Cannabinoid-induced mesenteric vasodilation through an endothelial site distinct from CB1 or CB2 receptors.
Cannabinoids, including the endogenous ligand arachidonyl ethanolamide (anandamide), elicit not only neurobehavioral but also cardiovascular effects. Two cannabinoid receptors, CB1 and CB2, have been cloned, and studies with the selective CB1 receptor antagonist SR141716A have implicated peripherally located CB1 receptors in the hypotensive action of cannabinoids. In rat mesenteric arteries, anandamide-induced vasodilation is inhibited by SR141716A, but other potent CB1 receptor agonists, such as HU-210, do not cause vasodilation, which implicates an as-yet-unidentified receptor in this effect. Here we show that "abnormal cannabidiol" (Abn-cbd) is a neurobehaviorally inactive cannabinoid that does not bind to CB1 receptors, yet causes SR141716A-sensitive hypotension and mesenteric vasodilation in wild-type mice and in mice lacking CB1 receptors or both CB1 and CB2 receptors. Hypotension by Abn-cbd is also inhibited by cannabidiol (20 microgram/g), which does not influence anandamide- or HU-210-induced hypotension. In the rat mesenteric arterial bed, Abn-cbd-induced vasodilation is unaffected by blockade of endothelial NO synthase, cyclooxygenase, or capsaicin receptors, but it is abolished by endothelial denudation. Mesenteric vasodilation by Abn-cbd, but not by acetylcholine, sodium nitroprusside, or capsaicine, is blocked by SR141716A (1 microM) or by cannabidiol (10 microM). Abn-cbd-induced vasodilation is also blocked in the presence of charybdotoxin (100 nM) plus apamin (100 nM), a combination of K(+)-channel toxins reported to block the release of an endothelium-derived hyperpolarizing factor (EDHF). These findings suggest that Abn-cbd and cannabidiol are a selective agonist and antagonist, respectively, of an as-yet-unidentified endothelial receptor for anandamide, activation of which elicits NO-independent mesenteric vasodilation, possibly by means of the release of EDHF.


A splicing mutation in the novel mitochondrial protein DNAJC11 causes motor neuron pathology associated with cristae disorganization, and lymphoid abnormalities in mice.
Mitochondrial structure and function is emerging as a major contributor to neuromuscular disease, highlighting the need for the complete elucidation of the underlying molecular and pathophysiological mechanisms. Following a forward genetics approach with N-ethyl-N-nitrosourea (ENU)-mediated random mutagenesis, we identified a novel mouse model of autosomal recessive neuromuscular disease caused by a splice-site hypomorphic mutation in a novel gene of unknown function, DnaJC11. Recent findings have demonstrated that DNAJC11 protein co-immunoprecipitates with proteins of the mitochondrial contact site (MICOS) complex involved in the formation of mitochondrial cristae and cristae junctions. Homozygous mutant mice developed locomotion defects, muscle weakness, spasticity, limb tremor, leucopenia, thymic and splenic hypoplasia, general wasting and early lethality. Neuropathological analysis showed severe vacuolation of the motor neurons in the spinal cord, originating from dilatations of the endoplasmic reticulum and notably from mitochondria that had lost their proper inner membrane organization. The causal role of the identified mutation in DnaJC11 was verified in rescue experiments by overexpressing the human ortholog. The full length 63 kDa isoform of human DNAJC11 was shown to localize in the periphery of the mitochondrial outer membrane whereas putative additional isoforms displayed differential submitochondrial localization. Moreover, we showed that DNAJC11 is assembled in a high molecular weight complex, similarly to mitofilin and that downregulation of mitofilin or SAM50 affected the levels of DNAJC11 in HeLa cells. Our findings provide the first mouse mutant for a putative MICOS protein and establish a link between DNAJC11 and neuromuscular diseases.


A reverse transcriptase-like activity of wheat (Triticum aestivum) embryo microsomal fraction.
A reverse transcriptase-like activity was isolated from germinating wheat (Triticum aestivum) embryos. The activity was found to be associated with a microsomal fraction (70,000 g pellet) of the embryo homogenate. The microsome-associated enzyme prefers homologous polyadenylated RNA to any other polynucleotides as template and requires all four deoxyribonucleoside triphosphates for maximal activity. The reaction product appears in the incubation mixture in the form of an RNA-DNA hybrid, which can be converted into single-stranded DNA by mild alkaline hydrolysis. These observations suggest that normal wheat embryo cells contain an enzyme which, functionally, is similar to retroviral reverse transcriptase.


The role of antibiotic therapy in the management of acute appendicitis.
Nonsurgical treatment with antibiotics has recently been proposed as the first line of treatment for noncomplicated appendicitis. This has met with considerable interest, illustrated by the number of reviews and meta-analyses, which exceed the number of original reports of the issue. The results in these studies are seriously biased due to inclusion of patients with resolving appendicitis. At a time when we need to reduce inappropriate use of antibiotics in the struggle against the increasing rate of antibiotics resistance, there must be strong requirements of a proven effect and an improved cost-benefit ratio before antibiotics treatment is introduced for a new group of patients. These requirements have not yet been met for nonsurgical treatment with antibiotics for assumed uncomplicated appendicitis. Due to the high rate of spontaneous resolution, a randomized placebo-controlled trial is needed that can compare the efficiency of antibiotics treatment and expectant management in this group of patients. Antibiotics treatment, however, remains indicated for treatment of perforated appendicitis with localized abscess or phlegmone and in selected surgical high-risk patients.


Neurodevelopmental Effects of Undernutrition and Placental Underperfusion in Fetal Growth Restriction Rabbit Models.
Chronic reduction of oxygen and nutrient delivery to the fetus has been related to neurodevelopmental problems. Placental underperfusion induces a significant reduction in oxygen and nutrient delivery, whereas maternal undernutrition causes mainly nutrient deficiency. A comparison of the neurodevelopmental effects of both situations in pregnant rabbits was performed. The placental underperfusion model was induced after uteroplacental vessel ligation at 25 days of pregnancy. The undernutrition model was induced after a reduction of 70% of the basal maternal intake at 22 days of pregnancy. Neurobehavioral tests were applied in the derived offspring at the neonatal period and over the long term. Structural brain differences were evaluated by brain networks obtained from diffusion magnetic resonance imaging. Birth weight was significantly lower in both cases. However, stillbirth was only increased in the placental underperfusion model. Cases from both models presented poorer neurobehavioral performance and network infrastructure, being more pronounced in the placental underperfusion model. Prenatal insults during the last third of gestation resulted in functional and structural disturbances. The degree of neurodevelopmental impairment and its association with structural brain reorganization seemed to be related to the type of the prenatal insult, showing stronger effects in the placental underperfusion model.


Childhood trauma and adult prostitution behavior in a multiethnic heterosexual drug-using population.
A cross-sectional study of the association between severity of childhood trauma and adult prostitution behaviors was conducted among 676 heterosexual drug addicts in San Antonio, Texas. Three hundred and fifty eight women and 338 men taking part in a national multisite program for AIDS prevention research completed the Childhood Trauma Questionnaire as part of a comprehensive risk behavior assessment. Women addicts in the sample were less educated, more likely to be in a common-law relationship, living with someone of the opposite sex or separated, and had lower incomes in comparison to men addicts. Among male subjects,higher educational levels and older age were positively associated with prostitution activities. Single female subjects were three times more likely to engage in selling sex than married subjects. Single women with higher incomes were more likely to be prostituting than single women with lower incomes. Black women reporting severe degrees of emotional abuse, emotional neglect, or physical neglect were more likely to engage in prostitution behavior than Hispanic or white women with similar levels of trauma. Black men with a history of childhood physical abuse were more likely to use prostitutes than Hispanic or white men.


Synthesis and transport of lipoprotein lipase in perfused guinea pig hearts.
Total lipoprotein lipase (LPL) activity did not differ significantly between hearts from fed or fasted guinea pigs. Incorporation of [35S]methionine into immunoprecipitable LPL was also the same. The rates at which perfused hearts from fed or fasted guinea pigs released LPL activity into the medium were, however, different (2 vs. 4 mU.g-1.min-1). These rates remained constant over 60 min of perfusion. Addition of heparin to the medium resulted in a peak of LPL activity during the first 2 min, followed by a shoulder of relatively high activity, which gradually declined to a constant rate from 30 min. The peak and shoulders were less with hearts from fed animals than with hearts from fasted animals, but the constant rates were similar. Cycloheximide added at the start of the perfusion had no effect on the peak or on the early part of the shoulder, but the LPL activity released from 30 min continuously decreased so that at 60 min it was less than half of that in controls. Studies in which the enzyme was pulse labeled by perfusion 15 min with medium containing [35S]methionine and then chased up to 75 min with unlabeled medium showed no differences in how LPL is transported and metabolized in hearts from fed vs. fasted guinea pigs. Thus the data suggest that factors outside the heart influence the disposition of heart LPL in vivo.


6-gingerdiols as the major metabolites of 6-gingerol in cancer cells and in mice and their cytotoxic effects on human cancer cells.
6-Gingerol, a major pungent component of ginger (Zingiber officinale Roscoe, Zingiberaceae), has been reported to have antitumor activities. However, the metabolic fate of 6-gingerol and the contribution of its metabolites to the observed activities are still unclear. In the present study, we investigated the biotransformation of 6-gingerol in different cancer cells and in mice, purified and identified the major metabolites from human lung cancer cells, and determined the effects of the major metabolites on the proliferation of human cancer cells. Our results show that 6-gingerol is extensively metabolized in H-1299 human lung cancer cells, CL-13 mouse lung cancer cells, HCT-116 and HT-29 human colon cancer cells, and in mice. The two major metabolites in H-1299 cells were purified and identified as (3R,5S)-6-gingerdiol (M1) and (3S,5S)-6-gingerdiol (M2) based on the analysis of their 1D and 2D NMR data. Both metabolites induced cytotoxicity in cancer cells after 24 h, with M1 having a comparable effect to 6-gingerol in H-1299 cells.


