 were identified utilizing the Gnansakthy et al., 2012 and 2016 publications. Labels were reviewed for inclusion of PRO data in the label and product summary basis of approval (SBA). Sponsor size and experience were measured for each year of product approval. 155 oncology products received initial approval between 2006-2020, of which only 7 contained PRO data in the label. More than half (53.5%) of products had PRO data described in the SBA. Over time, PRO information has increasingly been included in the product marketing application. Sponsors utilizing PRO data tend to be experienced in oncology development and larger in size. There has been a small increase in inclusion of PRO data in oncology product labeling over the past 15 years. Utilization and analysis of appropriate PRO tools and data remains a challenge to sponsors. Further collaboration with FDA is needed for the development of disease specific PRO tools that provide meaningful data to the targeted patient population.


A guide to reading health care news stories.
From April 16, 2006, through May 30, 2013, a team of reviewers from HealthNewsReview.org, many of whom were physicians, evaluated the reporting by US news organizations on new medical treatments, tests, products, and procedures. After reviewing 1889 stories (approximately 43% newspaper articles, 30% wire or news services stories, 15% online pieces [including those by broadcast and magazine companies], and 12% network television stories), the reviewers graded most stories unsatisfactory on 5 of 10 review criteria: costs, benefits, harms, quality of the evidence, and comparison of the new approach with alternatives. Drugs, medical devices, and other interventions were usually portrayed positively; potential harms were minimized, and costs were ignored. Our findings can help journalists improve their news stories and help physicians and the public better understand the strengths and weaknesses of news media coverage of medical and health topics.


Iterative soft-minimum mean-square error equalization for digital nonlinear page-oriented memories.
Digital page-oriented volume holographic memory (POVHM) is a promising candidate for next-generation ultrahigh capacity optical data storage technology. As the capacity of the POVHMs increases, the bit error rate performance of the system is degraded due to increased interpixel interference (IPI) and noise. To improve the system performance under these adverse effects and to increase the capacity, joint iterative soft equalization-detection and error correction decoding might be attractive. To address that, by considering the nonlinearity inherent in the channel, an iterative soft equalization method that is optimized in the minimum mean-square error (MMSE) sense, called the iterative soft-MMSE (ISMMSE) equalization, is devised. The performance of the ISMMSE is evaluated by use of numerical experiments under different amounts of IPI and optical noise. Simulation results suggest that the ISMMSE is a good candidate for an ultrahigh capacity POVHM, which employs joint iterative equalization-detection and decoding.


Inflammation, macrophage in cancer progression and chinese herbal treatment.
Inflammation is associated with cancer development, and has been recognized as the seventh hallmarks of the cancer. Cancer-related inflammation can be activated by genetic or epigenetic changes in cancer cells (intrinsic pathway) or mediated by tumor-infiltrating immune cells (extrinsic pathway). Immune cells involved in cancer-related inflammation mainly including tumor-associated macrophages or M2 macrophages, neutrophils, dendritic cells, mast cells, and lymphocytes. As major players of the cancer-related inflammation, M2 macrophages, secreting various of growth factors, immunomodulatory cytokines and matrix metalloproteinases, participate in remodeling of extracellular matrix, contribute to cancer invasion and metastasis, angiogenesis, and inhibit anti-cancer immunity. Inflammation has been considered as an important target for cancer therapy. Some Chinese herbal ingredients have been confirmed to be effective in inhibit inflammation related gene expression in cancer cells, such as COX-2 and NF-B. However, there is a shortage of study on Chinese herb or herbal ingredient against extrinsic cancer inflammation, especially in tumor-associated macrophages. Related studies may provide new insight into cancer treatment.


High level of viral suppression and low switch rate to second-line antiretroviral therapy among HIV-infected adult patients followed over five years: retrospective analysis of the DART trial.
In contrast to resource-rich countries, most HIV-infected patients in resource-limited countries receive treatment without virological monitoring. There are few long-term data, in this setting, on rates of viral suppression or switch to second-line antiretroviral therapy. The DART trial compared clinically driven monitoring (CDM) versus routine laboratory (CD4/haematology/biochemistry) and clinical monitoring (LCM) in HIV-infected adults initiating therapy. There was no virological monitoring in either study group during follow-up, but viral load was measured in Ugandan participants at trial closure. Two thousand three hundred and seventeen (2317) participants from this country initiated antiretroviral therapy with zidovudine/lamivudine plus tenofovir (n = 1717), abacavir (n = 300), or nevirapine (n = 300). Of 1896 (81.8%) participants who were alive and in follow-up at trial closure (median 5.1 years after therapy initiation), 1507 (79.5%) were on first-line and 389 (20.5%) on second-line antiretroviral therapy. The overall switch rate after the first year was 5.6 per 100 person-years; the rate was substantially higher in participants with low baseline CD4 counts (<50 cells/mm3). Among 1207 (80.1%) first-line participants with viral load measured, HIV RNA was <400 copies/ml in 963 (79.8%), 400-999 copies/ml in 37 (3.1%), 1,000-9,999 copies/ml in 110 (9.1%), and ≥10,000 copies/ml in 97 (8.0%). The proportion with HIV RNA <400 copies/ml was slightly lower (difference 7.1%, 95% CI 2.5 to 11.5%) in CDM (76.3%) than in LCM (83.4%). Among 252 (64.8%) second-line participants with viral load measured (median 2.3 years after switch), HIV RNA was <400 copies/ml in 226 (89.7%), with no difference between monitoring strategies. Low switch rates and high, sustained levels of viral suppression are achievable without viral load or CD4 count monitoring in the context of high-quality clinical care. ISRCTN13968779.


Alterations of lipid synthesis in the normal and dysmyelinating trembler mouse sciatic nerve by heavy metals (Hg, Pb, Mn, Cu, Ni).
Lipid synthesis from 14C-labeled acetate was dramatically reduced by inorganic mercury in the mouse sciatic nerve in vitro (IC50 was 10 microM). The dysmyelinating trembler mutant was less affected (IC50 was 40 microM). Under the same conditions, lipid synthesis was less inhibited by inorganic lead but was increased 3 times by manganese, copper and nickel at less than 2 mM. Although the synthesis of all lipids is dramatically reduced by inorganic mercury, their relative proportions vary in the presence of this metal:cholesterol synthesis was inhibited most strongly, phosphatidylcholine synthesis was also reduced, whereas synthesis of other lipids was relatively unchanged (phosphatidylserine, phosphatidyl inositol, sphingomyelin, cholesterol esters, cerebrosides). Incorporation of [14C]acetate into free fatty acids was enhanced by a factor of 4 in the presence of inorganic mercury.


