The relationship between family functioning and caregiving appraisal of dementia family caregivers: caregiving self-efficacy as a mediator.
The aim of this study was to explore caregiving self-efficacy as a mediator for the association between family functioning and caregiving appraisal of dementia family caregivers in Taiwan. This study adopted a cross-sectional correlational design. Purposive sampling was used to recruit 115 dyads of dementia patients and family caregivers from the outpatient neurological clinics of two hospitals in northern Taiwan. Data were gathered through interviews with a structured questionnaire, which included demographic characteristics for caregivers and patients, family functioning, caregiving self-efficacy, as well as positive and negative aspects of caregiving appraisal. Family functioning, patients' activities of daily living score, Neuropsychiatric Inventory caregiver distress, and three domains of self-efficacy were significantly associated with caregiver burden. Hierarchical multiple regression analyses indicated that self-efficacy for obtaining respite (SE-OR) significantly explained 20.5% of the variance in caregiver esteem. Caregiver perceived worsened health status, family functioning, and SE-OR significantly explained 59% of the variance in caregiver burden. The mediation test only supported the partially mediating role of SE-OR on the relationship between family functioning and caregiver burden, while the mediating effect of self-efficacy for responding to disruptive behaviours and controlling upsetting thoughts were insignificant. Our findings provided preliminary evidence for health professionals recommending that future studies should assess the family dynamic and health problems of caregivers, and develop appropriate family-centred interventions that focus on strengthening interfamily support and respite services to alleviate caregiver burden.


Conformational changes in serpins: II. The mechanism of activation of antithrombin by heparin.
Antithrombin, uniquely among plasma serpins acting as proteinase inhibitors in the control of the blood coagulation cascade, circulates in a relatively inactive form. Its activation by heparin, and specifically by a pentasaccharide core of heparin, has been shown to involve release of the peptide loop containing the reactive centre from partial insertion in the A sheet of the molecule. Here we compare the structures of the circulating inactive form of antithrombin with the activated structure in complex with heparin pentasaccharide. We show that the rearrangement of the reactive centre loop that occurs upon activation is part of a widespread conformational change involving a realignment of the two major domains of the molecule. We also examine natural mutants that possess high affinity for heparin pentasaccharide, and relate the kinetics of their interaction with heparin pentasaccharide to the structural transitions occuring in the activation process.


Contrasting roles for Myc and Mad proteins in cellular growth and differentiation.
The positive effects of Myc on cellular growth and gene expression are antagonized by activities of another member of the Myc superfamily, Mad. Characterization of the mouse homolog of human mad on the structural level revealed that domains shown previously to be required in the human protein for anti-Myc repression, sequence-specific DNA-binding activity, and dimerization with its partner Max are highly conserved. Conservation is also evident on the biological level in that both human and mouse mad can antagonize the ability of c-myc to cooperate with ras in the malignant transformation of cultured cells. An analysis of c-myc and mad gene expression in the developing mouse showed contrasting patterns with respect to tissue distribution and developmental stage. Regional differences in expression were more striking on the cellular level, particularly in the mouse and human gastrointestinal system, wherein c-Myc protein was readily detected in immature proliferating cells at the base of the colonic crypts, while Mad protein distribution was restricted to the postmitotic differentiated cells in the apex of the crypts. An increasing gradient of Mad was also evident in the more differentiated subcorneal layers of the stratified squamous epithelium of the skin. Together, these observations support the view that both downregulation of Myc and accumulation of Mad may be necessary for progression of precursor cells to a growth-arrested, terminally differentiated state.


Carotid thromboses in the early postoperative period after carotid endarterectomy.
The paper analyzes 10 cases of internal carotid artery (ICA) thrombosis, which occurred after 635 carotid endarterectomies (CEA), carried out from January 1997 to July 2004. CEA procedures included 346 (54.5%) open CEAs with PTFE patch angioplasty and 289 (45.5%) eversion CEAs. Patients with thromboses in the reconstructed area (n=10) had the profiles of comorbidities and cerebrovascular insufficiency grade similar to the total CEA group (p>0.1). Differences concerned the higher rate of atrial fibrillation cases (10%), diabetes mellitus (30%) and contralateral ICA occlusions (20%) in the group of thromboses (p>0.1). Among these 10 patients, 5 underwent eversion CEA (1.44% of the total eversion CEA group) and 5 - open CEA with patch angioplasty (1.73%). In 8 patients thromboses manifested as local neurological symptoms in the area supplied by the operated carotid artery. One patient demonstrated the progression of general cerebral symptomatology, while in the tenth patient thrombosis was accompanied with cerebral coma. Urgent ultrasonography in 7 patients failed to detect a blood flow in ICA, that proved the diagnosis of ICA thrombosis; in 3 patients ultrasonography showed a mural thrombosis. Nine patients underwent reoperation in emergency, while for the tenth patient an intervention was contraindicated due to the thrombosis of middle cerebral artery, unstable clinical state (cerebral coma) and progression of neurological symptomatology. PTFE arterial graft was implantedin 7 reoperated patients with ICA thrombosis, thrombectomy from ICA was carried out in 1 patient and in another one patient a thrombectomy from reconstructed segment with PTFE patch angioplasty of the arteriotomy defect was fulfilled. In 3 reoperated patients a complete resolution of neurological symptoms was evident 24 hours after intervention, in 2 patients neurological symptomatology regressed over 2-4 weeks. In another 3 reoperated patients local neurological symptoms persisted; 1 patient demonstrated postoperative progression of neurological symptomatology and coma with fatal outcome. The last patient, for whom a reoperation was contraindicated, died due to aggravation of cerebral coma. Dynamic follow-up in early postoperative period after CEA, especially in the first 6 hours, as well as an urgent duplex scanning of the reconstructed area in case of neurological symptoms development, provides timely diagnosis of postoperative thrombosis and the success of reoperations. Adjustments to patient's cardiac status and appropriate intraoperative heparinization can decrease the risk of this complication. An urgent reoperation aimed at the restoration of carotid blood flow, immediately after the diagnosis of thrombosis (provided the absence of deep coma), promotes a complete or partial regression of neurological symptoms and helps to prevent a progression of cerebrovascular disturbances.


Molecular epidemiology of breast cancers in northern and southern Japan: the frequency, clustering, and patterns of p53 gene mutations differ among these two low-risk populations.
Comparison of acquired mutations in the p53 tumor suppressor gene can illuminate factors contributing to carcinogenesis among cancer cohorts. Japan has an ethnically homogeneous population with a low incidence of breast cancer. Previously we reported an unusual frequency, allelic status, and clustering of mutations in breast cancers from the northern part of the main Japanese island. To extend these findings, exons 2-11 and adjacent intronic sequences were analysed in tumors of women from northern (Hokkaido) and southern (Tokushima) Japan. The frequency of breast cancers with p53 gene mutations in the Hokkaido group is the highest reported (81%) while that in Tokushima (28%) is similar to most other populations. Thirteen of the 19 mutations (68.4%) in the Hokkaido cohort were heterozygous, an unusually high frequency for p53 mutations in any tumor type. There were three missense mutations at codon 175, a known hotspot for alterations in the p53 gene, and three missense mutations at codon 179, a rare site for p53 changes. In addition, the patterns of p53 gene mutation differed between the two Japanese cohorts (P=0.04). The multiple differences in acquired p53 mutations suggest unsuspected biological differences among breast cancers in northern and southern Japan. In addition, the high frequency of p53 mutations in breast cancers from Hokkaido predicted a poorer prognosis for this population which was confirmed on examination of mortality data.