An essential nuclear envelope integral membrane protein, Brr6p, required for nuclear transport.
Despite rapid advances in our understanding of the function of the nuclear pore complex in nuclear transport, little is known about the role the nuclear envelope itself may play in this critical process. A small number of integral membrane proteins specific to the envelope have been identified in budding yeast, however, none has been reported to affect transport. We have identified an essential gene, BRR6, whose product, Brr6p, behaves like a nuclear envelope integral membrane protein. Notably, the brr6-1 mutant specifically affects transport of mRNA and a protein reporter containing a nuclear export signal. In addition, Brr6p depletion alters nucleoporin distribution and nuclear envelope morphology, suggesting that the protein is required for the spatial organization of nuclear pores. BRR6 interacts genetically with a subset of nucleoporins, and Brr6-green fluorescent protein (GFP) localizes in a punctate nuclear rim pattern, suggesting location at or near the nuclear pore. However, Brr6-GFP fails to redistribute in a (Delta)nup133 mutant, distinguishing Brr6p from known proteins of the pore membrane domain. We hypothesize that Brr6p is located adjacent to the nuclear pore and interacts functionally with the pore and transport machinery.


Effects of menopausal status on circulating calcitonin gene-related peptide and adipokines: implications for insulin resistance and cardiovascular risks.
To determine, first, the effects of menopausal status on circulating calcitonin gene-related peptide (CGRP) levels and, second, the correlation between circulating CGRP levels and biomarkers for cardiovascular disease. Cross-sectional study of healthy premenopausal and postmenopausal women volunteers and women admitted for elective benign abdominal surgery in a district general hospital. All women were non-smokers, had no history of endocrinological problems and were not receiving any hormone therapy. Fasting blood samples (premenopausal (n = 45): follicle stimulating hormone (FSH) < 20 IU/l, estradiol (mean +/- SEM) 440.33 +/- 51.82 pmol/l; postmenopausal women (n = 28): FSH > 20 IU/l, estradiol 93.79 +/- 17.40 pmol/l) were analyzed for CGRP, resistin, leptin, adiponectin, insulin and lipids using ELISA and immunoassays. Mean circulating CGRP levels were higher in the postmenopausal women compared with premenopausal women (pre: 41.79 +/- 9.01 pg/ml, post: 138.14 +/- 45.75 pg/ml; p = 0.047). Among women who were experiencing hot flushes, the postmenopausal women had significantly higher CGRP levels than the premenopausal women (pre: 21.98 +/- 4.95 pg/ml, post: 171.08 +/- 61.80 pg/ml; p = 0.028). Serum CGRP levels positively correlated with serum insulin levels (r = 0.652, p = 0.016) and HOMA index (r = 0.54, p < 0.001). These data show that circulating CGRP levels are influenced by menopausal status and suggest additional mechanisms through which increased risk of hyperinsulinemia and cardiovascular disease may arise in postmenopausal women.


Human leucine zipper protein promotes hepatic steatosis via induction of apolipoprotein A-IV.
The molecular mechanism of stress-induced hepatic steatosis is not well known. Human leucine zipper protein (LZIP) regulates the expression of genes involved in inflammation, cell migration, and stress response. The aim of this study was to determine the regulatory role of LZIP in stress-induced hepatic steatosis. We used a microarray analysis to identify LZIP-induced genes involved in hepatic lipid metabolism. LZIP increased the expression of apolipoprotein A-IV (APOA4) mRNA. In the presence of stress inducer, APOA4 promoter analysis was performed, and LZIP-induced lipid accumulation was monitored in mouse primary cells and human tissues. Under Golgi stress conditions, LZIP underwent proteolytic cleavage and was phosphorylated by AKT to protect against proteasome degradation. The stabilized N-terminal LZIP was translocated to the nucleus, where it directly bound to the APOA4 promoter, leading to APOA4 induction. LZIP-induced APOA4 expression resulted in increased absorption of surrounding free fatty acids. LZIP also promoted hepatic steatosis in mouse liver. Both LZIP and APOA4 were highly expressed in human steatosis samples. Our findings indicate that LZIP is a novel modulator of APOA4 expression and hepatic lipid metabolism. LZIP might be a therapeutic target for developing treatment strategies for hepatic steatosis and related metabolic diseases.-Kang, M., Kim, J., An, H.-T., Ko, J. Human leucine zipper protein promotes hepatic steatosis via induction of apolipoprotein A-IV.


Initial development and psychometric testing of an instrument to measure the quality of children's end-of-life care.
The field of pediatric palliative care is hindered by the lack of a well-defined, reliable, and valid method for measuring the quality of end-of-life care. The study purpose was to develop and test an instrument to measure mothers' perspectives on the quality of care received before, at the time of, and following a child's death. In Phase 1, key components of quality end-of-life care for children were synthesized through a comprehensive review of research literature. These key components were validated in Phase 2 and then extended through focus groups with bereaved parents. In Phase 3, items were developed to assess structures, processes, and outcomes of quality end-of-life care then tested for content and face validity with health professionals. Cognitive testing was conducted through interviews with bereaved parents. In Phase 4, bereaved mothers were recruited through 10 children's hospitals/hospices in Canada to complete the instrument, and psychometric testing was conducted. Following review of 67 manuscripts and 3 focus groups with 10 parents, 141 items were initially developed. The overall content validity index for these items was 0.84 as rated by 7 health professionals. Based on feedback from health professionals and cognitive testing with 6 parents, a 144-item instrument was finalized for further testing. In Phase 4, 128 mothers completed the instrument, 31 of whom completed it twice. Test-retest reliability, internal consistency, and construct validity were demonstrated for six subscales: Connect With Families, Involve Parents, Share Information With Parents, Share Information Among Health Professionals, Support Parents, and Provide Care at Death. Additional items with content validity were grouped in four domains: Support the Child, Support Siblings, Provide Bereavement Follow-up, and Structures of Care. Forty-eight items were deleted through psychometric testing, leaving a 95-item instrument. There is good initial evidence for the reliability and validity of this new quality of end-of-life care instrument as a mechanism for evaluative feedback to health professionals, health systems, and policy makers to improve children's end-of-life care.


Routine pathologic parameters can predict Oncotype DX recurrence scores in subsets of ER positive patients: who does not always need testing?
Oncotype DX(TM) is an RT-PCR-based assay used to predict chemotherapy benefit in patients with estrogen receptor (ER) positive breast cancers. We were interested if routinely available pathologic parameters could predict Oncotype DX Recurrence Scores (RS) in subsets of patients. We identified 173 breast cancers with available RSs and used 104 of these as a test set and 69 cases as a validation set. Pathologic characteristics including size, histologic type, Nottingham grade, and lymphatic invasion were recorded. Test set cases were stained for ER, progesterone receptor (PR), HER2, Ki67, CyclinD1, BCL2, D2-40, and P53. Statistical correlations with RS and regression tree analysis were performed. The validation set was subjected to analysis on the basis of grade, PR, and Ki67. In the test set, grade, PR levels and Ki67 had the strongest correlation with RS (P = 0.0002-0.0007). Regression tree analysis showed grade and PR as factors that could segregate cases into RS categories, with Ki67 adding value in certain subsets. A subset of cancers with a high likelihood of having a low RS (0-18) was identified with the following characteristics: grade 1, strong PR expression (Allred score ≥ 5) and Ki67 ≤ 10%. No cases with these characteristics had a high RS (≥ 31) and 73% had a low RS. Cancers highly likely to have a high RS were grade 3, low to absent PR expression (Allred score <5) and Ki67 > 10%. 80% of cases with these characteristics had a high RS and no cases had a low RS. Our validation set had similar findings in these two subsets. In conclusion, When cost and time are a consideration and the added value of Oncotype DX(TM) testing is in question, it may be reasonable to assume the results of this test in two specific subsets of breast cancers: (1) grade 1, high PR, low Ki67 cancers (low RS), and (2) grade 3, low PR, high Ki67 cancers (high RS).


Profiling lipid-protein interactions using nonquenched fluorescent liposomal nanovesicles and proteome microarrays.
Fluorescent liposomal nanovesicles (liposomes) are commonly used for lipid research and/or signal enhancement. However, the problem of self-quenching with conventional fluorescent liposomes limits their applications because these liposomes must be lysed to detect the fluorescent signals. Here, we developed a nonquenched fluorescent (NQF)1 liposome by optimizing the proportion of sulforhodamine B (SRB) encapsulant and lissamine rhodamine B-dipalmitoyl phosphatidylethanol (LRB-DPPE) on a liposomal surface for signal amplification. Our study showed that 0.3% of LRB-DPPE with 200 μm of SRB provided the maximal fluorescent signal without the need to lyse the liposomes. We also observed that the NQF liposomes largely eliminated self-quenching effects and produced greatly enhanced signals than SRB-only liposomes by 5.3-fold. To show their application in proteomics research, we constructed NQF liposomes that contained phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2) and profiled its protein interactome using a yeast proteome microarray. Our profiling led to the identification of 162 PI(3,5)P2-specific binding proteins (PI(3,5)P2-BPs). We not only recovered many proteins that possessed known PI(3,5)P2-binding domains, but we also found two unknown Pfam domains (Pfam-B_8509 and Pfam-B_10446) that were enriched in our dataset. The validation of many newly discovered PI(3,5)P2-BPs was performed using a bead-based affinity assay. Further bioinformatics analyses revealed that the functional roles of 22 PI(3,5)P2-BPs were similar to those associated with PI(3,5)P2, including vesicle-mediated transport, GTPase, cytoskeleton, and kinase. Among the 162 PI(3,5)P2-BPs, we found a novel motif, HRDIKP[ES]NJLL that showed statistical significance. A docking simulation showed that PI(3,5)P2 interacted primarily with lysine or arginine side chains of the newly identified PI(3,5)P2-binding kinases. Our study showed that this new tool would greatly benefit profiling lipid-protein interactions in high-throughput studies.


