Capping Motif for Peptide Helix Formation.
It is known that a C-terminal lysine stabilizes helix formation in polyalanine peptides that have seven or more residues. Using a combination of cold ion spectroscopy and DFT calculations, we demonstrate that even a three-residue peptide, Ac-Phe-Ala-LysH(+), adopts a structure in which the lysine side chain forms three hydrogen bonds with backbone carbonyls, reproducing the capping motif of larger polyalanine helices. This is confirmed by comparison with Ac-Phe-(Ala)5-LysH(+), which forms a 310 helix containing the same structural feature. In both molecules, we identified the vibrational bands of the N- and C-terminal amide NH stretches, which lack strong hydrogen bonds with carbonyls and consequently appear in a characteristic region above 3400 cm(-1). A similar pattern is also present in the even longer peptide Ac-Phe-(Ala)10-LysH(+), illustrating the generality of this capping motif. The two longer peptides contain additional, characteristic amide NH stretch bands below 3400 cm(-1), which form the core of the helix.


HSP70 confers protection against indomethacin-induced lesions of the small intestine.
In line with improvements in diagnostic procedures to detect intestinal lesions, it has become clear that nonsteroidal anti-inflammatory drugs (NSAIDs) such as indomethacin induce lesions not only in the stomach but also in the small intestine. However, clinical protocols for the treatment of NSAID-induced lesions of the small intestine have not been established. It is known that heat shock proteins (HSPs), particularly HSP70, confer protection against various stressors, and more recently, the anti-inflammatory activity of HSP70 was revealed. In this study, we examined the effect of expression of HSP70 on indomethacin-induced lesions of the small intestine. The extent of indomethacin-induced lesions to the small intestine was reduced in transgenic mice expressing HSP70 compared with controls. Oral administration of indomethacin increased the expression of HSP70 in the small intestine. Administration of indomethacin also induced mucosal cell apoptosis and expression of proinflammatory cytokines in the small intestines of control mice, with both of these responses suppressed in the transgenic mice. Geranylgeranylacetone (GGA), a clinically used antiulcer drug, increased expression of HSP70 in the small intestine and suppressed indomethacin-induced lesions of the small intestines in wild-type mice. These results suggest that indomethacin-induced increase in HSP70 expression reduces the extent of lesions to the small intestine by suppressing mucosal cell apoptosis and inflammatory responses. The HSP-inducing activity of GGA seems to contribute to the protective effect of drug against the lesions. Based on these results, we propose that nontoxic HSP70-inducers, such as GGA, would be therapeutically beneficial for treating NSAID-induced lesions of the small intestine.


Digital holographic interferometry for simultaneous orthogonal radial vibration measurements along rotating shafts.
A digital holographic interferometry setup used to measure radial vibrations along a rotating shaft is presented. A continuous Nd:YAG laser and a high-speed digital camera are used for recording the holograms. The shaft was polished optically smooth to avoid speckle noise from the rotating surface. The light reflected from the shaft was directed onto a diffuser which in turn was imaged by the holographic system. Simultaneous measurements with a laser vibrometer were performed at one point and comparisons between the signals showed good agreement. It is shown that different vibration components of a rotating shaft can be simultaneously measured with this technique.


Mental health provision in schools: approaches and interventions in 10 European countries.
The role of schools in providing community-based support for children's mental health and well-being is widely accepted and encouraged. Research has mainly focused on designing and evaluating specific interventions and there is little data available regarding what provision is available, the focus and priorities of schools and the professionals involved in providing this support. The current study presents these data from schools in 10 European countries. Online survey of 1466 schools in France, Germany, Ireland, Netherlands, Poland, Serbia, Spain, Sweden, UK and Ukraine. The participating countries were chosen based on their geographical spread, diversity of political and economic systems, and convenience in terms of access to the research group and presence of collaborators. Schools reported having more universal provision than targeted provision and there was greater reported focus on children who already have difficulties compared with prevention of problems and promotion of student well-being. The most common interventions implemented related to social and emotional skills development and anti-bullying programmes. Learning and educational support professionals were present in many schools with fewer schools reporting involvement of a clinical specialist. Responses varied by country with 7.4-33.5% between-country variation across study outcomes. Secondary schools reported less support for parents and more for staff compared with primary schools, with private schools also indicating more staff support. Schools in rural locations reported less student support and professionals involved than schools in urban locations. The current study provides up-to-date and cross-country insight into the approaches, priorities and provision available for mental health support in schools; highlighting what schools prioritise in providing mental health support and where coverage of provision is lacking.


Adrenergic vasoconstriction augments tissue O2 extraction during reductions in O2 delivery.
When systemic O2 delivery is reduced, increases in systemic O2 extraction are facilitated by sympathetically mediated increases in vascular resistance that limit blood flow to regions with low metabolic demand. Local metabolic vasodilation competes with this vasoconstriction, thereby effecting a balance between tissue O2 supply and demand. This study examined the role of sympathetically mediated vasoconstriction on the critical level of O2 extraction in hindlimb and whole body during progressive reductions in O2 delivery. In anesthetized dogs, the left hindlimb was vascularly isolated and its O2 delivery was decreased in stages by reducing the speed of an occlusive pump. In a normovolemic group (n = 6), blood volume was maintained to minimize sympathetic tone while flow to the hindlimb was reduced. In a hypovolemic group (n = 6), blood volume was removed in stages to augment sympathetic tone progressively while flow to the limb was reduced simultaneously. A phenoxybenzamine group (n = 6) was identical to the hypovolemic group, except that alpha-adrenergic effects were inhibited with phenoxybenzamine (3 mg/kg). The systemic critical O2 extraction ratio in the phenoxybenzamine group (0.60 +/- 0.06) was less than for the hypovolemic group (0.71 +/- 0.04; P = 0.004). In the hindlimb, critical O2 extractions were significantly less in the normovolemic (0.46 +/- 0.17) and phenoxybenzamine (0.49 +/- 0.10) groups compared with the hypovolemic group (0.72 +/- 0.10; P < or = 0.008).(ABSTRACT TRUNCATED AT 250 WORDS)


Palliative care for AIDS at a large urban teaching hospital: program description and preliminary outcomes.
Despite major advances in therapy, acquired immune deficiency syndrome (AIDS) remains an important cause of morbidity and mortality in young adult populations. As AIDS has been converted into a chronic disease, it has resulted for some patients in a more protracted course of symptomatic illness. Comprehensive care for late-stage human immunodeficiency virus (HIV) disease now involves an increasingly complex mixture of disease-specific and palliative therapies, requiring coordination and collaboration between AIDS and palliative care services. We describe the experience of developing a palliative care consultation service for patients with AIDS at a large urban teaching hospital, funded by the Health Resources and Services Administration as one of six national demonstration projects for the integration of HIV and palliative care. An 1100-bed medical center in the Bronx, New York. The multidisciplinary consultation team included a physician, nurse practitioner, social worker, chaplain, outreach worker, psychiatrist, and ethicist. Patients were referred from inpatient AIDS services and outpatient care sites. Patients underwent standardized assessment with clinical case review, Memorial Symptom Assessment Scale (MSAS), Mini-Mental Status Examination (MMSE), Karnofsky score, and Rapid Disability Rating Scale (RDRS). Interventions and follow-up outcomes were recorded and categorized. All deaths were analyzed and predictors of mortality were determined by bivariate and logistic regression analysis. Program referrals have been steady, with 132 patients followed by the consultation service from July 2000 through October 2001; 73% were referred from inpatient services (representing 12% of all AIDS inpatients admitted to the hospital during the study period); 57% of patients were male, 36% African American, 55% Hispanic; 44% had a history of injection drug use. Median baseline values included: CD4+ T-lymphocyte count = 35/mm3, HIV viral load = 53,813 copies per milliliter, Karnofsky = 40, MMSE = 0 (with a median score of 24 for those able to complete the examination); number of severe symptoms reported by MSAS = 4; 71% had one or more serious impairments in activities of daily living (ADL) by RDRS. In addition to AIDS, 20% of patients had malignancies and 13% had end-stage liver disease. Presenting problems and priority issues identified at consultation included: care decisions/goals of care (68%), pain (40%), psychosocial issues (31%), depression (23%), anxiety (19%), nausea/vomiting (14%), insomnia (13%), and patient/family/team conflict (13%); these problems were fully or partially resolved in 68-91% of cases. 63 patients died (median days enrolled = 35); leading causes of death included AIDS (38%), sepsis (19%), cancer (19%), and liver failure/cirrhosis (17%). Death was predicted only by baseline functional status (Karnofsky, MMSE, ADL impairment), and not by CD4+ count, viral load, or any AIDS-specific variables. Results suggest an important and ongoing need for palliative care services for patients with advanced HIV/AIDS, whose needs are likely to increase as AIDS evolves into more of a chronic disease. Patients were readily referred from predominantly inpatient settings, with very advanced disease; problems included a mix of medical and psychosocial issues, and were readily resolved by the consultation team in most cases. Death was predicted only by baseline functional status, not by traditional HIV disease markers. Mortality reflected both AIDS-related and non-AIDS-specific causes. Further studies are needed to identify more specific prognostic variables and to continue to improve palliative care treatment outcomes in late-stage patients with AIDS.


Maintenance of potential spermatogonial stem cells in vitro by GDNF treatment in a chondrichthyan model (Scyliorhinus canicula L.).
Previous work in dogfish, Scyliorhinus canicula, has identified the testicular germinative area as the spermatogonial stem cell niche. In the present study, an in vitro co-culture system of spermatogonia and somatic cells from the germinative area was developed. Long-term maintenance of spermatogonia has been successful, and addition of GDNF has promoted the development of clones of spermatogonia expressing stem cell characteristics such as alkaline phosphatase activity and has allowed maintenance of self-renewal in spermatogonia for at least 5 mo under culture conditions, notably by decreasing cell apoptosis. Furthermore, clones of spermatogonia expressed the receptor of GDNF, GFRalpha1, which is consistent with the effect of GDNF on cells despite the lack of identification of a GDNF sequence in the dogfish's transcriptome. However, a sequence homologous to artemin has been identified, and in silico analysis supports the hypothesis that artemin could replace GDNF in the germinative area in dogfish. This study, as the first report on long-term in vitro maintenance of spermatogonia in a chondrichthyan species, suggests that the GFRalpha1 signaling function in self-renewal of spermatogonial stem cells is probably conserved in gnathostomes.