Why we need confidence intervals.
The estimation approach to statistical analysis aims to quantify the effect of interest as an "estimate" of a clinically relevant quantity and to quantify the uncertainty in this estimate by means of a confidence interval (CI). As such, results expressed in this form are much more informative than results presented just as p values. This article focuses on the principles rather than the mathematics of CIs and discusses interpretation of CIs and some common misuses. CIs can be constructed for almost all analyses. They are especially useful for avoiding misinterpretation of nonsignificant results of small studies. CIs should be provided routinely for the main results of trials and observational studies.


Age at breast feeding start and postneonatal growth and survival.
Colostrum has important anti-infective properties. It may also somehow promote the development of the child's immunological system. Discarding colostrum, as practised in some cultures, could thus have adverse health consequences beyond the neonatal period. To test this hypothesis, the age at breast feeding start of 734 healthy newborns in urban Guinea-Bissau was ascertained. The children were then prospectively followed up to 3 years of age. Eighty nine deaths occurred during the study. The probability of death in the age interval 28 days to 3 years was about 20%. The child's age at breast feeding start had no statistical impact on postneonatal growth or survival. As a single measure, early breast feeding start is not likely to make much difference for the long term growth or survival of children living under material poverty conditions.


ZnT5 variant B is a bidirectional zinc transporter and mediates zinc uptake in human intestinal Caco-2 cells.
Zinc is an essential micronutrient, so it is important to elucidate the molecular mechanisms of zinc homeostasis, including the functional properties of zinc transporters. Mammalian zinc transporters are classified in two major families: the SLC30 (ZnT) family and the SLC39 family. The prevailing view is that SLC30 family transporters function to reduce cytosolic zinc concentration, either through efflux across the plasma membrane or through sequestration in intracellular compartments, and that SLC39 family transporters function in the opposite direction to increase cytosolic zinc concentration. We demonstrated that human ZnT5 variant B (ZnT5B (hZTL1)), an isoform expressed at the plasma membrane, operates in both the uptake and the efflux directions when expressed in Xenopus laevis oocytes. We measured increased activity of the zinc-responsive metallothionein 2a (MT2a) promoter when ZnT5b was co-expressed with an MT2a promoter-reporter plasmid construct in human intestinal Caco-2 cells, indicating increased total intracellular zinc concentration. Increased cytoplasmic zinc concentration mediated by ZnT5B, in the absence of effects on intracellular zinc sequestration by the Golgi apparatus or endoplasmic reticulum, was also confirmed by a dramatically enhanced signal from the zinc fluorophore Rhodzin-3 throughout the cytoplasm of Caco-2 cells overexpressing ZnT5B at the plasma membrane when compared with control cells. Our findings demonstrate clearly that, in addition to mediating zinc efflux, ZnT5B at the plasma membrane can function to increase cytoplasmic zinc concentration, thus indicating a need to reevaluate the current paradigm that SLC30 family zinc transporters operate exclusively to decrease cytosolic zinc concentration.


Human nocturnal frontal lobe epilepsy: pharmocogenomic profiles of pathogenic nicotinic acetylcholine receptor beta-subunit mutations outside the ion channel pore.
Certain mutations in specific parts of the neuronal nicotinic acetylcholine receptor (nAChR) subunit genes CHRNA4, CHRNB2, and probably CHRNA2, can cause autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). All but one of the known causative mutations are located in the second transmembrane region (TM2), which serves as the major ion poreforming domain of the receptor. Functional characterization of these ADNFLE mutations has shown that although each mutant exhibits specific properties, they all confer a gain of function with increased sensitivity to acetylcholine. In this work, we characterize the second and third ADNFLE-associated mutations that are external to TM2 but affect different amino acid residues within the third transmembrane region (TM3). The two new CHRNB2 mutations were identified in three families of Turkish Cypriot, Scottish, and English origin. These TM3 mutations elicit the same gain of function pathomechanism as observed for the TM2 mutations with enhanced acetylcholine sensitivity, despite their unusual localization within the gene. Electrophysiological experiments, including single channel measurements, revealed that incorporation of these new mutant subunits does not affect the conductance of the ionic pore but increases the probability of opening. Determination of the sensitivity to nicotine for nAChRs carrying mutations in TM2 and TM3 showed clear differences in the direction and the extent to which the window current for nicotine sensitivity was shifted for individual mutations, indicating differences in pharmacogenomic properties that are not readily correlated with increased ACh affinity.


On the CUSP: the politics and prospects of scaling social norms change programming.
In the past decades, donors and development actors have been increasingly mindful of the evidence to support long-term, dynamic social norms change. This paper draws lessons and implications on scaling social norms change initiatives for gender equality to prevent violence against women and girls (VAWG) and improve sexual and reproductive health and rights (SRHR), from the Community for Understanding Scale Up (CUSP). CUSP is a group of nine organisations working across four regions with robust experience in developing evidence-based social norms change methodologies and supporting their scale-up across various regions and contexts. More specifically, the paper elicits learning from methodologies and experiences from five CUSP members - GREAT, IMAGE, SASA!, Stepping Stones, and Tostan. The discussion raises political questions around the current donor landscape including those positioned to assume leadership to take such methodologies to scale, and the current evaluation paradigm to measure social norms change at scale. CUSP makes the following recommendations for donors and implementers to scale social norms initiatives effectively and ethically: invest in longer-term programming, ensure fidelity to values of the original programmes, fund women's rights organisations, prioritise accountability to their communities and demands, critically examine the government and marketplace's role in scale, and rethink evaluation approaches to produce evidence that guides scale-up processes and fully represents the voices of activists and communities from the Global South.


The gluteus maximus muscle-splitting myocutaneous flap for treatment of sacral and coccygeal pressure ulcers.
We describe a modified technique using the gluteus maximum muscle as a splitting myocutaneous flap to close specifically low sacral and coccygeal pressure ulcers. Twenty-eight patients with sacral or coccygeal stage IV pressure ulcers (average size 4 x 4 cm) underwent a gluteus maximus muscle-splitting myocutaneous flap when conservative treatment failed to heal the ulcer. Twenty-seven of the 28 patients had complete healing of the pressure ulcer site at an average follow-up of 15 months (range 2 to 40 months). Complications occurred in 7 patients, requiring revision of the flap in 2 patients. The advantages of this technique include reduced blood loss, preservation of most of the gluteus maximus for future use, and retained function of the gluteus maximus for stair climbing and single-limb support in the ambulatory patient. We recommend the gluteus maximus muscle-splitting myocutaneous flap as the procedure of choice for closure of small low sacral or coccygeal ulcers in both the ambulatory and nonambulatory patient.


CPEB and miR-15/16 Co-Regulate Translation of Cyclin E1 mRNA during Xenopus Oocyte Maturation.
Cell cycle transitions spanning meiotic maturation of the Xenopus oocyte and early embryogenesis are tightly regulated at the level of stored inactive maternal mRNA. We investigated here the translational control of cyclin E1, required for metaphase II arrest of the unfertilised egg and the initiation of S phase in the early embryo. We show that the cyclin E1 mRNA is regulated by both cytoplasmic polyadenylation elements (CPEs) and two miR-15/16 target sites within its 3'UTR. Moreover, we provide evidence that maternal miR-15/16 microRNAs co-immunoprecipitate with CPE-binding protein (CPEB), and that CPEB interacts with the RISC component Ago2. Experiments using competitor RNA and mutated cyclin E1 3'UTRs suggest cooperation of the regulatory elements to sustain repression of the cyclin E1 mRNA during early stages of maturation when CPEB becomes limiting and cytoplasmic polyadenylation of repressed mRNAs begins. Importantly, injection of anti-miR-15/16 LNA results in the early polyadenylation of endogenous cyclin E1 mRNA during meiotic maturation, and an acceleration of GVBD, altogether strongly suggesting that the proximal CPEB and miRNP complexes act to mutually stabilise each other. We conclude that miR-15/16 and CPEB co-regulate cyclin E1 mRNA. This is the first demonstration of the co-operation of these two pathways.