Antibacterial modification of an injectable, biodegradable, non-cytotoxic block copolymer-based physical gel with body temperature-stimulated sol-gel transition and controlled drug release.
Biomaterials are being extensively used in various biomedical fields; however, they are readily infected with microorganisms, thus posing a serious threat to the public health care. We herein presented a facile route to the antibacterial modification of an important A-B-A type biomaterial using poly (ethylene glycol) methyl ether (mPEG)- poly(ε-caprolactone) (PCL)-mPEG as a typical model. Inexpensive, commercial bis(2-hydroxyethyl) methylammonium chloride (DMA) was adopted as an antibacterial unit. The effective synthesis of the antibacterial copolymer mPEG-PCL-∼∼∼-PCL-mPEG (where ∼∼∼ denotes the segment with DMA units) was well confirmed by FTIR and (1)H NMR spectra. At an appropriate modification extent, the DMA unit could render the copolymer mPEG-PCL-∼∼∼-PCL-mPEG highly antibacterial, but did not largely alter its fascinating intrinsic properties including the thermosensitivity (e.g., the body temperature-induced sol-gel transition), non-cytotoxicity, and controlled drug release. A detailed study on the sol-gel-sol transition behavior of different copolymers showed that an appropriate extent of modification with DMA retained a sol-gel-sol transition, despite the fact that a too high extent caused a loss of sol-gel-sol transition. The hydrophilic and hydrophobic balance between mPEG and PCL was most likely broken upon a high extent of quaternization due to a large disturbance effect of DMA units at a large quantity (as evidenced by the heavily depressed PCL segment crystallinity), and thus the micelle aggregation mechanism for the gel formation could not work anymore, along with the loss of the thermosensitivity. The work presented here is highly expected to be generalized for synthesis of various block copolymers with immunity to microorganisms. Light may also be shed on understanding the phase transition behavior of various multiblock copolymers.


A stakeholder evaluation of the impact of the palliative care clinical nurse specialist upon doctors and nurses, within an acute hospital setting.
There has been an expansion in the number of palliative care teams based in the acute hospital setting. Although organization of these teams varies both in structure and approach, the clinical nurse specialist is one of the key members. The last decade has seen an escalation in the UK of clinical nurse specialists, and following the Calman-Hine Report and the more recent NHS Cancer Plan, it is anticipated that the number of clinical nurse specialists in palliative and cancer care is likely to grow. This paper presents the qualitative findings of an evaluation study designed to investigate the impact of the clinical nurse specialist within a palliative care team based in an acute hospital setting. A stakeholder evaluation that encompassed 31 tape-recorded semi-structured interviews with senior nurses, consultants, junior doctors and nurses representing different grades were performed. The data was analysed for emerging themes utilising a case and cross case analysis methodology. The results suggested the presence of the clinical nurse specialists is seen as beneficial to both medical and nursing staff. This paper will focus upon exploring these potential benefits, included providing support, advice and education.


[The organization of germinal material and dynamics of mother sporocyst reproduction in the genus Echinostoma (Trematoda: Echinostomatidae)].
The reproduction in the first parthenogenetic generation--mother sporocyst (MS) in two species of echinostomes (E. caproni, E. paraensei) is investigated. A group of densely packed cells, which noticeably differ from others, occupies the posterior part of the miracidium. They are characterized by large sizes and a large bubble-shaped nucleus with heterogeneous nucleolus and strong dispersed chromatin. The use of histological and electron microscopic methods has shown that with observed similarity these cells are classified in two tyoes and have a completely different origin. First of all, large secretory cells stand out. In E. caproni miracidia their number averages 6.8 +/- 0.2 and linear sizes is 10-12 microns. Secretory cells possess a large bubble-shaped nucleus. The caryoplasm looks optically empty because of strong dispersion of chromatin. A large nucleolus occupies a bit eccentric position. Eosinophilic cytoplasm contains poorly noticeable at light-optical level accumulation of small granules. The second group of cells is represented by typical germinal cells (GC). The number of GC does not exceed six. Their polymorphy is well above that of secretory cells. The sizes of GC vary from 5.4 to 9 microns. The largest cells (8.1-9 microns) occupy the front position and usually are located between secretory cells. Intensively basophilic cytoplasm surrounds bubble-shaped nucleus with a large nucleolus like border with uniforming thickness. The heterochromatin is evenly distributed over the caryoplasm. Its content of nuclei is more than that in nuclei of secretory cells. That is why they do not look optically empty. So, it is "mature" germinal cells. Four or five cells are located directly behind "mature" cells. Their sizes are gradually decrease towards the posterior of the miracidium (the diameter of the smallest cells reaches 5.4 microns). Nuclei with a centrally located nucleolus are characterized by larger amount and more condensation of the heterochromatin than those in "mature" cells. Meanwhile, they concern to nuclei of bubble-shaped type. In general, all cells of second type represent the primary germinal cells distinguished by the stage of their differentiation. Also, 2-3 undifferentiated cells occupy the most posterior part of the miracidium. Their sizes average 5.55 +/- 0.18 microns. The nucleus contains a lot of densely packed heterochromatin. On parasitic phase of MS development undifferentiated cells give rise to secondary GC. Electron microscopic data in details confirm the situation described above. The essentially similar results were received during the investigation of E. Paraensei miracidia. The differences are observed in parameterical characteristics of germinal material and in small variability of the extent of germinal material development. With E. paraensei, germinal material may be represented by not only GC and undifferentiated cells, but one germ as well. So, our investigation has shown that germinal material of echinostomes represents typical germinal mass. The germinal material condition does not change on parasitic phase of E. caproni MS development during the first day of post infection (PI). The activization of germinal material coincides in the time with the beginning of schizocoel formation in 2 Days PI. On the 3rd day of PI, the proliferation of undifferentiated cells begins and the first germs are free to float in enormous schizocoel. After 8 days of PI, MS release the first rediae. During the following 2-3 days the other rediae formed by primary GC left MS. The release of rediae derived from secondary GC was observed later. So, E. caproni MS give rise to 12-16 rediae which is much less than the number of GC formed in MS. The earlier release of the first E. paraensei rediae by MS is predetermined by the difference in the structure of germinal mass in E. paraensei miracidia. Therefore, Echinostomatidae is intermediate between two groups of trematodes. The first group has MS that completely realize reproductive function in the time of miracidial formation; but the second group includes higher trematodes characterized by the transfer of reproductive time on a parasitic phase of MS development. The question concerning to so-called "pedogenetic larvae of trematodes" is discussed.


A Vaccine Platform against Arenaviruses Based on a Recombinant Hyperattenuated Mopeia Virus Expressing Heterologous Glycoproteins.
Several Old World and New World arenaviruses are responsible for severe endemic and epidemic hemorrhagic fevers, whereas other members of the Arenaviridae family are nonpathogenic. To date, no approved vaccines, antivirals, or specific treatments are available, except for Junín virus. However, protection of nonhuman primates against Lassa fever virus (LASV) is possible through the inoculation of the closely related but nonpathogenic Mopeia virus (MOPV) before challenge with LASV. We reasoned that this virus, modified by using reverse genetics, would represent the basis for the generation of a vaccine platform against LASV and other pathogenic arenaviruses. After showing evidence of exoribonuclease (ExoN) activity in NP of MOPV, we found that this activity was essential for multiplication in antigen-presenting cells. The introduction of multiple mutations in the ExoN site of MOPV NP generated a hyperattenuated strain (MOPVExoN6b) that is (i) genetically stable over passages, (ii) has increased immunogenic properties compared to those of MOPV, and (iii) still promotes a strong type I interferon (IFN) response. MOPVExoN6b was further modified to harbor the envelope glycoproteins of heterologous pathogenic arenaviruses, such as LASV or Lujo, Machupo, Guanarito, Chapare, or Sabia virus in order to broaden specific antigenicity while preserving the hyperattenuated characteristics of the parental strain. Our MOPV-based vaccine candidate for LASV, MOPEVACLASV, was used in a one-shot immunization assay in nonhuman primates and fully protected them from a lethal challenge with LASV. Thus, our hyperattenuated strain of MOPV constitutes a promising new live-attenuated vaccine platform to immunize against several, if not all, pathogenic arenaviruses.IMPORTANCE Arenaviruses are emerging pathogens transmitted to humans by rodents and responsible for endemic and epidemic hemorrhagic fevers of global concern. Nonspecific symptoms associated with the onset of infection make these viruses difficult to distinguish from other endemic pathogens. Moreover, the unavailability of rapid diagnosis in the field delays the identification of the virus and early care for treatment and favors spreading. The vaccination of exposed populations would be of great help to decrease morbidity and human-to-human transmission. Using reverse genetics, we generated a vaccine platform for pathogenic arenaviruses based on a modified and hyperattenuated strain of the nonpathogenic Mopeia virus and showed that the Lassa virus candidate fully protected nonhuman primates from a lethal challenge. These results showed that a rationally designed recombinant MOPV-based vaccine is safe, immunogenic, and efficacious in nonhuman primates.


Early caregiving stress exposure moderates the relation between respiratory sinus arrhythmia reactivity at 1 month and biobehavioral outcomes at age 3.
There is a growing scientific interest in the psychophysiological functioning of children living in low-socioeconomic status (SES) contexts, though this research is complicated by knowledge that physiology-behavior relations often operate differently in these environments among adults. Importantly, such research is made more difficult because SES may be a proxy for a wide range of risk factors including poor caregiving and exposure to parental substance use. We used factor analysis to organize risk-exposure data collected from 827 children-many of whom were raised in low-SES contexts and exposed to substances prenatally-into dissociable components including economic stress, caregiving stress (e.g., stress the caregiver may experience, including parental psychopathology), and postnatal substance exposure. These factors, along with respiratory sinus arrhythmia (RSA) reactivity at age 1 month, were used to predict behavior dysregulation and resting RSA at age 3 years. A significant RSA Reactivity × Caregiving Stress interaction indicated that infants who exhibited high RSA reactivity at 1 month experienced the greatest behavior dysregulation at 3 years, but only when they were exposed to high levels of caregiving stress. Among African Americans, the highest resting RSA at 3 years was found in infants with less RSA reactivity, but only if they also experienced less caregiving stress. Our work is consistent with biological sensitivity to context, adaptive calibration, and allostatic load models, and highlights the importance of studying Physiology × Environment interactions in low-SES contexts for predicting behavior and resting RSA.