Randomized trial of adjunctive topiramate therapy in infants with refractory partial seizures.
To evaluate the efficacy and safety of adjunctive topiramate (sprinkle capsules or oral liquid) in reducing daily rates of partial-onset seizures (POS) in infants with refractory POS. In this double-blind, placebo-controlled, parallel-group, international study, infants (n = 149) with clinical or EEG evidence of refractory POS were randomly allocated (1:1:1:1) to receive adjunctive topiramate 5, 15, or 25 mg/kg/d or placebo for 20 days. The primary variable was the median percentage reductions in daily POS rate from baseline to final assessment as recorded on a 48-hour video-EEG. Of the 149 infants (mean age 12 months) included in the intent-to-treat analysis set, 130 completed the study. Median percentage reduction from baseline in daily POS rate was not significantly different (p = 0.97) between topiramate 25 mg/kg (20.4%) and placebo (13.1%). Lower doses were not formally tested, but nominal p values for comparisons with placebo were not significant (15-mg/kg/d dose: p = 0.97; 5-mg/kg/d dose: p = 0.91). Treatment-emergent fever, diarrhea, vomiting, anorexia, weight decrease, somnolence, and viral infection occurred more frequently (> or = 10% difference) with topiramate than with placebo. In infants aged 1-24 months, topiramate 5, 15, or 25 mg/kg/d was not effective as adjunctive treatment for refractory partial-onset seizures. No new safety concerns associated with topiramate use were noted. This interventional study provides Class I evidence that topiramate 5, 15, or 25 mg/kg/d compared with placebo does not significantly reduce seizure rates in infants aged 1 month to 2 years with refractory partial-onset seizures.


Genomic patterns of adaptive divergence between chromosomally differentiated sunflower species.
Understanding the genetic mechanisms of speciation and basis of species differences is among the most important challenges in evolutionary biology. Two questions of particular interest are what roles divergent selection and chromosomal differentiation play in these processes. A number of recently proposed theories argue that chromosomal rearrangements can facilitate the development and maintenance of reproductive isolation and species differences by suppressing recombination within rearranged regions. Reduced recombination permits the accumulation of alleles contributing to isolation and adaptive differentiation and protects existing differences from the homogenizing effects of introgression between incipient species. Here, we examine patterns of genetic diversity and divergence in rearranged versus collinear regions in two widespread, extensively hybridizing sunflower species, Helianthus annuus and Helianthus petiolaris, using sequence data from 77 loci distributed throughout the genomes of the two species. We find weak evidence for increased genetic divergence near chromosomal break points but not within rearranged regions overall. We find no evidence for increased rates of adaptive divergence on rearranged chromosomes; in fact, collinear chromosomes show a far greater excess of fixed amino acid differences between the two species. A comparison with a third sunflower species indicates that much of the nonsynonymous divergence between H. annuus and H. petiolaris probably occurred during or soon after their formation. Our results suggest a limited role for chromosomal rearrangements in genetic divergence, but they do document substantial adaptive divergence and provide further evidence of how species integrity and genetic identity can be maintained at many loci in the face of extensive hybridization and gene flow.


Breathing exercises for adults with asthma.
Asthma is a common long-term condition that remains poorly controlled in many people despite the availability of pharmacological interventions, evidence-based treatment guidelines and care pathways.(1) There is considerable public interest in the use of non-pharmacological approaches for the treatment of asthma.(2) A survey of people with asthma reported that many have used complementary and alternative medicine, often without the knowledge of their clinical team.(3) Such interventions include breathing techniques, herbal products, homeopathy and acupuncture. The role of breathing exercises within the management of asthma has been controversial, partly because early claims of effectiveness were exaggerated.(4) UK national guidance and international guidelines on the management of asthma have included the option of breathing exercise programmes as an adjuvant to pharmacological treatment.(5,6) Here we discuss the types of breathing exercises used and review the evidence for their effectiveness.


Perfluoroalkylated Substance Effects in Xenopus laevis A6 Kidney Epithelial Cells Determined by ATR-FTIR Spectroscopy and Chemometric Analysis.
The effects of four perfluoroalkylated substances (PFASs), namely, perfluorobutanesulfonate (PFBS), perfluorooctanoic acid (PFOA), perfluorooctanesulfonate (PFOS), and perfluorononanoic acid (PFNA) were assessed in Xenopus laevis A6 kidney epithelial cells by attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy and chemometric analysis. Principal component analysis-linear discriminant analysis (PCA-LDA) was used to visualize wavenumber-related alterations and ANOVA-simultaneous component analysis (ASCA) allowed data processing considering the underlying experimental design. Both analyses evidenced a higher impact of low-dose PFAS-treatments (10(-9) M) on A6 cells forming monolayers, while there was a larger influence of high-dose PFAS-treatments (10(-5) M) on A6 cells differentiated into dome structures. The observed dose-response PFAS-induced effects were to some extent related to their cytotoxicity: the EC50-values of most influential PFAS-treatments increased (PFOS < PFNA < PFOA ≪ PFBS), and higher-doses of these chemicals induced a larger impact. Major spectral alterations were mainly attributed to DNA/RNA, secondary protein structure, lipids, and fatty acids. Finally, PFOS and PFOA caused a decrease in A6 cell numbers compared to controls, whereas PFBS and PFNA did not significantly change cell population levels. Overall, this work highlights the ability of PFASs to alter A6 cells, whether forming monolayers or differentiated into dome structures, and the potential of PFOS and PFOA to induce cell death.


First Report of Pineapple Heart Rot Caused by Phytophthora nicotianae in Hainan Province, China.
Pineapple (Ananas comosus) is an economically important tropical fruit in Hainan Province, China. During September to November 2011, heart rot disese of pineapple was found in Ledong and Wangning of Hainan Province. A survey of 150 ha producing areas of pineapple revealed that the fields were affected at an incidence ranging from 25% to 30%. Infected plants showed water-soaked lesions and soft rot on the base of heart leaves near the soil surface. Heart leaves of infected plants were easily pulled out. As the disease progressed, plants collapsed and died. Diseased tissue fragments (2 × 2 mm) were surface-disinfected for 10 min with 0.3% NaClO, then rinsed three times in sterile water, and plated to 10% V8 juice agar (4). Inoculated dishes were incubated at 26°C in the dark. After 5 days, Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were isolated from the tissue cultures, which has aseptate hyphae. Sporangia were papillate, noncaducous, oval or spherical, and 34.5 to 58.2 μm. Clamydospores, both terminal and intercalary, were also spherical, and were 23.4 to 34.0 μm (2). The ITS region of rDNA was amplified using primers ITS4/ITS5, and the 927-bp product of the ITS showed 99% sequence identity to Phytophthora nicotianae (GenBank Accession No. JF792540), and the sequence was accessed to NCBI (JX978446). Pathogenicity tests were confirmed by irrigating the wounded stem bases of 10 2-month-old pineapple plants with 50 ml of P. nicotionae zoospore solution (15,000 zoospores/ml), and another 10 plants of the same cultivar inoculated with sterile water served as controls. Plants were placed in pots in a greenhouse at 28°C and 90% relative humidity. After 9 days, soft rot was observed clearly on the base of heart leaves of all 10 inoculated plants, while the control plants appeared normal. P. nicotianae was reisolated from the infected pineapple plants, and confirmed to be the same as the inoculated pathogen by conducting a ITS rDNA sequence comparison and morphological characteristics. P. nicotianae was previously reported as the causal agent of heart rot of pineapple in Hawaii, U.S.A. (3) and Guangdong Province of China (1). To our knowledge, this is the first report of P. nicotianae on pineapple in Hainan Province, China. References: (1) J. Z. Chen et al. J. Yunnan Agric. Univ. 8:134, 2003. (2) H. H. Ho. Mycologia 73:705, 1981.(3) K. W. Howard et al. Plant Dis. Rep. 48:848, 1964. (4) X. B. Zheng. Page 81 in: Phytophthora and its Research Technology. China Agricultural Press, Beijing, 1997.


[Substance use, risk situations, and HIV seroprevalence among individuals seeking free HIV testing in Porto Alegre, Brazil].
To describe substance use, HIV seroprevalence, and risk of exposure to HIV infection in individuals seeking HIV testing in two screening centers (a municipal one and a state one) in the city of Porto Alegre, Rio Grande do Sul, Brazil. Using a cross-sectional design, we enrolled a convenience sample of 1 026 men and women between 15 and 60 years of age. We included all the subjects who, after an initial triage, reported any drug use or any behavior or situation with risk of HIV transmission. A Brazilian-Portuguese version of the Risk Assessment Battery was used to assess exposure to risk situations. Blood samples were tested for anti-HIV antibodies, using enzyme-linked immunosorbent assay. Before the blood test, individuals participated in a group counseling session (with a maximum of 20 participants) on HIV and AIDS, in line with the criteria of the Ministry of Health of Brazil. Among the sample, a high overall HIV seroprevalence was found, 15.1%. Drug use did not explain all of the risk exposure. In comparison to those who reported no injecting drug use, the odds ratio (OR) for being HIV-positive for those who reported injecting drug use at some point in their life was 7.6 (95% confidence interval (CI) = 4.4 to 13.0). However, only 10.3% of the sample reported any injecting drug use. The variables that were associated with HIV seropositivity were: male gender (OR = 1.8; 95% CI = 1.1 to 2.8), monthly family income below three times the minimum-salary amount (OR = 2.1; 95% CI = 1.3 to 3.5), age over 25 (OR = 1.7; 95% CI = 1.1 to 2.7), and having had sexual relations with a partner who was possibly HIV-positive (OR = 1.8; 95% CI = 1.1 to 3.2). Even irregular or occasional drug use increases the odds of seropositivity, particularly if intravenous drugs are used. Sexual transmission played an important role in seropositivity in this sample, indicating that drug use can affect the judgment of risk and thus contribute to HIV transmission.


Direct imaging of DNA in living cells reveals the dynamics of chromosome formation.
Individual chromosomes are not directly visible within the interphase nuclei of most somatic cells; they can only be seen during mitosis. We have developed a method that allows DNA strands to be observed directly in living cells, and we use it to analyze how mitotic chromosomes form. A fluorescent analogue (e.g., Cy5-dUTP) of the natural precursor, thymidine triphosphate, is introduced into cells, which are then grown on the heated stage of a confocal microscope. The analogue is incorporated by the endogenous enzymes into DNA. As the mechanisms for recognizing and removing the unusual residues do not prevent subsequent progress around the cell cycle, the now fluorescent DNA strands can be followed as they assemble into chromosomes, and segregate to daughters and granddaughters. Movies of such strands in living cells suggest that chromosome axes follow simple recognizable paths through their territories during G2 phase, and that late replicating regions maintain their relative positions as prophase chromosomes form. Quantitative analysis confirms that individual regions move little during this stage of chromosome condensation. As a result, the gross structure of an interphase chromosome territory is directly related to that of the prophase chromosome.