Determinants of confrontation naming performance.
Confrontation naming tests are commonly employed in neuropsychological assessment. Surprisingly little work has been done, however, to determine how various demographic, linguistic, and disease status variables influence patterns of performance on these tests. The present study examined data on the Boston Naming Test (BNT) from a total of 1,131 subjects, including 719 normals, 325 patients with Alzheimer's disease, and 87 patients with temporal lobe epilepsy. The effects of age, education, gender, and diagnostic group were examined with respect to overall scores, the influence of phonemic cuing, and performance on individual items. Profiles of scores on individual items were similar across diagnostic groups, suggesting that anomia is characterized by quantitative rather than qualitative changes in naming performance. Age and education systematically influenced scores. There was a significant effect of gender across diagnostic groups (males scoring higher than females), which appears to be due to performance on specific items. Phonemic cuing effects were similar across groups. The results are discussed with respect to the neuropsychological construct of confrontation naming and the clinical interpretation of performance on the BNT.


The impact of antenatal care on pregnancy performance between adolescent girls and older women.
To investigate pregnancy performance and newborn outcome between adolescents and older women receiving adequate and similar antenatal care. Four hundred and twenty-five women attending the antenatal clinic at the University Hospital of the West Indies, Kingston, Jamaica, participated in a prospective study. Recruitment included women 19 years and younger (adolescents) and 20 years and older (older women). Anthropometric measurements of the women and their newborn were made. During the pregnancy, all admissions to hospital and the diagnoses were recorded. Neonatal admissions were also recorded. Three hundred and sixty-one women each (84.9%) delivered a live infant of which 175 (87.5%) women were less than 20 years old and comprised the adolescent group and 186 (82.7%) of these women were 20 years and older and comprised the women in the older age group. Thirty-nine (9.2%) had early pregnancy losses; twelve among the adolescents (6%) and twenty-seven (12%) among the older women (p = 0.03). There was one intrauterine death and one stillbirth in the adolescent group and two intrauterine deaths and no stillbirth in the older group. The attendance rate at the antenatal clinic for both groups was high throughout the study with each group having greater than 96% attendance. There were 53 (26.5%) admissions among the adolescent girls and 70 (31.1%) admissions among the older women. This was not statistically significant. There was a significant difference in the occurrence of urinary tract infections where 17% of the adolescents had a urinary tract infection compared to 1% of older women (p < 0.0001). There was a lower Caesarean section rate among the adolescent. No significant difference between the number of neonatal admissions for the two groups (p = 0.19) was seen. This study showed that with similar and adequate antenatal care there were minimal differences in pregnancy performance between the two groups with only an increased rate of urinary tract infections and a lower rate of Caesarean section in the adolescents.


Articulating tactical traction splint use on pulseless forearm fracture.
An articulating tactical traction splint (REEL™ Tactical Traction Splint), commonly issued to U.S. military personnel, was used to maintain traction in a pulseless forearm fracture during a confined space rescue, with good peripheral perfusion maintained through transport. This enabled a single rescuer to focus attention and provide care for other critical aspects of a multisystem trauma patient.


Application of the Transtheoretical Model to Physical Activity in Deaf Individuals.
The current study used the transtheoretical model (TTM) as a guiding theoretical framework to assess differences in processes of change, decisional balance, and self-efficacy among deaf individuals with different levels of physical activity. Overall, 146 participants (M age = 26.4 yr, SD = 4.28) completed anonymous questionnaires assessing the dimensions of the TTM, stages of change, processes of change, decisional balance, and self-efficacy. Analysis of variance showed that both experiential and behavioral processes of change were higher in the preparation, action, and maintenance stages than in the other stages. Accordingly, the benefits of physical activity participation were stronger in the preparation stage, whereas the costs were more evident in the precontemplation stage. Finally, self-efficacy at the preparation stage was higher than in the other stages. The findings revealed how different stages of physical activity participation can be explained through the TTM, and the implications for physical activity intervention are discussed.


Juvenile exposure to bisphenol A promotes ovarian differentiation but suppresses its growth - Potential involvement of pituitary follicle-stimulating hormone.
Bisphenol A (BPA), a plastic monomer and plasticizer, is commonly used in plastics industry, and it has been well documented to be an estrogenic endocrine disrupter. In the present study, we investigated the effect of early (juvenile) exposure to BPA on the hypothalamic-pituitary-gonad (HPG) axis in the zebrafish. Estradiol (E2) and testosterone (T) were also included as positive and negative controls respectively. Juvenile zebrafish were exposed to BPA (1 and 10μM), E2 (10nM) and T (10nM) from 20 to 40 dpf (days post-fertilization), the period of sex/gonadal differentiation, followed by histological and expression analyses at 40 dpf. The ovary and hepatic proteomes were also analyzed by mass spectrometry. Our results showed that 20day exposure to BPA and E2 increased the ratio of females; however, they both significantly suppressed ovarian growth. Meanwhile, BPA and E2 significantly suppressed fshb but stimulated lhb expression in the pituitary. These effects did not seem to involve the hypothalamus because neither BPA nor E2 altered the expression of kiss1, kiss2, gnrh2 and gnrh3 in the hypothalamus. At the ovary level, BPA and E2 both decreased lhcgr expression. Interestingly, E2 and BPA displayed different effects in the liver. E2 induced a significant hepatic hypertrophy; however, BPA had no such effect. Analysis of hepatic proteomes revealed distinct protein profiles in the E2 group as compared with the others, especially fructose-bisphospahte aldolase B. These results indicated that BPA has estrogenic effects on female reproduction, but it does not mimic all E2 actions. Our data in the zebrafish suggest that sex differentiation involves estrogens and it is a sensitive window for evaluating estrogenic activities of compounds and their impacts on wildlife reproduction.


Medicaid Expansions and Cervical Cancer Screening for Low-Income Women.
Medicaid coverage for low-income women may play an important role in ensuring access to preventive care. This study examines how Medicaid eligibility expansions to nonelderly adults impact cervical cancer screening among low-income women. We use data from the Behavioral Risk Factor Surveillance System from 2000 to 2010. The primary outcome of interest is whether women in the relevant guideline consistent age range reported having a Pap test in the previous year. We use a difference-in-differences approach with matched treatment and comparison states and a simulated eligibility approach based on a continuous measure of Medicaid generosity. Our results indicate that cervical cancer screening increased among low-income women in expansion states relative to comparison states. Increases in screening rates are largest among low-income Hispanic women. Medicaid expansions during the period from 2000 to 2010 were associated with improved cervical cancer screening rates, which is critical for early cervical cancer detection and prevention of cancer morbidity and mortality in women. The results suggest that more widespread Medicaid expansions may have positive effects on preventive health care for women.