Axonopathy in the Central Nervous System Is the Hallmark of Mice with a Novel Intragenic Null Mutation of Dystonin.
Dystonia musculorum is a neurodegenerative disorder caused by a mutation in the dystonin gene. It has been described in mice and humans where it is called hereditary sensory autonomic neuropathy. Mutated mice show severe movement disorders and die at the age of 3-4 weeks. This study describes the discovery and molecular, clinical, as well as pathological characterization of a new spontaneously occurring mutation in the dystonin gene in C57BL/6N mice. The mutation represents a 40-kb intragenic deletion allele of the dystonin gene on chromosome 1 with exactly defined deletion borders. It was demonstrated by Western blot, mass spectrometry, and immunohistology that mice with a homozygous mutation were entirely devoid of the dystonin protein. Pathomorphological lesions were restricted to the brain stem and spinal cord and consisted of swollen, argyrophilic axons and dilated myelin sheaths in the white matter and, less frequently, total chromatolysis of neurons in the gray matter. Axonal damage was detected by amyloid precursor protein and nonphosphorylated neurofilament immunohistology. Axonopathy in the central nervous system (CNS) represents the hallmark of this disease. Mice with the dystonin mutation also showed suppurative inflammation in the respiratory tract, presumably due to brain stem lesion-associated food aspiration, whereas skeletal muscles showed no pathomorphological changes. This study describes a novel mutation in the dystonin gene in mice leading to axonopathy in the CNS. In further studies, this model may provide new insights into the pathogenesis of neurodegenerative diseases and may elucidate the complex interactions of dystonin with various other cellular proteins especially in the CNS.


Effect of board game activities on cognitive function improvement among older adults in adult day care centers.
Stimulating leisure activities are considered as possible protective factors against dementia and cognitive decline in older adults, particularly due to the enhancement of cognitive reserve. This study tested the effectiveness of board game activities improving the cognitive function of older adults in adult day care centers. This was a quasi-experimental study. A purposive sampling strategy was used to select 82 subjects who were aged 65 and above with intact mental functions and currently residing in adult day care centers. 41 subjects who participated in a selection of 12 board game activities were assigned to the experimental group and 41 subjects who adhered to their ordinary activities were allocated to the control group. Structured questionnaires of the board game programs were used for data collection. The board game programs showed promising effects in the cognitive function of older adults living in adult day care centers. A possible beneficial effect of board game playing on the risk of dementia could be mediated by a less cognitive decline in older adults. Board game activities may benefit the cognitive function of older adults. Incorporating board game activities into social work care may help develop long-term care into a more diverse, unique and innovative direction.


Subjective well-being and human welfare around the world as reflected in the Gallup World Poll.
We present data on well-being and quality of life in the world, including material quality of life such as not going hungry, physical health quality of life such as longevity, social quality of life such as social support, environmental health such as clean water, equality in income and life satisfaction, and levels of subjective well-being (SWB). There are large differences between nations in SWB, and these are predicted not only by economic development, but also by environmental health, equality and freedom in nations. Improving trends in SWB are seen in many countries, but declining SWB is evident in a few. Besides average differences in SWB between nations, there are also large disparities within many countries. We discuss the policy opportunities provided by national accounts of SWB, which are increasingly being adopted by many societies. They provide the opportunity to inform policy deliberations with well-being information that reflects not only economic development, but also other facets of quality of life as well. National accounts of SWB reflect the quality of life in areas such as health, social relationships and the natural environment, and therefore capture a broader view of societal well-being than afforded by measures of economic progress alone.


Measurement properties of oral health assessments for non-dental healthcare professionals in older people: a systematic review.
Regular inspection of the oral cavity is required for prevention, early diagnosis and risk reduction of oral- and general health-related problems. Assessments to inspect the oral cavity have been designed for non-dental healthcare professionals, like nurses. The purpose of this systematic review was to evaluate the content and the measurement properties of oral health assessments for use by non-dental healthcare professionals in assessing older peoples' oral health, in order to provide recommendations for practice, policy, and research. A systematic search in PubMed, EMBASE.com, and Cinahl (via Ebsco) has been performed. Search terms referring to 'oral health assessments', 'non-dental healthcare professionals' and 'older people (60+)' were used. Two reviewers individually performed title/abstract, and full-text screening for eligibility. The included studies have investigated at least one measurement property (validity/reliability) and were evaluated on their methodological quality using "The Consensus-based Standards for the selection of health Measurement Instruments" (COSMIN) checklist. The measurement properties were then scored using quality criteria (positive/negative/indeterminate). Out of 879 hits, 18 studies were included in this review. Five studies showed good methodological quality on at least one measurement property and 14 studies showed poor methodological quality on some of their measurement properties. None of the studies assessed all measurement properties of the COSMIN. In total eight oral health assessments were found: the Revised Oral Assessment Guide (ROAG); the Minimum Data Set (MDS), with oral health component; the Oral Health Assessment Tool (OHAT); The Holistic Reliable Oral Assessment Tool (THROAT); Dental Hygiene Registration (DHR); Mucosal Plaque Score (MPS); The Brief Oral Health Screening Examination (BOHSE) and the Oral Assessment Sheet (OAS). Most frequently assessed items were: lips, mucosa membrane, tongue, gums, teeth, denture, saliva, and oral hygiene. Taken into account the scarce evidence of the proposed assessments, the OHAT and ROAG are most complete in their included oral health items and are of best methodological quality in combination with positive quality criteria on their measurement properties. Non-dental healthcare professionals, policymakers and researchers should be aware of the methodological limitations of the available oral health assessments and realize that the quality of the measurement properties remains uncertain.


A uniform bacterial growth potential assay for different water types.
The bacterial growth potential is important to understand and manage bacterial regrowth-related water quality concerns. Bacterial growth potential depends on growth promoting/limiting compounds, therefore, nutrient availability is the key factor governing bacterial growth potential. Selecting proper tools for bacterial growth measurement is essential for routine implementation of the growth potential measurement. This study proposes a growth potential assay that is universal and can be used for different water types and soil extract without restrictions of pure culture or cultivability of the bacterial strain. The proposed assay measures the sample bacterial growth potential by using the indigenous community as inocula. Flow cytometry (FCM) and adenosine tri-phosphate (ATP) were used to evaluate the growth potential of six different microbial communities indigenous to the sample being analyzed, with increasing carbon concentrations. Bottled mineral water, non-chlorinated tap water, seawater, river water, wastewater effluent and a soil organic carbon extract were analyzed. Results showed that indigenous bacterial communities followed normal batch growth kinetics when grown on naturally present organic carbon. Indigenous bacterial growth could detect spiked organic carbon concentrations as low as 10 μg/L. The indigenous community in all samples responded proportionally to the increase in acetate-carbon and proportional growth could be measured with both FCM and ATP. Bacterial growth was proportional to the carbon concentration but not the same proportion factor for the different water samples tested. The effect of inoculating the same water with different indigenous microbial communities on the growth potential was also examined. The FCM results showed that the highest increase in total bacterial cell concentration was obtained with bacteria indigenous to the water sample. The growth potential assay using indigenous bacterial community revealed consistent results of bacterial growth in all the different samples tested and therefore providing a fast, more stable, and accurate approach for monitoring the biological stability of waters compared to the previously developed assays. The growth potential assay can be used to aid in detecting growth limitations by compounds other than organic carbon.


Developmental neurotoxicity of different pesticides in PC-12 cells in vitro.
The detection of developmental neurotoxicity (DNT) of chemicals has high relevance for protection of human health. However, DNT of many pesticides is only little known. Furthermore, validated in vitro systems for assessment of DNT are not well established. Here we employed the rat phaeochromocytoma cell line PC-12 to evaluate DNT of 18 frequently used pesticides of different classes, including neonicotinoids, pyrethroids, organophosphates, organochlorines, as well as quaternary ammonium compounds, the organic compound used in pesticides, piperonyl butoxide, as well as the insect repellent diethyltoluamide (DEET). We determined the outgrowth of neurites in PC-12 cells co-treated with nerve growth factor and different concentrations of biocides for 5days. Furthermore, we determined transcriptional alterations of selected genes that may be associated with DNT, such as camk2α and camk2β, gap-43, neurofilament-h, tubulin-α and tubulin-β. Strong and dose- dependent inhibition of neurite outgrowth was induced by azamethiphos and chlorpyrifos, and dieldrin and heptachlor, which was correlated with up-regulation of gap-43. No or only weak effects on neurite outgrowth and transcriptional alterations occurred for neonicotinoids acetamiprid, clothianidin, imidacloprid and thiamethoxam, the pyrethroids λ-cyhalothrin, cyfluthrin, deltamethrin, and permethrin, the biocidal disinfectants C12-C14-alkyl(ethylbenzyl)dimethylammonium (BAC), benzalkonium chloride and barquat (dimethyl benzyl ammonium chloride), and piperonyl butoxide and DEET. Our study confirms potential developmental neurotoxicity of some pesticides and provides first evidence that azamethiphos has the potential to act as a developmental neurotoxic compound. We also demonstrate that inhibition of neurite outgrowth and transcriptional alterations of gap-43 expression correlate, which suggests the employment of gap-43 expression as a biomarker for detection and initial evaluation of potential DNT of chemicals.


mTORC1 and mTORC2 have largely distinct functions in Purkinje cells.
The mammalian target of rapamycin (mTOR) is a key regulator of cellular growth which associates with other proteins to form two multi-protein complexes called mTORC1 and mTORC2. Dysregulation of mTORC1 signalling in brain is implicated in neuropathological conditions such as autism spectrum or neurodegenerative disorders. Accordingly, allosteric mTOR inhibitors are currently in clinical trials for the treatment of such disorders. Here, we ablated either mTORC1 or mTORC2 conditionally in Purkinje cells of the mouse cerebellum to dissect their role in the development, function and survival of these neurons. We find that the two mouse models largely differ from each other by phenotype and cellular responses. Inactivation of mTORC2, but not of mTORC1, led to motor coordination deficits at an early age. This phenotype correlated with developmental deficits in climbing fibre elimination and impaired dendritic self-avoidance in mTORC2-deficient Purkinje cells. In contrast, inactivation of mTORC1, but not of mTORC2, affected social interest of the mice and caused a progressive loss of Purkinje cells due to apoptosis. This cell loss was paralleled by age-dependent motor deficits. Comparison of mTORC1-deficient Purkinje cells with those deficient for the mTORC1 inhibitor TSC1 revealed a striking overlap in Purkinje cell degeneration and death, which included neurofilamentopathy and reactive gliosis. Altogether, our study reveals distinct roles of mTORC1 and mTORC2 in Purkinje cells for mouse behaviour and the survival of neurons. Our study also highlights a convergence between the phenotypes of Purkinje cells lacking mTORC1 activity and those expressing constitutively active mTORC1 due to TSC1 deficiency.