Mercury in wild fish from high-altitude aquatic ecosystems in the Tibetan Plateau.
Our understanding of the biogeochemistry of mercury (Hg) in high-altitude aquatic environments remains limited. The Tibetan Plateau (TP) is one of the Earth's most significant continental-scale high lands, yet much remains unknown about the Hg bioaccumulation and biomagnification in these pristine ecosystems. In this study, 166 wild fish samples of 13 species were collected from 13 rivers and lakes across the southern TP. Total Hg (THg) and methyl-Hg (MeHg) concentrations in the axial muscle of fish ranged from 25.1 to 1218 ng g(-1) of wet weight (median ± average deviation of 100.5 ± 149.2 ng g(-1)) and from 24.9 to 1196 ng g(-1) of wet weight (median ± average deviation of 90.7 ± 137.0 ng g(-1)), respectively. Hg concentrations varied greatly within and between species. The fish Hg concentrations were then linked to the limited available environmental Hg data and special geochemical characteristics in the region, such as Hg loading, pH, low temperature, and high ultraviolet (UV). The long lifespan and slow growth of the fish under the low-productivity environments may be the major biological factors that help to build up the fish Hg levels comparable to those observed in wild fish growing in human-impacted areas. δ(13)C signals suggested that pelagic fish had higher Hg concentrations, but no relationship was found between the Hg concentrations and the trophic levels. Zooplankton and benthic amphipods had typically higher percentages of MeHg compared to the previously reported values, suggesting the efficient transfer of MeHg from the base of the aquatic food web. This study sheds some light on the geochemical and biological controls of Hg bioaccumulation in fish and biomagnification in the aquatic food web in arid high-altitude environments.


Ordered ionic liquid structure observed at terraced graphite interfaces.
Reflection high-energy electron diffraction is presented as a contactless, surface-specific method to probe the ion organization and layering at the ionic liquid-solid interfaces. Three regimes can be identified for the structure of 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([EMIM][Tf2N]) on highly oriented pyrolytic graphite, which is strongly dependent on the distances of ions from the surface. Direct observations showed that the ultrathin ionic liquid (IL) assembly can exhibit bulk-like phase-transition behaviours as a result of the structural matching between the IL and graphite layers and the confinement template effect due to the surface topography of graphite. The present study illustrates the opportunities for conducting further studies of the structures and ultrafast dynamics of IL-solid interfaces.


Evolution of three human GPCRs for higher expression and stability.
We recently developed a display method for the directed evolution of integral membrane proteins in the inner membrane of Escherichia coli for higher expression and stability. For the neurotensin receptor 1, a G-protein-coupled receptor (GPCR), we had evolved a mutant with a 10-fold increase in functional expression that largely retains wild-type binding and signaling properties and shows higher stability in detergent-solubilized form. We have now evolved three additional human GPCRs. Unmodified wild-type receptor cDNA was subjected to successive cycles of mutagenesis and fluorescence-activated cell sorting, and functional expression could be increased for all three GPCR targets. We also present a new stability screening method in a 96-well assay format to quickly identify evolved receptors showing increased thermal stability in detergent-solubilized form and rapidly evaluate them quantitatively. Combining the two methods turned out to be very powerful; even for the most challenging GPCR target--the tachykinin receptor NK(1), which is hardly expressed in E. coli and cannot be functionally solubilized--receptor mutants that are functionally expressed at 1 mg/l levels in E. coli and are stable in detergent solution could be quickly evolved. The improvements result from cumulative small changes in the receptor sequence. This combinatorial approach does not require preconceived notions for designing mutations. Our results suggest that this method is generally applicable to GPCRs. Existing roadblocks in structural and biophysical studies can now be removed by providing sufficient quantities of correctly folded and stable receptor protein.


Radiofrequency heating of painful annular disruptions: one-year outcomes.
Although several studies have reported on outcomes following heating of annular tears with a thermoresistive catheter (SpineCATH), no data are available on the efficacy of thermal treatment with a flexible radiofrequency electrode (discTRODE). A prospective case-control study was conducted to determine the efficacy of radiofrequency heating of painful annular tears in the lumbar spine. After at least 6 months of conservative treatment, 46 patients were studied for the presence of single-level painful annular tears with magnetic resonance imaging and provocative discography. Thirty-one patients underwent heating of their annular tears with a flexible radiofrequency electrode placed across the posterior annulus. The remaining 15 patients, who mostly could not obtain funding for the procedure, continued with conservative management and acted as a control group. The Visual Analogue Scale (VAS), Oswestry Disability Index (ODI), and Medication Quantification Score (MQS) were obtained before and at 3-monthly intervals after treatment. The VAS decreased significantly after the radiofrequency treatment, and this decrease persisted at 12 months of follow-up. The VAS did not change over 12 months in untreated control subjects. The decrease in VAS was significantly greater in the treated patients than the control subjects. The ODI also decreased in treated patients but not in control subjects. The MQS did not change in either group over the 12-month follow-up period. Radiofrequency heating of annular tears can lead to an improvement in the pain of internal disc disruption. The improvement gained by this treatment method is significantly better than that obtained from conservative management.


Identification of NY-ESO-1 epitopes presented by human histocompatibility antigen (HLA)-DRB4*0101-0103 and recognized by CD4(+) T lymphocytes of patients with NY-ESO-1-expressing melanoma.
NY-ESO-1 is a member of the cancer-testis family of tumor antigens that elicits strong humoral and cellular immune responses in patients with NY-ESO-1-expressing cancers. Since CD4(+) T lymphocytes play a critical role in generating antigen-specific cytotoxic T lymphocyte and antibody responses, we searched for NY-ESO-1 epitopes presented by histocompatibility leukocyte antigen (HLA) class II molecules. Autologous monocyte-derived dendritic cells of cancer patients were incubated with recombinant NY-ESO-1 protein and used in enzyme-linked immunospot (ELISPOT) assays to detect NY-ESO-1-specific CD4(+) T lymphocyte responses. To identify possible epitopes presented by distinct HLA class II alleles, overlapping 18-mer peptides derived from NY-ESO-1 were synthetized and tested for recognition by CD4(+) T lymphocytes in autologous settings. We identified three NY-ESO-1-derived peptides presented by DRB4*0101-0103 and recognized by CD4(+) T lymphocytes of two melanoma patients sharing these HLA class II alleles. Specificity of recognition was confirmed by proliferation assays. The characterization of HLA class II-restricted epitopes will be useful for the assessment of spontaneous and vaccine-induced immune responses of cancer patients against defined tumor antigens. Further, the therapeutic efficacy of active immunization using antigenic HLA class I-restricted peptides may be improved by adding HLA class II-presented epitopes.


The role of inferior petrosal sinus sampling in the diagnostic localization of Cushing's disease.
Cushing's syndrome can present a complex problem of differential diagnosis. Of cases in which hypercortisolemia results from an adrenocorticotropic hormone (ACTH)-dependent process, approximately 80% are due to a pituitary adenoma (Cushing's disease [CD]), 10% are due to adrenal lesions, and the remaining 10% are secondary to ectopic ACTH secretion. For patients with CD, surgical removal of the pituitary adenoma is the treatment of choice. Thus, localization of the source of ACTH secretion is critical in guiding timely treatment decisions. Inferior petrosal sinus sampling (IPSS) is considered to be the gold standard for confirming the origin of ACTH secretion in patients with Cushing's syndrome. The authors present an overview of IPSS--both the technique and its interpretation--as well as a summary of recent studies. A number of other techniques are discussed including sampling from the cavernous sinus, the jugular vein, and multiple sites to aid the diagnosis and lateralization of ACTH-producing pituitary adenomas. Management is best undertaken by a comprehensive multidisciplinary team taking into account the results of all the biochemical and imaging studies available, to provide the best advice in patient treatment decisions.


EVA: a new theoretically based molecular descriptor for use in QSAR/QSPR analysis.
A new descriptor of molecular structure, EVA, for use in the derivation of robustly predictive QSAR relationships is described. It is based on theoretically derived normal coordinate frequencies, and has been used extensively and successfully in proprietary chemical discovery programmes within Shell Research. As a result of informal dissemination of the methodology, it is now being used successfully in related areas such as pharmaceutical drug discovery. Much of the experimental data used in development remain proprietary, and are not available for publication. This paper describes the method and illustrates its application to the calculation of nonproprietary data, log P(ow), in both explanatory and predictive modes. It will be followed by other publications illustrating its application to a range of data derived from biological systems.


[MtDNA and Y-chromosome lineages in the Yakut population].
The structure of female (mtDNA) and male (Y-chromosome haplotypes) lineages in the Yakut population was examined. To determine mtDNA haplotypes, sequencing of hypervariable segment I and typing of haplotype-specific point substitutions in the other parts of the mtDNA molecule were performed. Y haplogroups were identified through typing of biallelic polymorphisms in the nonrecombining part of the chromosome. Haplotypes within haplogroups were analyzed with seven microsatellite loci. Mitochondrial gene pool of Yakuts is mainly represented by the lineages of eastern Eurasian origin (haplogroups A, B, C, D, G, and F). In Yakuts haplogroups C and D showing the total frequency of almost 80% and consisting of 12 and 10 different haplopypes, respectively, were the most frequent and diverse. The total part of the lineages of western Eurasian origin ("Caucasoid") was about 6% (4 haplotypes, haplogroups H, J, and U). Most of Y chromosomes in the Yakut population (87%) belonged to haplogroup N3 (HG16), delineated by the T-C substitution at the Tat locus. Chromosomes of haplogroup N3 displayed the presence of 19 microsatellite haplotypes, the most frequent of which encompassed 54% chromosomes of this haplogroup. Median network of haplogroup N3 in Yakuts demonstrated distinct "starlike phylogeny". Male lineages of Yakuts were shown to be closest to those of Eastern Evenks.