Right versus left prefrontal transcranial magnetic stimulation for obsessive-compulsive disorder: a preliminary investigation.
There is preliminary evidence that repetitive transcranial magnetic stimulation (rTMS) may be useful for the treatment of obsessive-compulsive disorder (OCD), but no definitive study has been published, and the effect of laterality of stimulation is uncertain. Subjects (N = 12) with resistant OCD were allocated randomly to either right or left prefrontal rTMS daily for 2 weeks and were assessed by an independent rater at 1 and 2 weeks and 1 month later. Subjects had an overall significant improvement in the obsessions (p < .01), compulsions (p < .01), and total (p < .01) scores on the Yale-Brown Obsessive Compulsive Scale (Y-BOCS) after 2 weeks and at 1-month follow-up. This improvement was significant for obsessions (p < .05) and tended to significance for total Y-BOCS scores (p = .06) after correction for changes in depression scores on the Montgomery-Asberg Depression Rating Scale. There was no significant difference between right- and left-sided rTMS on any of the parameters examined. Two subjects (33%) in each group showed a clinically significant improvement that persisted at I month but with relapse later in I subject. A proportion (about one quarter) of patients with resistant OCD appear to respond to rTMS to either prefrontal lobe, although in the absence of a sham treatment group in this study, we cannot rule out the possibility of this being a placebo response. This treatment warrants further investigation to better establish its efficacy and examine the best parameters for response.


Identification of SNPs associated with variola virus virulence.
Decades after the eradication of smallpox, its etiological agent, variola virus (VARV), remains a threat as a potential bioweapon. Outbreaks of smallpox around the time of the global eradication effort exhibited variable case fatality rates (CFRs), likely attributable in part to complex viral genetic determinants of smallpox virulence. We aimed to identify genome-wide single nucleotide polymorphisms associated with CFR. We evaluated unadjusted and outbreak geographic location-adjusted models of single SNPs and two- and three-way interactions between SNPs. Using the data mining approach multifactor dimensionality reduction (MDR), we identified five VARV SNPs in models significantly associated with CFR. The top performing unadjusted model and adjusted models both revealed the same two-way gene-gene interaction. We discuss the biological plausibility of the influence of the SNPs identified these and other significant models on the strain-specific virulence of VARV. We have identified genetic loci in the VARV genome that are statistically associated with VARV virulence as measured by CFR. While our ability to infer a causal relationship between the specific SNPs identified in our analysis and VARV virulence is limited, our results suggest that smallpox severity is in part associated with VARV strain variation and that VARV virulence may be determined by multiple genetic loci. This study represents the first application of MDR to the identification of pathogen gene-gene interactions for predicting infectious disease outbreak severity.


Cortisol secretion and rate of bone loss in a population-based cohort of elderly men and women.
Although excessive glucocorticoids are a well-recognized cause of osteoporosis, little is known about the role of endogenous glucocorticoids in determining skeletal mass. We have performed a detailed study of the hypothalamic-pituitary-adrenal (HPA) axis to explore the relationships between cortisol secretion and adult bone mass in 151 healthy men and 96 healthy women aged 61 to 73 years. At baseline and 4-year follow-up, bone mineral density (BMD) was measured by dual energy X-ray absorptiometry (DXA) at the lumbar spine and proximal femur; a lifestyle questionnaire was completed; and height, weight, and waist and hip circumferences were measured. At follow-up subjects underwent a very low-dose (0.25 mg) dexamethasone suppression test, a low-dose (1 microg) short synacthen test, and a 24-hour urine collection for measurement of cortisol and its metabolites. In men, elevated peak plasma cortisol was associated with accelerated loss of mineral density in the lumbar spine (r = 0.16, P = 0.05). This relationship remained significant after adjustment for testosterone, estradiol, 25-hydroxyvitamin D, and parathyroid hormone levels (r = 0.22, P = 0.01) and after additional adjustment for age, (BM), activity, cigarette and alcohol consumption, and Kellgren/Lawrence score (r = 0.19, P = 0.03). In contrast in women, elevated peak plasma cortisol was associated with lower baseline BMD at the femoral neck (r = -0.23, P = 0.03) and greater femoral neck loss rate (r = 0.24, P = 0.02). There was no association between plasma cortisol concentrations after dexamethasone or urinary total cortisol metabolite excretion and bone density or bone loss rate at any site. These data provide evidence that circulating endogenous glucocorticoids influence the rate of involutional bone loss in healthy individuals.


Glycosylphosphatidylinositol-anchored Ecm33p influences conidial cell wall biosynthesis in Aspergillus fumigatus.
ECM33 encodes a glycosylphosphatidylinositol-anchored protein whose orthologs in yeast are essential for sporulation. Aspergillus fumigatus Ecm33p is unique and has an apparent mass of 55 kDa. Disruption of A. fumigatus ECM33 results in a mutant with several morphogenetic aberrations, including the following: (i) a defect in conidial separation, (ii) an increase in the diameter of the conidia of the mutant associated with an increase in the concentration of the cell wall chitin, (iii) conidia that were sensitive to the absence of aeration during long-term storage, and (iv) conidia that were more resistant to killing by phagocytes, whereas the mycelium was more easily killed by neutrophils.


Development and characterization of a non-natural nucleoside that displays anticancer activity against solid tumors.
Nucleoside analogs are an important class of anticancer agent that historically show better efficacy against hematological cancers versus solid tumors. This report describes the development and characterization of a new class of nucleoside analog that displays anticancer effects against both hematological and adherent cancer cell lines. These new analogs lack canonical hydrogen-bonding groups yet are effective nucleotide substrates for several high-fidelity DNA polymerases. Permutations in the position of the non-hydrogen-bonding functional group greatly influence the kinetic behavior of these nucleosides. One particular analog designated 4-nitroindolyl-2'-deoxynucleoside triphosphate (4-NITP) is unique as it is incorporated opposite C and T with high catalytic efficiencies. In addition, this analog functions as a nonobligate chain terminator of DNA synthesis, since it is poorly elongated. Consistent with this mechanism, the corresponding nucleoside, 4-nitroindolyl-2'-deoxynucleoside (4-NIdR), produces antiproliferative effects against leukemia cells. 4-NIdR also produces cytostatic and cytotoxic effects against several adherent cancer cell lines, especially those that are deficient in mismatch repair and p53. Cell death in this case appears to occur via mitotic catastrophe, a specialized form of apoptosis. Mass spectroscopy experiments performed on nucleic acid isolated from cells treated with 4-NIdR validate that the non-natural nucleoside is stably incorporated into DNA. Xenograft mouse studies demonstrate that administration of 4-NIdR delays tumor growth without producing adverse side effects such as anemia and thrombocytopenia. Collectively, the results of in vitro, cell-based, and animal studies provide evidence for the development of a novel nucleoside analog that shows enhanced effectiveness against solid tumors.