Allosuppression: evidence for the involvement of both "noncytotoxic" and cytotoxic T cells.
Allogeneic suppressor cells were generated by priming to a minor histocompatibility antigen. C57BL/10 female mice were primed to the male-specific minor transplantation antigen, H-Y. After boosting, anti-male-primed cells could potently suppress the secondary antibody response of male spleen cells in vitro. Anti-male suppressor cells were H-2-restricted, radiation-resistant T cells which could act on either T or B cells in the responding population. Mapping the restriction elements for anti-male suppressor cells revealed two distinct subpopulations of effector cells. The majority subpopulation was restricted to H-Y in the context of the H-2Db molecule. These cells were probably cytotoxic T cells as they were inhibited by culture of the suppressor cells in pyrilamide (a histamine receptor antagonist, which prevents the maturation of cytotoxic T cells from their precursors). The second subpopulation of suppressor cells was restricted to H-Y in the context of H-2K or I region-coded molecules. as neither H-2Kb nor H-2Ib molecules can act as restriction elements for anti-male-specific cytotoxic T cells, this subpopulations was almost certainly not composed of conventional cytotoxic cells. Furthermore, these cells were not affected by culture in pyrilamide. Together these two populations act to completely inhibit the anti-trinitrophenyl (TNP) plaque-forming cell response of male spleen cells in vitro.


The Effects of Tocotrienols Added to Canola Oil on Microalbuminuria, Inflammation, and Nitrosative Stress in Patients with Type 2 Diabetes: A Randomized, Double-blind, Placebo-controlled Trial.
Tocotrienols (T3) were neglected in the past; today, get attentions due to their antioxidant and none-antioxidant activity. The objective of this study was to evaluate the effects of the daily intake of 200 mg T3 added in canola oil over 8 weeks on microalbuminuria, inflammation, and nitrosative stress in type 2 diabetic patients. This study was a double-blinded, placebo-controlled, randomized trial. A total of 50 patients with T2DM and FBS >126 mg/dl treated by non-insulin hypoglycemic drugs were randomly assigned to receive either 15 ml T3-enriched canola oil (200 mg/day T3) or pure canola oil for 8 weeks. Urine microalbumin, volume and creatinine levels, serum hs-CRP, and nitric oxide (NO) levels were measured before and after intervention. From 50 patients participated in this study, 44 completed the study. There were no significant differences in baseline characteristics, dietary intake, and physical activity between groups. Urine microalbumin and serum hs-CRP were declined significantly in T3-treated group. At the end of the study, patients who treated with T3 had lower urine microalbumin (11 (9, 25) vs. 22 (15, 39.75) nmol/dl, P = 0.003) and hs-CRP changes (-10.91 ± 15.5 vs. -9.88 ± 27.5 Pg/ml, P = 0.048) than control group. A non-significant decrease was also observed in serum NO level in T3-treated group with no changes in urine volume and creatinine levels. These findings indicate that T3 leads to ameliorate proteinuria and can protect the kidney against inflammation (hs-CRP) and nitrosative stress (NO).


Twenty years of work-related injury and illness among union carpenters in Washington State.
Individuals who work in the construction industry are at high risk of occupational injury. Robust surveillance systems are needed to monitor the experiences of these workers over time. We updated important surveillance data for a unique occupational cohort of union construction workers to provide information on long-term trends in their reported work-related injuries and conditions. Combining administrative data sources, we identified a dynamic cohort of union carpenters who worked in Washington State from 1989 through 2008, their hours worked by month, and their workers' compensation claims. Incidence rates of reported work-related injuries and illnesses were examined. Poisson regression was used to assess risk by categories of age, gender, time in the union, and calendar time contrasting medical only and paid lost time claims. Over the 20-year study period, 24,830 carpenters worked 192.4 million work hours. Work-related injuries resulting in medical care or paid lost time (PLT) from work occurred at a rate of 24.3 per 200,000 hr worked (95% CI: 23.5-25.0). Medical only claims declined 62% and PLT claims declined 77%; more substantive declines were seen for injuries resulting from being struck and falls to a lower level than from overexertion with lifting. Differences in risk based on union tenure and age diminished over time as well. Significant declines in rates of reported work-related injuries and illnesses were observed over the 20-year period among these union carpenters. Greater declines were observed among workers with less union tenure and for claims resulting in PLT.


Changes in arterial plasma potassium and ventilation during exercise in man.
We have investigated the relationship between arterial plasma potassium ( [K+]a) and ventilation (VE) in man because hyperkalaemia has been shown to increase VE in the anaesthetized cat by direct stimulation of the arterial chemoreceptors. Six healthy male volunteers undertook about of sub-maximal (100 W) and maximal (sprint ca. 350 W) exercise on a cycle ergometer. VE was measured breath-by-breath and arterial blood was sampled at regular intervals from a catheter inserted into a brachial artery for measurement of [K+]a and base excess. Changes in [K+]a closely mirrored changes in VE during exercise and recovery. At 100 W [K+]a increased from 4 mM to 5 mM, and during the sprint [K+]a increased to ca. 7 mM. Base excess did not mirror VE in that it reached its nadir 1-3 min after exercise had stopped, when [K+]a and VE were both falling. The increases in [K+]a seen here are probably sufficient to enhance the arterial chemoreceptor drive during exercise. Furthermore, the close temporal relationship between [K+]a and VE suggests that it is possible that exercise hyperkalaemia may contribute to the control of breathing in exercise.


Single-mode delivery of 250 nm light using a large mode area photonic crystal fiber.
We demonstrate that large mode area (LMA) photonic crystal fibers (PCFs) can be used as single-mode patch-cords for 250 nm laser light. We have studied the transmission of the 250 nm output beam of a frequency-quadrupled diode laser through a triangular structure LMA PCF with 10 microm core. We have achieved single-mode output with coupling loss of 1.8 +/- 0.6 dB and transmission loss of 1.5 +/- 0.2 dB/m. The critical bend loss radius is approximately 6 cm. The transmission loss is compared with published bulk silica measurements. Effects of optically induced damage were observed after prolonged operation and have been studied as function of laser power and time. The optical damage occurs primarily at the fiber input and can be partly ameliorated by cleaving the fiber input. For input power levels of < approximately 0.3 mW stable operation can be achieved for periods of >40 hours which is sufficient for many laboratory based applications. The results show the utility of these fibers for single-mode beam delivery in a spectral region where step-index single-mode fibers are not readily available.


Comparative proteomics reveals signature metabolisms of exponentially growing and stationary phase marine bacteria.
Much of the phenotype of a microorganism consists of its repertoire of metabolisms and how and when its proteins are deployed under different growth conditions. Hence, analyses of protein expression could provide important understanding of how bacteria adapt to different environmental settings. To characterize the flexibility of proteomes of marine bacteria, we investigated protein profiles of three important marine bacterial lineages - Oceanospirillaceae (Neptuniibacter caesariensis strain MED92), Roseobacter (Phaeobacter sp. MED193) and Flavobacteria (Dokdonia sp. MED134) - during transition from exponential to stationary phase. As much as 59-80% of each species' total proteome was expressed. Moreover, all three bacteria profoundly altered their expressed proteomes during growth phase transition, from a dominance of proteins involved in translation to more diverse proteomes, with a striking appearance of enzymes involved in different nutrient-scavenging metabolisms. Whereas the three bacteria shared several overarching metabolic strategies, they differed in important details, including distinct expression patterns of membrane transporters and proteins in carbon and phosphorous metabolism and storage compounds. These differences can be seen as signature metabolisms - metabolisms specific for lineages. These findings suggest that quantitative proteomics can inform about the divergent ecological strategies of marine bacteria in adapting to changes in environmental conditions.


Climate change as the dominant control on glacial-interglacial variations in C3 and C4 plant abundance.
Although C4 plant expansions have been recognized in the late Miocene, identification of the underlying causes is complicated by the uncertainties associated with estimates of ancient precipitation, temperature, and partial pressure of atmospheric carbon dioxide (PCO2). Here we report the carbon isotopic compositions of leaf wax n-alkanes in lake sediment cores from two sites in Mesoamerica that have experienced contrasting moisture variations since the last glacial maximum. Opposite isotopic trends obtained from these two sites indicate that regional climate exerts a strong control on the relative abundance of C3 and C4 plants and that in the absence of favorable moisture and temperature conditions, low PCO2 alone is insufficient to drive an expansion of C4 plants.


Gender-Related Differential Effects of Obesity on Health-Related Quality of Life via Obesity-Related Comorbidities: A Mediation Analysis of a French Nationwide Survey.
Negative effects of obesity on health-related quality of life (HRQoL) have been reported, especially in women, but the relative contribution of cardiometabolic and other obesity-related comorbidities to such effects remains unclear. Our objective was to model the association by sex between body mass index and HRQoL and to precisely quantify the indirect effects mediated by obesity-related comorbidities. Data were drawn from the latest French Decennial Health Survey, a nationwide cross-sectional study conducted in 2003 (21 239 adults aged 25-64 years analyzed). HRQoL was measured by the 36-item short-form health survey questionnaire. A mediation analysis based on the counterfactual framework was performed to quantify the proportion of obesity effects on HRQoL mediated by related comorbidities, including cardiometabolic risk factors (diabetes mellitus, hypertension, dyslipidemia) and diseases (ischemic heart disease, cerebrovascular, and peripheral vascular disease), musculoskeletal disorders, and asthma. After multiple linear regression, inverse associations were found between increasing body mass index category and physically oriented and most mentally oriented 36-item short-form health survey dimensions, with evidence of greater effects in women. Mediation analysis revealed that obesity effects were significantly mediated by several comorbidities, more apparently in men (eg, proportion of obesity class II total effect mediated via cardiometabolic factors: general health 27.0% [men] versus 13.6% [women]; proportion of obesity class II total effect mediated via total count of comorbidities: physical functioning 17.8% [men] versus 7.7% [women] and general health 37.1% [men] versus 20.3% [women]). Women have a greater overall impact of obesity on HRQoL, but with proportionally lower effects mediated by cardiometabolic and other obesity-related conditions, suggesting the possible role of other specific psychosocial processes.


