Global target profile of the kinase inhibitor bosutinib in primary chronic myeloid leukemia cells.
The detailed molecular mechanism of action of second-generation BCR-ABL tyrosine kinase inhibitors, including perturbed targets and pathways, should contribute to rationalized therapy in chronic myeloid leukemia (CML) or in other affected diseases. Here, we characterized the target profile of the dual SRC/ABL inhibitor bosutinib employing a two-tiered approach using chemical proteomics to identify natural binders in whole cell lysates of primary CML and K562 cells in parallel to in vitro kinase assays against a large recombinant kinase panel. The combined strategy resulted in a global survey of bosutinib targets comprised of over 45 novel tyrosine and serine/threonine kinases. We have found clear differences in the target patterns of bosutinib in primary CML cells versus the K562 cell line. A comparison of bosutinib with dasatinib across the whole kinase panel revealed overlapping, but distinct, inhibition profiles. Common among those were the SRC, ABL and TEC family kinases. Bosutinib did not inhibit KIT or platelet-derived growth factor receptor, but prominently targeted the apoptosis-linked STE20 kinases. Although in vivo bosutinib is inactive against ABL T315I, we found this clinically important mutant to be enzymatically inhibited in the mid-nanomolar range. Finally, bosutinib is the first kinase inhibitor shown to target CAMK2G, recently implicated in myeloid leukemia cell proliferation.


Plasma Oxytocin Level and Sexual Dysfunction in Depressed Women Treated by Either Fluoxetine or Citalopram: a Pilot Clinical Trial.
Sexual dysfunction is a common cause of selective serotonin reuptake inhibitor (SSRI) withdrawal. Various studies indicate that decreased oxytocin is involved as a mechanism of delayed ejaculation induced by SSRIs. The aim of the present pilot study was to evaluate and compare sexual dysfunction and oxytocin levels in women being treated with either fluoxetine or citalopram. Thirty-nine women with the diagnosis of major depressive disorder were enrolled in the study. A baseline blood sample was collected and each participant was given either fluoxetine 20 mg/d or citalopram 20 mg/d. After 1 month, a second blood sample was collected and sexual dysfunction was evaluated via the Female Sexual Function Index (FSFI) questionnaire. Twenty-three women completed the study (12 and 11 in the fluoxetine and citalopram groups, respectively). After 1 month, the FSFI scores were 22.8 ± 7.8 and 22.5 ± 4.8 in the fluoxetine and citalopram groups, respectively. The oxytocin levels were 187.8 ± 38.8 pg/mL and 214.6 ± 23.1 pg/mL in the fluoxetine and citalopram groups, respectively. Statistical analysis did not reveal any difference in the FSFI score between the two groups after 1 month (p = 0.89). However, the oxytocin levels were significantly lower in the fluoxetine group than in the citalopram group (p = 0.05). We also observed a positive relationship between the FSFI score and oxytocin level at 1 month after starting fluoxetine or citalopram (r = 0.43, p = 0.04).A positive relationship between the oxytocin level and FSFI score supports the hypothesis that the oxytocin level plays a role in sexual dysfunction induced by SSRIs.


Large-scale in silico mapping of complex quantitative traits in inbred mice.
Understanding the genetic basis of common disease and disease-related quantitative traits will aid in the development of diagnostics and therapeutics. The processs of gene discovery can be sped up by rapid and effective integration of well-defined mouse genome and phenome data resources. We describe here an in silico gene-discovery strategy through genome-wide association (GWA) scans in inbred mice with a wide range of genetic variation. We identified 937 quantitative trait loci (QTLs) from a survey of 173 mouse phenotypes, which include models of human disease (atherosclerosis, cardiovascular disease, cancer and obesity) as well as behavioral, hematological, immunological, metabolic, and neurological traits. 67% of QTLs were refined into genomic regions <0.5 Mb with approximately 40-fold increase in mapping precision as compared with classical linkage analysis. This makes for more efficient identification of the genes that underlie disease. We have identified two QTL genes, Adam12 and Cdh2, as causal genetic variants for atherogenic diet-induced obesity. Our findings demonstrate that GWA analysis in mice has the potential to resolve multiple tightly linked QTLs and achieve single-gene resolution. These high-resolution QTL data can serve as a primary resource for positional cloning and gene identification in the research community.


Recognition of a cell-surface oligosaccharide of pathogenic Salmonella by an antibody Fab fragment.
The 2.05 angstrom (A) resolution crystal structure of a dodecasaccharide-Fab complex revealed an unusual carbohydrate recognition site, defined by aromatic amino acids and a structured water molecule, rather than the carboxylic acid and amide side chains and a structured water molecule, rather than the carboxylic acid and amide side chains that are features of transport and other carbohydrate binding proteins. A trisaccharide epitope of a branched bacterial lipopolysaccharide fills this hydrophobic pocket (8 A deep by 7 A wide) in an entropy-assisted association (association constant = 2.05 x 10(5) liters per mole, enthalpy = -20.5 +/- 1.7 kilojoules per mole, and temperature times entropy = +10.0 +/- 2.9 kilojoules per mole). The requirement for the complementarity of van der Waals surfaces and the requirements of saccharide-saccharide and protein-saccharide hydrogen-bonding networks determine the antigen conformation adopted in the bound state.


Acute Fatty Liver Disease of Pregnancy: Updates in Pathogenesis, Diagnosis, and Management.
Acute fatty liver of pregnancy (AFLP) is an obstetric emergency characterized by maternal liver failure and may have complications for the mother and fetus, including death. This review examines recent literature on the epidemiology, pathogenesis, diagnosis, and treatment of acute fatty liver of pregnancy. Pathogenesis of this disease has been linked to defects in fatty acid metabolism during pregnancy, especially in the setting of fetal genetic defects in fatty acid oxidation. The value of screening all patients for these genetic defects remains to be determined. Distinguishing AFLP from other high-risk liver diseases of pregnancy that have overlap features, such as HELLP and preeclampsia, can be challenging. Although sensitive diagnostic tools such as the Swansea criteria have been developed, further work is needed to diagnose AFLP more quickly. Although survival rates have improved in the past 30 years, delay in diagnosis and treatment of AFLP has life-threatening consequences; an algorithmic approach to AFLP may be a valuable resource for clinicians. Future epidemiological and long-term studies will improve our prediction of women at risk for developing AFLP and determine the long-term consequences of this condition.


Changes in semantic fluency across childhood: Normative data from Australian-English speakers.
Verbal fluency tests are often used as part of an assessment battery to investigate children's lexical knowledge as well as executive function skills. To date, however, issues surrounding consistency of measurement cloud comparisons across studies, with the developmental performance of Australian-English speaking children also currently lacking. This study tracked verbal fluency development as measured by two semantic fluency tasks that included coding of fluency, clustering and switching type responses. Participants included 355 typically developing Australian-English speaking children (4-10 years) and 46 young adults. Total fluency was determined by the number of words produced for each category (Animals or Food), minus repetitions and rule violations. Semantic clusters (words generated within a subcategory) were coded while switches between single words or subcategories were differentiated and coded as either hard or cluster switches. Fluency showed consistent improvement over age. Cluster Switches and Hard Switches showed some evidence of a plateau in performance relative to fluency, but in opposite direction. Other measures showed no strong trends over age. Results were similar for both semantic categories. Our results highlight the rich information available within a semantic fluency task and the importance of differentiating hard and cluster switches in paediatric samples.


Treatment of osseous defects using Vicryl mesh (polyglactin 910) and the Brånemark implant: a case report.
The effectiveness of using Vicryl mesh (polyglactin 910) in combination with a Brånemark titanium implant is described. A maxillary central incisor with an apical osseous defect resulting from endodontic failure was treated with the Brånemark method of osseointegration for single tooth replacement. Vicryl mesh was used over the osseous defect site and uncovered 5 months later. New bone formation filling the defect and around the implant was observed.


[Clinical evaluation of tulobuterol patch in patients with mild or moderate persistent bronchial asthma-effects of long-term treatment on airway inflammation and hypersensitivity].
The tulobuterol transdermal therapeutic system (TTS) is the world's first commercially available transdermal preparation of tulobuterol, a beta-2 stimulant, that can maintain effective blood tulobuterol levels for 24 hours when applied once daily. In the present study, a total of 36 adult patients with mildly persistent (Step 2) or moderately persistent (Step 3) bronchial asthma 19 who were using inhalational steroids and 17 who were not used tulobuterol TTS for one year and underwent measurement of peak expiratory flow (PEF) once daily. Peripheral eosinophil count, serum eosinophil cationic protein (ECP) level and airway responsiveness (Dmin) were evaluated at 6 months and 1 year after the start of the study. PEF exhibited significant improvements at 6 months and 1 year in patients treated with or without inhalational steroids, while serum ECP was improved significantly only in the patients on inhalational steroids. Patients not using inhalational steroids exhibited no significant exacerbation of Dmin at either 6 months or 1 year: One-year treatment with tulobuterol TTS did not appear to cause tachyphylaxis. The significant improvements in Dmin at 6 months and 1 year in the patients using inhalational steroids suggested that inhalational steroids offer beneficial effects in controlling airway inflammation. Tulobuterol TTS is considered quite beneficial in improving quality of life (QOL) in patients with bronchial asthma because its incidence of adverse effects including palpitations and shivering is significantly lower than those of oral preparations, because of its remarkable improvement of pulmonary function and symptoms of airway obstruction without increasing airway responsiveness even after repeated use, and because it is simple to use and offers excellent clinical efficacy.