Nano-scale morphology of cardiomyocyte t-tubule/sarcoplasmic reticulum junctions revealed by ultra-rapid high-pressure freezing and electron tomography.
Detailed knowledge of the ultrastructure of intracellular compartments is a prerequisite for our understanding of how cells function. In cardiac muscle cells, close apposition of transverse (t)-tubule (TT) and sarcoplasmic reticulum (SR) membranes supports stable high-gain excitation-contraction coupling. Here, the fine structure of this key intracellular element is examined in rabbit and mouse ventricular cardiomyocytes, using ultra-rapid high-pressure freezing (HPF, omitting aldehyde fixation) and electron microscopy. 3D electron tomograms were used to quantify the dimensions of TT, terminal cisternae of the SR, and the space between SR and TT membranes (dyadic cleft). In comparison to conventional aldehyde-based chemical sample fixation, HPF-preserved samples of both species show considerably more voluminous SR terminal cisternae, both in absolute dimensions and in terms of junctional SR to TT volume ratio. In rabbit cardiomyocytes, the average dyadic cleft surface area of HPF and chemically fixed myocytes did not differ, but cleft volume was significantly smaller in HPF samples than in conventionally fixed tissue; in murine cardiomyocytes, the dyadic cleft surface area was higher in HPF samples with no difference in cleft volume. In both species, the apposition of the TT and SR membranes in the dyad was more likely to be closer than 10 nm in HPF samples compared to CFD, presumably resulting from avoidance of sample shrinkage associated with conventional fixation techniques. Overall, we provide a note of caution regarding quantitative interpretation of chemically-fixed ultrastructures, and offer novel insight into cardiac TT and SR ultrastructure with relevance for our understanding of cardiac physiology.


Lorazepam Challenge for Individuals at Varying Genetic Risk for Alzheimer Disease.
This study set out to clarify the differential acute cognitive impact of lorazepam based on varying genetic risk for Alzheimer disease. Fifty-seven cognitively unimpaired individuals aged 51 to 88, genotyped according to apolipoprotein E (APOE) and translocase of outer mitochondrial membrane (40 homolog) poly-T lengths, completed cognitive testing before, 2.5 and 5 hours after receiving a 1 mg dose of lorazepam. Post-lorazepam, there were significant (P<0.05) declines from baseline in memory, psychomotor processing speed, and executive function. At 2.5 hours, the magnitude of this lorazepam-induced cognitive change was significantly greater in the APOE3/4 group than in the APOE3/3 group for tests of working memory and visuospatial memory/executive function. At 5 hours postchallenge, verbal memory and working memory deficits persisted in the APOE3/4 group compared with the APOE3/3 group. At 5 hours after lorazepam challenge, as compared with the very long/very long group, the short/short group performed slightly worse on a test of working memory (P<0.05), but no other differences were observed among translocase of the outer mitochondrial membrane 40 homolog poly-T variant groups. The lorazepam challenge may be unmasking presymptomatic cognitive dysfunction associated with APOE4 carriage.


Inositol trisphosphate-independent agonist-stimulated calcium influx in rat pancreatic acinar cells.
CCK-JMV-180 is a cholecystokinin analog that stimulates digestive enzyme secretion from pancreatic acinar cells but does not cause either generation of inositol 1,4,5-trisphosphate or depletion of the inositol 1,4,5-trisphosphate-sensitive intracellular Ca2+ storage pool. We report that CCK-JMV-180 can accelerate Ca2+ influx into fura-2-loaded dispersed rat pancreatic acini and single acinar cells. Furthermore, CCK-JMV-180 accelerates Ca2+ influx into cells microinjected with the inositol 1,4,5-trisphosphate receptor antagonist heparin and into acini loaded with the Ca(2+)-chelating agent BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid). These results indicate that agonist-stimulated Ca2+ influx can occur (a) without depletion of the inositol 1,4,5-trisphosphate-sensitive intracellular Ca2+ storage pool, (b) without a rise in cytoplasmic free Ca2+ concentrations, and (c) after blockade of inositol 1,4,5-trisphosphate receptors. They suggest that depletion of an inositol 1,4,5-trisphosphate-independent intracellular Ca2+ storage pool and/or generation of a non-inositol 1,4,5-trisphosphate second messenger by CCK-JMV-180 may be a sufficient signal for acceleration of Ca2+ influx into rat pancreatic acinar cells.


Definition of the unique human extraocular muscle allotype by expression profiling.
The extraocular muscles (EOMs) are a unique group of specialized muscles that are anatomically and physiologically distinct from other skeletal muscles. Perhaps the most striking characteristic of the EOMs is their differential sensitivity to disease. EOMs are spared in Duchenne's muscular dystrophy (DMD) despite widespread involvement of other skeletal muscles. Conversely, they are early and prominent targets in myasthenia gravis and mitochondrial myopathies. It is unclear how EOMs achieve such specialization or a differential response to diseases; however, this has been attributed to a unique, group-specific pattern of gene expression or "allotype." To begin to address these issues as well as define the human EOM allotype, we analyzed the human EOM transcriptome using oligonucleotide-based expression profiling. Three hundred thirty-eight genes were found to be differentially expressed in EOM compared with quadriceps femoris limb muscle, using a twofold cutoff. Functional characterization revealed expression patterns corresponding to known metabolic and structural properties of EOMs such as expression of EOM-specific myosin heavy chain (MYH13) and high neural, vascular, and mitochondrial content, suggesting that the profiling was sensitive and specific. Genes related to myogenesis, stem cells, and apoptosis were detected at high levels in normal human EOMs, suggesting that efficient and continuous regeneration and/or myogenesis may be a mechanism by which the EOMs remain clinically and pathologically spared in diseases such as DMD. Taken together, this study provides insight into how human EOMs achieve their unique structural, metabolic, and pathophysiological properties.


Categorical selection of the antibody repertoire in splenic B cells.
In the bone marrow, the passage of developing B cells through critical checkpoints of differentiation is associated with a reduction of specific categories of CDR3 of the Ig heavy chain (CDR-H3), particularly those with excessive hydrophobic or charged amino acids and those with a length of eight or fewer residues. To gain insight into the role of CDR-H3 content in the development of B cells in the spleen, we compared the sequences of V(H)7183DJCmu transcripts from sorted transitional T1, marginal zone, and follicular B cell subsets to those expressed by immature IgM(+)IgD(-) and mature IgM(lo)IgD(hi) B cells in the bone marrow. Although differences in V(H) utilization were noted, the T1 CDR-H3 repertoire showed extensive similarity to that of immature bone marrow B cells, and the follicular CDR-H3 repertoire most resembled that of mature bone marrow B cells. Unlike the splenic follicular and bone marrow mature B cell CDR-H3 repertoires, the marginal zone B cell CDR-H3 repertoire retained both short and highly charged amino acid motifs, including those with two arginines. Our findings suggest that antigen binding sites containing specific categories of CDR-H3 sequence content may inhibit, permit, or even facilitate passage of the host B cell through critical checkpoints in peripheral as well as central development.


Insulin-like growth factor-I is an essential regulator of the differentiation of 3T3-L1 adipocytes.
Murine 3T3-L1 preadipocytes proliferate normally in medium containing fetal calf serum depleted of insulin, growth hormone, and insulin-like growth factor-I (IGF-I). However, the cells do not differentiate into adipocytes in the presence of the hormone-depleted serum. Supplementation of the growth medium with 10-20 nM IGF-I or 2 microM insulin restores the ability of 3T3-L1 cells to develop into adipocytes. The cells acquire an adipocyte morphology, accumulate triglycerides, and express a 450-fold increase in the activity of the lipogenic enzyme glycerol-3-phosphate dehydrogenase. The increase in glycerol-3-phosphate dehydrogenase activity is paralleled by the accumulation of glycerol-3-phosphate dehydrogenase mRNA and mRNA for the myelin P2-like protein aP2, another marker for fat cell development. IGF-I or insulin-stimulated adipogenesis in 3T3-L1 cells is not dependent on growth hormone. Occupancy of preadipocyte IGF-I receptors by IGF-I (or insulin) is implicated as a central step in the differentiation process. The IGF-I receptor binds insulin with a 70-fold lower affinity than IGF-I, and 30-70-fold higher levels of insulin are required to duplicate the effects of an optimal amount of IGF-I. The effects of 10-20 nM IGF-I are likely to be mediated by high affinity (KD = 5 nM) IGF-I receptors that are expressed at a density of 13,000 sites/preadipocyte. In undifferentiated cells the IGF-I receptor concentration is twice that of the insulin receptor. After adipocyte differentiation is triggered, the number and affinity of IGF-I receptors remain constant while insulin receptor number increases approximately 25-fold as developing adipocytes become responsive to insulin at the level of metabolic regulation. Thus, preadipocytes have the potential for a maximal response to IGF-I, whereas the accumulation of more than 95% of adipocyte insulin receptors and the appearance of responsiveness to insulin are consequences of differentiation. IGF-I or insulin is essential for the induction of a variety of abundant and nonabundant mRNAs characteristic of 3T3-L1 adipocytes.