Reducing Superfluous Opioid Prescribing Practices After Brain Surgery: It Is Time to Talk About Drugs.
Opioids are prescribed routinely after cranial surgery despite a paucity of evidence regarding the optimal quantity needed. Overprescribing may adversely contribute to opioid abuse, chronic use, and diversion. To evaluate the effectiveness of a system-wide campaign to reduce opioid prescribing excess while maintaining adequate analgesia. A retrospective cohort study of patients undergoing a craniotomy for tumor resection with home disposition before and after a 2-mo educational intervention was completed. The educational initiative was composed of directed didactic seminars targeting senior staff, residents, and advanced practice providers. Opioid prescribing patterns were then assessed for patients discharged before and after the intervention period. A total of 203 patients were discharged home following a craniotomy for tumor resection during the study period: 98 who underwent surgery prior to the educational interventions compared to 105 patients treated post-intervention. Following a 2-mo educational period, the quantity of opioids prescribed decreased by 52% (median morphine milligram equivalent per day [interquartile range], 32.1 [16.1, 64.3] vs 15.4 [0, 32.9], P < .001). Refill requests also decreased by 56% (17% vs 8%, P = .027) despite both groups having similar baseline characteristics. There was no increase in pain scores at outpatient follow-up (1.23 vs 0.85, P = .105). A dramatic reduction in opioids prescribed was achieved without affecting refill requests, patient satisfaction, or perceived analgesia. The use of targeted didactic education to safely improve opioid prescribing following intracranial surgery uniquely highlights the ability of simple, evidence-based interventions to impact clinical decision making, lessen potential patient harm, and address national public health concerns.


What are the roles of the many different types of potassium channel expressed in cerebellar granule cells?
Potassium (K) channels have a key role in the regulation of neuronal excitability. Over a hundred different subunits encoding distinct K channel subtypes have been identified so far. A major challenge is to relate these many different channel subunits to the functional K currents observed in native neurons. In this review, we have concentrated on cerebellar granule neurons (CGNs). We have considered each of the three principal super families of K channels in turn, namely, the six transmembrane domain, voltage-gated super family, the two transmembrane domain, inward-rectifier super family and the four transmembrane domain, leak channel super family. For each super family, we have identified the subunits that are expressed in CGNs and related the properties of these expressed channel subunits to the functional currents seen in electrophysiological recordings from these neurons. In some cases, there are strong molecular candidates for proteins underlying observed currents. In other cases the correlation is less clear. We show that at least 26 potassium channel alpha subunits are moderately or strongly expressed in CGNs. Nevertheless, a good empirical model of CGN function has been obtained with just six distinct K conductances. The transient KA current in CGNs, seems due to expression of Kv4.2 channels or Kv4.2/4.3 heteromers, while the KCa current is due to expression of large-conductance slo channels. The G-protein activated KIR current is probably due to heteromeric expression of KIR3.1 and KIR3.2. Perhaps KIR2.2 subunits underlie the KIR current when it is constitutively active. The leak conductance can be attributed to TASK-1 and or TASK-3 channels. With less certainty, the IK-slow current may be due to expression of one or more members of the KCNQ or EAG family. Lastly, the delayed-rectifier Kv current has as many as six different potential contributors from the extensive Kv family of alpha subunits. Since many of these subunits are highly regulated by neurotransmitters, physiological regulators and, often, auxiliary subunits, the resulting electrical properties of CGNs may be highly dynamic and subject to constant fine-tuning.


Microplastics profile along the Rhine River.
Microplastics result from fragmentation of plastic debris or are released to the environment as pre-production pellets or components of consumer and industrial products. In the oceans, they contribute to the 'great garbage patches'. They are ingested by many organisms, from protozoa to baleen whales, and pose a threat to the aquatic fauna. Although as much as 80% of marine debris originates from land, little attention was given to the role of rivers as debris pathways to the sea. Worldwide, not a single great river has yet been studied for the surface microplastics load over its length. We report the abundance and composition of microplastics at the surface of the Rhine, one of the largest European rivers. Measurements were made at 11 locations over a stretch of 820 km. Microplastics were found in all samples, with 892,777 particles km (-2) on average. In the Rhine-Ruhr metropolitan area, a peak concentration of 3.9 million particles km (-2) was measured. Microplastics concentrations were diverse along and across the river, reflecting various sources and sinks such as waste water treatment plants, tributaries and weirs. Measures should be implemented to avoid and reduce the pollution with anthropogenic litter in aquatic ecosystems.


Characterization of quantitative trait loci for the age of first foraging in honey bee workers.
Identifying the basis of quantitative trait loci (QTL) remains challenging for the study of complex traits, such as behavior. The honey bee is a good model combining interesting social behavior with a high recombination rate that facilitates this identification. Several studies have focused on the pollen hoarding syndrome, identifying multiple QTL as the genetic basis of its behavioral components. One component, the age of first foraging, is central for colony organization and four QTL were previously described without identification of their genomic location. Enabled by the honey bee genome project, this study provides data from multiple experiments to scrutinize these QTL, including individual and pooled SNP mapping, sequencing of AFLP markers, and microsatellite genotyping. The combined evidence confirms and localizes two of the previous QTL on chromosome four and five, dismisses the other two, and suggests one novel genomic region on chromosome eleven to influence the age of first foraging. Among the positional candidates the Ank2, PKC, Erk7, and amontillado genes stand out due to corroborating functional evidence. This study thus demonstrates the power of combined, genome-based approaches to enable targeted studies of a manageable set of candidate genes for natural behavioral variation in the important, complex social trait "age of first foraging".


Melatonin based therapies for delirium and dementia.
Melatonin levels have been shown to decline with aging. Melatonin and its analogs in addition to their effect on sleep promotion, has been shown to have multiple pleiotropic effects. It can also help with neuroprotection through different mechanisms. Evidence in animal and human studies suggests that low levels of melatonin have been linked to delirium, mild cognitive impairment, dementia, and with certain behavioral problems. Recent clinical trials have showed that both melatonin and its analogs may be useful in the prevention, treatment of delirium, and the management of dementia. These medications seem to have the advantage of less side effects and better safety profile when compared to antipsychotics and sedatives like benzodiazepines. These medications are available over the counter in North America, Europe, and Asia, and some of these medications are approved by FDA. This manuscript will discuss the promising role of these melatonergic medications alone or in combination with other medications for the management of Geriatric Psychiatric diseases like delirium and dementia.


Prospective study of seroreactivity to JC virus T-antigen and risk of colorectal cancers and adenomas.
John Cunningham virus (JCV) is a common polyomavirus classified as a possible carcinogen by the International Agency for Research on Cancer. JCV may play a role in colorectal carcinogenesis, although we previously reported no association between JCV capsid antibodies and colorectal cancer. No studies have examined the role of seroreactivity to JCV T-antigen (T-Ag) oncoprotein in colorectal cancer. A case-control study nested within a community-based prospective cohort (CLUE II) was conducted. In 1989, 25,080 residents of Washington County, Maryland, were enrolled in CLUE II, completing baseline questionnaires and providing blood samples. At follow-up, 257 incident colorectal cancer cases were identified by linkage to population-based cancer registries through 2006 and matched to controls on age, sex, race, and date of blood draw. One hundred and twenty-three colorectal adenoma cases were identified through self-report during follow-up and matched to controls on age, sex, race, date of blood draw, and colorectal cancer screening. Baseline serum samples were tested for seroreactivity to JCV T-Ag. Associations between JCV T-Ag seroreactivity and colorectal cancer/adenomas were evaluated using conditional logistic regression models. Overall, seroreactivity to JCV T-Ag was not statistically significantly associated with the risk of either colorectal cancer [OR, 1.34; 95% confidence interval (CI), 0.89-2.01] or adenoma (OR, 1.30; 95% CI, 0.70-2.42), while a borderline association with colorectal cancer was observed among women (OR, 1.82; 95% CI, 1.00-3.31). Our past evaluation of JCV capsid seropositivity, combined with current findings, does not support a notable etiologic role for JCV infection in colorectal cancer.


Silhouette estimation.
Silhouettes arise in a variety of imaging scenarios. Pristine silhouettes are often degraded via blurring, detector sampling, and detector noise. We present a maximum a posteriori estimator for the restoration of parameterized facial silhouettes. Extreme dealiasing and dramatic superresolution, well beyond the diffraction limit, are demonstrated through the use of strong prior knowledge.


High throughput crystallography of TB drug targets.
Tuberculosis (TB) infects one-third of the world population. Despite 50 years of available drug treatments, TB continues to increase at a significant rate. The failure to control TB stems in part from the expense of delivering treatment to infected individuals and from complex treatment regimens. Incomplete treatment has fueled the emergence of multi-drug resistant (MDR) strains of Mycobacterium tuberculosis (Mtb). Reducing non-compliance by reducing the duration of chemotherapy will have a great impact on TB control. The development of new drugs that either kill persisting organisms, inhibit bacilli from entering the persistent phase, or convert the persistent bacilli into actively growing cells susceptible to our current drugs will have a positive effect. We are taking a multidisciplinary approach that will identify and characterize new drug targets that are essential for persistent Mtb. Targets are exposed to a battery of analyses including microarray experiments, bioinformatics, and genetic techniques to prioritize potential drug targets from Mtb for structural analysis. Our core structural genomics pipeline works with the individual laboratories to produce diffraction quality crystals of targeted proteins, and structural analysis will be completed by the individual laboratories. We also have capabilities for functional analysis and the virtual ligand screening to identify novel inhibitors for target validation. Our overarching goals are to increase the knowledge of Mtb pathogenesis using the TB research community to drive structural genomics, particularly related to persistence, develop a central repository for TB research reagents, and discover chemical inhibitors of drug targets for future development of lead compounds.


Longitudinal analysis of CD8 T-cell responses to HIV and hepatitis C virus in a cohort of co-infected haemophiliacs.
To investigate CD8 T-cell responses to HIV and hepatitis C virus (HCV) over time in a group of co-infected children with haemophilia to assess the influence of the virus infections on each other and on clinical outcome. The HIV and HCV CD8 T-cell response of HLA-A2 co-infected individuals in the cohort were analysed at two time points, looking at the frequency and phenotype of HIV-specific T cells and assessing overall responses to the two viruses. Peripheral blood mononuclear cells (PBMC) from 72 HLA-A2 co-infected individuals were analysed using an HIV HLA-A2 tetramer and by IFN-gamma ELISpot using a panel of HIV and HCV antigens. PBMC from a group of 26 HLA-A2 HIV mono-infected adults were also analysed as a comparison. We identified two distinct patterns of response: some patients had a limited response to either virus whilst others made responses to a range of HIV epitopes. HCV responses were detected only in those who made multiple responses to HIV epitopes (P<0.0001). HCV infection had an influence on the phenotype of HIV-specific CD8 T cells, with a reduction in relative perforin and CD57 expression. Lack of functional or tetramer-positive HIV-specific T cells was associated with a decline in absolute CD4 T-cell counts between the time points (up to 7 years; P = 0.005). HCV infection has an impact on the phenotype of HIV-specific CD8 T cells. In this well-defined cohort, failure to maintain effective CD8 T-cell responses against HIV may contribute to disease progression.