PhosTryp: a phosphorylation site predictor specific for parasitic protozoa of the family trypanosomatidae.
Protein phosphorylation modulates protein function in organisms at all levels of complexity. Parasites of the Leishmania genus undergo various developmental transitions in their life cycle triggered by changes in the environment. The molecular mechanisms that these organisms use to process and integrate these external cues are largely unknown. However Leishmania lacks transcription factors, therefore most regulatory processes may occur at a post-translational level and phosphorylation has recently been demonstrated to be an important player in this process. Experimental identification of phosphorylation sites is a time-consuming task. Moreover some sites could be missed due to the highly dynamic nature of this process or to difficulties in phospho-peptide enrichment. Here we present PhosTryp, a phosphorylation site predictor specific for trypansomatids. This method uses an SVM-based approach and has been trained with recent Leishmania phosphosproteomics data. PhosTryp achieved a 17% improvement in prediction performance compared with Netphos, a non organism-specific predictor. The analysis of the peptides correctly predicted by our method but missed by Netphos demonstrates that PhosTryp captures Leishmania-specific phosphorylation features. More specifically our results show that Leishmania kinases have sequence specificities which are different from their counterparts in higher eukaryotes. Consequently we were able to propose two possible Leishmania-specific phosphorylation motifs.We further demonstrate that this improvement in performance extends to the related trypanosomatids Trypanosoma brucei and Trypanosoma cruzi. Finally, in order to maximize the usefulness of PhosTryp, we trained a predictor combining all the peptides from L. infantum, T. brucei and T. cruzi. Our work demonstrates that training on organism-specific data results in an improvement that extends to related species. PhosTryp is freely available at http://phostryp.bio.uniroma2.it.


Reflected Light from Giant Planets in Habitable Zones: Tapping into the Power of the Cross-Correlation Function.
The direct detection of reflected light from exoplanets is an excellent probe for the characterization of their atmospheres. The greatest challenge for this task is the low planet-to-star flux ratio, which even in the most favourable case is of the order of 10-4 in the optical. This ratio decreases even more for planets in their host's habitable zone, typically lower than 10-7. To reach the signal-to-noise level required for such detections, we propose to unleash the power of the Cross Correlation Function in combination with the collecting power of next generation observing facilities. The technique we propose has already yielded positive results by detecting the reflected spectral signature of 51 Pegasi b (see Martins et al. 2015). In this work, we attempted to infer the number of hours required for the detection of several planets in their host's habitable zone using the aforementioned technique from theoretical EELT observations. Our results show that for 5 of the selected planets it should be possible to directly recover their reflected spectral signature.


A PAX1 enhancer locus is associated with susceptibility to idiopathic scoliosis in females.
Idiopathic scoliosis (IS) is a common paediatric musculoskeletal disease that displays a strong female bias. By performing a genome-wide association study (GWAS) of 3,102 individuals, we identify significant associations with 20p11.22 SNPs for females (P=6.89 × 10(-9)) but not males (P=0.71). This association with IS is also found in independent female cohorts from the United States of America and Japan (overall P=2.15 × 10(-10), OR=1.30 (rs6137473)). Unexpectedly, the 20p11.22 IS risk alleles were previously associated with protection from early-onset alopecia, another sexually dimorphic condition. The 174-kb associated locus is distal to PAX1, which encodes paired box 1, a transcription factor involved in spine development. We identify a sequence in the associated locus with enhancer activity in zebrafish somitic muscle and spinal cord, an activity that is abolished by IS-associated SNPs. We thus identify a sexually dimorphic IS susceptibility locus, and propose the first functionally defined candidate mutations in an enhancer that may regulate expression in specific spinal cells.


Comparison of AngioJet rheolytic pharmacomechanical thrombectomy versus AngioJet rheolytic thrombectomy in a porcine peripheral arterial model.
Rheolytic thrombectomy using the AngioJet catheter for arterial thrombosis has been shown to be effective in restoring blood flow. Additional infusion of thrombolytic agents via the AngioJet catheter results in combined rheolytic pharmacomechanical thrombolysis (PMT), which further enhances thrombectomy efficacy. However, the histologic response to rheolytic PMT therapy remains unclear. This study compares the acute and chronic vessel wall response and hemolysis due to conventional AngioJet rheolytic thrombectomy (RT) and AngioJet PMT in the porcine peripheral arterial model. A total of 19 juvenile pigs were divided into acute and chronic groups. In the acute group (n = 6), bilateral common carotid, femoral, and iliac arteries ranging from 3 to 6 mm in diameter were randomized to the control RT group or to PMT therapy. Vessels were analyzed 4 days following interventions. In the chronic group (n = 5), bilateral common carotid, femoral, and iliac arteries ranging from 3 to 6 mm in diameter were randomized to the control RT group or to PMT therapy. Vessels were analyzed at 30 days following interventions. Hemolytic evaluation was performed in additional eight pigs, which were randomized to either RT or PMT intervention. In the acute group, similar histologic injury grades were noted between the RT- and PMT-treated femoral and iliac vessels. Endothelial denudation in the RT and PMT vessels were 43% and 39% (NS), respectively. Vessels with intact internal elastic lamina (IEL) in the RT and PMT groups were 54% and 57% (NS), respectively. In vessels < 4 mm in diameter, fractured IEL in the AT and PMT groups occurred in 23% and 27% (NS), respectively. The degrees of smooth muscle cell (SMC) loss were similar for the RT- and PMT-treated vessels (45% and 40%, respectively; NS). In the chronic group, no differences were seen between the RT and PMT groups with respect to endothelial denudation, IEL fracture rate, or SMC loss. Similar degrees of medial thickening or intimal hyperplasia were noted in the RT and PMT groups (49% and 43%, respectively; NS). No difference in hemolytic effect was noted in the treatment groups. AngioJet rheolytic pharmacomechanical thrombectomy treatment incurs an equivalent safety profile in medium-caliber peripheral arteries when compared to rheolytic thrombectomy treatment. No difference in hemolytic reaction occurred in either group. The observed clinical efficacy of rheolytic pharmacomechanical thrombectomy does not result in untoward vessel injury compared to conventional rheolytic thrombectomy therapy.


Ovarian stromal cells as a source of cancer-associated fibroblasts in human epithelial ovarian cancer: A histopathological study.
Fibroblasts are a major component of cancer tissue and known to contribute to cancer progression. However, it remains unknown whether they are derived from local fibroblasts or of other origin. This study was designed to identify the contribution of local stromal cells to cancer stroma in human epithelial ovarian cancer. Seventy-six cases of surgically resected primary ovarian carcinoma (48 cases confined to the ovaries and 28 cases with distant metastases) and 17 cases of secondary ovarian tumor (e.g. colon cancer metastasized to the ovary) were enrolled in this study. The tissues were immunostained for forkhead box protein L2 (FOXL2), a transcription factor crucial for ovarian development and function, and markers for cancer-associated fibroblasts (CAFs) and inflammatory cells. Under normal condition, FOXL2 expression was restricted to ovarian stromal cells and some other types of cells in female genital tracts and never found in other sites of the body. FOXL2-positive cells were found in all primary and secondary tumors in the ovary, and were the dominant stromal cells in most cases. In contrast, only a few FOXL2-positive cells were found in peritoneal seeding sites of four serous carcinoma cases, and all the other tumors at extraovarian sites had no FOXL2-positive cells. FOXL2-positive cells in the ovarian lesion variably expressed CAFs markers, such as alpha-smooth muscle actin and fibroblast activating protein, as determined by double immunostaining. Background inflammation, but not histological subtype or origin of the neoplasm seemed to correlate with the proportion of FOXL2-positive cells. These results suggest that ovarian stromal cells are the main source of cancer stroma in the ovary but do not seem to move to distant sites via circulation together with tumor cells. Our results also support the hypothesis that cancer-associated fibroblasts may originate locally, which was previously demonstrated using animal models.


The epidemiology of burn wound infections: then and now.
Burn wound infections are a serious complication of thermal injury. Although pneumonia is now the most important infection in patients with burns, burn wound infection remains a serious complication unique to the burn recipient. The methods for managing thermal injury have evolved during the past 50 years. This evolution has been accompanied by changes in the etiology, epidemiology, and approach to prevention of burn wound infections. In the 1950s, 1960s, and 1970s and into the mid-1980s, burn wounds were treated by the exposure method, with application of topical antimicrobials to the burn wound surface and gradual debridement with immersion hydrotherapy. As early burn wound excision and wound closure became the focal point of burn wound management, accompanied by a change from immersion hydrotherapy to showering hydrotherapy, the rate of burn wound infection appeared to decrease. Few epidemiologic studies have been done since this change in the approach to management of thermal injury. There are few data on the epidemiology of burn wound infections from the era of early excision and closure. Data are needed on infection rates for excised and closed burn wounds, the etiologies of these infections, and the epidemiology and the prevention of such infections. Additional studies are needed on the indications for topical and antimicrobial prophylaxis and selective decontamination of the digestive tract.


Human immunodeficiency virus type 1 genomic RNA sequences in the female genital tract and blood: compartmentalization and intrapatient recombination.
Investigation of human immunodeficiency virus type 1 (HIV-1) in the genital tract of women is crucial to the development of vaccines and therapies. Previous analyses of HIV-1 in various anatomic sites have documented compartmentalization, with viral sequences from each location that were distinct yet phylogenetically related. Full-length RNA genomes derived from different compartments in the same individual, however, have not yet been studied. Furthermore, although there is evidence that intrapatient recombination may occur frequently, recombinants comprising viruses from different sites within one individual have rarely been documented. We compared full-length HIV-1 RNA sequences in the plasma and female genital tract, focusing on a woman with high HIV-1 RNA loads in each compartment who had been infected heterosexually and then transmitted HIV-1 by the same route. We cloned and sequenced 10 full-length HIV-1 RNA genomes from her genital tract and 10 from her plasma. We also compared viral genomes from the genital tract and plasma of four additional heterosexually infected women, sequencing 164 env and gag clones obtained from the two sites. Four of five women, including the one whose complete viral sequences were determined, displayed compartmentalized HIV-1 genomes. Analyses of full-length, compartmentalized sequences made it possible to document complex intrapatient HIV-1 recombinants that were composed of alternating viral sequences characteristic of each site. These findings demonstrate that the genital tract and blood harbor genetically distinct populations of replicating HIV-1 and provide evidence that recombination between strains from the two compartments contributes to rapid evolution of viral sequence variation in infected individuals.


Immune evasion of microsatellite unstable colorectal cancers.
Colorectal cancers (CRC) develop through 2 major pathways of genetic instability. In contrast to the majority of CRCs, which are characterized by chromosomal instability, high-level microsatellite unstable (MSI-H) CRCs arise as a consequence of the loss of DNA mismatch repair (MMR) functions and show accumulation of insertion and deletion mutations particularly in microsatellite sequences. MSI-H occurs in about 15% of CRCs, and virtually all CRCs occurring in the context of the hereditary cancer-predisposing Lynch syndrome. These tumors are characterized by a comparably good prognosis and a low frequency of distant metastases. Because of the expression of a defined set of tumor-specific antigens, MSI-H CRCs elicit a strong local and systemic antitumoral immune response of the host and therefore use different strategies to evade the control of the immune system. In this review, we will summarize novel molecular mechanisms that at the same time drive pathogenesis, immunogenicity and immune evasion during the development and progression of MSI-H CRCs. We will focus on the current knowledge about alterations in human leukocyte antigen (HLA) antigen presentation and discuss how immune evasion-while offering protection against local antitumoral immune responses-paradoxically might interfere with the ability of the tumor to form distant organ metastases.