Demonstration that a mouse immunoglobulin light chain messenger RNA hybridizes exclusively with unique DNA.
32P-labeled light chain messenger RNA was prepared from mouse MOPC 21 myeloma cells. The messenger RNA was hybridized to purified repetitive nuclear DNA and both the hybridized (repetitive 32P-RNA) and nonhybridized (nonrepetitive 32P-RNA) fractions were isolated. Only the nonhybridized RNA gave a T1 ribonuclease fingerprint showing oligonucleotides derived from the variable and constant regions of the light chain messenger RNA. In addition, this fingerprint showed oligonucleotides derived from the untranslated regions of the light chain messenger RNA. The nonrepetitive 32P-RNA was shown to rehybridize only with the unique fraction of total nuclear DNA. The rapidly hybridizing part of the unfractionated 32P-RNA preparation, therefore, is not a component of the light chain messenger RNA itself. Complementary DNA was prepared with reverse transcriptase using unlabeled light chain messenger RNA as template, and the transcripts were fractionated into various size classes. Complementary DNA molecules greater than 900 bases in length hybridized with both the initial messenger RNA and with the nonrepetitive 32P-RNA but failed to hybridize with excess purified repetitive 32P-RNA. The rapidly hybridizing component of the messenger RNA fraction, therefore, does not appear to be transcribed by reverse transcriptase. It is concluded that, under the experimental conditions used, the light chain messenger RNA hybridizes exclusively with unique DNA.


Dietary supplement hymecromone and sorafenib: a novel combination for the control of renal cell carcinoma.
Current treatments for metastatic renal cell carcinoma do not extend survival beyond a few months. Sorafenib is a targeted drug approved for metastatic renal cell carcinoma but it has modest efficacy. Hymecromone is a nontoxic dietary supplement with some antitumor activity at high doses of 450 to 3,000 mg per day. Hymecromone inhibits the synthesis of hyaluronic acid, which promotes tumor growth and metastasis. We recently noted that the hyaluronic acid receptors CD44 and RHAMM are potential predictors of metastatic renal cell carcinoma. In the current study we examined the antitumor properties of hymecromone, sorafenib and the combination in renal cell carcinoma models. Using proliferation, clonogenic and apoptosis assays, we examined the effects of hymecromone (0 to 32 μg/ml), sorafenib (0 to 3.2 μg/ml) and hymecromone plus sorafenib in Caki-1, 786-O, ACHN and A498 renal cell carcinoma cells, and HMVEC-L and HUVEC endothelial cells. A Boyden chamber was used for motility and invasion assays. Apoptosis indicators, hyaluronic acid receptors, epidermal growth factor receptor and c-Met were evaluated by immunoblot. The efficacy of hymecromone, sorafenib and hymecromone plus sorafenib was assessed in the sorafenib resistant Caki-1 xenograft model. Hymecromone plus sorafenib synergistically inhibited proliferation (greater than 95%), motility/invasion (65%) and capillary formation (76%) in renal cell carcinoma and/or endothelial cells, and induced apoptosis eightfold (p <0.001). Hymecromone plus sorafenib inhibited hyaluronic acid synthesis and adding hyaluronic acid reversed the cytotoxicity of hymecromone plus sorafenib. Hymecromone plus sorafenib up-regulated pro-apoptotic indicators and down-regulated Mcl-1, CD44, RHAMM, phospho-epidermal growth factor receptor and phospho-cMet. In all assays hymecromone and sorafenib alone were ineffective. Oral administration of hymecromone (50 to 200 mg/kg) plus sorafenib (30 mg/kg) eradicated Caki-1 tumor growth without toxicity. Hymecromone and sorafenib alone were ineffective. To our knowledge this is the first study to show that the combination of sorafenib and the nontoxic dietary supplement hymecromone is highly effective for controlling renal cell carcinoma.


Self-focusing of femtosecond diffraction-resistant vortex beams in water.
We report experiments on self-focusing of femtosecond diffraction-resistant vortex beams in water. These beams are higher-order Bessel beams with weak azimuthal modulation of the transverse intensity patterns. The modulation overrides the self-focusing dynamics and results in the formation of regular bottlelike filament distributions. The peak-power thresholds for filamentation, at a particular distance, are relatively accurately estimated by the adaptation of the Marburger formula derived earlier for Gaussian beams. The nonlinear conversion of the incident conical waves into the localized spatial wave packets propagating near the beam axis is observed.


Chromosome painting in the African four-striped mouse Rhabdomys pumilio: detection of possible murid specific contiguous segment combinations.
Fluorescence in-situ hybridization was used to construct a comparative chromosome map between the laboratory mouse, Mus musculus and the African four-striped mouse, Rhabdomys pumilio. A high degree of homology between the species was detected using both FISH and G-banding. Ten mouse chromosomes (2-4, 7, 14-16, 18, 19 and the X) were retained as chromosomal arms or intact chromosome blocks. Six mouse chromosome painting probes that correspond to mouse autosomes 5, 6, 8, 11, 12 and 13, produced double signals; the remaining four painting probes (1, 9, 10 and 17) hybridized to three or more R. pumilio chromosomes respectively. In total, the 20 mouse chromosome paints revealed 40 segments of conserved synteny in the R. pumilio genome. Most of the mouse chromosomes that produced single signals in R. pumilio have previously been shown to be conserved in the Black and Norwegian rats and the Chinese hamster. Eight contiguous segment associations appear to be R. pumilio specific, two were shared by R. pumilio and the Black and Norwegian rats, but to the exclusion of the Chinese hamster. Our data suggest that mouse chromosomes 1, 10, and 17 have undergone extensive rearrangements during genome evolution in the murids and may be useful markers for enhancing our understanding of the mode and tempo of chromosome evolution in rodents.


Use of age-specific hospital catchment populations to investigate geographical variation in inpatient admissions for children and young people in England: retrospective, cross-sectional study.
To develop a method for calculating age-specific hospital catchment populations (HCPs) for children and young people (CYP) in England. To show how these methods allow geographical variation in hospital activity to be investigated and addressed more effectively. Retrospective, secondary analysis of existing national datasets. Inpatient care of CYP (0-18 years) in England. Hospital Episode Statistics (HES) data were accessed for all inpatient admissions (elective and emergency) for CYP from birth to 18 years, 364 days, for 2011/2012-2014/2015. In 2014/2015, 857 112 admissions were analysed, from an eligible population of approximately 11.9 million CYP. For each hospital Trust, the catchment population of CYP was calculated; Trust-level admission rates per thousand per year were then calculated for admissions due to (1) any diagnostic code, (2) primary diagnosis of epilepsy and (3) epilepsy listed as primary diagnosis or comorbidity. Estimated 2014/2015 HCPs for CYP ranged from 268 558 for Barts Health NHS Trust to around 30 000 for the smallest acute general paediatric services and below 10 000 for many Trusts providing specialist services. As expected, the composition of HCPs was fairly consistent for age breakdown but levels of deprivation varied widely. After standardising for population characteristics, admission rates with a primary diagnosis of epilepsy ranged from 14.3 to 157.7 per 100 000 per year (11.0-fold variation) for Trusts providing acute general paediatric services. All-cause admission rates showed less variation, ranging from 4033 to 11 681 per 100 000 per year (2.9-fold variation). Use of age-specific catchment populations allows variation in hospital activity to be linked to specific teams and care pathways. This provides an evidence base for initiatives to tackle unwarranted variation in healthcare activity and health outcomes.


Sea ice, rain-on-snow and tundra reindeer nomadism in Arctic Russia.
Sea ice loss is accelerating in the Barents and Kara Seas (BKS). Assessing potential linkages between sea ice retreat/thinning and the region's ancient and unique social-ecological systems is a pressing task. Tundra nomadism remains a vitally important livelihood for indigenous Nenets and their large reindeer herds. Warming summer air temperatures have been linked to more frequent and sustained summer high-pressure systems over West Siberia, Russia, but not to sea ice retreat. At the same time, autumn/winter rain-on-snow (ROS) events have become more frequent and intense. Here, we review evidence for autumn atmospheric warming and precipitation increases over Arctic coastal lands in proximity to BKS ice loss. Two major ROS events during November 2006 and 2013 led to massive winter reindeer mortality episodes on the Yamal Peninsula. Fieldwork with migratory herders has revealed that the ecological and socio-economic impacts from the catastrophic 2013 event will unfold for years to come. The suggested link between sea ice loss, more frequent and intense ROS events and high reindeer mortality has serious implications for the future of tundra Nenets nomadism.


Spatial coherence effects on second- and fourth-order temporal interference.
We report the results of two experiments performed with two-photon light, produced via collinear degenerate optical spontaneous parametric downconversion (SPDC), in which both second-order (one-photon) and fourth-order (two-photon) interferograms are recorded in a Mach-Zehnder interferometer (MZI). In the first experiment, high-visibility fringes are obtained for both the second- and fourth-order interferograms. In the second experiment, the MZI is modified by the removal of a mirror from one of its arms; this leaves the fourth-order interferogram unchanged, but extinguishes the second-order interferogram. A theoretical model that takes into consideration both the temporal and spatial degrees-of-freedom of the two-photon state successfully explains the results. While the temporal interference in the MZI is independent of the spatial coherence of the source, that of the modified MZI is not.


Attitudes and Beliefs of Pathology Residents Regarding the Subspecialty of Clinical Chemistry: Results of a Survey.
-Previous studies suggest that training in pathology residency programs does not adequately prepare pathology residents to become competent in clinical chemistry. -To define the beliefs of pathology residents in the United States regarding their preparation for practicing clinical chemistry in their career, their attitude toward the discipline, and the attractiveness of clinical chemistry as a career. -The residents of all pathology residency programs in the United States were given the opportunity to participate in an online survey. -Three hundred thirty-six pathology residents responded to the survey. Analysis of the survey results indicates that pathology residents are more likely to believe that their income may be lower if they select a career that has a clinical chemistry focus and that their faculty do not value clinical chemistry as much as the anatomic pathology part of the residency. Residents also report that clinical chemistry is not as enjoyable as anatomic pathology rotations during residency or preferable as a sole career path. A large proportion of residents also believe that they will be slightly prepared or not prepared to practice clinical chemistry by the end of their residency and that they do not have enough background and/or time to learn clinical chemistry during their residency programs to be able to practice this specialty effectively post graduation. -Our survey results suggest that many pathology residents do not have a positive attitude toward clinical chemistry and do not experience a supportive learning environment with an expectation that they will become competent in clinical chemistry with a residency alone.