Vaginal delivery in the 30+4 weeks of pregnancy and organ donation after brain death in early pregnancy.
A 28-year-old woman suffered a traffic accident resulting in severe head injuries with deleterious prognosis. Diagnostics further revealed a hitherto unknown pregnancy, at suspected week 9. Based on the patient's wish to donate organs, brain death protocol confirmed irreversible loss of brain function. Yet, vital pregnancy rendered organ transplantation impossible. Multiple ethical and legal issues arose, from invalidation of established legal care after brain death to the delivery of a healthy child after trauma and long-term critical care. After medicolegal and ethical counselling, pregnancy was sustained, and the goal of organ donation postponed. Critical care focused on foetal homeostasis. At 30+4 weeks, a viable girl was born via assisted vaginal delivery. Postpartal organ donation resulted in heart, kidney and pancreas transplantation. The case emphasises the medical, legal and ethical challenges to combine two apparently diametrical goals: the successful full-term pregnancy and the fulfilment of a patient's wish to donate organs.


Transferrin receptor polarity and recycling accuracy in "tight" and "leaky" strains of Madin-Darby canine kidney cells.
We have characterized the polarity of the transferrin receptor in the epithelial Madin-Darby canine kidney (MDCK) cell line. The receptor is present in approximately 165,000 copies per cell, migrates as a diffuse band upon SDS gel electrophoresis with Mr 90,000, displays a dissociation constant for diferritransferrin at neutral pH of approximately 2 nM, and is active in essentially all of the cells of the population. Transferrin-mediated 55Fe uptake was used to measure the polarity of active transferrin receptors in filter-grown MDCK cells. The ratio of basolateral to apical receptors was approximately 800:1 for the high resistance strain I MDCK cells (typically greater than 2,000 ohm X cm2) and approximately 300:1 for the lower resistance strain II cells (less than 350 ohm X cm2). In combination with morphometric data this shows that a difference in resistance between these two strains is not reflected in a significant difference in cell surface polarity. We used the recycling of transferrin receptor in filter-grown MDCK cells to evaluate the accuracy of the sorting of a basolateral protein during endocytosis. Monitoring the amount of apically released 125I-labeled transferrin after application of 55Fe- and 125I-labeled transferrin to the basolateral surface provided a sensitive assay of the accuracy of sorting during recycling of the receptor from endosomes to the plasma membrane. The accuracy of transferrin receptor sorting (greater than 99.88%) during a single cycle of transit between the endosome and the plasma membrane is sufficient to maintain the high level of polarity of the cell.


Interferometric measurement of the degree of polarization and control of the contrast of intensity fluctuations.
We introduce a technique for determining the polarimetric characteristics of light by measuring the contrast of the intensity fluctuations in an interferometric setup. The method permits simultaneous measurement of the degree of polarization and of the second normalized Stokes component, which is related to the ellipsometric parameters azimuth and ellipticity, based on only two measurements. We also show that by using. phase modulation we can increase the signal-to-noise ratio as much as 40% under certain conditions.


Wired to freedom: Life science, public politics, and the case of Cochlear Implantation.
Cochlear Implantation is now regarded as the most successful medical technology. It carries promises to provide deaf/hearing impaired individuals with a technological sense of hearing and an access to participate on a more equal level in social life. In this article, we explore the adoption of cochlear implantations among Danish users in order to shed more light on their social and political implications. We situate cochlear implantation in a framework of new life science advances, politics, and user experiences. Analytically, we draw upon the notion of social imaginary and explore the social dimension of life science through a notion of public politics adopted from the political theory of John Dewey. We show how cochlear implantation engages different social imaginaries on the collective and individual levels and we suggest that users share an imaginary of being "wired to freedom" that involves new access to social life, continuous communicative challenges, common practices, and experiences. In looking at their lives as "wired to freedom," we hope to promote a wider spectrum of civic participation in the benefit of future life science developments within and beyond the field of Cochlear Implantation. As our empirical observations are largely based in the Scandinavian countries (notably Denmark), we also provide some reflections on the character of the technology-friendly Scandinavian welfare states and the unintended consequences that may follow in the wake of rapid technology implementation of life science in society.


Neurosurgical Injuries Caused by the 2011 Van Earthquake: The Experience at the Van Regional Training and Research Hospital.
On October 23, 2011, a devastating earthquake, magnitude 7.2 on the Richter Scale, jolted the province of Van in the east of Turkey and led to hundreds of casualties. In this study, we aimed to present our clinical experience in the management of patients with cranial and spinal injuries who were admitted to the Van Regional Training and Research Hospital. The retrospective study included 44 (77.2%) patients who were referred to the neurosurgery department after being diagnosed with spinal and cranial injuries due to earthquake at the emergency department between October 23 and 27, 2011. The patients comprised 32 male (72.7%) and 12 (27.3%) female patients with a mean age of 23.5 years. The injuries included scalp injury (n = 16), burst fracture (n=7), compression fracture (n=3), epidural hematoma (n=9), subdural hematoma (n=3), contusion (n=1), traumatic subarachnoid hemorrhage (n=2), depressed skull fracture (n=3), linear fracture (n=9), cervical fracture (n=2), and pneumocephalus (n=1). Most of the patients (90.9%) had isolated injuries and the others (9.1%) presented with combined cranial and spinal injuries. At discharge, the 3 patients with spinal fractures were paraplegic, and of the 2 patients who were operatively treated due to subdural hematoma, 1 was hemiparesic and the other was hemiplegic. No mortality occurred in our patients. The results of this study demonstrated that, in the aftermath of a natural disaster, conducting correct triage procedures and performing a prompt intervention with appropriate and qualified equipment play key roles in reducing morbidity and mortality.


Scaling and universality in urban economic diversification.
Understanding cities is central to addressing major global challenges from climate change to economic resilience. Although increasingly perceived as fundamental socio-economic units, the detailed fabric of urban economic activities is only recently accessible to comprehensive analyses with the availability of large datasets. Here, we study abundances of business categories across US metropolitan statistical areas, and provide a framework for measuring the intrinsic diversity of economic activities that transcends scales of the classification scheme. A universal structure common to all cities is revealed, manifesting self-similarity in internal economic structure as well as aggregated metrics (GDP, patents, crime). We present a simple mathematical derivation of the universality, and provide a model, together with its economic implications of open-ended diversity created by urbanization, for understanding the observed empirical distribution. Given the universal distribution, scaling analyses for individual business categories enable us to determine their relative abundances as a function of city size. These results shed light on the processes of economic differentiation with scale, suggesting a general structure for the growth of national economies as integrated urban systems.


Multiple sclerosis patients' benefit-risk preferences: serious adverse event risks versus treatment efficacy.
The aim of this study is to estimate the willingness of multiple sclerosis (MS) patients to accept life-threatening adverse event risks in exchange for improvements in their MS related health outcomes. MS patients completed a survey questionnaire that included a series of choice-format conjoint tradeoff tasks. Patients chose hypothetical treatments from pairs of treatment alternatives with varying levels of clinical efficacy and associated risks. Among the 651 patients who completed the survey, delay in years to disability progression was the most important factor in treatment preferences. In return for decreases in relapse rates from 4 to 1 and increases in delay in progression from 3 to 5 years, patients were willing to accept a 0.38% annual risk of death or disability from PML, a 0.39% annual risk of death from liver failure or a 0.48% annual risk of death from leukemia. Medical interventions carry risks of adverse outcomes that must be evaluated against their clinical benefits. Most MS patients indicated they are willing to accept risks in exchange for clinical efficacy. Patient preferences for potential benefits and risks can assist in decision-making.


Ocean warming compresses the three-dimensional habitat of marine life.
Vertical migration to reach cooler waters is a suitable strategy for some marine organisms to adapt to ocean warming. Here, we calculate that realized vertical isotherm migration rates averaged -6.6 + 18.8 m dec-1 across the global ocean between 1980 and 2015. Throughout this century (2006-2100), surface isotherms are projected to deepen at an increasing rate across the globe, averaging -32.3 m dec-1 under the representative concentration pathway (RCP)8.5 'business as usual' emissions scenario, and -18.7 m dec-1 under the more moderate RCP4.5 scenario. The vertical redistribution required by organisms to follow surface isotherms over this century is three to four orders of magnitude less than the equivalent horizontal redistribution distance. However, the seafloor depth and the depth of the photic layer pose ultimate limits to the vertical migration possible by species. Both limits will be reached by the end of this century across much of the ocean, leading to a rapid global compression of the three-dimensional (3D) habitat of many marine organisms. Phytoplankton diversity may be maintained but displaced toward the base of the photic layer, whereas highly productive benthic habitats, especially corals, will have their suitable 3D habitat rapidly reduced.


The effects of the smart environment on the information divide experienced by people with disabilities.
Information and Communication Technology (ICT) is connected with every aspect of social, cultural, economic, educational, and commercial activity. Smart devices in particular have changed society and are necessary goods for modern people. Smart device usage is rapidly growing in everyday life, so the ability to use a smart device is increasingly important, yet there is little data supporting increased digital inclusion of people with disabilities in mobile device use. This study investigates the effects of the smart environment on the information divide experienced by people with disabilities. Data from the 2013 Information Divide Index Data of the National Information Society Agency was analyzed regarding three aspects: access, skill, and competence. The accessibility difference was investigated by comparing access to a PC or smart device in two groups. The effects of a smart environment on the information divide were analyzed using General Linear Modeling (GLM). The access rate was higher for the general group than for that of those with disabilities, and this difference appeared to be greater in the smart environment. The results of the GLM showed that disability and device access had statistically significant effects on skill and all aspects of competence. These results provide evidence that the smart environment further creates the information divide for people with disabilities. Strategies should be formed to reduce this divide, particularly within smart environments.