High diagnostic value in rheumatoid arthritis of antibodies to the stratum corneum of rat oesophagus epithelium, so-called 'antikeratin antibodies'.
Serum antibodies to the stratum corneum of rat oesophagus epithelium, so-called 'antikeratin antibodies', have been largely demonstrated in rheumatoid arthritis (RA). IgM and IgG antibodies to this epithelium were studied by semiquantitative immunofluorescence in 528 patients with perfectly characterised rheumatic diseases, including 178 with classical or definite RA. Histological analysis of IgG antibodies showed that only antibodies which produce a linear laminated pattern restricted to the stratum corneum (IgG antikeratin antibodies) are highly specific for RA; all the other labelling patterns are not disease specific. By a semiquantitative evaluation of the stratum corneum fluorescence intensity it was shown that the diagnostic value of IgG antikeratin antibodies closely depends on their titre and it was established in objective conditions that the sensitivity is 43.26% when the specificity reaches 99.14%. A high titre of IgG antikeratin antibodies was actually pathognomonic for RA. Both the histological and semi-quantitative analyses showed that IgM antibodies to rat oesophagus epithelium, though frequently detected, are of no diagnostic value, either for RA or for any other rheumatic disease that was studied. From a review of all the international reports on IgG antikeratin antibodies it was found that, to date, 4080 patients, including 1694 with RA, have been assayed for antikeratin antibodies by 11 different research groups. Analysis of all the results obtained under comparable technical conditions showed that IgG antikeratin antibodies constitute the most specific serological criterion for the diagnosis of RA. Furthermore, it was found that their incidence does not depend on disease duration: they are present in one third of rheumatoid factor negative patients with RA, and they seem to be related to disease severity or activity, or both. Their detection in the diagnosis of rheumatic diseases should become systematic.


Methylmercury in freshwater fish linked to atmospheric mercury deposition.
A connection between accumulation of methylmercury (MeHg) in wild fish populations and atmospheric mercury deposition has not been made. Large databases for both MeHg in fish and atmospheric mercury deposition have been assimilated from monitoring efforts spanning the contiguous United States. Here, we compare results of these data sets and show that state-wide average concentrations of MeHg in a cosmopolitan freshwater fish, the largemouth bass Micropterus salmoides, are related positively to wet atmospheric Hg fluxes among most of the 25 states that are analyzed, which span a 5-fold range in Hg deposition. Differences in largemouth bass MeHg concentrations among states are unrelated to average precipitation depth, wet atmospheric acid deposition, or interstate variations in the type of water body (river, lake, reservoir) from which the fish were sampled. There are modest correlations between MeHg in bass and surface water pH, temperature, and wet atmospheric deposition of sulfate. However, when fish and atmospheric mercury results are combined at the state level, wet atmospheric Hg deposition accounts for about two-thirds of the variation in bass MeHg among most states, and stepwise multiple regression analysis reveals thatthese variables do not improve the linear model significantly. This suggests the accumulation of MeHg in wild fish populations is linked to atmospheric Hg loadings, two-thirds of which are estimated to be from anthropogenic sources.


Exterospecific component of the motion parallax field.
For the egocentric orientation of observers moving with respect to a plane (e.g., pilots and automobile drivers), the movement parallax field provides the main cue. The parallax field is split into a lamellar and a solenoidal part, and it is shown that the solenoidal part is purely propriospecific. For instance, it can be shown that this component can be completely canceled by an appropriate eye movement. Thus all exterospecific information is contained in the lamellar part, and this part is completely determined by the divergence of the parallax field. Thus the measure of expansion of the visual field as a function of direction of gaze is sufficient to provide all information available for egocentric orientation. It is further shown that the widely used focus of expansion, as introduced by Gibson, is not invariant against eye movements and does not (in general) correspond to extrema of the divergence.


Aminopeptidase N (CD13) as a target for cancer chemotherapy.
The enzyme aminopeptidase N (APN, also known as CD13) is a Zn(2+) dependent membrane-bound ectopeptidase that degrades preferentially proteins and peptides with a N-terminal neutral amino acid. Aminopeptidase N has been associated with the growth of different human cancers and suggested as a suitable target for anti-cancerous therapy. Different approaches have been used to develop new drugs directed to this target, including enzyme inhibitors as well as APN-targeted carrier constructs. This review discusses the prevalence and possible function of APN in malignant diseases, mainly solid tumors, as well as its "drugability" evaluated in preclinical in vivo models, and also provides a brief overview of current clinical trials focused on APN.


Treatment of Non-Infectious Corneal Injury: Review of Diagnostic Agents, Therapeutic Medications, and Future Targets.
Corneal injuries can occur secondary to traumatic, chemical, inflammatory, metabolic, autoimmune, and iatrogenic causes. Ocular infection may frequently occur concurrent to corneal injury; however, antimicrobial agents are excluded from this present review. While practitioners may primarily rely on clinical examination techniques to assess these injuries, several pharmacological agents, such as fluorescein, lissamine green, and rose bengal, can be used to formulate a diagnosis and develop effective treatment strategies. Practitioners may choose from several analgesic medications to help with patient comfort without risking further injury or delaying ocular healing. Atropine, cyclopentolate, scopolamine, and homatropine are among the most frequently used medications for this purpose. Additional topical analgesic agents may be used judiciously to augment patient comfort to facilitate diagnosis. Steroidal anti-inflammatory agents are frequently used as part of the therapeutic regimen. A variety of commonly used agents, including prednisolone acetate, loteprednol, difluprednate, dexamethasone, fluorometholone, and methylprednisolone are discussed. While these medications are effective for controlling ocular inflammation, side effects, such as elevated intraocular pressure and cataract formation, must be monitored by clinicians. Non-steroidal medications, such as ketorolac, bromfenac, nepafenac, and diclofenac, are additionally used for their efficacy in controlling ocular inflammation without incurring side effects seen with steroids. However, these agents have their own respective side effects, warranting close monitoring by clinicians. Additionally, ophthalmologists routinely employ several agents in an off-label manner for supplementary control of inflammation and treatment of corneal injuries. Patients with corneal injuries not infrequently have significant ocular surface disease, either as a concurrent pathology or as an exacerbation of previously existing disease. Several agents used in the management of ocular surface disease have also been found to be useful as part of the therapeutic armamentarium for treatment of corneal injuries. For example, several antibiotics, such as doxycycline and macrolides, have been used for their anti-inflammatory effects on specific cytokines that are upregulated during acute injuries. There has been a recent wave of interest in amniotic membrane therapies (AMTs), including topical, cryopreserved and dehydrated variants. AMT is particularly effective in ocular injuries with violation of corneal surface integrity due to its ability to promote re-epithelialization of the corneal epithelium. Blood-based therapies, including autologous serum tears, plasma-enriched growth factor eyedrops and autologous blood drops, have additionally been explored in small case series for effectiveness in challenging and recalcitrant cases. Protection of the ocular surface is also a vital component in the treatment of corneal injuries. Temporary protective methods, such as bandage contact lenses and mechanical closure of the eyelids (tarsorrhaphy) can be particularly helpful in selective cases. Glue therapies, including biologic and non-biologic variants, can also be used in cases of severe injury and risk of corneal perforation. Finally, there are a variety of recently introduced and in-development agents that may be used as adjuvant therapies in challenging patient populations. Neurotrophic corneal disease may occur as a result of severe or chronic injury. In such cases, recombinant human nerve growth factor (cenegermin), topical insulin, and several other novel agents may be an alternate and effective option for clinicians to consider.


A microRNA-7 binding site polymorphism in HOXB5 leads to differential gene expression in bladder cancer.
To investigate the biological function of HOXB5 in human bladder cancer and explore whether the HOXB5 3'-UTR SNP (1010A/G), which is located within the microRNA-7 binding site, was correlated with clinical features of bladder cancer. Expression of HOXB5 in 35 human bladder cancer tissues and 8 cell lines were examined using real-time PCR and immunohistochemistry. Next, we explored the biological function of HOXB5 in vitro using cell proliferation, migration and colony formation assays. Using bioinformatics, a SNP (1010A/G) was found located within the microRNA-7 binding site in the 3'-UTR of HOXB5. Real-time PCR was used to test HOXB5 expression affected by different alleles. Finally, multivariate logistic regression analysis was used to determine the relationship between SNP (1010A/G) frequency and clinical features in 391 cases. HOXB5 was frequently over-expressed both in bladder cancer tissues and cell lines. Inhibition of HOXB5 suppressed the oncogenic function of cancer cells. Next, we demonstrated that a SNP (1010A/G), located within the microRNA-7 binding site in the 3'-UTR of HOXB5, could affect HOXB5 expression in bladder cancer mainly by differential binding activity of microRNA-7 and SNP-related mRNA stability. Finally, we also showed the frequency of 1010G genotype was higher in cancer group compared to normal controls and correlated with the risk of high grade and high stage. HOXB5 is overexpressed in bladder cancer. A miRNA-binding SNP (1010A/G) located within 3'-UTR of HOXB5 is associated with gene expression and may be a promising prognostic factor for bladder cancer.


A Novel Photoelectrochemical Biosensor for Tyrosinase and Thrombin Detection.
A novel photoelectrochemical biosensor for step-by-step assay of tyrosinase and thrombin was fabricated based on the specific interactions between the designed peptide and the target enzymes. A peptide chain with a special sequence which contains a positively charged lysine-labeled terminal, tyrosine at the other end and a cleavage site recognized by thrombin between them was designed. The designed peptide can be fixed on surface of the CdTe quantum dots (QDs)-modified indium-tin oxide (ITO) electrode through electrostatic attraction to construct the photoelectrochemical biosensor. The tyrosinase target can catalyze the oxidization of tyrosine by oxygen into ortho-benzoquinone residues, which results in a decrease in the sensor photocurrent. Subsequently, the cleavage site could be recognized and cut off by another thrombin target, restoring the sensor photocurrent. The decrease or increase of photocurrent in the sensor enables us to assay tyrosinase and thrombin. Thus, the detection of tyrosinase and thrombin can be achieved in the linear range from 2.6 to 32 μg/mL and from 4.5 to 100 μg/mL with detection limits of 1.5 μg/mL and 1.9 μg/mL, respectively. Most importantly, this strategy shall allow us to detect different classes of enzymes simultaneously by designing various enzyme-specific peptide substrates.