Evaluation of effects of a novel probiotic feed supplement on the quality of broiler meat.
A local microorganism-based probiotic has been developed as an alternative to using antibiotic growth promoter and its effect on broiler meat quality has been studied, when supplemented with poultry feed at different concentrations. This study aimed to understand the effect of local microorganism-based probiotic sourced from cattle rumen and chicken intestine and added as feed supplement at different concentrations on broiler meat quality. The local microorganism-based probiotic made from cattle rumen and chicken intestine contained Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., and Bacillus spp. The experiments followed a completely randomized design. Treatments in the study were: P0, i.e., control (without probiotic), P1 (probiotic administered at 5 ml/kg feed), P2 (probiotic administered at 10 ml/kg feed), and P3 (probiotic administered at 15 ml/kg feed). Each treatment was repeated 5 times. Parameters examined in this study were pH, meat tenderness, fat content, and meat protein content. Based on a total of 200 chickens, the percentage of meat protein content in treatments P1, P2, and P3 showed an increase of 19.34%, 19.42%, and 19.64%, respectively, when compared with P0 that showed a protein content increase of 19.14%. The fat content of meat for P1, P2, and P3 was 21.54%, 21.46%, and 21.30%, respectively, which was less than the value for P0 (21.69%). The treatments did not significantly affect pH or meat tenderness when compared with the control. The usage of this novel probiotic as a feed supplement resulted in an increase in meat protein content and a decrease in fat content. This study indicates that using the local microorganism-based probiotic sourced from cattle rumen and chicken intestine to supplement poultry feed did not have a significantly different effect (p>0.05) on meat pH; however, it had a significantly different (p<0.05) on protein and fat content of broiler meat.


Evolutionary theory in letters to the editor.
This research note presents the results of a content analysis of 234 letters to the editors that discuss evolutionary theory and were published in American newspapers. We find that letters to the editor both support and hinder the cause of teaching evolutionary theory in American secondary schools. On the one hand, anti-evolutionary theory messages are marginalized in the letters section. This marginalization signals a low level of legitimacy for creationism. It might also contribute to the sense of tension that sustains creationist identities. On the other hand, relatively few letters explicitly note the fact that scientists or the scientific community accept evolution. Interestingly, the obscuration of the scientific community's support for evolutionary theory occurs both in letters supporting and opposing evolutionary theory.


Bacterial isolates degrading ritalinic acid-human metabolite of neuro enhancer methylphenidate.
The consumption of nootropic drugs has increased tremendously in the last decade, though the studies on their environmental fate are still scarce. Nootropics are bioactive compounds designed to alter human's physiology therefore the adverse effects towards wildlife can be expected. In order to understand their environmental impact, the knowledge on their transformation pathways is necessary. Methylphenidate belongs to the most prescribed neuro-enhancers and is among the most favored smart drugs used in non-medical situations. It is metabolized in human liver and excreted as ritalinic acid. Here, we showed for the first time that ritalinic acid can be biodegraded and used as a sole carbon and nitrogen source by various microbial strains originating from different environmental samples. Five axenic strains were isolated and identified as: Arthrobacter sp. strain MW1, MW2 and MW3, Phycicoccus sp. strain MW4 and Nocardioides sp. strain MW5. Our research provides the first insight into the metabolism of ritalinic acid and suggests that it may differ depending on the strain and growth conditions, especially on availability of nitrogen. The isolates obtained in this study can serve as model organisms in further studies on the catabolism of ritalinic acid and methylphenidate but potentially also other compounds with similar structures. Our findings have important implication for the sewage epidemiology. We demonstrated that ritalinic acid is subject to quick and efficient biodegradation thus its use as a stable biomarker should be reconsidered.


Financing of health care in four Caribbean territories: a comparison with reforms in Europe.
This paper considers health care finance in four Caribbean territories and plans for reform in comparison with developments in European countries, to which these territories are historically linked. European health care reforms are aimed at making resource allocation in health care more efficient and more responsive to consumers' demands and preferences. These reforms in Europe have been continuing without appearing to have influenced the developments in the Caribbean very much, except in Martinique. In Trinidad and Tobago current reform entails delegation of responsibility for providing services to four regional health authorities and no purchaser/provider split at the regional or facility level as in the UK has been implemented. In the Bahamas, managed care arrangements are likely to emerge given the proximity of the United States. Recent universal coverage reform in Martinique was aimed at harmonisation of finance by bringing social security and social aid functions together under one management structure and may provide more opportunities for contracting and other initiatives towards greater efficiency. The first priority in Suriname is to restore proper functioning of the current system. Reforms in the four Caribbean territories have a largely administrative character and affect the organisation of the third party role in health care rather than fundamentally changing the relationship between this third party and the various other parties in health care.


The development of infants' circadian rest-activity rhythm and mothers' rhythm.
In order to know when and how infants obtain their circadian sleep-wake rhythm, infants' developing circadian rest-activity rhythm and mothers' circadian rhythm in the postpartum period were examined using actigraph monitoring. The subjects were 11 primipara and their infants. Actigraphic recordings for the infants and their mothers were made over 3-5 continuous days during the 3rd, 6th, 9th and 12th weeks after birth. A 24-h peak on a mean autocorrelogram of the infants' movements was detected at the 3rd week. The infants' circadian rest-activity rhythm already existed in the 3rd week. The amplitude of this 24-h peak gradually increased from the 6th to 12th week. This may be useful as an index of the development of infants' circadian rest-activity rhythm. An 11-h peak was also observed at the 3rd week. This 11-h peak was thought to be a semi-circadian rhythm. Regarding the mothers, the amplitude of the 24-h peak on the mean autocorrelogram at the 3rd week was the smallest of all other weeks, and it became larger from the 3rd to 12th week. This meant that the mothers' circadian rhythm at the 3rd week was influenced by their interrupted sleep at night to take care of their infants. The mother-infant synchronization is probably the 1st factor in the entrainment of infants' circadian sleep-wake rhythm. In this study, we also propose a novel method for compensating for missing data in autocorrelogram analysis.