Both PU.1 and nuclear factor-kappa B mediate lipopolysaccharide- induced HIV-1 long terminal repeat transcription in macrophages.
We recently reported that LPS stimulation of monocytic cells leads to the activation of PU.1, a member of the Ets family of transcription factors. Phosphorylation of PU.1 by protein kinase CK2 was found to up-regulate its trans-activation function, but not its DNA binding activity. Previous studies suggested that Ets proteins could bind to NF-kappa B motifs at the tetrameric core sequence TTCC. In macrophages, LPS-inducible HIV-1 gene expression is mediated in part by binding of NF-kappa B to identical tandem binding sites located within the long terminal repeat (LTR). Thus, we performed additional studies to determine whether PU.1 also played a role in regulating HIV-1 gene expression in macrophages. Our functional studies revealed that activation of the HIV-1 LTR in LPS-stimulated cells requires both NF-kappa B and PU.1. Extensive mutagenesis of the HIV-1 LTR revealed that PU.1-dependent activation requires the Ets motif within the upstream NF-kappa B site, whereas NF-kappa B itself binds to the downstream site. We also found that insertion of five additional nucleotides between the NF-kappa B sites abolished LPS inducibility, suggesting a direct interaction between factors that bind these sites. Lastly, we found that mutation of PU.1 at serine 148, which prevents its phosphorylation by CK2, blocked its ability to activate the HIV-1 LTR in response to LPS. These effects were promoter specific because PU.1 did not affect LPS-inducible activation of a distinct NF-kappa B-dependent promoter. While these data do not demonstrate direct binding of PU.1 to the HIV-1 LTR, they illustrate a novel role for PU.1 in activation of the HIV-1 LTR by LPS.


Complementary and alternative medicine (CAM) use in an Italian cohort of pediatric headache patients: the tip of the iceberg.
The use of complementary alternative medicine (CAM) in paediatric populations is considerably increased, especially for pain and chronic conditions, as demonstrated by epidemiological surveys both in Europe and in the USA. In our study, CAM was used in 76 % patients of a cohort of 124 children affected by headache (age 4-16 years; 67 % female; 70 % migraine without aura, 12 % migraine with aura, 18 % tensive headache according to IHS criteria) consecutively recruited at a Pediatric Headache University Center. CAM was used as preventive treatment in 80 % cases. The main reasons for seeking CAM were: the wish of avoiding chronic use of drugs with their related side effects, the desire of an integrated approach, the reported inefficacy of conventional medicine, and a more suitable children disposition to CAM than to pharmacological compound. Female gender, younger age, migraine without aura, parents' higher educational status, maternal use of CAM and other associated chronic conditions, correlated with CAM use (p < 0.05). 73 % patients chose CAM also to treat other diseases (i.e. allergies, colitis, asthma, insomnia, muscle-scheletric disorders and dysmenorrhoea). The most assumed CAM were: herbal remedies (64 %) such as Valeriana, Ginkgo biloba, Boswellia serrata, Vitex agnus-castus, passion flower, Linden tree; vitamins/minerals supplements (40 %) with magnesium, 5-Hydroxytryptophan, vitamin B6 or B12, Multivitamin compounds; Homeopathy (47 %) with Silicea, Ignatia Amara, Pulsatilla, Aconitum, Nux Vomica, Calcarea phosphorica; physical treatment (45 %) such as Ayurvedic massage, shiatsu, osteopathy; yoga (33 %); acupuncture (11 %). CAM-often integrated with conventional care-was auto-prescribed in 30 % of the cases, suggested by non-physician in 22 %, by the General Practitioner in 24 % and by paediatrician in 24 %. Both general practitioners and neurologists were mostly unaware of their patients' CAM use. In conclusion, neurologists should inquire for CAM use and be prepared to learn about CAM therapies or to directly interact with CAM trained experts, in order to coordinate an integrative approach to health, as especially required in paediatric headache patients and their parents. Further studies are required to investigate safety and efficacy of CAM in pediatric headache, as a possible side-medicine to conventional pharmacological approach.


Prospective randomized controlled multi-centre trial of cuffed or uncuffed endotracheal tubes in small children.
The use of cuffed tracheal tubes (TTs) in small children is still controversial. The aim of this study was to compare post-extubation morbidity and TT exchange rates when using cuffed vs uncuffed tubes in small children. Patients aged from birth to 5 yr requiring general anaesthesia with TT intubation were included in 24 European paediatric anaesthesia centres. Patients were prospectively randomized into a cuffed TT group (Microcuff PET) and an uncuffed TT group (Mallinckrodt, Portex, Rüsch, Sheridan). Endpoints were incidence of post-extubation stridor and the number of TT exchanges to find an appropriate-sized tube. For cuffed TTs, minimal cuff pressure required to seal the airway was noted; maximal cuff pressure was limited at 20 cm H(2)O with a pressure release valve. Data are mean (SD). A total of 2246 children were studied (1119/1127 cuffed/uncuffed). The age was 1.93 (1.48) yr in the cuffed and 1.87 (1.45) yr in the uncuffed groups. Post-extubation stridor was noted in 4.4% of patients with cuffed and in 4.7% with uncuffed TTs (P=0.543). TT exchange rate was 2.1% in the cuffed and 30.8% in the uncuffed groups (P<0.0001). Minimal cuff pressure required to seal the trachea was 10.6 (4.3) cm H(2)O. The use of cuffed TTs in small children provides a reliably sealed airway at cuff pressures of <or=20 cm H(2)O, reduces the need for TT exchanges, and does not increase the risk for post-extubation stridor compared with uncuffed TTs.


HIV prevention programs of nongovernmental organizations in Latin America and the Caribbean: the Global AIDS Intervention Network project.
The objective of this paper is to describe HIV prevention programs conducted by nongovernmental organizations (NGO) that are meeting this challenge. One NGO undertaking HIV prevention programs was evaluated in each of the 23 countries participating in the Global AIDS Intervention Network (GAIN) Project throughout Latin America and the Caribbean. A two-stage selection process was used: (1) a search in databases and other information sources; (2) identification of NGOs that were best established and most actively engaged in HIV prevention activity. Executive directors were questioned about staffing, budget issues, populations served and barriers faced by these entities. The 23 NGOs conducted 58 direct-service programs and had been conducting HIV prevention activities for a mean of 8 years (SD=4.45; range 1-18 years). Average annual program budget was US $205,393 (range: US $10,000 to US $1,440,000). The NGOs reported a mean of 4.5 full-time employees (range 0-15, SD=4.7). Many relied on volunteers (median=10, mean=51, range 0-700, SD=150) to conduct HIV prevention activities. The NGOs provided prevention services for the general community (82.6%), children and adolescents (34.8%) and men who have sex with men (30.4%). Activities conducted by NGOs included train-the-trainer activities (43.5%) and face-to-face prevention activities (34.8%). Obstacles cited included lack of funding (60.9%) and HIV-related stigma and discrimination (56.5%). The strategies used by NGOs to overcome barriers to prevention are a testament to their ingenuity and commitment, and serve as examples for NGOs in other world regions.