Respiratory and ocular symptoms in workers exposed to potassium aluminium-tetrafluoride soldering flux.
Exposure to aluminium compounds, such as fluorides in gaseous and particulate form, places people who work in potrooms at risk for respiratory symptoms. Workers in potrooms, however, also are exposed to a number of other air contaminants. In this study, we present the first report of a dose-response relationship after exposure to potassium aluminium tetrafluoride (KAlF(4)) and the influence of smoking and atopy. All workers (308) from an industrial plant that used KAlF as soldering flux were invited to participate in the study. In all, 289 workers participated and 118 employees not exposed to chemicals in their professional work served as an unexposed group. In the first step, all subjects answered a questionnaire concerning respiratory symptoms and work history, and participated in a lung function examination. In a second step, all workers who reported work-related complaints from lower respiratory airways were invited to participate in medical examination, methacholine test, screening test of respiratory allergy, and skin prick test against KAlF(4). The exposed subjects had more symptoms than the unexposed group; dry cough odds ratio (OR): 5.17 (confidence interval 1.79-15.0), stuffy nose: 2.3 (1.25-4.22), nose bleeding: 10.7 (3.26-35.3) and ocular symptoms 5.01 (1.92-13.1) except for chest tightening and wheezing, and shortness of breath. The symptoms appeared in a dose response-like manner although the ORs between high and low exposed were significant for only chest tightening and wheezing, 2.62 (1.30-5.26) and stuffy nose 2.1 (1.22-3.66). Smokers and atopics did not report more frequent work-related symptoms. Smokers were significantly less hyperreactive than non-smokers, indicating a healthy-worker effect. No one showed a positive skin prick test against KAlF(4). In spite of exposure levels of KAlF(4 )well below the new Swedish threshold limit, value frequent respiratory and ocular symptoms were reported. No evidence of IgE mediated allergy was found.


The siting of UK nuclear reactors.
Choosing a suitable site for a nuclear power station requires the consideration and balancing of several factors. Some 'physical' site characteristics, such as the local climate and the potential for seismic activity, will be generic to all reactors designs, while others, such as the availability of cooling water, the area of land required and geological conditions capable of sustaining the weight of the reactor and other buildings will to an extent be dependent on the particular design of reactor chosen (or alternatively the reactor design chosen may to an extent be dependent on the characteristics of an available site). However, one particularly interesting tension is a human and demographic one. On the one hand it is beneficial to place nuclear stations close to centres of population, to reduce transmission losses and other costs (including to the local environment) of transporting electricity over large distances from generator to consumer. On the other it is advantageous to place nuclear stations some distance away from such population centres in order to minimise the potential human consequences of a major release of radioactive materials in the (extremely unlikely) event of a major nuclear accident, not only in terms of direct exposure but also concerning the management of emergency planning, notably evacuation.This paper considers the emergence of policies aimed at managing this tension in the UK. In the first phase of nuclear development (roughly speaking 1945-1965) there was a highly cautious attitude, with installations being placed in remote rural locations with very low population density. The second phase (1965-1985) saw a more relaxed approach, allowing the development of AGR nuclear power stations (which with concrete pressure vessels were regarded as significantly safer) closer to population centres (in 'semi-urban' locations, notably at Hartlepool and Heysham). In the third phase (1985-2005) there was very little new nuclear development, Sizewell B (the first and so far only PWR power reactor in the UK) being colocated with an early Magnox station on the rural Suffolk coast. Renewed interest in nuclear new build from 2005 onward led to a number of sites being identified for new reactors before 2025, all having previously hosted nuclear stations and including the semi-urban locations of the 1960s and 1970s. Finally, some speculative comments are made as to what a 'fifth phase' starting in 2025 might look like.


Stimulation of Ras guanine nucleotide exchange activity of Ras-GRF1/CDC25(Mm) upon tyrosine phosphorylation by the Cdc42-regulated kinase ACK1.
Ras-GRF1 is a brain-specific guanine nucleotide exchange factor (GEF) for Ras, whose activity is regulated in response to Ca(2+) influx and G protein-coupled receptor signals. In addition, Ras-GRF1 acts as a GEF for Rac when tyrosine-phosphorylated following G protein-coupled receptor stimulation. However, the mechanisms underlying the regulation of Ras-GRF1 functions remain incompletely understood. We show here that activated ACK1, a nonreceptor tyrosine kinase that belongs to the focal adhesion kinase family, causes tyrosine phosphorylation of Ras-GRF1. On the other hand, kinase-deficient ACK1 exerted no effect. GEF activity of Ras-GRF1 toward Ha-Ras, as defined by in vitro GDP binding and release assays, was augmented after tyrosine phosphorylation by ACK1. In contrast, GEF activity toward Rac1 remained latent, implying that ACK1 does not represent a tyrosine kinase that acts downstream of G protein-coupled receptors. Consistent with enhanced Ras-GEF activity, accumulation of the GTP-bound form of Ras within the cell was shown through the use of Ras-binding domain pull-down assays. Furthermore, Ras-dependent activation of ERK2 by Ras-GRF1 was enhanced following co-expression of activated ACK1. These results implicate ACK1 as an upstream modulator of Ras-GRF1 and suggest a signaling cascade consisting of Cdc42, ACK1, Ras-GRF1, and Ras in neuronal cells.


Learning better by repetition or variation? Is transfer at odds with task specific training?
Transfer of motor skills is the ultimate goal of motor training in rehabilitation practice. In children with Developmental Coordination Disorder (DCD), very little is known about how skills are transferred from training situations to real life contexts. In this study we examined the influence of two types of practice on transfer of motor skills acquired in a virtual reality (VR) environment. One hundred and eleven children with DCD and their typically developing (TD) peers, aged 6-10 years (M = 8.0 SD = 1.0) were randomly assigned to either variable (n = 56) or repetitive practice (n = 55). Participants in the repetitive practice played the same exergame (ski slalom) twice weekly for 20 minutes, over a period of 5 weeks, while those in the variable group played 10 different games. Motor skills such as balance tasks (hopping), running and agility tasks, ball skills and functional activities were evaluated before and after 5 weeks of training. ANOVA repeated measures indicated that both DCD and TD children demonstrated transfer effects to real life skills with identical and non-identical elements at exactly the same rate, irrespective of the type of practice they were assigned to. Based on these findings, we conclude that motor skills acquired in the VR environment, transfers to real world contexts in similar proportions for both TD and DCD children. The type of practice adopted does not seem to influence children's ability to transfer skills acquired in an exergame to life situations but the number of identical elements does.


TRIENNIAL GROWTH AND DEVELOPMENT SYMPOSIUM: Factors influencing bovine intramuscular adipose tissue development and cellularity.
Appearance, distribution, and amount of intramuscular fat (IMF), often referred to as marbling, are highly variable and depend on environmental and genetic factors. On the molecular level, the concerted action of several drivers, including hormones, receptors, transcription factors, etc., determines where clusters of adipocytes arise. Therefore, the aim of future studies remains to identify such factors as biological markers of IMF to increase the ability to identify animals that deposit IMF early in age to increase efficiency of high-quality meat production. In an attempt to unravel the cellular development of marbling, we investigated the abundance of markers for adipogenic differentiation during fattening of cattle and the transcriptome of muscle and dissected IMF. Markers of different stages of adipogenic differentiation are well known from cell culture experiments. They are usually transiently expressed, such as delta-like homolog 1 (DLK1) that is abundant in preadipocytes and absent during differentiation to mature adipocytes. It is even a greater challenge to detect those markers in live animals. Within skeletal muscles, hyperplasia and hypertrophy of adipocytes can be observed throughout life. Therefore, development of marbling requires, on the cellular level, recruitment, proliferation, and differentiation of adipogenic cells to store excess energy in the form of lipids in new cells. In a recent study, we investigated the localization and abundance of early markers of adipogenic differentiation, such as DLK1, in bovine muscle tissue. An inverse relationship between IMF content and number of DLK1-positive cells in bovine muscle was demonstrated. Considering the cellular environment of differentiating adipocytes in muscle and the secretory action of adipocytes and myocytes, it becomes obvious that cross talk between cells via adipokines and myokines may be important for IMF development. Secreted proteins can act on other cells, inhibiting or stimulating their function via autocrine and paracrine actions. Such factors with potential influence on IMF, among them, agouti signaling protein and thrombospondin 4, were identified in transcriptome analyses and further investigated. Furthermore, results from transcriptome analysis indicate involvement of genes that are not directly related to adipogenesis and lipid metabolism, providing new candidates for future research.


Catheter therapy of Swiss cheese ventricular septal defects using the Amplatzer muscular VSD occluder.
The medical and surgical management of patients with multiple muscular ventricular septal defects (VSDs) is associated with morbidity and mortality. Three children with Swiss cheese VSDs were treated with transcatheter occlusion of their multiple defects using the Amplatzer muscular VSD occluder. Seventeen defects were closed in five catheterization procedures. One patient had three devices placed in two procedures, the second had five defects closed in one procedure, and the third had nine defects closed in two procedures. Two patients had previously been treated with pulmonary artery banding and required subsequent surgical band removal. There was immediate reduction in the left-to-right shunting and clinical improvement in all patients. Complications included the need for blood transfusion during the two longest procedures and tricuspid valve regurgitation in one. Transcatheter occlusion of multiple VSDs is an acceptable alternative or adjunct to surgical therapy for these complex patients.