Apolipoprotein E2 accentuates postprandial inflammation and diet-induced obesity to promote hyperinsulinemia in mice.
Genetic studies have revealed the association between the ε2 allele of the apolipoprotein E (apoE) gene and greater risk of metabolic diseases. This study compared C57BL/6 mice in which the endogenous mouse gene has been replaced by the human APOE2 or APOE3 gene (APOE2 and APOE3 mice) to identify the mechanism underlying the relationship between ε2 and obesity and diabetes. In comparison with APOE3 mice, the APOE2 mice had elevated fasting plasma lipid and insulin levels and displayed prolonged postprandial hyperlipidemia accompanied by increased granulocyte number and inflammation 2 h after being fed a lipid-rich meal. In comparison with APOE3 mice, the APOE2 mice also showed increased adiposity when maintained on a Western-type, high-fat, high-cholesterol diet. Adipose tissue dysfunction with increased macrophage infiltration, abundant crown-like structures, and inflammation were also observed in adipose tissues of APOE2 mice. The severe adipocyte dysfunction and tissue inflammation corresponded with the robust hyperinsulinemia observed in APOE2 mice after being fed the Western-type diet. Taken together, these data showed that impaired plasma clearance of apoE2-containing, triglyceride-rich lipoproteins promotes lipid redistribution to neutrophils and adipocytes to accentuate inflammation and adiposity, thereby accelerating the development of hyperinsulinemia that will ultimately lead to advanced metabolic diseases.


Caffeic Acid Phenethyl Ester from the Twigs of Cinnamomum cassia Inhibits Malignant Cell Transformation by Inducing c-Fos Degradation.
The twigs of Cinnamomum cassia, commonly referred to as Cinnamomi Ramulus, are widely used as one of the primary ingredients in Chinese/Korean traditional medicines that have anticancer effects. However, the active constituents responsible for its anticancer effects and their molecular mechanisms still remain to be elucidated. Caffeic acid phenethyl ester (CAPE) and caffeic acid (CA) were isolated for the first time from C. cassia using LC-MS-guided phytochemical isolation methods. CAPE significantly suppressed EGF- and TPA-induced cell transformation of JB6 P+ cells at sub-micromolar concentrations, whereas CA, a structurally similar compound to CAPE, had no such effect. The antiproliferative and chemopreventive activity of CAPE was found to arise through the inhibition of AP-1 transcriptional activity via the promotion of c-Fos degradation. These findings demonstrate that CAPE may contribute to the chemopreventive/chemotherapeutic effects of C. cassia through downregulating c-Fos.


Segmental, Domain-Selective Perdeuteration and Small-Angle Neutron Scattering for Structural Analysis of Multi-Domain Proteins.
Multi-domain proteins play critical roles in fine-tuning essential processes in cellular signaling and gene regulation. Typically, multiple globular domains that are connected by flexible linkers undergo dynamic rearrangements upon binding to protein, DNA or RNA ligands. RNA binding proteins (RBPs) represent an important class of multi-domain proteins, which regulate gene expression by recognizing linear or structured RNA sequence motifs. Here, we employ segmental perdeuteration of the three RNA recognition motif (RRM) domains in the RBP TIA-1 using Sortase A mediated protein ligation. We show that domain-selective perdeuteration combined with contrast-matched small-angle neutron scattering (SANS), SAXS and computational modeling provides valuable information to precisely define relative domain arrangements. The approach is generally applicable to study conformational arrangements of individual domains in multi-domain proteins and changes induced by ligand binding.


Laparoscopic-assisted tension-free vaginal mesh: an innovative approach to placing synthetic mesh: transvaginally for surgical correction of pelvic organ prolapse.
Polypropylene mesh implants for the correction of pelvic organ prolapse (POP) are now available in Japan. We developed an innovative approach for correcting POP by placing polypropylene mesh transvaginally with laparoscopic assistance. From June 2007 through March 2010, sixteen consecutive patients with symptomatic stage 2 or 3 pelvic organ prolapse underwent the laparoscopic-assisted tension-free vaginal mesh procedure at Okayama University Hospital. All patients were evaluated before and at 1, 3, 6, and 12 months after surgery. Female sexual function was also evaluated with the Female Sexual Function Index (FSFI). The procedure was performed successfully without significant complications. Fifteen of 16 patients were considered anatomically cured (93.8%) at 12 months postoperatively. One patient with a recurrent stage 3 vaginal vault prolapse required sacral colpopexy six months postoperatively. Total FSFI scores improved significantly from 10.3 ± 1.3 at baseline to 18.0 ± 1.2 at 12 months after surgery. The laparoscopic-assisted trans-vaginal mesh is a safe, effective, and simple procedure for POP repairs. The procedure not only restores anatomic relationships but also improves sexual function.


Identification of a point mutation in the thyrotropin receptor of the hyt/hyt hypothyroid mouse.
The hyt/hyt hypothyroid mouse has an autosomal recessive, fetal-onset, severe hypothyroidism related to TSH hyporesponsiveness and associated with elevated TSH. Our previous work has suggested that the hypothyroidism and TSH hyporesponsiveness may result from a mutation in the hyt/hyt TSH receptor (TSHr) of the thyroid gland. Based on DNA sequencing of the entire coding region of the TSHr gene from the wild-type BALB/cBY +/+ mouse, the +/+ TSHr is 92% and 94% identical at the nucleotide and amino acid residue levels, respectively, compared to the rat TSHr gene. The coding region of the hyt/hyt TSHr, compared to that of the +/+ TSHr, has a single base change, CCG to CTG, at nucleotide position 1666, which leads to the replacement of a highly conserved proline at amino acid position 556 with a leucine in transmembrane domain IV. This mutation was introduced by site-directed mutagenesis into the wild-type human TSHr and transiently expressed in COS-7 cells. Although the size and abundance of the mutant TSHr mRNA suggested that there was no effect on the nature of the mRNA, TSH binding and the response to TSH in transfected cells were abolished. Further studies are necessary to clarify how the Pro to Leu replacement interferes with receptor expression on the cell surface or influences TSH binding. These functional consequences of the mutation appear to account for the observed TSH hyporesponsiveness and hypothyroidism in the hyt/hyt mouse.


Engineering the interface between lipid membranes and nanoporous gold: A study by quartz crystal microbalance with dissipation monitoring.
Nanoporous gold (np-Au) is a nanostructured metal with many desirable attributes. Despite the growing number of applications of nanoporous materials, there are still open questions regarding their fabrication and subsequent surface functionalization. For example, the hydrophobic nature of gold surfaces makes the formation of planar supported lipid layers challenging. Here, the authors engineer the interface between np-Au and 1,2-dioleoyl-sn-glycero-3-phosphocholine lipid layers using well-differentiated approaches based on vesicle adsorption and solvent exchange methods. The results reveal that the nanotopography of the np-Au surface plays a clear role in the vesicle adsorption process. Compared to vesicle adsorption, the solvent exchange method proves successful in the formation of planar supported lipid bilayers in both np-Au and planar Au surfaces, being less sensitive to the surface morphological features. The influence of nanostructured surfaces on lipid layer formation is determined by the driving mechanisms behind each process, i.e., the balance of adhesion and cohesion forces in vesicle adsorption and lyotropic lipid phase transitions in solvent exchange, respectively. A better understanding of such interactions will contribute to the development of a variety of applications, from electrochemical biosensors to drug screening and delivery systems, using nanoporous gold coated with stimuli-responsive lipid layers.