Multispectral processing without spectra.
It is often the case that multiplications of whole spectra, component by component, must be carried out,for example when light reflects from or is transmitted through materials. This leads to particularly taxing calculations, especially in spectrally based ray tracing or radiosity in graphics, making a full-spectrum method prohibitively expensive. Nevertheless, using full spectra is attractive because of the many important phenomena that can be modeled only by using all the physics at hand. We apply to the task of spectral multiplication a method previously used in modeling RGB-based light propagation. We show that we can often multiply spectra without carrying out spectral multiplication. In previous work [J. Opt. Soc. Am. A 11, 1553 (1994)] we developed a method called spectral sharpening, which took camera RGBs to a special sharp basis that was designed to render illuminant change simple to model. Specifically, in the new basis, one can effectively model illuminant change by using a diagonal matrix rather than the 3 x 3 linear transform that results from a three-component finite-dimensional model [G. Healey and D. Slater, J. Opt. Soc. Am. A 11, 3003 (1994)]. We apply this idea of sharpening to the set of principal components vectors derived from a representative set of spectra that might reasonably be encountered in a given application. With respect to the sharp spectral basis, we show that spectral multiplications can be modeled as the multiplication of the basis coefficients. These new product coefficients applied to the sharp basis serve to accurately reconstruct the spectral product. Although the method is quite general, we show how to use spectral modeling by taking advantage of metameric surfaces, ones that match under one light but not another, for tasks such as volume rendering. The use of metamers allows a user to pick out or merge different volume structures in real time simply by changing the lighting.


Precarious employment, business performance and occupational injuries: a study protocol of a register-based Swedish project.
There is uncertainty regarding the trends in occupational injuries (OIs) in Sweden due to a significant and increasing problem with under-reporting to injury registers. Under-reporting, in general, is likely to be exacerbated by the rise in precarious employment (PE), a set of unfavourable employment characteristics that would benefit from formal definition and study. PE and global trends are believed also to affect companies and their commitment to health and safety. The present study attempts to bridge these knowledge gaps and presents a study protocol for planned studies, with three main objectives: first, to review the literature for definitions of PE emphasising those that are multidimensional and operationalise components in routinely collected register data; second, to estimate the under-reporting of OI in Swedish registers and third, using results from the first objective, to conduct large, register-based prospective studies, designed to measure effect sizes and interactions between PE, business performance and OI. First, a scientific literature review will be conducted, including scientific databases and grey literature. Second, data from two major OI registers will be used to estimate the magnitude of under-reporting using capture-recapture methodology. Finally, all residents aged 18-65 in Sweden with any registered income during 2003-2015 will be included. Data sources encompass Swedish population and labour market registers with linkage to both the main OI register with national coverage and hospital records. Trends in PE and OI will be explored, together with risk of OI associated with PE and business performance. The project has been approved by the Regional Ethics Committee, Stockholm (dnr: 2016/2325-31; 2017/2173-32). Dissemination of study results will include a series of peer-reviewed papers, at least one PhD thesis and one report in Swedish, engaging relevant stakeholders. Results will be presented in national and international conferences and through press releases to mass media.


[Long-acting antipsychotics: The QAAPAPLE algorithm review].
Eight years ago, a committee of experts from 4 Quebec university psychiatry departments has provided the QAAPAPLE algorithm in order to guide clinicians in their use of long-acting antipsychotics (LAAP) for patients with psychotic disorders. Update the QAAPAPLE algorithm. Using a qualitative and selective literature review, the experts have focused on several aspects related to the use of LAAP and the relevance of modifying the algorithm: 1) new data on LAAP (including polypharmacy and co-prescription with clozapine, dose frequency/interval); 2) perception and attitude regarding algorithms and evidence; 3) difficulties in implementing algorithms; 4) polypharmacy involving LAAP and co-prescriptions with clozapine; 5) partner patients perspective on the algorithm. Thirteen meta-analysis were published and completed observational studies (including those on national registries), confirming the LAAP benefits. Literature adds specifications about using some drug associations as well as dose frequency and interval. Therefore, scientific advances have been considered to modify the algorithm. Interacting with Quebec psychiatrists, we have examined changes in prescription and literature to better understand the use of algorithm. The committee has updated the QAAPAPLE algorithm to guide clinicians in using LAAP along the path of patients with psychosis as early as the first episode and through different clinical settings (including treatment resistance) in order to have a more flexible and user-friendly treatment.


In vitro and in vivo evaluation of novel ciprofloxacin-releasing silicone hydrogel contact lenses.
The purpose of this study was to evaluate ciprofloxacin-releasing silicone hydrogel contact lens materials in vitro and in vivo for the treatment of microbial keratitis. Model silicone hydrogel contact lens materials were manufactured using a molecular imprinting technique to modify ciprofloxacin release kinetics. Various contact lens properties, including light transmission and surface wettability, were determined, and the in vitro ciprofloxacin release kinetics elucidated using fluorescence spectrophotometry. The materials then were evaluated for their ability to inhibit Pseudomonas aeruginosa growth in vitro and in an in vivo rabbit model of microbial keratitis. Synthesized lenses had similar material properties to commercial contact lens materials. There was a decrease in light transmission in the shorter wavelengths due to incorporation of the antibiotic, but over 80% light transmission between 400 and 700 nm. Modified materials released for more than 8 hours, significantly longer than unmodified controls (P < 0.05). In vivo, there was no statistically significant difference between the number of colony-forming units (CFU) recovered from corneas treated with eye drops and those treated with one of two modified contact lenses (P > 0.05), which is significantly less than corneas treated with unmodified control lenses or those that received no treatment at all (P < 0.05). These novel contact lenses designed for the extended release of ciprofloxacin may be beneficial to supplement or augment future treatments of sight-threatening microbial keratitis.


Sublobar resection for node-negative lung cancer 2-5 cm in size.
Sublobar resection (SLR) is an alternative to lobectomy for non-small-cell lung cancer (NSCLC). Outcomes following SLR for tumours >2 cm are not well described. We sought to determine the utilization of SLR for stage I tumours >2-5 cm in size and to determine predictors of outcome. We utilized the Surveillance, Epidemiology and End Results Program (SEER)-Medicare database to identify NSCLC patients with primary lung cancer ≥66 years old with stage I cancers >2-5 cm in size. We evaluated overall survival and cancer-specific survival among cohorts undergoing lobectomy versus SLR. Propensity score matching was performed. We compared patient characteristics and survival between groups. For the study time period (2007-2012), among patients with tumours >2 cm and ≤5 cm (n = 4582), 3890 lobectomies (85%) and 692 SLR (15%) were performed. Patients undergoing SLR were older, had smaller tumours and more comorbidities. Patients undergoing lobectomy were much more likely to have any lymph nodes removed (95.6% vs 65.6%, P < 0.001) and to have >10 nodes removed (29.6% vs 7.5%, P < 0.001). All-cause mortality [hazard ratio (HR) 1.65, confidence interval (CI) 1.48-1.85] and cancer-specific (HR 1.63, CI 1.29-2.06) mortality were higher following SLR. At 3 years, overall survival (60.9%, CI 57.0-64.6% vs 54.4%, CI 50.4-58.2%) and cancer-specific survival (87.3%, CI 83.5-90.3% vs 76.5%, CI 71.0-81.1%) favoured lobectomy over SLR. In propensity-matched groups, both all-cause (HR 1.27, CI 1.10-1.47) and cancer-specific (HR 1.54, CI 1.11-2.16) mortality rates were higher with SLR. In pathologically staged patients, SLR appears inferior to lobectomy for stage I NSCLC 2-5 cm in size. SLR is associated with less extensive lymphadenectomy and with worse survival than lobectomy in this cohort of patients. However, the 76.5% 3-year cancer-specific survival in patients undergoing SLR may exceed that of other localized treatment options for NSCLC. As such, SLR may be an appropriate option for high-risk patients with carefully staged 2-5 cm N0 tumours.


The multifactorial nature of Alzheimer's disease for developing potential therapeutics.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder with several target proteins contributing to its aetiology. Pathological, genetic, biochemical, and modeling studies all point to a critical role of Aβ aggregation in AD. Though there are still many enigmatic aspects of the Aβ cascade, none of the gaps invalidate the hypothesis. The amyloid hypothesis determines that the production, aggregation and accumulation of Aβ in the brain gives rise to a cascade of neurotoxic events that proceed to neuronal degeneration. Different targets of the disease include APP pathogenic cleavage, cytoskeletal destabilization, neurotransmitter and ion dyshomeostasis, metal ion accumulation, protein misfolding, oxidative stress, neuronal death and gene mutations. Thus, disease-modifying treatments for AD must interfere with the pathogenic steps responsible for the clinical symptoms: the deposition of extracellular Aβ plaques, the intracellular neurofibrillary tangles, inflammation, oxidative stress, iron deregulation, among others. The observations supporting the development of multifunctional compounds in association with the perception that several dual binding site AChEIs were able to reach different targets guided the development of a new drug design strategy, the multi-target-directed-ligand (MTDL) approach. This may be regarded as the buildup of hybrid molecules composed of distinct pharmacophores of different drugs. Thus, each pharmacophore of the new hybrid drug would preserve the capacity of interacting with their specific sites on the targets and, therefore, generate multiple specific pharmacological responses which would enable the treatment of multi-factorial diseases. This review summarizes a few current therapeutic trends on MTDL strategy intended to halt or revert the progression of the disease.


Creatine monohydrate increases strength in patients with neuromuscular disease.
Creatine monohydrate has been shown to increase strength in studies of young healthy subjects and in a few studies with patients. Creatine monohydrate (10 g daily for 5 days to 5 g daily for 5 days) was administered to patients with neuromuscular disease in a pilot study (Study 1; n = 81), followed by a single-blinded study (Study 2; n = 21). Body weight, handgrip, dorsiflexion, and knee extensor strength were measured before and after treatment. Creatine administration increased all measured indices in both studies. Short-term creatine monohydrate increased high-intensity strength significantly in patients with neuromuscular disease.


Potassium channels as anti-epileptic drug targets.
It is estimated that up to 30% of epilepsy patients are poorly treated with available anti-epileptic drugs (AEDs). Thus, there is a medical need for new AEDs with novel mechanisms of action to serve as alternate or adjunct therapy for the treatment of drug-resistant or refractory epilepsy. One potential anti-epileptic mechanism that has not yet been exploited is K(+) channel opening. Considerable genetic, molecular, physiological and pharmacological evidence now exists to support a role for K(+) channels such as KCNQ2/Q3, Kv1.1, KATP and GIRK2 in the control of neuronal excitability and epileptogenesis. Evidence supporting these K(+) channels as novel AED targets will be reviewed in the following article.


Intrinsically disordered CsoS2 acts as a general molecular thread for α-carboxysome shell assembly.
Carboxysomes are a paradigm of self-assembling proteinaceous organelles found in nature, offering compartmentalisation of enzymes and pathways to enhance carbon fixation. In α-carboxysomes, the disordered linker protein CsoS2 plays an essential role in carboxysome assembly and Rubisco encapsulation. Its mechanism of action, however, is not fully understood. Here we synthetically engineer α-carboxysome shells using minimal shell components and determine cryoEM structures of these to decipher the principle of shell assembly and encapsulation. The structures reveal that the intrinsically disordered CsoS2 C-terminus is well-structured and acts as a universal "molecular thread" stitching through multiple shell protein interfaces. We further uncover in CsoS2 a highly conserved repetitive key interaction motif, [IV]TG, which is critical to the shell assembly and architecture. Our study provides a general mechanism for the CsoS2-governed carboxysome shell assembly and cargo encapsulation and further advances synthetic engineering of carboxysomes for diverse biotechnological applications.