A novel calmodulin-like protein from the liver fluke, Fasciola hepatica.
An 18.2 kDa protein from the liver fluke, Fasciola hepatica has been identified and characterised. The protein shows strongest sequence similarity to egg antigen proteins from Schistosoma mansoni, Schistosoma japonicum and Clonorchis sinensis. The protein is predicted to adopt a calmodulin-like fold; it thus represents the third calmodulin-like protein to be characterised in F. hepatica and has been named FhCaM3. Compared to the classical calmodulin structure there are some variations. Most noticeably, the central, linker helix is disrupted by a cysteine residue. Alkaline native gel electrophoresis showed that FhCaM3 binds calcium ions. This binding event increases the ability of the protein to bind the hydrophobic fluorescent probe 8-anilinonaphthalene-1-sulphonate, consistent with an increase in surface hydrophobicity as seen in other calmodulins. FhCaM3 binds to the calmodulin antagonists trifluoperazine and W7, but not to the myosin regulatory light chain binding compound praziquantel. Immunolocalisation demonstrated that the protein is found in eggs and vitelline cells. Given the critical role of calcium ions in egg formation and hatching this suggests that FhCaM3 may play a role in calcium signalling in these processes. Consequently the antagonism of FhCaM3 may, potentially, offer a method for inhibiting egg production and thus reducing the spread of infection.


Laser capture microdissection of human pancreatic islets reveals novel eQTLs associated with type 2 diabetes.
Genome wide association studies (GWAS) for type 2 diabetes (T2D) have identified genetic loci that often localise in non-coding regions of the genome, suggesting gene regulation effects. We combined genetic and transcriptomic analysis from human islets obtained from brain-dead organ donors or surgical patients to detect expression quantitative trait loci (eQTLs) and shed light into the regulatory mechanisms of these genes. Pancreatic islets were isolated either by laser capture microdissection (LCM) from surgical specimens of 103 metabolically phenotyped pancreatectomized patients (PPP) or by collagenase digestion of pancreas from 100 brain-dead organ donors (OD). Genotyping (> 8.7 million single nucleotide polymorphisms) and expression (> 47,000 transcripts and splice variants) analyses were combined to generate cis-eQTLs. After applying genome-wide false discovery rate significance thresholds, we identified 1,173 and 1,021 eQTLs in samples of OD and PPP, respectively. Among the strongest eQTLs shared between OD and PPP were CHURC1 (OD p-value=1.71 × 10-24; PPP p-value = 3.64 × 10-24) and PSPH (OD p-value = 3.92 × 10-26; PPP p-value = 3.64 × 10-24). We identified eQTLs in linkage-disequilibrium with GWAS loci T2D and associated traits, including TTLL6, MLX and KIF9 loci, which do not implicate the nearest gene. We found in the PPP datasets 11 eQTL genes, which were differentially expressed in T2D and two genes (CYP4V2 and TSEN2) associated with HbA1c but none in the OD samples. eQTL analysis of LCM islets from PPP led us to identify novel genes which had not been previously linked to islet biology and T2D. The understanding gained from eQTL approaches, especially using surgical samples of living patients, provides a more accurate 3-dimensional representation than those from genetic studies alone.


High-Throughput Feasible Screening Tool for Determining Enzyme Stabilities against Organic Solvents Directly from Crude Extracts.
Biotransformations in organic chemistry frequently suffer from limitations caused by low water-solubility of substrates and product inhibition. Both, usually are addressed by the addition of organic cosolvents, which often accompanies at the expense of enzyme stability. A common method for measuring enzyme stability is to determine the melting temperature (Tm ) of the enzyme. However, current methods are limited to the application of purified enzymes. Herein, for the first time, an easy and fast (<1 h) high-throughput feasible method to determine enzyme stabilities directly from crude extracts is reported. In pure buffer, the Tm value measured in the crude extract was identical to that obtained for the purified enzyme. Through the addition of different organic compounds, the Tm values in the crude extract differed by up to 2.4 °C from that of the purified enzymes due to the presence of the host-cell proteins. Thus, the measurement of enzyme stabilities in crude extracts appears to represent conditions in whole-cell catalysts even better. The applied nano differential scanning fluorimetry technology is further proven to be suitable for whole-cell catalysts with two overexpressed enzymes; thus representing a tool for the rapid screening of natural and mutant enzyme libraries in terms of process stability for challenging biotransformations.


ACCURATE KAP METER CALIBRATION AS A PREREQUISITE FOR OPTIMISATION IN PROJECTION RADIOGRAPHY.
Modern X-ray units register the air kerma-area product, PKA, with a built-in KAP meter. Some KAP meters show an energy-dependent bias comparable with the maximum uncertainty articulated by the IEC (25 %), adversely affecting dose-optimisation processes. To correct for the bias, a reference KAP meter calibrated at a standards laboratory and two calibration methods described here can be used to achieve an uncertainty of <7 % as recommended by IAEA. A computational model of the reference KAP meter is used to calculate beam quality correction factors for transfer of the calibration coefficient at the standards laboratory, Q0, to any beam quality, Q, in the clinic. Alternatively, beam quality corrections are measured with an energy-independent dosemeter via a reference beam quality in the clinic, Q1, to beam quality, Q Biases up to 35 % of built-in KAP meter readings were noted. Energy-dependent calibration factors are needed for unbiased PKA Accurate KAP meter calibration as a prerequisite for optimisation in projection radiography.


Novel strategy to create hypoallergenic peanut protein-polyphenol edible matrices for oral immunotherapy.
Peanut allergy is an IgE-mediated hypersensitivity. Upon peanut consumption by an allergic individual, epitopes on peanut proteins bind and cross-link peanut-specific IgE on mast cell and basophil surfaces triggering the cells to release inflammatory mediators responsible for allergic reactions. Polyphenolic phytochemicals have high affinity to bind proteins and form soluble and insoluble complexes with unique functionality. This study investigated the allergenicity of polyphenol-fortified peanut matrices prepared by complexing various polyphenol-rich plant juices and extracts with peanut flour. Polyphenol-fortified peanut matrices reduced IgE binding to one or more peanut allergens (Ara h 1, Ara h 2, Ara h 3, and Ara h 6). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) suggested changes in secondary protein structure. Peanut protein-cranberry polyphenol fortified matrices triggered significantly less basophil degranulation than unmodified flour in an ex vivo assay using human blood and less mast cell degranulation when used to orally challenge peanut-allergic mice. Polyphenol fortification of peanut flour resulted in a hypoallergenic matrix with reduced IgE binding and degranulation capacity, likely due to changes in protein secondary structure or masking of epitopes, suggesting potential applications for oral immunotherapy.


Regulated efflux of photoreceptor outer segment-derived cholesterol by human RPE cells.
Genetic studies have linked age-related macular degeneration (AMD) to genes involved in high-density lipoprotein (HDL) metabolism, including ATP-binding cassette transporter A1 (ABCA1). The retinal pigment epithelium (RPE) handles large amounts of lipids, among others cholesterol, partially derived from internalized photoreceptor outer segments (OS) and lipids physiologically accumulate in the aging eye. To analyze the potential function of ABCA1 in the eye, we measured cholesterol efflux, the first step of HDL generation, in RPE cells. We show the expression of selected genes related to HDL metabolism in mouse and human eyecups as well as in ARPE-19 and human primary RPE cells. Immunofluorescence staining revealed localization of ABCA1 on both sides of polarized RPE cells. This was functionally confirmed by directional efflux to apolipoprotein AI (ApoA-I) of 3H-labeled cholesterol given to the cells via serum or via OS. ABCA1 expression and activity was modulated using a liver-X-receptor (LXR) agonist and an ABCA1 neutralizing antibody, demonstrating that the efflux was ABCA1-dependent. We concluded that the ABCA1-mediated lipid efflux pathway, and hence HDL biosynthesis, is functional in RPE cells towards both the basal (choroidal) and apical (subretinal) space. Impaired activity of the pathway might cause age-related perturbations of lipid homeostasis in the outer retina and thus may contribute to disease development and/or progression.


Gain-of-function RAF1 mutations cause Noonan and LEOPARD syndromes with hypertrophic cardiomyopathy.
Noonan and LEOPARD syndromes are developmental disorders with overlapping features, including cardiac abnormalities, short stature and facial dysmorphia. Increased RAS signaling owing to PTPN11, SOS1 and KRAS mutations causes approximately 60% of Noonan syndrome cases, and PTPN11 mutations cause 90% of LEOPARD syndrome cases. Here, we report that 18 of 231 individuals with Noonan syndrome without known mutations (corresponding to 3% of all affected individuals) and two of six individuals with LEOPARD syndrome without PTPN11 mutations have missense mutations in RAF1, which encodes a serine-threonine kinase that activates MEK1 and MEK2. Most mutations altered a motif flanking Ser259, a residue critical for autoinhibition of RAF1 through 14-3-3 binding. Of 19 subjects with a RAF1 mutation in two hotspots, 18 (or 95%) showed hypertrophic cardiomyopathy (HCM), compared with the 18% prevalence of HCM among individuals with Noonan syndrome in general. Ectopically expressed RAF1 mutants from the two HCM hotspots had increased kinase activity and enhanced ERK activation, whereas non-HCM-associated mutants were kinase impaired. Our findings further implicate increased RAS signaling in pathological cardiomyocyte hypertrophy.


The photolysis of α-hydroperoxycarbonyls.
In this work, we theoretically elucidated the mechanism and predicted the major products of the photolysis of α-hydroperoxycarbonyls, known to be products of the atmospheric oxidation of biogenic volatile organic compounds (BVOC) and components of secondary organic aerosol (SOA) in rural and remote areas. Using 2-hydroperoxypropanal OCHCH(OOH)CH3 as a model compound, we show that the likely major photolysis mechanism is a fast 1,5 H-shift in the initially excited singlet S1 state followed by spontaneous elimination of singlet oxygen to yield an enol HOCH[double bond, length as m-dash]CHCH3, while intersystem crossing (ISC) to the triplet T1 state and C-C scission into HC˙O + HOOC˙HCH3 followed by expulsion of a hydroxyl radical from the unstable HOOC˙HCH3 is another product channel. The direct S1 reaction was found to occur at such a high rate that the quantum yield in atmospheric conditions is expected to approach unity. In the atmosphere, the enol should generally react with OH radicals or tautomerize into the more stable carbonyl O[double bond, length as m-dash]CH-CH2CH3. Vinylalcohol is shown to be a major end product of the photolysis of hydroperoxyacetaldehyde, an isoprene oxidation product. Taking into account also the important enhancement of the absorption cross sections over those of the constituent monofunctional compounds as observed for the analogous β-ketohydroperoxides, (F. Jorand et al., J. Photochem. Photobiol. A: Chem., 2000, 134, 119-125) the atmospheric photolysis rate of α-hydroperoxycarbonyls was estimated to be in the range of (1 to 5) × 10-4 s-1, generally faster than the rate of their OH reactions.