Pre-B-cell leukemias in Btk/Slp65-deficient mice arise independently of ongoing V(D)J recombination activity.
The adapter protein Slp65 and Bruton's tyrosine kinase (Btk) are key components of the precursor-B (pre-B) cell receptor (pre-BCR) signaling pathway. Slp65-deficient mice spontaneously develop pre-B-cell leukemia, expressing high levels of the pre-BCR on their cell surface. As leukemic Slp65-deficient pre-B cells express the recombination activating genes (Rag)1 and Rag2, and manifest ongoing immunoglobulin (Ig) light-chain rearrangement, it has been hypothesized that deregulated recombinase activity contributes to malignant transformation. In this report, we investigated whether Rag-induced DNA damage is involved in oncogenic transformation of Slp65-deficient B cells. We employed Btk/Slp65 double-deficient mice carrying an autoreactive 3-83μδ BCR transgene. When developing B cells in their bone marrow express this BCR, the V(D)J recombination machinery will be activated, allowing for secondary Ig light-chain gene rearrangements to occur. This phenomenon, called receptor editing, will rescue autoreactive B cells from apoptosis. We observed that 3-83μδ transgenic Btk/Slp65 double-deficient mice developed B-cell leukemias expressing both the 3-83μδ BCR and the pre-BCR components λ5/VpreB. Importantly, such leukemias were found at similar frequencies in mice concomitantly deficient for Rag1 or the non-homologous end-joining factor DNA-PKcs. We therefore conclude that malignant transformation of Btk/Slp65 double-deficient pre-B cells is independent of deregulated V(D)J recombination activity.


Development of a theoretical screening tool to assess caries risk in Nevada youth.
One objective of this study was to determine the prevalence and severity of caries among Nevada youth, subsequently comparing these data with national statistics. A second objective was to identify the risk factors associated with caries prevalence and severity in order to develop and tailor a theoretical screening tool for this cohort for future validation. Researchers computed the prevalence rates of dental caries (D-score) and severity rates of decayed, missing, and filled teeth indices in a cohort of 9202 students, 13 to 18 years of age, attending public/private schools in the 2005/2006 academic year. Multiple regression established which of the 13 variables significantly contributed to caries risk, subsequently using logistic regression to ascertain the weight of contribution and odds ratios of significant variables. Living in counties with no municipal water fluoridation, increased exposure to environmental smoke, minority race, living in rural communities, and increasing age were the largest significant contributors (respectively). Exposure to tobacco, being female, lack of dental insurance, increased body mass index risk, and lack of dental sealants were also significant, but to a lesser extent. Nonsignificant factors included socioeconomic status, ethnicity, and family history of diabetes. This study confirmed high caries prevalence and severity and identified significant risk factors for inclusion in a theoretical risk screening tool for future validation and translation for use in the early detection of caries risk in Nevada youth.


Intensive Hemodialysis and Treatment Complications and Tolerability.
Hemodialysis (HD) treatment can be difficult to tolerate. Common complications are intradialytic hypotension (IDH) and long time to recovery after an HD session. IDH, as defined by nadir systolic blood pressure < 90mmHg and intradialytic decline > 30mmHg, occurs in almost 8% of HD sessions. IDH may be caused by aggressive ultrafiltration in response to interdialytic weight gain, can lead to myocardial stunning and cardiac arrhythmias, and is associated with increased risk for death. Long recovery time after a treatment session is also common. In DOPPS (Dialysis Outcomes and Practice Patterns Study), recovery time was 2 to 6 hours for 41% of HD patients and longer than 6 hours for 27%; recovery time was linearly associated with increased risks for death and hospitalization. Importantly, both decreases in blood pressure and feeling washed out or drained have been identified by patients as more important outcomes than death or hospitalization. Intensive HD likely reduces the likelihood of IDH. In the Frequent Hemodialysis Network trial, short daily and nocturnal schedules reduced the per-session probability of IDH by 20% and 68%, respectively, relative to 3 sessions per week. Due to lower ultrafiltration volume and/or rate, intensive HD may reduce intradialytic blood pressure variability. In a cross-sectional study, short daily and nocturnal schedules were associated with slower ultrafiltration and less dialysis-induced myocardial stunning than 3 sessions per week. In FREEDOM (Following Rehabilitation, Economics, and Everyday-Dialysis Outcome Measurements), a prospective cohort study of short daily HD, recovery time was reduced after 12 months from 8 hours to 1 hour, according to per-protocol analysis. Recovery time after nocturnal HD may be minutes. In conclusion, intensive HD can improve the tolerability of HD treatment by reducing the risk for IDH and decreasing recovery time after HD. These changes may improve the patient centeredness of end-stage renal disease care.


Simulation of propagation along an isolated skeletal muscle fiber in an isotropic volume conductor.
This paper describes a model of the frog skeletal muscle fiber that includes the effects of the transverse tubular system (T system) on propagation. Uniform propagation on an isolated fiber suspended in Ringer's solution or in air is simulated by placing the cylindrical fiber model in a concentric three-dimensional isotropic volume conductor. The current through the T system outlets at the sarcolemmal surface is comparable in magnitude to the sarcolemmal current density, but is of opposite polarity. When it is added to the sarcolemmal current, the resulting triphasic waveform has a 100% increase in the leading positive peak, a 50% reduction in the negative peak, and more than 60% reduction in the trailing positive peak. As a result the tubular output current causes a reduction in the conduction velocity, a decrease in the maximum rate of rise of the action potential. and an important modification of the extracellular potential. Compared to an isolated fiber in a large volume of Ringer's solution, uniform propagation within a 2-micron-thick volume conductor annulus is slowed down from 1.92 to 0.72 m/s, and the extracellular potential is increased from 1 to 108 mV peak to peak, in agreement with published experimental measurements.