The stimulatory effect of growth hormone, prolactin, and placental lactogen on beta-cell proliferation is not mediated by insulin-like growth factor-I.
The effects of GH, PRL, and placental lactogen (PL) on the proliferation of pancreatic beta-cells in vitro were studied as well as the possible effect of insulin-like growth factor-I (IGF-I) in mediating this effect. Proliferating beta-cells were identified by staining with a monoclonal antibody to bromodeoxyuridine (BrdU) after cells were incubated for 1 h in the presence of 10 microM BrdU. By double staining with insulin antibodies it was found that 6.3% of the beta-cells had incorporated BrdU when cultured for 7 days in the presence of 1 microgram/ml human GH (hGH) compared to 0.6% when cultured in the absence of hGH. Similar results were obtained using rat GH. The half-maximal effect of hGH on beta-cell proliferation was observed at 10 ng/ml, and the maximal effect at 100 ng/ml. Islet cells cultured in the presence of PRL or PL caused a dose-dependent increase in beta-cell proliferation similar to that caused by hGH. GH, PRL, and PL had no effect on the proliferation of glucagon- or somatostatin-producing cells. The addition of 100 ng/ml IGF-I to either control or GH-stimulated islet cells did not affect the labeling index. When GH-stimulated proliferation of beta-cells was measured in the presence of neutralizing concentrations of a rabbit IGF-I antiserum, the percentage of beta-cells incorporating BrdU was unaffected. Using Northern blot analysis, no IGF-I transcripts could be detected in RNA from GH-stimulated islets, whereas IGF-I transcripts were readily detected in RNA isolated from rat liver tissue. These data suggest that the stimulatory effect of GH, PRL, and PL on beta-cell proliferation is not mediated by IGF-I, but, rather, is a direct mitogenic effect on the beta-cell.


Changes in climate extremes, fresh water availability and vulnerability to food insecurity projected at 1.5°C and 2°C global warming with a higher-resolution global climate model.
We projected changes in weather extremes, hydrological impacts and vulnerability to food insecurity at global warming of 1.5°C and 2°C relative to pre-industrial, using a new global atmospheric general circulation model HadGEM3A-GA3.0 driven by patterns of sea-surface temperatures and sea ice from selected members of the 5th Coupled Model Intercomparison Project (CMIP5) ensemble, forced with the RCP8.5 concentration scenario. To provide more detailed representations of climate processes and impacts, the spatial resolution was N216 (approx. 60 km grid length in mid-latitudes), a higher resolution than the CMIP5 models. We used a set of impacts-relevant indices and a global land surface model to examine the projected changes in weather extremes and their implications for freshwater availability and vulnerability to food insecurity. Uncertainties in regional climate responses are assessed, examining ranges of outcomes in impacts to inform risk assessments. Despite some degree of inconsistency between components of the study due to the need to correct for systematic biases in some aspects, the outcomes from different ensemble members could be compared for several different indicators. The projections for weather extremes indices and biophysical impacts quantities support expectations that the magnitude of change is generally larger for 2°C global warming than 1.5°C. Hot extremes become even hotter, with increases being more intense than seen in CMIP5 projections. Precipitation-related extremes show more geographical variation with some increases and some decreases in both heavy precipitation and drought. There are substantial regional uncertainties in hydrological impacts at local scales due to different climate models producing different outcomes. Nevertheless, hydrological impacts generally point towards wetter conditions on average, with increased mean river flows, longer heavy rainfall events, particularly in South and East Asia with the most extreme projections suggesting more than a doubling of flows in the Ganges at 2°C global warming. Some areas are projected to experience shorter meteorological drought events and less severe low flows, although longer droughts and/or decreases in low flows are projected in many other areas, particularly southern Africa and South America. Flows in the Amazon are projected to decline by up to 25%. Increases in either heavy rainfall or drought events imply increased vulnerability to food insecurity, but if global warming is limited to 1.5°C, this vulnerability is projected to remain smaller than at 2°C global warming in approximately 76% of developing countries. At 2°C, four countries are projected to reach unprecedented levels of vulnerability to food insecurity.This article is part of the theme issue 'The Paris Agreement: understanding the physical and social challenges for a warming world of 1.5°C above pre-industrial levels'.


A multistep kinase-based sertoli cell autocrine-amplifying loop regulates prostaglandins, their receptors, and cytokines.
In Sertoli epithelial cells, the IL-1beta induces prostaglandins (PG) PGE(2), PGF(2alpha) and PGI(2) (7-, 11-, and 2-fold, respectively), but not PGD(2), production. Cyclohexamide pretreatment inhibiting protein synthesis prevents IL-1beta increases in PG levels, indicating that induction requires de novo protein synthesis. IL-1beta-regulated PGE(2) and PGF(2alpha) production and cytokine expression require activation of cyclooxygenase-2 (COX-2) and c-Jun NH(2)-terminal kinase, as shown using specific enzyme inhibition. PGE(2) and PGF(2alpha) stimulate expression of IL-1alpha, -1beta, and -6, findings consistent with PG involvement in IL signaling within the seminiferous tubule. PGE(2) and PGF(2alpha) reverse COX-2-mediated inhibition of IL-1beta induction of cytokine expression and PG production. Sertoli PG receptor expression was determined; four known E-prostanoid receptor (EP) subtypes (1-4) and the F-prostanoid and prostacyclin prostanoid receptors were demonstrated using RNA and protein analyses. Pharmacological characterization of Sertoli PG receptors associated with cytokine regulation was ascertained by quantitative real-time RT-PCR analyses. IL-1beta regulates both EP(2) mRNA and protein levels, data consistent with a regulatory feedback loop. Butaprost (EP(2) agonist) and 11-deoxy PGE(1) (EP(2) and EP(4) agonist) treatments show that EP(2) receptor activation stimulates Sertoli cytokine expression. Consistent with EP(2)-cAMP signaling, protein kinase A inhibition blocks both IL-1beta- and PGE(2)-induced cytokines. Together, the data indicate an autocrine-amplifying loop involving IL-1beta-regulated Sertoli function mediated by COX-2-induced PGE(2) and PGF(2alpha) production. PGE(2) activates EP(2) and/or EP(4) receptor(s) and the protein kinase A-cAMP pathway; PGF(2alpha) activates F-prostanoid receptor-protein kinase C signaling. Further identification of the molecular mechanisms subserving these mediators may offer new insights into physiological events as well as proinflammatory-mediated pathogenesis in the testis.


Krh1p and Krh2p act downstream of the Gpa2p G(alpha) subunit to negatively regulate haploid invasive growth.
The yeast G(alpha) subunit Gpa2p and its coupled receptor Gpr1p function in a signaling pathway that is required for the transition to pseudohyphal and invasive growth. A two-hybrid screen using a constitutively active allele of GPA2 identified the KRH1 gene as encoding a potential binding partner of Gpa2p. Strains containing deletions of KRH1 and its homolog KRH2 were hyper-invasive and displayed a high level of expression of FLO11, a gene involved in pseudohyphal and invasive growth. Therefore, KRH1 and KRH2 encode negative regulators of the invasive growth pathway. Cells containing krh1Delta krh2Delta mutations also displayed increased sensitivity to heat shock and decreased sporulation efficiency, indicating that Krh1p and Krh2p regulate multiple processes controlled by the cAMP/PKA pathway. The krh1Delta krh2Delta mutations suppressed the effect of a gpa2Delta mutation on FLO11 expression and eliminated the effect of a constitutively active GPA2 allele on induction of FLO11 and heat shock sensitivity, suggesting that Krh1p and Krh2p act downstream of Gpa2p. The Sch9p kinase was not required for the signal generated by deletion of KRH1 and KRH2; however, the cAMP-dependent kinase Tpk2p was required for generation of this signal. These results support a model in which activation of Gpa2p relieves the inhibition exerted by Krh1p and Krh2p on components of the cAMP/PKA signaling pathway.


The World Federation of Societies of Biological Psychiatry (WFSBP) Guidelines for the Biological Treatment of Bipolar Disorders: Update 2010 on the treatment of acute bipolar depression.
These guidelines are based on a first edition that was published in 2002, and have been edited and updated with the available scientific evidence until September 2009. Their purpose is to supply a systematic overview of all scientific evidence pertaining to the treatment of acute bipolar depression in adults. The data used for these guidelines have been extracted from a MEDLINE and EMBASE search, from the clinical trial database clinicaltrials.gov, from recent proceedings of key conferences, and from various national and international treatment guidelines. Their scientific rigor was categorised into six levels of evidence (A-F). As these guidelines are intended for clinical use, the scientific evidence was finally assigned different grades of recommendation to ensure practicability. We identified 10 pharmacological monotherapies or combination treatments with at least limited positive evidence for efficacy in bipolar depression, several of them still experimental and backed up only by a single study. Only one medication was considered to be sufficiently studied to merit full positive evidence. Although major advances have been made since the first edition of this guideline in 2002, there are many areas which still need more intense research to optimize treatment. The majority of treatment recommendations is still based on limited data and leaves considerable areas of uncertainty.


Recurrence after abdominal surgery for Crohn's disease: relationship to disease site and surgical procedure.
We investigated the hypothesis that there is an "aggressive" subtype of Crohn's disease characterized by early recurrence and that disease location and surgical procedure are associated with differing patterns of recurrence. We analyzed 280 patient records totaling 482 major abdominal operations from a prospectively compiled Crohn's disease database. Patterns of recurrence, as defined by reoperation, were analyzed by Kaplan-Meier plots and log-rank tests for the group as a whole, as well as according to disease location and operation performed using log-rank and Cox regression analysis. The overall survival curve followed a simple curve with no apparent early rise in recurrence. There was a significantly higher recurrence rate for ileal disease than for ileocolic or colic disease (median reoperation-free survival, 37.8 vs. 47.8 and 54.7 months, respectively; log-rank test = 13.6; P = 0.001), and there was a significantly shorter reoperation-free survival for those patients treated by strictureplasty alone or stricture-plasty combined with resection than for those treated by resection alone (41.7 and 48.6 vs. 51 months, respectively; log-rank test = 12; P = 0.002), but only disease site was confirmed as an independent risk factor for recurrence by multiple regression analysis. These data suggest that there is no evidence for the existence of a separate, early recurring, aggressive disease type. Shorter reoperation-free survival after strictureplasty may reflect patterns of recurrence in ileal disease.