Shear stress induces expression, intracellular reorganization and enhanced Notch activation potential of Jagged1.
Notch signaling and blood flow regulate vascular formation and maturation, but how shear stress affects the different components of the Notch pathway in endothelial cells is poorly understood. We show that laminar shear stress results in a ligand specific gene expression profile in endothelial cells (HUVEC). JAG1 expression increases while DLL4 expression decreases. Jagged1 shows a unique response by clustering intracellularly six to nine hours after the onset of flow. The formation of the Jagged1 clusters requires protein production, ER export and endocytosis. Clustering is associated with reduced membrane levels but is not affected by Notch signaling activity. Jagged1 relocalization is reversible, the clusters disappear and membrane levels increase upon removal of shear stress. We further demonstrate that the signaling potential of endothelial cells is enhanced after exposure to shear stress. Together we demonstrate a Jagged1 specific shear stress response for Notch signaling in endothelial cells.


VING: a software for visualization of deep sequencing signals.
Next generation sequencing (NGS) data treatment often requires mapping sequenced reads onto a reference genome for further analysis. Mapped data are commonly visualized using genome browsers. However, such software are not suited for a publication-ready and versatile representation of NGS data coverage, especially when multiple experiments are simultaneously treated. We developed 'VING', a stand-alone R script that takes as input NGS mapping files and genome annotations to produce accurate snapshots of the NGS coverage signal for any specified genomic region. VING offers multiple viewing options, including strand-specific views and a special heatmap mode for representing multiple experiments in a single figure. VING produces high-quality figures for NGS data representation in a genome region of interest. It is available at http://vm-gb.curie.fr/ving/. We also developed a Galaxy wrapper, available in the Galaxy tool shed with installation and usage instructions.


Oligodendroglia: metabolic supporters of neurons.
Oligodendrocytes are glial cells that populate the entire CNS after they have differentiated from oligodendrocyte progenitor cells. From birth onward, oligodendrocytes initiate wrapping of neuronal axons with a multilamellar lipid structure called myelin. Apart from their well-established function in action potential propagation, more recent data indicate that oligodendrocytes are essential for providing metabolic support to neurons. Oligodendrocytes transfer energy metabolites to neurons through cytoplasmic "myelinic" channels and monocarboxylate transporters, which allow for the fast delivery of short-carbon-chain energy metabolites like pyruvate and lactate to neurons. These substrates are metabolized and contribute to ATP synthesis in neurons. This Review will discuss our current understanding of this metabolic supportive function of oligodendrocytes and its potential impact in human neurodegenerative disease and related animal models.


The unseen invaders: introduced earthworms as drivers of change in plant communities in North American forests (a meta-analysis).
Globally, biological invasions can have strong impacts on biodiversity as well as ecosystem functioning. While less conspicuous than introduced aboveground organisms, introduced belowground organisms may have similarly strong effects. Here, we synthesize for the first time the impacts of introduced earthworms on plant diversity and community composition in North American forests. We conducted a meta-analysis using a total of 645 observations to quantify mean effect sizes of associations between introduced earthworm communities and plant diversity, cover of plant functional groups, and cover of native and non-native plants. We found that plant diversity significantly declined with increasing richness of introduced earthworm ecological groups. While plant species richness or evenness did not change with earthworm invasion, our results indicate clear changes in plant community composition: cover of graminoids and non-native plant species significantly increased, and cover of native plant species (of all functional groups) tended to decrease, with increasing earthworm biomass. Overall, these findings support the hypothesis that introduced earthworms facilitate particular plant species adapted to the abiotic conditions of earthworm-invaded forests. Further, our study provides evidence that introduced earthworms are associated with declines in plant diversity in North American forests. Changing plant functional composition in these forests may have long-lasting effects on ecosystem functioning.


Hooked on heart regeneration: the zebrafish guide to recovery.
While humans lack sufficient capacity to undergo cardiac regeneration following injury, zebrafish can fully recover from a range of cardiac insults. Over the past two decades, our understanding of the complexities of both the independent and co-ordinated injury responses by multiple cardiac tissues during zebrafish heart regeneration has increased exponentially. Although cardiomyocyte regeneration forms the cornerstone of the reparative process in the injured zebrafish heart, recent studies have shown that this is dependent on prior neovascularization and lymphangiogenesis, which in turn require epicardial, endocardial, and inflammatory cell signalling within an extracellular milieu that is optimized for regeneration. Indeed, it is the amalgamation of multiple regenerative systems and gene regulatory patterns that drives the much-heralded success of the adult zebrafish response to cardiac injury. Increasing evidence supports the emerging paradigm that developmental transcriptional programmes are re-activated during adult tissue regeneration, including in the heart, and the zebrafish represents an optimal model organism to explore this concept. In this review, we summarize recent advances from the zebrafish cardiovascular research community with novel insight into the mechanisms associated with endogenous cardiovascular repair and regeneration, which may be of benefit to inform future strategies for patients with cardiovascular disease.


Selection of soybean pods by the stink bugs, Nezara viridula and Piezodorus guildinii.
Different biological parameters of the stink bugs, Nezara viridula L. and Piezodorus guildinii Westwood (Hemiptera: Pentatomidae), are affected by the developmental stage of the soybean (Glycine max Merrill) pods they feed on. These effects of the soybean on the stink bugs could represent a selection pressure leading to the ability of these species to discriminate the phenological stage of soybean pods, and, therefore, to exhibit feeding preferences. We designed three studies: (1) Distant detection of soybean pods through an olfactometer; (2) Free choice tests to evaluate preferences for soybean pods of different developmental stages; (3) No choice tests to study effects of soybean pod development on feeding time and number of probes. Stink bugs showed no differential response to olfactometer arms with or without soybean pods, suggesting an inability to detect soybean volatiles. Free choice tests showed no species effects on pods selection, but significant differences among fifth instar nymphs, adult male, and adult females. Fifth instar nymphs fed more frequently on soybean pods of advanced development stages compared to female adults, despite previous evidence showing poor development of stink bugs fed pods of the same stage. No choice tests showed significant effects of stink bug species, stink bug stage and sex, and soybean pod phenology. N. viridula expressed shorter feeding times and higher numbers of probes than P. guildinii. The highest numbers of probes of both species were observed when they were fed soybean pods in early phenological stages. When placed in direct contact with food, fifth instar nymphs prefered to feed on more developed pods, despite these pods being suboptimal food items. These results suggest that for the ecological time framework of soybean-stink bugs coexistence, around thirty-five years in Argentina, the selection pressure was not enough for stink bugs to evolve food preferences that match their performance on soybean pods of different development stages.


The effect of educational attainment on cognition of older adults: results from the Mexican Health and Aging Study 2001 and 2012.
Objective: This paper seeks to document changes in the effect of educational attainment on cognitive function of older adults in Mexico, and measure gender differences using data from two time periods. Methods: The data come from the Mexican Health and Aging Study (MHAS), taking the cross-sections of adults aged 60 years or older interviewed in 2001 and 2012. We perform an OLS regression using standardized z-scores for five individual cognitive domains and for total cognition. Results: Total cognitive scores and educational attainment were higher for men than women in both years. When cognitive components were analyzed separately, women had higher verbal memory and verbal recall scores than men. The gender gap in overall cognition score was smaller in 2012 compared to 2001, while the gender gap in educational attainment was larger in 2012 than in 2001. Even though men had higher educational attainment than women, the effect of educational attainment on cognition was higher for women. Similarly, the difference between total scores for each task for men compared to women decreased between 2012 and 2001, except for verbal learning and verbal recall where the gender difference widened. Conclusions: If younger cohorts of women continue to progressively achieve higher levels of education, the gender gap in old-age cognition should close. Additional work should determine the mechanisms through which added formal education seems to translate into higher cognitive gains for women compared to men.


Gender-based power and couples' HIV risk in Uttar Pradesh and Uttarakhand, north India.
Gender inequality is a long-recognized driver of the HIV epidemic. However, few studies have investigated the association between gender-based power and HIV risk in India, which has the world's third largest HIV epidemic. Population-based data collected in 2003 from 3,385 couples residing in Uttar Pradesh and Uttarakhand, North India, were used to examine associations between gender-based power (wife's autonomy and husband's inequitable gender attitudes) and indicators of couples' HIV risk (whether the husband had had premarital sex with someone other than his eventual spouse, extramarital sex in the past year or STI symptoms in the past year). Structural equation modeling was used to create composite variables for the gender-based power measures and test their associations with HIV risk measures. Twenty-four percent of husbands had had premarital sex, 7% had had extramarital sex in the past year and 6% had had STI symptoms in the past year. Structural equation models indicated that wives who reported higher levels of autonomy were less likely than other wives to have husbands who had had extramarital sex in the past year (direct association) or STI symptoms in the past year (indirect association). Moreover, husbands who endorsed more inequitable gender attitudes were more likely than others to report having had premarital sex with someone other than their spouse, which in turn was associated with having had extramarital sex and STI symptoms in the past year. If the associations identified in this study reflect a causal relationship between gender-based power and HIV risk behavior, then HIV prevention programs that successfully address inequitable gender roles may reduce HIV risks in North India.


Motion perception by a moving observer in a three-dimensional environment.
Perceiving three-dimensional object motion while moving through the world is hard: not only must optic flow be segmented and parallax resolved into shape and motion, but also observer motion needs to be taken into account in order to perceive absolute, rather than observer-relative motion. In order to simplify the last step, it has recently been suggested that if the visual background is stationary, then foreground object motion, computed relative to the background, directly yields absolute motion. A series of studies with immobile observers and optic flow simulating observer movement have provided evidence that observers actually utilize this so-called "flow parsing" strategy (Rushton & Warren, 2005). We test this hypothesis by using mobile observers (as well as immobile ones) who judge the motion in depth of a foreground object in the presence of a stationary or moving background. We find that background movement does influence motion perception but not as much as predicted by the flow-parsing hypothesis. Thus, we find evidence that, in order to perceive absolute motion, observers partly use flow-parsing but also compensate egocentric motion by a global self-motion estimate.