Chemomechanical Coupling in Hexameric Protein-Protein Interfaces Harnesses Energy within V-Type ATPases.
ATP synthase is the most prominent bioenergetic macromolecular motor in all life forms, utilizing the proton gradient across the cell membrane to fuel the synthesis of ATP. Notwithstanding the wealth of available biochemical and structural information inferred from years of experiments, the precise molecular mechanism whereby vacuolar (V-type) ATP synthase fulfills its biological function remains largely fragmentary. Recently, crystallographers provided the first high-resolution view of ATP activity in Enterococcus hirae V1-ATPase. Employing a combination of transition-path sampling and high-performance free-energy methods, the sequence of conformational transitions involved in a functional cycle accompanying ATP hydrolysis has been investigated in unprecedented detail over an aggregate simulation time of 65 μs. Our simulated pathways reveal that the chemical energy produced by ATP hydrolysis is harnessed via the concerted motion of the protein-protein interfaces in the V1-ring, and is nearly entirely consumed in the rotation of the central stalk. Surprisingly, in an ATPase devoid of a central stalk, the interfaces of this ring are perfectly designed for inducing ATP hydrolysis. However, in a complete V1-ATPase, the mechanical property of the central stalk is a key determinant of the rate of ATP turnover. The simulations further unveil a sequence of events, whereby unbinding of the hydrolysis product (ADP + Pi) is followed by ATP uptake, which, in turn, leads to the torque generation step and rotation of the center stalk. Molecular trajectories also bring to light multiple intermediates, two of which have been isolated in independent crystallography experiments.


Increased postvoid residual volume after measuring the isovolumetric bladder pressure using the noninvasive condom catheter method.
To test, in an ongoing noninvasive longitudinal study in healthy men, whether the condom catheter method (a noninvasive urodynamic test to assess bladder function and bladder outlet obstruction) inhibits bladder function and whether this affects the reliability of the measured isovolumetric bladder pressure (P(ves.iso)). Subjects (754, aged 40-79 years) voided three times, i.e. one free void and two condom measurements. The postvoid residual volume (PVR) was measured after each void using transabdominal ultrasonography. The statistical significance of differences was tested using Wilcoxon rank test and the Mann-Whitney U-test. After free voiding the median (interquartile range) PVR was 18 (37) mL, and independent of the amount of fluid intake. In a subgroup of volunteers, when the free void was done last, the PVR was no different (P = 0.25), suggesting that the bladder did not become exhausted during the protocol. The PVR after two subsequent condom measurements was significantly higher than after free voiding, at 45 (78) and 57 (88) (both P < 0.05), independent of the number of interruptions in voiding. After supplementary fluid intake before the condom measurements, the PVR was double that with a normal fluid intake (P = 0.03). The median P(ves.iso) was 3 cmH(2)O higher in the second condom measurement than in the first (P < 0.05), although this small difference was not clinically relevant. The condom measurement is associated with a significantly higher PVR, partly caused by supplementary fluid intake. This effect was only temporary and did not affect the measured P(ves.iso).


Addressing childhood overweight through schools.
Rates of childhood obesity in have reached alarming proportions in many countries. Sixteen percent of school-aged children and adolescents in the U.S. are overweight. Legislation implemented in 2004 in the U.S. requires local education agencies (LEAs) that sponsor school meal programs to establish local wellness policies to address childhood obesity. Project PA, a collaboration between a state agency and a university providing school-based interventions focuses on the school environment and policy changes. Interventions have targeted foodservice personnel, administrators, teachers, parents and students. In two recent projects schools assessed their school nutrition environments, developed nutrition policies, and implemented strategies to encourage healthier food selections. Schools identified weaknesses in the areas of marketing and communication of policies. Media attention on the childhood obesity facilitated policy changes. Time and cost were identified as barriers to policy development and there were concerns about weak enforcement of policies. These themes are discussed.


Oct4 mediates tumor initiating properties in oral squamous cell carcinomas through the regulation of epithelial-mesenchymal transition.
Overexpression of Oct4, an important transcription factor of embryonic stem cells (ESC), has been reported in several cancers. The aim of this study was to determine the emerging role of Oct4 in oral squamous cell carcinoma (OSCC) both in vitro and in vivo. Tumourigenic activity and molecular mechanisms of Oct4 overexpression or knockdown by lentiviral infection in OSCC was investigated in vitro and in vivo. Initially, we demonstrated that Oct4 expression was increased in OSCC cell lines as compared to a normal oral epithelial cell line SG. Overexpression of Oct4 was demonstrated to enhance cell proliferation, invasiveness, anchorage-independent growth and xenotransplantation tumourigenicity. These findings were coupled with epithelial-mesenchymal transition (EMT) transformation in OSCCs. In contrast, the silence of Oct4 significantly blocked the xenograft tumorigenesis of OSCC-derived cancer stem cells (OSCC-CSCs) and significantly improved the recipient survival. Clinically, the level of Oct4 expression was higher in recurrent and metastatic OSCC specimens but lower in primary OSCC specimens. Our results suggest that Oct4-mediated tumorigenecity is associated with the regulation of EMT. Oct4 might be a therapeutic target for OSCC.


Is autologous chondrocyte implantation (ACI) an adequate treatment option for repair of cartilage defects in paediatric patients?
Cartilage lesions in the knee of juvenile patients require an effective repair to regain life-long functional activity of the joint. Autologous chondrocyte implantation (ACI) is discussed to be advantageous over other methods for cartilage repair regarding long-term outcome. ACI has successfully been applied in juvenile patients, although currently recommended for patients ≥18 years of age. Only few controlled clinical trials present evidence of efficacy and safety of ACI in adolescent patients. ACI products have to undergo the process of a marketing authorisation application, including the submission of a paediatric investigation plan (PIP). Data from prospective clinical studies or retrospective collection of long-term data in paediatric patients should be submitted for risk-benefit evaluation by the Paediatric Committee (PDCO).


Factors associated with American Board of Medical Specialties member board certification among US medical school graduates.
Certification by an American Board of Medical Specialties (ABMS) member board is emerging as a measure of physician quality. To identify demographic and educational factors associated with ABMS member board certification of US medical school graduates. Retrospective study of a national cohort of 1997-2000 US medical school graduates, grouped by specialty choice at graduation and followed up through March 2, 2009. In separate multivariable logistic regression models for each specialty category, factors associated with ABMS member board certification were identified. ABMS member board certification. Of 42,440 graduates in the study sample, 37,054 (87.3%) were board certified. Graduates in all specialty categories with first-attempt passing scores in the highest tertile (vs first-attempt failing scores) on US Medical Licensing Examination Step 2 Clinical Knowledge were more likely to be board certified; adjusted odds ratios (AORs) varied by specialty category, with the lowest odds for emergency medicine (87.4% vs 73.6%; AOR, 1.82; 95% CI, 1.03-3.20) and highest odds for radiology (98.1% vs 74.9%; AOR, 13.19; 95% CI, 5.55-31.32). In each specialty category except family medicine, graduates self-identified as underrepresented racial/ethnic minorities (vs white) were less likely to be board certified, ranging from 83.5% vs 95.6% in the pediatrics category (AOR, 0.44; 95% CI, 0.33-0.58) to 71.5% vs 83.7% in the other nongeneralist specialties category (AOR, 0.79; 95% CI, 0.64-0.96). With each $50,000 unit increase in debt (vs no debt), graduates choosing obstetrics/gynecology were less likely to be board certified (AOR, 0.89; 95% CI, 0.83-0.96), and graduates choosing family medicine were more likely to be board certified (AOR, 1.13; 95% CI, 1.01-1.26). Demographic and educational factors were associated with board certification among US medical school graduates in every specialty category examined; findings varied among specialty categories.