Nationwide survey of comorbidity, use, and costs of all medications in Finnish diabetic individuals.
To investigate comorbidity and overall use and costs of medication for all Finnish individuals with diabetes treated with drugs compared with sex- and age-matched control subjects. According to a cross-sectional population study using national registries, 116,224 individuals purchased antidiabetic medications in Finland in 1995. The same number of nondiabetic individuals matched for sex, age, and area of residence were chosen as control subjects. Age at onset of diabetes was used as a criterion for distinguishing between type 1 and type 2 diabetes. The criterion could be applied in 74% of cases. A total of 16,955 individuals were defined as having type 1 diabetes, and 68,517 were defined as having type 2 diabetes. Comorbidity was determined by linkage with a national register including all individuals entitled to special reimbursement for drug treatment for a range of chronic diseases. Data on use and costs of all medications prescribed were obtained from drug purchase records. Cardiovascular diseases and uremia were, as expected, the chronic diseases most closely associated with diabetes. Use of almost all kinds of medication was significantly greater in individuals with type 1 and type 2 diabetes than in control subjects. The greatest differences were observed in relation to cardiovascular drugs and antibiotics. Unexpectedly low use of antiasthmatics was observed in individuals with both types of diabetes, low use of neuroleptics was observed in type 1 diabetic individuals, and low use of hormone replacement therapy was observed in women with type 2 diabetes. Total costs of medications for individuals with diabetes were 3.5 times greater than those for nondiabetic control subjects. The higher costs were mostly attributable to insulin therapy for individuals with type 1 diabetes. The higher costs for individuals with type 2 diabetes were related to the cost of medications other than antidiabetic medication. The possible selection bias in omitting diabetic individuals treated with diet only and individuals in whom diabetes type could not be determined must be considered in interpreting the results. Greater use by and costs of medications for individuals with diabetes than for nondiabetic individuals is related not only to antidiabetic treatment but also to all other kinds of medications. Although drug treatment and the prevalence of several chronic conditions were overall greater in individuals with diabetes versus other individuals, some exceptions merit further study


Bond strength of glass-ionomer cement and composite resin combinations.
The tensile bond strength to dentin was measured for three glass-ionomer cement and composite resin combinations: two light-curing glass-ionomer cements (Vitrebond and XR-Ionomer) and one traditional glass-ionomer cement (Ketac-Bond), two adhesive systems (Scotchbond, and XR-Bonding System), and a corresponding composite resin. The bond strength of this "sandwich" was also compared with that of the same cements used in bulk. Vitrebond showed a significantly higher bond strength in bulk than did the other two cements. Of the sandwiches, the XR-Ionomer and XR-Bond combination showed a bond strength significantly higher than that of the Vitrebond and Scotchbond or Ketac-Bond and Scotchbond combination. The fracture of the bond was mainly adhesive for Vitrebond, cohesive for XR-Ionomer when used in bulk and adhesive-cohesive when used in a sandwich, and cohesive for Ketac-Bond.


Medical Informatics and Information Technology Supporting Oral Medicine.
Electronic healthcare documentation is the key element of electronic healthcare (eHealth). Electronic oral health record (EOHR) supporting oral medicine is discussed. To provide dentists with a methodology and instrument to create oral health documentation in more efficient way, support information exchange and integration in dental domain and to ease dental decision-making and forensic dentistry identification tasks.The proposed methodology is used to model lifelong EOHR based on a small specific ontology where the use of other classification systems and nomenclatures, e.g. SNODENT, is possible. EOHRwith Lifelong DentCross user interface was developed and it has been supporting dental care at the University Hospital in Prague-Motol. The user interface is working in four languages and controlled by voice or keyboard. Lifelong DentCross user interface is reflecting the way of the work in dentistry and the EOHR can provide both structured and free text information to oral medicine.


Molecular layering of fluorinated ionic liquids at a charged sapphire (0001) surface.
Room-temperature ionic liquids (RTILs) are promising candidates for a broad range of "green" applications, for which their interaction with solid surfaces plays a crucial role. In this high-energy x-ray reflectivity study, the temperature-dependent structures of three ionic liquids with the tris(pentafluoroethyl)trifluorophosphate anion in contact with a charged sapphire substrate were investigated with submolecular resolution. All three RTILs show strong interfacial layering, starting with a cation layer at the substrate and decaying exponentially into the bulk liquid. The observed decay length and layering period point to an interfacial ordering mechanism, akin to the charge inversion effect, which is suggested to originate from strong correlations between the unscreened ions. The observed layering is expected to be a generic feature of RTILs at charged interfaces.


Caffeine increases whole-body fat oxidation during 1 h of cycling at Fatmax.
The ergogenic effect of caffeine on exercise of maximum intensity has been well established. However, there is controversy regarding the effect of caffeine on shifting substrate oxidation at submaximal exercise. The aim of this study was to investigate the effect of acute caffeine ingestion on whole-body substrate oxidation during 1 h of cycling at the intensity that elicits maximal fat oxidation (Fatmax). In a double-blind, randomized, and counterbalanced experiment, 12 healthy participants (VO2max = 50.7 ± 12.1 mL/kg/min) performed two acute experimental trials after ingesting either caffeine (3 mg/kg) or a placebo (cellulose). The trials consisted of 1 h of continuous cycling at Fatmax. Energy expenditure, fat oxidation rate, and carbohydrate oxidation rate were continuously measured by indirect calorimetry. In comparison to the placebo, caffeine increased the amount of fat oxidized during the trial (19.4 ± 7.7 vs 24.7 ± 9.6 g, respectively; P = 0.04) and decreased the amount of carbohydrate oxidized (94.6 ± 30.9 vs 73.8 ± 32.4 g; P = 0.01) and the mean self-perception of fatigue (Borg scale = 11 ± 2 vs 10 ± 2 arbitrary units; P = 0.05). In contrast, caffeine did not modify total energy expenditure (placebo = 543 ± 175; caffeine = 559 ± 170 kcal; P = 0.60) or mean heart rate (125 ± 13 and 127 ± 9 beats/min; P = 0.30) during exercise. Before exercise, caffeine increased systolic and diastolic blood pressure whilst it increased the feelings of nervousness and vigour after exercise (P < 0.05). These results suggest that a moderate dose of caffeine (3 mg/kg) increases the amount of fat oxidized during 1 h of cycling at Fatmax. Thus, caffeine might be used as an effective strategy to enhance body fat utilization during submaximal exercise. The occurrence of several side effects should be taken into account when using caffeine to reduce body fat in populations with hypertension or high sensitivity to caffeine.


Projected impacts of climate change on environmental suitability for malaria transmission in West Africa.
Climate change is expected to affect the distribution of environmental suitability for malaria transmission by altering temperature and rainfall patterns; however, the local and global impacts of climate change on malaria transmission are uncertain. We assessed the effect of climate change on malaria transmission in West Africa. We coupled a detailed mechanistic hydrology and entomology model with climate projections from general circulation models (GCMs) to predict changes in vectorial capacity, an indication of the risk of human malaria infections, resulting from changes in the availability of mosquito breeding sites and temperature-dependent development rates. Because there is strong disagreement in climate predictions from different GCMs, we focused on the GCM projections that produced the best and worst conditions for malaria transmission in each zone of the study area. Simulation-based estimates suggest that in the desert fringes of the Sahara, vectorial capacity would increase under the worst-case scenario, but not enough to sustain transmission. In the transitional zone of the Sahel, climate change is predicted to decrease vectorial capacity. In the wetter regions to the south, our estimates suggest an increase in vectorial capacity under all scenarios. However, because malaria is already highly endemic among human populations in these regions, we expect that changes in malaria incidence would be small. Our findings highlight the importance of rainfall in shaping the impact of climate change on malaria transmission in future climates. Even under the GCM predictions most conducive to malaria transmission, we do not expect to see a significant increase in malaria prevalence in this region.