Clinical Significance of Guanylate Cyclase Stimulator, Riociguat, on Right Ventricular Functional Improvement in Patients with Pulmonary Hypertension.
Riociguat is a soluble guanylate cyclase stimulator that improves hemodynamics in patients with pulmonary hypertension (PH). Accumulating evidence implicates the additional effect of riociguat on the increase in cardiac output. However, its mechanisms have not been fully understood. This study aimed to investigate whether riociguat could ameliorate right ventricular (RV) contraction as well as hemodynamics. We studied 45 patients with pulmonary arterial hypertension (14) or chronic thromboembolic pulmonary hypertension (31) and evaluated hemodynamics, using right-sided heart catheterization, before and after the administration of riociguat. RV function was assessed by echocardiography, including speckle-tracking echocardiography. Riociguat significantly improved the WHO functional class and reduced the mean pulmonary arterial pressure and vascular resistance. In addition, the cardiac index increased. RV remodeling was ameliorated after riociguat administration as assessed by the echocardiographic parameters, such as RV diameter and RV area index. RV function, including RV fractional area change and RV global longitudinal strain, also significantly improved, and their improvement was even observed in patients with mild PH after pulmonary endarterectomy or balloon pulmonary angioplasty. Furthermore, covariance analysis revealed that RV global longitudinal strain and RV fractional area change improved after riociguat administration, even with the same mean pulmonary arterial pressure, implicating the improvement of RV contractile function by riociguat, regardless of RV loading. Riociguat not only improved the hemodynamics of patients with PH but also ameliorated the echocardiographic parameters with RV function. RV strain could detect the subtle improvement in mild PH, and riociguat may have a benefit even after intervention, as assessed by speckle-tracking echocardiography.


Screening of Pleural Mesothelioma Cell Lines for Kinase Activity May Identify New Mechanisms of Therapy Resistance in Patients Receiving Platin-Based Chemotherapy.
Malignant pleural mesothelioma (MPM) is a rare, predominantly asbestos-related and biologically highly aggressive tumor associated with a dismal prognosis. Multimodal therapy consisting of platinum-based chemotherapy is the treatment of choice. The reasons underlying the rather poor efficacy of platinum compounds remain largely unknown. Kinase activity might influence cellular response to these regimens. For this exploratory study, we screened MPM cell lines (NCI-H2452, NCI-H2052, and MSTO-211H) differing in response to cisplatin and benign control fibroblasts (MRC-5) for overall phosphorylation patterns as well as kinase activity with respect to cellular response to cisplatin-based therapeutics. We analysed the cell lines for cellular kinases in a high-throughput manner using the highly innovative technique PamGene. Cell state analysis including apoptosis, necrosis, and cell viability was performed by using enzyme activity and fluorescent-based assays. Cisplatin alters cellular phosphorylation patterns affecting cell cycle, migration, adhesion, signal transduction, immune modulation, and apoptosis. In cisplatin-responsive cell lines, phosphorylation of AKT1 and GSK3B was decreased but could not be influenced in cisplatin-resistant NCI-H2452 cells. Cisplatin-responsive cell lines showed increased phosphorylation levels of JNK1/2/3 but decreased phosphorylation in cisplatin-resistant NCI-H2452 cells. Kinase phosphorylation and activity might play a crucial role in cellular response to cytostatic agents. Cisplatin influences phosphorylation patterns with distinct features in cisplatin-resistant cells. These alterations exert a significant impact on cell cycle, migration, adhesion, signal transduction, immune modulation, and apoptosis of the respective tumor cells. Based on our results, the induction of p38 or JNK1/3, or inhibition of AKT1 by, for example, BIA-6, might offer a positive synergistic effect by induction of an apoptotic response to cisplatin-based treatment, thus potentially enhancing the clinical outcome of MPM patients.


Lactobacillus plantarum alleviate aflatoxins (B1 and M1) induced disturbances in the intestinal genes expression and DNA fragmentation in mice.
This study aimed to assess the disturbances in intestinal genes expression and DNA fragmentation in mice treated orally with aflatoxin B1 (AFB1) or aflatoxin M1 (AFM1) and the protective activity of Lactobacillus plantarum (LP). Male Balb/c mice were divided into 6 groups including the control group, the group treated with 2 mg/kg b.w of LP (2 × 109 cfu/mL), the groups treated with AFB1 or AFM1 (100 μg/kg b.w), and the groups treated with AFB1 or AFM1 during, after or before LP. Small intestines were collected for the determination of DNA fragmentation, gene expression and target protein content. The results showed that AFB1 or AFM1 increased DNA fragmentation, down regulated the expressions of caspase-3, caspase-9, CYP3A13, Bax and p53 as well as up-regulated the expression of TNF-α and Bcl-2 and their target proteins. LP succeeded to alleviate the disturbances in DNA fragmentation and the expression of these genes. The improvement was more pronounced in the group co-administered with the toxins plus LP. It could be concluded that AFB1 and AFM1 induced disturbances in intestinal function via the disturbances in DNA fragmentation and genes expression. LP induced a potential protective effect and is considered a promising agent against the genotoxicity induced by these mycotoxins.


Control of mammary epithelial differentiation: basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity.
Functional differentiation in mammary epithelia requires specific hormones and local environmental signals. The latter are provided both by extracellular matrix and by communication with adjacent cells, their action being intricately connected in what appears to be a cascade of events leading to milk production. To distinguish between the influence of basement membrane and that of cell-cell contact in this process, we developed a novel suspension culture assay in which mammary epithelial cells were embedded inside physiological substrata. Single cells, separated from each other, were able to assimilate information from a laminin-rich basement membrane substratum and were induced to express beta-casein. In contrast, a stromal environment of collagen I was not sufficient to induce milk synthesis unless accompanied by cell-cell contact. The expression of milk proteins did not depend on morphological polarity since E-cadherin and alpha 6 integrin were distributed evenly around the surface of single cells. In medium containing 5 microM Ca2+, cell-cell interactions were impaired in small clusters and E-cadherin was not detected at the cell surface, yet many cells were still able to produce beta-casein. Within the basement membrane substratum, signal transfer appeared to be mediated through integrins since a function-blocking anti-integrin antibody severely diminished the ability of suspension-cultured cells to synthesize beta-casein. These results provide evidence for a central role of basement membrane in the induction of tissue-specific gene expression.


Comparing mothers' reports on the content of prenatal care received with recommended national guidelines for care.
The Public Health Service's Expert Panel on the Content of Prenatal Care Report in 1989 provided detailed guidelines for the components of each prenatal visit. However, the extent to which women were receiving the recommended care when the guidelines were being formulated has yet to be determined. The 1988 National Maternal and Infant Health Survey results permit an examination of the proportion of women who reported receiving some of the recommended procedures. Women were asked if they received six of the recommended procedures (blood pressure measurement, urine test, blood test, weight and height taken, pelvic examination, and pregnancy history) in the first two visits, and whether they received seven types of advice or counseling (nutrition; vitamin use; smoking, alcohol, and drug use cessation; breastfeeding; and maternal weight gain) any time during their pregnancy. Only 56 percent of the respondents said they received all of the recommended procedures in the first two visits, and only 32 percent of the respondents said they received advice in all of the areas. Logistic regression analysis indicated that women receiving their care from private offices were significantly less likely to receive all the procedures and advice than women at publicly funded sites of care. This study suggests that recommendations of the Public Health Service's expert panel were not being met.


Epigenetic history of an Arabidopsis trans-silencer locus and a test for relay of trans-silencing activity.
Meiotically heritable epimutations affecting transgene expression are not well understood, even and in particular in the plant model species, Arabidopsis thaliana. The Arabidopsis trans-silencer locus, C73, which encodes a fusion protein between the repressor of photomorphogenesis, COP1, and green fluorescent protein (GFP-COP1), heritably modifies the expression pattern and cop1-like cosuppression phenotypes of multiple GFP-COP1 target loci by transcriptional gene silencing. Here we describe three additional features of trans-silencing by the C73 locus. First, the silencing phenotype of C73 and of similar complex loci was acquired epigenetically over the course of no more than two plant generations via a stage resembling posttranscriptional silencing. Second, imprints imposed by the C73 locus were maintained heritably for at least five generations in the absence of the silencer with only sporadic spontaneous reversion. Third, the pairing of two other GFP-COP1 transgene loci, L91 and E82, showed an increased tendency for epigenetic modification when L91 carried an epigenetic imprint from C73, but not when E82 bore the imprint. The latter data suggest a transfer of trans-silencing activity from one transgene locus, C73, to another, namely L91. These results extend our operational understanding of interactions among transgenes in Arabidopsis.


Calcium-sensing receptor activation depresses synaptic transmission.
At excitatory synapses, decreases in cleft [Ca] arising from activity-dependent transmembrane Ca flux reduce the probability of subsequent transmitter release. Intense neural activity, induced by physiological and pathological stimuli, disturb the external microenvironment reducing extracellular [Ca] ([Ca](o)) and thus may impair neurotransmission. Increases in [Ca](o) activate the extracellular calcium sensing receptor (CaSR) which in turn inhibits nonselective cation channels at the majority of cortical nerve terminals. This pathway may modulate synaptic transmission by attenuating the impact of decreases in [Ca](o) on synaptic transmission. Using patch-clamp recording from isolated cortical terminals, cortical neuronal pairs and isolated neuronal soma we examined the modulation of synaptic transmission by CaSR. EPSCs were increased on average by 88% in reduced affinity CaSR-mutant (CaSR(-/-)) neurons compared with wild-type. Variance-mean analysis indicates that the enhanced synaptic transmission was due largely to an increase in average probability of release (0.27 vs 0.46 for wild-type vs CaSR(-/-) pairs) with little change in quantal size (23 +/- 4 pA vs 22 +/- 4 pA) or number of release sites (11 vs 13). In addition, the CaSR agonist spermidine reduced synaptic transmission and increased paired-pulse depression at physiological [Ca](o). Spermidine did not affect quantal size, consistent with a presynaptic mechanism of action, nor did it affect voltage-activated Ca channel currents. In summary, reduced CaSR function enhanced synaptic transmission and CaSR stimulation had the opposite effect. Thus CaSR provides a mechanism that may compensate for the fall in release probability that accompanies decreases in [Ca](o).