Localized Wounding by Heat Initiates the Accumulation of Proteinase Inhibitor II in Abscisic Acid-Deficient Plants by Triggering Jasmonic Acid Biosynthesis.
To test whether the response to electrical current and heat treatment is due to the same signaling pathway that mediates mechanical wounding, we analyzed the effect of electric-current application and localized burning on proteinase inhibitor II (Pin2) gene expression in both wild-type and abscisic acid (ABA)-deficient tomato (Lycopersicon esculentum Mill.) and potato (Solanum phureja) plants. Electric-current application and localized burning led to the accumulation of Pin2 mRNA in potato and tomato wild-type plants. Among the treatments tested, only localized burning of the leaves led to an accumulation of Pin2 mRNA in the ABA-deficient plants. Electric-current application, like mechanical injury, was able to initiate ABA and jasmonic acid (JA) accumulation in wild-type but not in ABA-deficient plants. In contrast, heat treatment led to an accumulation of JA in both wild-type and ABA-deficient plants. Inhibition of JA biosynthesis by aspirin blocked the heat-induced Pin2 gene expression in tomato wild-type leaves. These results suggest that electric current, similar to mechanical wounding, requires the presence of ABA to induce Pin2 gene expression. Conversely, burning of the leaves activates Pin2 gene expression by directly triggering the biosynthesis of JA by an alternative pathway that is independent of endogenous ABA levels.


Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study.
The peptidoglycan (PG) metabolic process is essential for bacterial growth. β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. For deglycosylation, both the hydrolysis and phosphorylation of the glycosyl-enzyme intermediate were explored. The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. This is the first computational report for understanding the phosphorylase activity of NagZ enzymes.


Turnover, age and length of service: a comparison of nurses and other staff in the National Health Service.
Using information collected from more than 100 district health authorities in England for the year 1989-1990, this paper examines turnover rates amongst a range of nursing and other staff groups in the National Health Service (NHS), and their relationship to the age and length of service characteristics of the labour force. The evidence collected suggests that the NHS employs a significantly younger workforce than is found in the economy as a whole. The age profile of nurses is even more skewed towards younger age groups than that of non-nursing staff working in the NHS. Nurses tend on average to have longer lengths of service than non-nursing staff groups, and it would seem that the average length of service has increased over the last 20 years, certainly amongst registered nurses. Overall, the study found an annual turnover rate amongst all NHS staff of 13.6%. Turnover rates were significantly higher among full-time staff than part-time, and amongst non-nursing staff groups compared with nurses. Broadly, turnover rates decline with age then rise close to retirement. However, there is a more complex relationship between length of service and turnover: turnover rates tend to be high in the first year of service, and to remain high or even rise during the second year of service, before declining. Turnover remains a poorly understood issue in the NHS. The evidence presented here should move some aspects of debate onto a more solid empirical foundation.


Relative contributions of infraspinatus and deltoid during external rotation in healthy shoulders.
Balanced forces around the shoulder are important for normal function; however, rehabilitation guidelines are not well defined because the muscle contributions and optimal exercise technique to recruit them are poorly understood. This study aimed to determine (1) the conditions of resisted isometric external rotation that optimized the contribution of infraspinatus and (2) the load of external rotation at which the adduction strategy was most effective at reducing deltoid contributions. Eighteen subjects with healthy shoulders (n = 36) performed resisted isometric external rotation at 3 increasing loads--10%, 40%, and 70% of their maximal resisted external rotation voluntary isometric contraction--with and without adduction. Surface electromyographic activity of the infraspinatus, posterior and middle deltoid, and pectoralis major was recorded and normalized against the average activity of all 4 muscles, representing each muscle's relative contribution to the task. To optimize the relative contribution of the infraspinatus with the least deltoid involvement during isometric external rotation, a load between 10% and 40% maximal voluntary isometric contraction is appropriate. At low loads, use of the adduction strategy during external rotation reduces middle deltoid involvement. In contrast, the posterior deltoid is activated in parallel with the infraspinatus at low loads and may even act as an adductor with the arm by the side. This study provides a useful guide to optimize rehabilitative exercises for rotator cuff dysfunction; in particular, highlighting that activation of the deltoid could be counterproductive to infraspinatus retraining.


Longitudinal analysis of censored medical cost data.
This paper applies the inverse probability weighted (IPW) least-squares method to estimate the effects of treatment on total medical cost, subject to censoring, in a panel-data setting. IPW pooled ordinary-least squares (POLS) and IPW random effects (RE) models are used. Because total medical cost might not be independent of survival time under administrative censoring, unweighted POLS and RE cannot be used with censored data, to assess the effects of certain explanatory variables. Even under the violation of this independency, IPW estimation gives consistent asymptotic normal coefficients with easily computable standard errors. A traditional and robust form of the Hausman test can be used to compare weighted and unweighted least squares estimators. The methods are applied to a sample of 201 Medicare beneficiaries diagnosed with lung cancer between 1994 and 1997.


Activity-dependent regulation of dendritic BC1 RNA in hippocampal neurons in culture.
Several neuronal RNAs have been identified in dendrites, and it has been suggested that the dendritic location of these RNAs may be relevant to the spatiotemporal regulation of mosaic postsynaptic protein repertoires through transsynaptic activity. Such regulation would require that dendritic RNAs themselves, or at least some of them, be subject to physiological control. We have therefore examined the functional regulation of somatodendritic expression levels of dendritic BC1 RNA in hippocampal neurons in culture. BC1 RNA, an RNA polymerase III transcript that is a component of a ribonucleoprotein particle, became first detectable in somatodendritic domains of developing hippocampal neurons at times of initial synapse formation. BC1 RNA was identified only in such neurons that had established synapses on cell bodies and/or developing dendritic arbors. When synaptic contact formation was initiated later in low-density cultures, BC1 expression was coordinately delayed. Inhibition of neuronal activity in hippocampal neurons resulted in a substantial but reversible reduction of somatodendritic BC1 expression. We conclude that expression of BC1 RNA in somatic and dendritic domains of hippocampal neurons is regulated in development, and is dependent upon neuronal activity. These results establish (for the first time to our knowledge) that an RNA polymerase III transcript can be subject to control through physiological activity in nerve cells.


Gliding flight in a jackdaw: a wind tunnel study.
We examined the gliding flight performance of a jackdaw Corvus monedula in a wind tunnel. The jackdaw was able to glide steadily at speeds between 6 and 11 m s(-1). The bird changed its wingspan and wing area over this speed range, and we measured the so-called glide super-polar, which is the envelope of fixed-wing glide polars over a range of forward speeds and sinking speeds. The glide super-polar was an inverted U-shape with a minimum sinking speed (V(ms)) at 7.4 m s(-1) and a speed for best glide (V(bg)) at 8.3 m s(-)). At the minimum sinking speed, the associated vertical sinking speed was 0.62 m s(-1). The relationship between the ratio of lift to drag (L:D) and airspeed showed an inverted U-shape with a maximum of 12.6 at 8.5 m s(-1). Wingspan decreased linearly with speed over the whole speed range investigated. The tail was spread extensively at low and moderate speeds; at speeds between 6 and 9 m s(-1), the tail area decreased linearly with speed, and at speeds above 9 m s(-1) the tail was fully furled. Reynolds number calculated with the mean chord as the reference length ranged from 38 000 to 76 000 over the speed range 6-11 m s(-1). Comparisons of the jackdaw flight performance were made with existing theory of gliding flight. We also re-analysed data on span ratios with respect to speed in two other bird species previously studied in wind tunnels. These data indicate that an equation for calculating the span ratio, which minimises the sum of induced and profile drag, does not predict the actual span ratios observed in these birds. We derive an alternative equation on the basis of the observed span ratios for calculating wingspan and wing area with respect to forward speed in gliding birds from information about body mass, maximum wingspan, maximum wing area and maximum coefficient of lift. These alternative equations can be used in combination with any model of gliding flight where wing area and wingspan are considered to calculate sinking rate with respect to forward speed.