Field application of farmstead runoff to vegetated filter strips: surface and subsurface water quality assessment.
Farmstead runoff poses significant environmental impacts to ground and surface waters. Three vegetated filter strips were assessed for the treatment of dairy farmstead runoff at the soil surface and subsurface at 0.3- or 0. 46-m and 0. 76-m depths for numerous storm events. A medium-sized Michigan dairy was retrofitted with two filter strips on sandy loam soil and a third filter strip was implemented on a small Michigan dairy with sandy soil to collect and treat runoff from feed storage, manure storage, and other impervious farmstead areas. All filter strips were able to eliminate surface runoff via infiltration for all storm events over the duration of the study, eliminating pollutant contributions to surface water. Subsurface effluent was monitored to determine the contributing groundwater concentrations of numerous pollutants including chemical oxygen demand (COD), metals, and nitrates. Subsurface samples have an average reduction of COD concentrations of 20, 11, and 85% for the medium dairy Filter Strip 1 (FS1), medium dairy Filter Strip 2 (FS2), and the small Michigan dairy respectively, resulting in average subsurface concentrations of 355, 3960, and 718 mg L COD. Similar reductions were noted for ammonia and total Kjeldahl nitrogen (TKN) in the subsurface effluent. The small Michigan dairy was able to reduce the pollutant leachate concentrations of COD, TKN, and ammonia over a range of influent concentrations. Increased influent concentrations in the medium Michigan dairy filter strips resulted in an increase in COD, TKN, and ammonia concentrations in the leachate. Manganese was leached from the native soils at all filter strips as evidenced by the increase in manganese concentrations in the leachate. Nitrate concentrations were above standard drinking water limits (10 mg L), averaging subsurface concentrations of 11, 45, and 25 mg L NO-N for FS1, FS2, and the small Michigan dairy, respectively.


Direct correlation between hyperoxaluria/oxalate stone disease and the absence of the gastrointestinal tract-dwelling bacterium Oxalobacter formigenes: possible prevention by gut recolonization or enzyme replacement therapy.
Oxalobacter formigenes is a specific oxalate-degrading, anaerobic bacterium inhabiting the gastrointestinal tracts of vertebrates, including humans. This bacterium maintains an important symbiotic relationship with its host by regulating oxalate homeostasis, primarily by preventing enteric absorption. Increased absorption of oxalate can lead to multiple complications associated with hyperoxaluria, especially recurrent calcium oxalate urolithiasis. Detection of O. formigenes in the gastrointestinal tract has attracted attention because the absence of this bacterium appears to be a risk factor for development of hyperoxaluria and/or recurrent calcium oxalate kidney stone disease. In the present study, epidemiologic studies with patients at high risk for calcium oxalate urolithiasis showed a direct correlation between the number of recurrent kidney stone episodes and the lack of O. formigenes colonization. As expected, the lack of O. formigenes revealed a clear association with prophylactic antibiotic therapy. To confirm the importance of O. formigenes in regulating hyperoxaluria, laboratory rats known to be noncolonized were colonized with live bacteria or treated with a preparation of oxalate-degrading enzymes derived from O. formigenes to determine any subsequent increased resistance to high oxalate challenge. Rats receiving either bacteria or enzyme replacement therapy excreted far lower levels of oxalate, did not develop the crystalluria observed with control rats, and resisted the formation of calcium oxalate crystals in their nephrons. These observations, taken together, support the concept that O. formigenes is important in maintaining oxalate homeostasis, that its absence from the gut increases the risk for hyperoxaluria and recurrent kidney stone disease, and that replacement therapy is an efficient procedure to prevent hyperoxaluria and its complications.


Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting….
Defining and manipulating specific neurons in the brain has garnered enormous interest in recent years, because such an approach is now widely recognized as crucial for deepening our understanding of how the brain works. When I started exploring the Cre-loxP recombination for brain research in the early 1990s, it was written off as a dead-end project by a young fool. Yet over the past 20 years, Cre-lox recombination-mediated neurogenetics has emerged as one of the most powerful and versatile technology platforms for cell-specific gene knockouts, transgenic overexpression, Brainbow imaging, neural pathway tracing with retrovirus and CLARITY, chemical genetics, and optogenetics. Its popularity and greater utility in neuroscience research is also largely thanks to the NIH's bold Blueprint for Neuroscience Research Initiative to launch several Cre-driver resource projects, as well as individual laboratories and private research organizations. With newly-discovered, genetically-encoded molecules that are capable of responding to sonar and magnetic stimulation, for sonogenetics or magnetogenetics, respectively, or detecting rapid voltage changes in neurons, Cre-lox neurogenetics will continue to aid brain research for years to come.


Transgenesis upgrades for Drosophila melanogaster.
Drosophila melanogaster is a highly attractive model system for the study of numerous biological questions pertaining to development, genetics, cell biology, neuroscience and disease. Until recently, our ability to manipulate flies genetically relied heavily on the transposon-mediated integration of DNA into fly embryos. However, in recent years significant improvements have been made to the transgenic techniques available in this organism, particularly with respect to integrating DNA at specific sites in the genome. These new approaches will greatly facilitate the structure-function analyses of Drosophila genes, will enhance the ease and speed with which flies can be manipulated, and should advance our understanding of biological processes during normal development and disease.


Survey of population coverage in cervical cancer screening in the Oxford region.
A postal survey of a random sample of 3307 women aged 18-64 years living in the Oxford region (the Oxford healthy life survey) revealed age and social class differences in the proportion of women reporting an up-to-date cervical smear test. Although 86% of women in the 25-34 years age group reported a test in -the previous five years, the proportion dropped to 53% among the 55-64 year olds. Women in social classes 4 and 5 were significantly less likely to report a recent smear test (P<0.01) than women in higher social classes. An audit of the general practice notes of 176 of the respondents suggested that these self-reported results are likely to be reasonably accurate.A reduction in the number of deaths from cervical cancer will largely depend on the efforts of primary health care teams to screen women who are at relatively high risk of developing the disease, namely older women and those of lower social class.