Thiamin supplementation does not reduce the frequency of adverse events after anti-malarial therapy among patients with falciparum malaria in southern Laos.
In a recent study one third of Lao patients presenting with uncomplicated Plasmodium falciparum malaria had biochemical evidence of thiamin deficiency, which was associated with a higher incidence of adverse events. Thiamin supplementation might, therefore, reduce adverse events in this population. An exploratory, double-blind, parallel group, placebo-controlled, superiority trial of thiamin supplementation in patients of all ages with uncomplicated and severe falciparum malaria was conducted in Xepon District, Savannakhet Province, southern Laos. Patients were randomly assigned to either oral thiamin 10 mg/day for 7 days immediately after standard anti-malarial treatment then 5 mg daily until day 42, or identical oral placebo. After interim analyses when 630 patients (314 in thiamin and 316 in placebo groups) had been recruited, the trial was discontinued on the grounds of futility. On admission biochemical thiamin deficiency (alpha ≥ 25%) was present in 27% of patients and 9% had severe deficiency (alpha > 31%). After 42 days of treatment, the frequency of thiamin deficiency was lower in the thiamin (2%, 1% severe) compared to the placebo (11%, 3% severe) groups (p < 0.001 and p = 0.05), respectively. Except for diarrhoea, 7% in the placebo compared to 3% in the thiamin group (p = 0.04), and dizziness on day 1 (33% vs 25%, p = 0.045), all adverse events were not significantly different between the groups (p > 0.05). Clinical, haematological, and parasitological responses to treatment did not differ significantly between the two groups. Thiamin supplementation reduced biochemical thiamin deficiency among Lao malaria patients following anti-malarial drug treatment, but it did not reduce the frequency of adverse events after anti-malarial therapy or have any detected clinical or parasitological impact. ISRCTN 85411059.


An entomological review of invasive mosquitoes in Europe.
Among the invasive mosquitoes registered all over the world, Aedes species are particularly frequent and important. As several of them are potential vectors of disease, they present significant health concerns for 21st century Europe. Five species have established in mainland Europe, with two (Aedes albopictus and Aedes japonicus) becoming widespread and two (Ae. albopictus and Aedes aegypti) implicated in disease transmission to humans in Europe. The routes of importation and spread are often enigmatic, the ability to adapt to local environments and climates are rapid, and the biting nuisance and vector potential are both an ecomonic and public health concern. Europeans are used to cases of dengue and chikungunya in travellers returning from the tropics, but the threat to health and tourism in mainland Europe is substantive. Coupled to that are the emerging issues in the European overseas territorities and this paper is the first to consider the impacts in the remoter outposts of Europe. If entomologists and public health authorities are to address the spread of these mosquitoes and mitigate their health risks they must first be prepared to share information to better understand their biology and ecology, and share data on their distribution and control successes. This paper focusses in greater detail on the entomological and ecological aspects of these mosquitoes to assist with the risk assessment process, bringing together a large amount of information gathered through the ECDC VBORNET project.


Modeling the image quality of enhanced reflectance x-ray multilayers as a surface power spectral density filter function.
Residual surface roughness over the entire range of relevant spatial frequencies must be specified and controlled in many high-performance optical systems. This is particularly true for enhanced reflectance multilayers if both high reflectance and high spatial resolution are desired. If we assume that the interfaces making up a multilayer coating are uncorrelated at high spatial frequencies (microroughness) and perfectly correlated at low spatial and midspatial frequencies, then the multilayer can be thought of as a surface power spectral density (PSD) filter function. Multilayer coatings thus behave as a low-pass spatial frequency filter acting on the substrate PSD, with the exact location and shape of this cutoff being material and process dependent. This concept allows us to apply conventional linear systems techniques to the evaluation of image quality and to the derivation of optical fabrication tolerances for applications utilizing enhanced reflectance x-ray multilayers.


Evolution of the core and pan-genome of Streptococcus: positive selection, recombination, and genome composition.
The genus Streptococcus is one of the most diverse and important human and agricultural pathogens. This study employs comparative evolutionary analyses of 26 Streptococcus genomes to yield an improved understanding of the relative roles of recombination and positive selection in pathogen adaptation to their hosts. Streptococcus genomes exhibit extreme levels of evolutionary plasticity, with high levels of gene gain and loss during species and strain evolution. S. agalactiae has a large pan-genome, with little recombination in its core-genome, while S. pyogenes has a smaller pan-genome and much more recombination of its core-genome, perhaps reflecting the greater habitat, and gene pool, diversity for S. agalactiae compared to S. pyogenes. Core-genome recombination was evident in all lineages (18% to 37% of the core-genome judged to be recombinant), while positive selection was mainly observed during species differentiation (from 11% to 34% of the core-genome). Positive selection pressure was unevenly distributed across lineages and biochemical main role categories. S. suis was the lineage with the greatest level of positive selection pressure, the largest number of unique loci selected, and the largest amount of gene gain and loss. Recombination is an important evolutionary force in shaping Streptococcus genomes, not only in the acquisition of significant portions of the genome as lineage specific loci, but also in facilitating rapid evolution of the core-genome. Positive selection, although undoubtedly a slower process, has nonetheless played an important role in adaptation of the core-genome of different Streptococcus species to different hosts.


Clusters of conserved beta cell marker genes for assessment of beta cell phenotype.
The aim of this study was to establish a gene expression blueprint of pancreatic beta cells conserved from rodents to humans and to evaluate its applicability to assess shifts in the beta cell differentiated state. Genome-wide mRNA expression profiles of isolated beta cells were compared to those of a large panel of other tissue and cell types, and transcripts with beta cell-abundant and -selective expression were identified. Iteration of this analysis in mouse, rat and human tissues generated a panel of conserved beta cell biomarkers. This panel was then used to compare isolated versus laser capture microdissected beta cells, monitor adaptations of the beta cell phenotype to fasting, and retrieve possible conserved transcriptional regulators. A panel of 332 conserved beta cell biomarker genes was found to discriminate both isolated and laser capture microdissected beta cells from all other examined cell types. Of all conserved beta cell-markers, 15% were strongly beta cell-selective and functionally associated to hormone processing, 15% were shared with neuronal cells and associated to regulated synaptic vesicle transport and 30% with immune plus gut mucosal tissues reflecting active protein synthesis. Fasting specifically down-regulated the latter cluster, but preserved the neuronal and strongly beta cell-selective traits, indicating preserved differentiated state. Analysis of consensus binding site enrichment indicated major roles of CREB/ATF and various nutrient- or redox-regulated transcription factors in maintenance of differentiated beta cell phenotype. Conserved beta cell marker genes contain major gene clusters defined by their beta cell selectivity or by their additional abundance in either neural cells or in immune plus gut mucosal cells. This panel can be used as a template to identify changes in the differentiated state of beta cells.


Cortical source of blink-related delta oscillations and their correlation with levels of consciousness.
Recently, blink-related delta oscillations (delta BROs) have been observed in healthy subjects during spontaneous blinking at rest. Delta BROs have been linked with continuous gathering of information from the surrounding environment, which is classically attributed to the precuneus. Furthermore, fMRI studies have shown that precuneal activity is reduced or missing when consciousness is low or absent. We therefore hypothesized that the source of delta BROs in healthy subjects could be located in the precuneus and that delta BROs could be absent or reduced in patients with disorders of consciousness (DOC). To test these hypotheses, electroencephalographic (EEG) activity at rest was recorded in 12 healthy controls and nine patients with DOC (four vegetative states, and five minimally conscious states). Three-second-lasting EEG epochs centred on each blink instance were analyzed in both time- (BROs) and frequency domains (event-related spectral perturbation or ERSP and intertrial coherence or ITC). Cortical sources of the maximum blink-related delta power, corresponding to the positive peak of the delta BROs, were estimated by standardized Low Resolution Electromagnetic Tomography. In control subjects, as expected, the source of delta BROs was located in the precuneus, whereas in DOC patients, delta BROs were not recognizable and no precuneal localization was possible. Furthermore, we observed a direct relationship between spectral indexes and levels of cognitive functioning in all subjects participating in the study. This reinforces the hypothesis that delta BROs reflect neural processes linked with awareness of the self and of the environment.


Age, treatment, and outcomes in high-risk non-ST-segment elevation acute coronary syndrome patients: insights from the EARLY ACS trial.
Elderly patients with acute coronary syndromes (ACS) are at high risk for death and recurrent thrombotic events. We evaluated the efficacy and safety of intensive treatment with glycoprotein IIb/IIIa inhibitors in an elderly population, and the relationships between age, timing of administration, and clinical outcomes. We used data from high-risk non-ST-segment elevation ACS patients randomized to early eptifibatide vs. delayed provisional use at percutaneous coronary intervention. In multivariable models, we included age×treatment interaction terms to assess whether treatment effect varied by age after adjusting for confounders. Of 9406 patients, 13.9% were aged <55 years; 27.6%, 55-64 years; 33.2%, 65-74 years; and 25.3%, ≥ 75 years. For each 10-year age increase, the adjusted odds ratio (OR) (95% confidence interval [CI]) for 96-hour death, myocardial infarction (MI), recurrent ischemia requiring urgent revascularization, or thrombotic bailout was 1.13 (1.04-1.23) and for 30-day death or MI was 1.13 (1.04-1.22). Increasing age was also associated with greater 1-year mortality (adjusted hazard ratio per 10 years: 1.44, 95% CI 1.30-1.60). There was no interaction between age and treatment (p interaction=0.99, 0.54, and 0.87, respectively). Increasing age was associated with more non-coronary artery bypass grafting-related TIMI major bleeding (adjusted OR and 95% CI per 10 years: 1.54 [1.24-1.92]), GUSTO moderate/severe bleeding (1.52 [1.33-1.75]), and transfusion (1.25 [1.07-1.45]). The amount by which TIMI major bleeding was increased with early vs. delayed provisional eptifibatide use was significantly greater with increasing age (p interaction=0.02), but the age×treatment interactions were not significant for GUSTO moderate/severe bleeding or transfusion (p interaction=0.33 and 0.54, respectively). Increasing age was associated with greater risk for ischemic events and more bleeding. Despite higher baseline ischemic risk in older patients, there was no preferential benefit of early vs. delayed provisional eptifibatide use for ischemic outcomes as age increased, but the incremental bleeding risk was amplified.