SOAP Opera: Self as Object and Agent in Prioritizing Attention.
A growing body of evidence has demonstrated that multiple sources of salience tune attentional sets toward aspects of the environment, including affectively and motivationally significant categories of stimuli such as angry faces and reward-associated target locations. Recent evidence further indicates that objects that have gained personal significance through ownership can elicit similar attentional prioritization. Here we discuss current research on sources of attentional prioritization that shape our awareness of the visual world from moment to moment and the underlying neural systems and contextualize what is known about attentional prioritization of our possessions within that research. We review behavioral and neuroimaging research on the influence of self-relevance and ownership on cognition and discuss challenges to this literature stemming from different modes of conceptualizing and operationalizing the self. We argue that ownership taps into both "self-as-object," which characterizes the self as an object with a constellation of traits and attributes, and "self-as-subject," which characterizes the self as an agentic perceiver and knower. Despite an abundance of research probing neural and behavioral indices of self-as-object and its effects on attention, there exists a paucity of research on the influence of self-relevance of attention when self is operationalized from the perspective of a first-person subject. To begin to address this gap, we propose the Self as Ownership in Attentional Prioritization (SOAP) framework to explain how ownership increases salience through attention to external representations of self-identity (i.e., self as object) and attention to contextually mediated permission to act (i.e., self as subject).


Triple combination of siRNAs targeting TGFβ1, TGFβR2, and CTGF enhances reduction of collagen I and smooth muscle actin in corneal fibroblasts.
Transforming growth factor β1 (TGFβ1), TGFβ receptor (TGFβR2), and connective tissue growth factor (CTGF) are key regulators of fibrosis in the cornea and in other tissues, including liver, skin, and kidney. We developed an antifibrotic treatment targeting these three critical scarring genes by using a combination of small interfering RNAs (siRNAs) and assessed its effect on downstream scarring genes, collagen I, and α smooth muscle actin (SMA). Up to six individual siRNAs for each of the three target gene mRNAs were transfected into cultures of rabbit corneal fibroblasts at concentrations from 15 to 90 nM. The knockdown of target gene proteins was measured by ELISA, and the two most effective siRNAs were tested in dual combinations. Knockdown percentages of both individual and dual siRNA combinations were analyzed for synergy by using combination index to predict "effective" and "ineffective" triple siRNA combinations. Effects of both triple siRNA combinations on target and downstream mRNAs were measured by using quantitative RT-PCR, and levels of SMA protein were assessed by immunohistochemistry. Single and dual siRNA combinations produced a wide range of protein knockdown of target genes (5%-80%). The effective triple siRNA combination significantly reduced mRNA levels of target genes (>80%) and downstream scarring genes (>85%), and of SMA protein (>95%), and significantly reduced cell migration without reducing cell viability. Simultaneous targeting of TGFβ1, TGFβR2, and CTGF genes by effective triple siRNA combination produced high knockdown of target and downstream scarring genes without cell toxicity, which may have clinical applications in reducing corneal fibrosis and scarring in other tissues.


The "Shaken Baby" syndrome: pathology and mechanisms.
The "Shaken Baby" syndrome (SBS) is the subject of intense controversy; the diagnosis has in the past depended on the triad of subdural haemorrhage (SDH), retinal haemorrhage and encephalopathy. While there is no doubt that infants do suffer abusive injury at the hands of their carers and that impact can cause catastrophic intracranial damage, research has repeatedly undermined the hypothesis that shaking per se can cause this triad. The term non-accidental head injury has therefore been widely adopted. This review will focus on the pathology and mechanisms of the three physiologically associated findings which constitute the "triad" and are seen in infants suffering from a wide range of non-traumatic as well as traumatic conditions. "Sub" dural bleeding in fact originates within the deep layers of the dura. The potential sources of SDH include: the bridging veins, small vessels within the dura itself, a granulating haemorrhagic membrane and ruptured intracranial aneurysm. Most neuropathologists do not routinely examine eyes, but the significance of this second arm of the triad in the diagnosis of Shaken Baby syndrome is such that it merits consideration in the context of this review. While retinal haemorrhage can be seen clinically, dural and subarachnoid optic nerve sheath haemorrhage is usually seen exclusively by the pathologist and only rarely described by the neuroradiologist. The term encephalopathy is used loosely in the context of SBS. It may encompass anything from vomiting, irritability, feeding difficulties or floppiness to seizures, apnoea and fulminant brain swelling. The spectrum of brain pathology associated with retinal and subdural bleeding from a variety of causes is described. The most important cerebral pathology is swelling and hypoxic-ischaemic injury. Mechanical shearing injury is rare and contusions, the hallmark of adult traumatic brain damage, are vanishingly rare in infants under 1 year of age. Clefts and haemorrhages in the immediate subcortical white matter have been assumed to be due to trauma but factors specific to this age group offer other explanations. Finally, examples of the most common causes of the triad encountered in clinical diagnostic and forensic practice are briefly annotated.


The production of alpha-cyclodextrin by enzymatic degradation of starch.
The maximum concentration of alpha-cyclodextrin for the enzymatic degradation of starch is limited to about 13.5 g X 1(-1). By addition of decanol, the equilibrium of the reaction system can be shifted towards an alpha-cyclodextrin yield of 50% even at high substrate concentrations. The main variables of the decanol process--pH, temperature, substrate quality, substrate, and enzyme concentration--have been studied. The cyclodextrin-glycosyltransferase from Klebsiella pneumoniae M5 al can preferentially be employed at pH 6 to 8, temperatures of 40 to 50 degrees C and a decanol concentration of 0.1 kg-1 starch. The dextrose equivalent of starch is important with respect to the maximum achievable starch concentration, but not with respect to the reaction. Under process conditions, the rate of alpha-cyclodextrin evolution is limited by the enzymatic reaction and not by mass transfer of decanol into the aqueous phase.


Myocardial infarction-induced acute stress and post-traumatic stress symptoms: the moderating role of an alexithymia trait - difficulties identifying feelings.
It has been acknowledged that medical life-threatening experiences such as an acute myocardial infarction (MI) often lead to acute stress disorder symptoms (ASS), which in turn can result in the development of post-traumatic stress symptoms (PTSS). Previous studies have suggested an association between various traumatic experiences and alexithymia. The association of alexithymia with ASS and PTSS in patients with MI is elusive. The aim of this study was to examine the association of alexithymia with MI-induced ASS and PTSS in patients at high risk of developing PTSD. Patients (N = 154) were examined twice, once within 48 hours, and then again three months after acute MI. All patients completed the self-rating Acute Stress Disorder Scale (ASDS) within 48 hours after the cardiac event. Three months after hospital discharge, all patients completed the Toronto Alexithymia Scale (TAS-20) and underwent the Clinician-Administered PTSD Scale (CAPS), a structured interview to assess the severity of PTSS. Descriptive statistics, correlations, multivariate linear regressions, and moderation analysis were conducted. The linear regression model explained 23% of the variance in MI-induced PTSS-symptoms (F(6.109) = 5.58, p < 0.001, R 2 = 0.23. ASS was significantly related to PTSS severity (r (152) = p < 0.001). The scores of the TAS-20 subscale difficulties identifying feelings (DIF) were found to significantly moderate this relationship (R2 = 0.03, p = 0.04). The scores of TAS-20 subscales DDF and EOT as well as the TAS-20 total score had no influence on the relationship between ASS and PTSS (p > 0.05). In MI patients with high levels of DIF, ASS predicted the development of PTSS. If replicated, the finding may inform emotion-oriented interventions to investigate whether increasing the capacity to identify feelings following acute MI could be beneficial in preventing the development of PTSS.


Bi2Se3 nanoplates for contrast-enhanced photoacoustic imaging at 1064 nm.
Photoacoustic (PA) imaging is a high-resolution biomedical imaging modality, which can be used to visualize biological tissues located beyond the limited penetration depth of existing optical imaging techniques. An optical wavelength of 1064 nm is of great interest in PA imaging due to low intrinsic absorption at this wavelength. Reduced absorption implies an increased depth of imaging, which enables several new clinical applications such as bladder imaging, gastrointestinal (GI) imaging, and sentinel lymph node (SLN) imaging. In addition, a 1064 nm Nd:YAG laser system enables a high power, cost-effective, and compact laser-based PA imaging system. However, at this wavelength, due to low intrinsic contrast, high absorption exogenous PA contrast agents are necessary for imaging. To this end, we present new Bi2Se3 nanoplates as PA contrast agents at 1064 nm wavelength for PA imaging. We successfully synthesized Bi2Se3 nanoplates and they exhibited relatively strong PA signals at 1064 nm. We confirmed the increased imaging depth of penetration by imaging the Bi2Se3-containing tube located 4.6 cm deep in biological tissues. We present in vivo PA imaging of the bladder, GI tract, and SLN in mice using a Bi2Se3 contrast agent establishing the clinical feasibility of these agents with a clinical photoacoustic/ultrasound imaging system. Our results confirm that Bi2Se3 nanoplates are promising PA contrast agents at 1064 nm that offer a high optical absorbance in the second NIR region providing a high contrast imaging and increased depth of penetration.


Zinc Finger 259 Gene Polymorphism rs964184 is Associated with Serum Triglyceride Levels and Metabolic Syndrome.
Metabolic syndrome (MetS) is characterized by a cluster of cardiovascular risk factors that include: abdominal obesity, dyslipidaemia, hypertension, insulin resistance and impaired glucose tolerance. Recent genome wide association studies have identified several susceptibility regions involved in lipid metabolism that are also associated with MetS. We have explored the association of 9 genetic polymorphisms involved in lipid metabolism and hypertension, including: MTHFR C677T, SELE L554F, FGB - 455G>A, GNB3 C825T, ZNF259 C>G, PSRC-1 A>G, CETP I405V, LPL S447X and LPA C>T in 97 subjects with MetS and 96 individuals without MetS who were recruited randomly from Mashhad stroke and heart atherosclerotic disorder (MASHAD) study using a stratified cluster random sampling technique. Anthropometric parameters and biochemical measurements were determined in all the subjects. Genotyping was carried out followed by univariate and multivariate analyses. The subjects with MetS had a higher triglyceride and lower HDL- C. CG+ GG genotypes of ZNF259 polymorphism (rs964184 C>G) and TT+CT genotypes of MTHFR C677T (rs1801133) were associated with MetS, and individuals carrying the G allele for ZNF259 or the T allele for MTHFR polymorphisms were associated with MetS (e.g, odds ratio (OR) for CG+GG genotypes vs. CC wild type: 2.52, CI=1.33-4.77; P=0.005). However, after multiple comparison adjustment, this relationship remained significant only for CG+ GG genotypes of ZNF259 polymorphism. Moreover, the ZNF259 CG+ GG genotypes were associated with increased serum concentrations of triglycerides and LDL-C, compared to the wild type. These data support the necessity for further studies in larger multicenter settings.