Coexistence between wildlife and humans at fine spatial scales.
Many wildlife species face imminent extinction because of human impacts, and therefore, a prevailing belief is that some wildlife species, particularly large carnivores and ungulates, cannot coexist with people at fine spatial scales (i.e., cannot regularly use the exact same point locations). This belief provides rationale for various conservation programs, such as resettling human communities outside protected areas. However, quantitative information on the capacity and mechanisms for wildlife to coexist with humans at fine spatial scales is scarce. Such information is vital, because the world is becoming increasingly crowded. Here, we provide empirical information about the capacity and mechanisms for tigers (a globally endangered species) to coexist with humans at fine spatial scales inside and outside Nepal's Chitwan National Park, a flagship protected area for imperiled wildlife. Information obtained from field cameras in 2010 and 2011 indicated that human presence (i.e., people on foot and vehicles) was ubiquitous and abundant throughout the study site; however, tiger density was also high. Surprisingly, even at a fine spatial scale (i.e., camera locations), tigers spatially overlapped with people on foot and vehicles in both years. However, in both years, tigers offset their temporal activity patterns to be much less active during the day when human activity peaked. In addition to temporal displacement, tiger-human coexistence was likely enhanced by abundant tiger prey and low levels of tiger poaching. Incorporating fine-scale spatial and temporal activity patterns into conservation plans can help address a major global challenge-meeting human needs while sustaining wildlife.


Brain-wide pathway for waste clearance captured by contrast-enhanced MRI.
The glymphatic system is a recently defined brain-wide paravascular pathway for cerebrospinal fluid (CSF) and interstitial fluid (ISF) exchange that facilitates efficient clearance of solutes and waste from the brain. CSF enters the brain along para-arterial channels to exchange with ISF, which is in turn cleared from the brain along para-venous pathways. Because soluble amyloid β clearance depends on glymphatic pathway function, we proposed that failure of this clearance system contributes to amyloid plaque deposition and Alzheimer's disease progression. Here we provide proof of concept that glymphatic pathway function can be measured using a clinically relevant imaging technique. Dynamic contrast-enhanced MRI was used to visualize CSF-ISF exchange across the rat brain following intrathecal paramagnetic contrast agent administration. Key features of glymphatic pathway function were confirmed, including visualization of para-arterial CSF influx and molecular size-dependent CSF-ISF exchange. Whole-brain imaging allowed the identification of two key influx nodes at the pituitary and pineal gland recesses, while dynamic MRI permitted the definition of simple kinetic parameters to characterize glymphatic CSF-ISF exchange and solute clearance from the brain. We propose that this MRI approach may provide the basis for a wholly new strategy to evaluate Alzheimer's disease susceptibility and progression in the live human brain.


Public preferences for the return of research results in genetic research: a conjoint analysis.
Recent policies specifying criteria about which individual research results to return leave considerable discretion to researchers. This study investigated the types of results that the public wants when participating in genetic research and whether preferences differ based on willingness to participate. A representative survey of US adults used conjoint analysis to measure priorities among eight principles of a results policy for a proposed large-cohort study. Policy preferences were measured using 12 tasks in which respondents chose between two groupings of the policy principles. Stratified analysis compared those self-identified as likely or unlikely to participate in genomic research. Of 1,515 respondents, 56% would participate in the proposed study. All eight principles were positively endorsed by participants (all P < 0.003), with priority placed on providing results at no cost and returning well-validated results for treatable and serious diseases. Providing detailed result reports was more highly valued than providing staff to explain results (P = 0.0005). Receiving results about major changes in risk was marginally disvalued by those unlikely to participate (P = 0.35). Public preferences for well-validated individual research results for serious, actionable diseases agree with emerging recommendations. However, because preferences for receiving individual research results vary, some choices should be offered to research participants.


Defining short and long sleep duration for future paediatric research: A systematic literature review.
Short and long sleep patterns in children have been associated with a range of poor health outcomes. However, there is no consensus regarding the definitions of these abnormal sleep parameters in childhood for use in paediatric research. Given that there is a clear lack of definitions for sleep duration throughout paediatric sleep literature, this review aimed to establish recommendations for standard cut-offs of short and long sleep for children aged 1-16 years to enable homogeneity in future studies of paediatric sleep duration. Four databases were systematically searched to identify prospective studies that defined short or long sleep patterns in children. Included papers (38) were assessed for methodological quality, and their definitions were extracted to examine the current applied cut-offs in the literature for short or long sleep duration. The definitions were analysed in a regression model to summarize applied cut-offs from subjective data into cut-offs for short and long sleep duration. These models were fitted to reference values of three commonly cited paediatric population studies to establish new definitions of sleep duration for future use in research. Across the age groups there was little consensus in applied cut-offs for short and long sleep duration. This study found the best compromise for short sleep was defined as the 2.5th centile (hours = 0.25*age + 11) and long sleep as the 97.5th centile (hours = 0.017*age2 - 0.68*age + 16) of sleep duration in children. Recommendations for the hourly cut-offs of short and long sleep duration based on these percentiles were described.


Piecing the puzzle together: case studies of international research in health-promoting sports clubs.
This paper seeks to review the current international health-promoting sports club (HPSC) research, drawing together findings based on case studies from various countries to illustrate the status of HPSCs. In addition, future challenges for HPSC research and implementation are considered. The review includes six case studies from five countries. In summary, there are two major research themes in this area, namely 'research into HPSC activity' and 'research into HPSC networks'. The first theme investigates the extent to which sports clubs and/or national sports organisations invest in health promotion (HP) - both in policy and practice. The latter theme is driven by an intention to widen the scope of HPSCs to reach novel internal actors, like parents, siblings, etc., and/or external non-sporting bodies, like communities, schools, etc. The future challenges for HPSC research require a better understanding of the motives, barriers and capacities of sports clubs and coaches. Sports organisations, clubs and coaches generally support the intent of the HPSC concept, but even with the best evidence- or theory-based HP programmes/guidelines/standards, nothing will happen in practice if the nature and capacities of sports clubs are not better acknowledged. Therefore, a call for embracing implementation science is finally made to enhance implementation.


Recombinant sterol esterase from Ophiostoma piceae: an improved biocatalyst expressed in Pichia pastoris.
The ascomycete Ophiostoma piceae produces a sterol esterase (OPE) with high affinity towards p-nitrophenol, glycerol and sterol esters. Its hydrolytic activity on natural mixtures of triglycerides and sterol esters has been proposed for pitch biocontrol in paper industry since these compounds produce important economic losses during paper pulp manufacture. Recently, this enzyme has been heterologously expressed in the methylotrophic yeast Pichia pastoris, and the hydrolytic activity of the recombinant protein (OPE*) studied. After the initial screening of different clones expressing the enzyme, only one was selected for showing the highest production rate. Different culture conditions were tested to improve the expression of the recombinant enzyme. Complex media were better than minimal media for production, but in any case the levels of enzymatic activity were higher (7-fold in the best case) than those obtained from O. piceae. The purified enzyme had a molecular mass of 76 kDa, higher than that reported for the native enzyme under SDS-PAGE (60 kDa). Steady-state kinetic characterization of the recombinant protein showed improved catalytic efficiency for this enzyme as compared to the native one, for all the assayed substrates (p-nitrophenol, glycerol, and cholesterol esters). Different causes for this were studied, as the increased glycosylation degree of the recombinant enzyme, their secondary structures or the oxidation of methionine residues. However, none of these could explain the improvements found in the recombinant protein. N-terminal sequencing of OPE* showed that two populations of this enzyme were expressed, having either 6 or 8 amino acid residues more than the native one. This fact affected the aggregation behaviour of the recombinant protein, as was corroborated by analytical ultracentrifugation, thus improving the catalytic efficiency of this enzyme. P. pastoris resulted to be an optimum biofactory for the heterologous production of recombinant sterol esterase from O. piceae, yielding higher activity levels than those obtained with the saprophytic fungus. The enzyme showed improved kinetic parameters because of its modified N-terminus, which allowed changes in its aggregation behaviour, suggesting that its hydrophobicity has been modified.


A comparison of the leaf gel extracts of Aloe ferox and Aloe vera in the topical treatment of atopic dermatitis in Balb/c mice.
Aloe vera gel is widely used in the treatment of an array of disturbances, especially skin disorders. The wound-healing effects have been attributed to its moisturizing and anti-inflammatory effects as well as its beneficial effect on the maturation of collagen. The aim of the present study is to compare the effects of topically applied extracts of Aloe ferox with that of Aloe vera on the symptoms as well as IgE levels of a mouse model of atopic dermatitis (AD). Mice were sensitized and challenged with 2,4-dinitrochlorobenzene and treated afterwards for 10 consecutive days with the gels of either A. ferox or A. vera applied topically to the affected areas. A placebo gel was used for the control mice. Blood was collected at the beginning and end of the treatment period to measure serum IgE levels. Although the gels of both the Aloe species inhibited the cutaneous inflammatory response as well as serum IgE levels in the rats, the extracts of A. ferox were superior to that of A. vera in reducing IgE levels. The gels of A. ferox and A. vera, applied topically, may be a safe and useful alternative to antihistamines and topical corticosteroids, for the treatment of patients suffering from recurring chronic AD.


WO3 nanosheets rich in oxygen vacancies for enhanced electrocatalytic N2 reduction to NH3.
The Haber-Bosch process for industrial-scale NH3 production suffers from harsh conditions and serious CO2 release. Electrochemical N2 reduction is an alternative approach to synthesize NH3 under ambient conditions, but it requires highly-efficient electrocatalysts for the N2 reduction reaction (NRR). In this Communication, we demonstrate that WO3 nanosheets rich in oxygen vacancies (R-WO3 NSs) exhibit greatly enhanced NRR performances. In 0.1 M HCl, such R-WO3 NSs achieve a large NH3 yield of 17.28 μg h-1 mgcat.-1 and a high faradaic efficiency of 7.0% at -0.3 V vs. a reversible hydrogen electrode, much superior to the WO3 nanosheets deficient in oxygen vacancies (6.47 μg h-1 mgcat.-1 and 1.02%). Remarkably, R-WO3 NSs also show high electrochemical stability.