Exercise adherence, cardiopulmonary fitness and anthropometric changes improve exercise self-efficacy and health-related quality of life.
Regular exercise increases exercise self-efficacy and health-related quality of life (HRQOL); however, the mechanisms are unknown. We examined the associations of exercise adherence and physiological improvements with changes in exercise self-efficacy and HRQOL. Middle-aged adults (N = 202) were randomized to 12 months aerobic exercise (360 minutes/week) or control. Weight, waist circumference, percent body fat, cardiopulmonary fitness, HRQOL (SF-36), and exercise self-efficacy were assessed at baseline and 12 months. Adherence was measured in minutes/day from activity logs. Exercise adherence was associated with reduced bodily pain, improved general health and vitality, and reduced role-emotional scores (P(trend) ≤ 0.05). Increased fitness was associated with improved physical functioning, bodily pain and general health scores (P(trend) ≤ 0.04). Reduced weight and percent body fat were associated with improved physical functioning, general health, and bodily pain scores (P(trend) < 0.05). Decreased waist circumference was associated with improved bodily pain and general health but with reduced role-emotional scores (P(trend) ≤ 0.05). High exercise adherence, increased cardiopulmonary fitness and reduced weight, waist circumference and percent body fat were associated with increased exercise self-efficacy (P(trend) < 0.02). Monitoring adherence and tailoring exercise programs to induce changes in cardiopulmonary fitness and body composition may lead to greater improvements in HRQOL and self-efficacy that could promote exercise maintenance.


Adenosine acting at an A1 receptor decreases N-type calcium current in mouse motoneurons.
The neuromodulator adenosine is known to decrease neurotransmitter release at the neuromuscular junction by activation of an A1 adenosine receptor coupled to a pertussis toxin-sensitive G protein. Among the mechanisms that could contribute to the depression of neurotransmitter release is reduced entry of calcium through channels located in the presynaptic terminal. In the present study, we have examined the effects of adenosine on high-voltage-activated (HVA) calcium currents in motoneurons, the presynaptic cells of the neuromuscular junction. The motoneurons were isolated from embryonic mice, placed in primary tissue culture for 16 hr, and analyzed by means of the whole-cell patch-clamp technique. Adenosine (40 microM) reduced both transient and sustained components of HVA calcium current. This effect was blocked by the A1 antagonist 8-cyclopentyltheophylline (CPT; 100 nM) and was mimicked by the A1 agonist N6-cyclohexyladenosine (CHA; 50 nM to 10 microM) but not by the A2a agonist 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamido adenosine (CGS-21680; 1 micron). Pretreatment with pertussis toxin (200 ng/ml, > 16 hr) abolished the depression of HVA calcium current by adenosine receptor activation. Brief (3 min) exposure of the cells to 10 microM omega-conotoxin GVIA irreversibly blocked a part of the HVA current, which can therefore be attributed to N-type channels; the remaining current was unaffected by adenosine receptor activation. Hence, it appears that adenosine decreases only the N-current portion of HVA current and that this inhibition occurs via an A1 receptor linked to a pertussis toxin-sensitive G protein. Other investigators have shown that N-type channels do not play a primary role in eliciting transmitter release at the mammalian neuromuscular junction. Thus, it is uncertain what motoneuronal functions are influenced by adenosine modulation of N-type channels.


Efficacy and honesty in communication between relatives.
The evolution of honest communication has recently become the focus of intense theoretical attention. However, strategic models dealing with honesty have largely ignored the implications of noise and perceptual error for signal evolution (just as models dealing with signal detection in the presence of noise ignore strategic issues). Here, I analyze an extended version of Maynard Smith's strategic model of signaling of need between relatives, the Philip Sidney game, that incorporates the possibility of perceptual error. I show that even in the presence of noise, there exists over a wide range of parameter values a unique, continuously stable signaling equilibrium, at which the signaler employs a costly display when needy but refrains from doing so when healthy. For a subset of this range, there also exists a second, lower cost signaling equilibrium that is not continuously stable. At the former equilibrium, predicted signal cost is inversely related to the coefficient of relatedness (r) between signaler and receiver. Cost is not, however, predicted to drop to zero even when r = 1 and there is no conflict of interest between the two (as is the case in errofree models), because it serves to enhance the efficacy of communication as well as to discourage deceit. Equilibrium signal cost is inversely related to the probability that the signaler is needy, and tends to increase with the level of noise. If noise becomes too great (i.e., if a detectable signal is too costly to produce), signaling is no longer stable; surprisingly, it is also unstable if the level of noise is too low (i.e., if a detectable signal is too cheap to produce).


Risk of adverse events in children completing treatment for acute lymphoblastic leukemia: St. Jude Total Therapy studies VIII, IX, and X.
We studied the frequency, causes, and predictors of adverse events in 624 patients who had completed treatment for acute lymphoblastic leukemia (ALL) in three consecutive total therapy studies (VII, IX, and X, 1972 to 1983). Event-free survival in study X was significantly better overall than that in studies VIII and IX (P less than .0001 by the log-rank test). In study X, 75% of the patients were electively taken off therapy, compared with 54% in studies VIII and IX. However, the risks of having an adverse event during the first 5 years after completion of therapy were remarkably similar: 22% (95% confidence interval, 17% to 29%) in study X versus 24% (20% to 29%) in studies VIII and IX. Bone marrow, testicular, and CNS relapses accounted for the majority of failures in both groups (85% in study X and 92% in studies VIII and IX). Late adverse events consisted largely of hematologic relapses and the development of solid tumors. Black race (P = .001) and leukemia without an anterior mediastinal mass (P = .05) were associated with an increased risk of failure after completion of treatment in the two earlier clinical trials, whereas a lower leukemic cell DNA content (DNA index less than 1.16) was the only predictor of late treatment failure in the more recent trial (P = .019). None of the other presenting features that were examined (eg, age, leukocyte count, and sex) had value as predictors of late failure. Thus, improved treatment altered the impact of specific prognostic factors and the distribution of sites of relapse, but it did not significantly affect the risk of delayed failure.


Going forward: Increasing the accessibility of imaging mass spectrometry.
The driving force behind the high and increasing popularity of imaging mass spectrometry is its demonstrated potential for the determination of new diagnostic/prognostic biomarkers and its ability to simultaneously trace the distributions of pharmaceuticals and their metabolites in tissues without the need to develop expensive radioactively-labeled analogues. Both of these applications would benefit from standardized methods, for the development of novel MS-based molecular histology tests and governmental-approved MS-based assays for pharmaceutical development. In addition, the broader scientific community would benefit from the increased accessibility of the technique. Currently imaging MS studies are individual endeavors, utilizing the individual expertise and infrastructure of a single laboratory and their immediate collaborators. A wide array of tissue preparation, data acquisition and data analysis techniques has been developed but lacks an international collaborative structure and data sharing capabilities. Such a collaborative framework would enable methodological exchange and detailed comparisons of analytical capabilities, to explore synergies between the different methods and result in the development of robust standardized methods. Here we describe the activities of a new European imaging MS network that will explicitly compare and contrast existing methods to provide best practice guidelines for the entire healthcare research community.