Dynamics of a stochastic excitable system with slowly adapting feedback.
We study an excitable active rotator with slowly adapting nonlinear feedback and noise. Depending on the adaptation and the noise level, this system may display noise-induced spiking, noise-perturbed oscillations, or stochastic bursting. We show how the system exhibits transitions between these dynamical regimes, as well as how one can enhance or suppress the coherence resonance or effectively control the features of the stochastic bursting. The setup can be considered a paradigmatic model for a neuron with a slow recovery variable or, more generally, as an excitable system under the influence of a nonlinear control mechanism. We employ a multiple timescale approach that combines the classical adiabatic elimination with averaging of rapid oscillations and stochastic averaging of noise-induced fluctuations by a corresponding stationary Fokker-Planck equation. This allows us to perform a numerical bifurcation analysis of a reduced slow system and to determine the parameter regions associated with different types of dynamics. In particular, we demonstrate the existence of a region of bistability, where the noise-induced switching between a stationary and an oscillatory regime gives rise to stochastic bursting.


Reuse of treated wastewater and sewage sludge for fertilization and irrigation.
The objective of the present work was to assess the short-term potential of treated wastewater and sewage sludge for ornamental lawn fertilization and irrigation. A field experiment was performed and the following treatments were considered: sewage sludge application + irrigation with public water; sewage sludge application + irrigation with treated wastewater; irrigation with public water; irrigation with treated wastewater (TW). Irrigation with treated wastewater showed a positive effect on lawn installation through higher growth of grass (1,667 cm) and higher dry matter yield (18,147 g m(-2)). These results represent a significant increase in the grass yield compared with public water irrigation. The grass height (2,606 cm) and dry matter yield (23,177 g m(-2)) increased even more, when sewage sludge produced in the wastewater treatment plant (WWTP) was applied to soil, which proves once more its benefits as an organic fertilizer. At the end of the experiment, an increase of some soil parameters (pH, electrical conductivity, organic matter, Ca2+, Na+, K+, Mg2+ and NH4+) was observed, indicating that treated wastewater irrigation can cause a soil sodization. This short-term study indicated that use of treated wastewater and sewage sludge for ornamental lawn fertilization and irrigation is an environmentally sustainable option for re-use of the WWTP by-products.


Dose- and ApoE isoform-dependent cognitive injury after cranial ⁵⁶Fe irradiation in female mice.
Apolipoprotein E (ApoE) plays an important role in lipid metabolism and neuronal repair. In humans, there are three major apoE isoforms: apoE2, apoE3 and apoE4. Compared to apoE3, apoE4 increases the risk to develop Alzheimer's disease, particularly in women, and of developing cognitive impairments after specific environmental challenges. ApoE isoform might also be a determinant of cognitive injury after cranial ⁵⁶Fe irradiation. To assess this possibility, in this study female apoE2, apoE3 and apoE4 mice were cranially irradiated with ⁵⁶Fe particles (600 MeV, 0, 1 or 2 Gy) and behaviorally tested 3 months later. Exploratory activity and measures of anxiety were also assessed as they can affect performance on cognitive tests. There were no effects of irradiation on exploratory activity or measures of anxiety in the open field or elevated zero maze. However, there were dose- and apoE isoform-dependent effects of irradiation on novel object recognition and spatial memory retention in the water maze. Compared to apoE2 and apoE3 mice, apoE4 mice were more sensitive to 2 Gy induced impairments in hippocampus-dependent spatial memory retention in the water maze after training to locate the first hidden platform location, but less sensitive to 2 Gy induced cortical impairments in novel object recognition. Conversely, of the irradiated mice, apoE4 mice irradiated with 1 Gy were the only group of mice that showed spatial memory retention for the second platform location after reversal learning in the water maze. Together, these data show that cranial ⁵⁶Fe irradiation causes dose- and apoE isoform-dependent cognitive impairments in female mice and that anatomical specificity might contribute to the relative sensitivity of apoE4 mice to develop space radiation-induced cognitive impairments.


Primary murine bone marrow cultures support continuous growth of infectious human trypanosomes.
The human parasite Trypanosoma brucei gambiense grew continuously at 37 degrees C in primary cultures of murine bone marrow. Cultured parasites remained virulent for mice. Rapid parasite growth coincided with the appearance of adherent adipocyte-epitheloid cell aggregates that also promoted hematopoiesis. This culture system should permit studies of host cell control of trypanosome proliferation, pathogenic effects of trypanosomes on blood cell development, and the relative trypanocidal and marrow suppressive activities of drugs.


Treatment-associated suicidal ideation and adverse effects in an open, multicenter trial of fluoxetine for major depressive episodes.
Some reports suggest that a subset of depressed patients may experience suicidality - that is increase or emergence of suicidal ideation (SI) or behavior--after initiation of an antidepressant. The time course and clinical correlates of this phenomenon have not been characterized in detail. We conducted a secondary analysis of a multicenter, prospective, open, 12-week trial of fluoxetine 20 mg in outpatients with nonpsychotic major depressive episodes. Adverse effects and other clinical features associated with the emergence of suicidality, defined using item 3 of the Hamilton Depression Rating Scale, were examined using Cox regression models. Among 414 subjects without SI at baseline, 59 (14.3%) reported SI on at least 1 postbaseline visit. In a Cox regression, emergence of activation and worsening of depression severity were independently associated with emergence of SI, along with female gender, younger age and having thoughts that life was not worth living prior to treatment. Treatment response and remission were significantly less likely among subjects who developed SI. New SI was relatively common in this trial of fluoxetine and associated with the emergence of activation and overall symptomatic worsening. Whether prophylaxis against or aggressive treatment of adverse events can decrease emergence of SI merits further study.


Enhanced insulin secretion and insulin sensitivity in young lambs with placental insufficiency-induced intrauterine growth restriction.
Intrauterine growth restriction (IUGR) is associated with persistent metabolic complications, but information is limited for IUGR infants. We determined glucose-stimulated insulin secretion (GSIS) and insulin sensitivity in young lambs with placental insufficiency-induced IUGR. Lambs with hyperthermia-induced IUGR (n = 7) were compared with control lambs (n = 8). GSIS was measured at 8 ± 1 days of age, and at 15 ± 1 days, body weight-specific glucose utilization rates were measured with radiolabeled d-glucose during a hyperinsulinemic-euglycemic clamp (HEC). IUGR lambs weighed 23% less (P < 0.05) than controls at birth. Fasting plasma glucose and insulin concentrations were not different between IUGR and controls for either study. First-phase insulin secretion was enhanced 2.3-fold in IUGR lambs compared with controls. However, second-phase insulin concentrations, glucose-potentiated arginine-stimulated insulin secretion, and β-cell mass were not different, indicating that IUGR β-cells have an intrinsic enhancement in acute GSIS. Compared with controls, IUGR lambs had higher body weight-specific glucose utilization rates and greater insulin sensitivity at fasting (1.6-fold) and hyperinsulinemic periods (2.4-fold). Improved insulin sensitivity for glucose utilization was not due to differences in skeletal muscle insulin receptor and glucose transporters 1 and 4 concentrations. Plasma lactate concentrations during HEC were elevated in IUGR lambs compared with controls, but no differences were found for glycogen content or citrate synthase activity in liver and muscle. Greater insulin sensitivity for glucose utilization and enhanced acute GSIS in young lambs are predicted from fetal studies but may promote conditions that exaggerate glucose disposal and lead to episodes of hypoglycemia in IUGR infants.


Automatised pharmacophoric deconvolution of plant extracts - application to Cinchona bark crude extract.
We present a development of the "Plasmodesma" dereplication method [Margueritte et al., Magn. Reson. Chem., 2018, 56, 469]. This method is based on the automatic acquisition of a standard set of NMR experiments from a medium sized set of samples differing by their bioactivity. From this raw data, an analysis pipeline is run and the data is analysed by leveraging machine learning approaches in order to extract the spectral fingerprints of the active compounds. The optimal conditions for the analysis are determined and tested on two different systems, a synthetic sample where a single active molecule is to be isolated and characterized, and a complex bioactive matrix with synergetic interactions between the components. The method allows the identification of the active compounds and performs a pharmacophoric deconvolution. The program is freely available on the Internet, with an interactive visualisation of the statistical analysis, at https://plasmodesma.igbmc.science.


Endothelial arginine resynthesis contributes to the maintenance of vasomotor function in male diabetic mice.
Argininosuccinate synthetase (ASS) is essential for recycling L-citrulline, the by-product of NO synthase (NOS), to the NOS substrate L-arginine. Here, we assessed whether disturbed arginine resynthesis modulates endothelium-dependent vasodilatation in normal and diabetic male mice. Endothelium-selective Ass-deficient mice (Assfl/fl/Tie2Cretg/- = Ass-KOTie2) were generated by crossing Assfl/fl mice ( = control) with Tie2Cre mice. Gene ablation in endothelial cells was confirmed by immunohistochemistry. Blood pressure (MAP) was recorded in 34-week-old male mice. Vasomotor responses were studied in isolated saphenous arteries of 12- and 34-week-old Ass-KOTie2 and control animals. At the age of 10 weeks, diabetes was induced in control and Ass-KOTie2 mice by streptozotocin injections. Vasomotor responses of diabetic animals were studied 10 weeks later. MAP was similar in control and Ass-KOTie2 mice. Depletion of circulating L-arginine by arginase 1 infusion or inhibition of NOS activity with L-NAME resulted in an increased MAP (10 and 30 mmHg, respectively) in control and Ass-KOTie2 mice. Optimal arterial diameter, contractile responses to phenylephrine, and relaxing responses to acetylcholine and sodium nitroprusside were similar in healthy control and Ass-KOTie2 mice. However, in diabetic Ass-KOTie2 mice, relaxation responses to acetylcholine and endothelium-derived NO (EDNO) were significantly reduced when compared to diabetic control mice. Absence of endothelial citrulline recycling to arginine did not affect blood pressure and systemic arterial vasomotor responses in healthy mice. EDNO-mediated vasodilatation was significantly more impaired in diabetic Ass-KOTie2 than in control mice demonstrating that endothelial arginine recycling becomes a limiting endothelial function in diabetes.