Economic burden of occupational injury and illness in the United States.
The allocation of scarce health care resources requires a knowledge of disease costs. Whereas many studies of a variety of diseases are available, few focus on job-related injuries and illnesses. This article provides estimates of the national costs of occupational injury and illness among civilians in the United States for 2007. This study provides estimates of both the incidence of fatal and nonfatal injuries and nonfatal illnesses and the prevalence of fatal diseases as well as both medical and indirect (productivity) costs. To generate the estimates, I combined primary and secondary data sources with parameters from the literature and model assumptions. My primary sources were injury, disease, employment, and inflation data from the U.S. Bureau of Labor Statistics (BLS) and the Centers for Disease Control and Prevention (CDC) as well as costs data from the National Council on Compensation Insurance and the Healthcare Cost and Utilization Project. My secondary sources were the National Academy of Social Insurance, literature estimates of Attributable Fractions (AF) of diseases with occupational components, and national estimates for all health care costs. Critical model assumptions were applied to the underreporting of injuries, wage-replacement rates, and AFs. Total costs were calculated by multiplying the number of cases by the average cost per case. A sensitivity analysis tested for the effects of the most consequential assumptions. Numerous improvements over earlier studies included reliance on BLS data for government workers and ten specific cancer sites rather than only one broad cancer category. The number of fatal and nonfatal injuries in 2007 was estimated to be more than 5,600 and almost 8,559,000, respectively, at a cost of $6 billion and $186 billion. The number of fatal and nonfatal illnesses was estimated at more than 53,000 and nearly 427,000, respectively, with cost estimates of $46 billion and $12 billion. For injuries and diseases combined, medical cost estimates were $67 billion (27% of the total), and indirect costs were almost $183 billion (73%). Injuries comprised 77 percent of the total, and diseases accounted for 23 percent. The total estimated costs were approximately $250 billion, compared with the inflation-adjusted cost of $217 billion for 1992. The medical and indirect costs of occupational injuries and illnesses are sizable, at least as large as the cost of cancer. Workers' compensation covers less than 25 percent of these costs, so all members of society share the burden. The contributions of job-related injuries and illnesses to the overall cost of medical care and ill health are greater than generally assumed.


DNA polymerase X from Deinococcus radiodurans possesses a structure-modulated 3'-->5' exonuclease activity involved in radioresistance.
Recently a family X DNA polymerase (PolXDr) was identified in the radioresistant bacterium Deinococcus radiodurans. Knockout cells show a delay in double-strand break repair (DSBR) and an increased sensitivity to gamma-irradiation. Here we show that PolXDr possesses 3'-->5' exonuclease activity that stops cutting close to a loop. PolXDr consists of a DNA polymerase X domain (PolXc) and a Polymerase and Histidinol Phosphatase (PHP) domain. Deletion of the PHP domain abolishes only the structure-modulated but not the canonical 3'-->5' exonuclease activity. Thus, the exonuclease resides in the PolXc domain, but the structure-specificity requires additionally the PHP domain. Mutation of two conserved glycines in the PolXc domain leads to a specific loss of the structure-modulated exonuclease activity but not the exonuclease activity in general. The PHP domain itself does not show any activity. PolXDr is the first family X DNA polymerase that harbours an exonuclease activity. The wild-type protein, the glycine mutant and the two domains were expressed separately in DeltapolXDr cells. The wild-type protein could restore the radiation resistance, whereas intriguingly the mutant proteins showed a significant negative effect on survival of gamma-irradiated cells. Taken together our in vivo results suggest that both PolXDr domains play important roles in DSBR in D. radiodurans.


Impaired mitochondrial oxidative phosphorylation in the peroxisomal disease X-linked adrenoleukodystrophy.
X-linked adrenoleukodystrophy (X-ALD) is an inherited metabolic disorder of the nervous system characterized by axonopathy in spinal cords and/or cerebral demyelination, adrenal insufficiency and accumulation of very long-chain fatty acids (VLCFAs) in plasma and tissues. The disease is caused by malfunction of the ABCD1 gene, which encodes a peroxisomal transporter of VLCFAs or VLCFA-CoA. In the mouse, Abcd1 loss causes late onset axonal degeneration in the spinal cord, associated with locomotor disability resembling the most common phenotype in patients, adrenomyeloneuropathy. We have formerly shown that an excess of the VLCFA C26:0 induces oxidative damage, which underlies the axonal degeneration exhibited by the Abcd1(-) mice. In the present study, we sought to investigate the noxious effects of C26:0 on mitochondria function. Our data indicate that in X-ALD patients' fibroblasts, excess of C26:0 generates mtDNA oxidation and specifically impairs oxidative phosphorylation (OXPHOS) triggering mitochondrial ROS production from electron transport chain complexes. This correlates with impaired complex V phosphorylative activity, as visualized by high-resolution respirometry on spinal cord slices of Abcd1(-) mice. Further, we identified a marked oxidation of key OXPHOS system subunits in Abcd1(-) mouse spinal cords at presymptomatic stages. Altogether, our results illustrate some of the mechanistic intricacies by which the excess of a fatty acid targeted to peroxisomes activates a deleterious process of oxidative damage to mitochondria, leading to a multifaceted dysfunction of this organelle. These findings may be of relevance for patient management while unveiling novel therapeutic targets for X-ALD.


Validating the use of intrinsic markers in body feathers to identify inter-individual differences in non-breeding areas of northern fulmars.
Many wildlife studies use chemical analyses to explore spatio-temporal variation in diet, migratory patterns and contaminant exposure. Intrinsic markers are particularly valuable for studying non-breeding marine predators, when direct methods of investigation are rarely feasible. However, any inferences regarding foraging ecology are dependent upon the time scale over which tissues such as feathers are formed. In this study, we validate the use of body feathers for studying non-breeding foraging patterns in a pelagic seabird, the northern fulmar. Analysis of carcasses of successfully breeding adult fulmars indicated that body feathers moulted between September and March, whereas analyses of carcasses and activity patterns suggested that wing feather and tail feather moult occurred during more restricted periods (September to October and September to January, respectively). By randomly sampling relevant body feathers, average values for individual birds were shown to be consistent. We also integrated chemical analyses of body feather with geolocation tracking data to demonstrate that analyses of δ13C and δ15N values successfully assigned 88 % of birds to one of two broad wintering regions used by breeding adult fulmars from a Scottish study colony. These data provide strong support for the use of body feathers as a tool for exploring non-breeding foraging patterns and diet in wide-ranging, pelagic seabirds.


Foraging range of honey bees, Apis mellifera, in alfalfa seed production fields.
A study was conducted in 2006 and 2007 designed to examine the foraging range of honey bees, Apis mellifera (Hymenoptera: Apidae), in a 15.2 km(2) area dominated by a 128.9 ha glyphosate-resistant Roundup Ready® alfalfa seed production field and several non-Roundup Ready alfalfa seed production fields (totaling 120.2 ha). Each year, honey bee self-marking devices were placed on 112 selected honey bee colonies originating from nine different apiary locations. The foraging bees exiting each apiary location were uniquely marked so that the apiary of origin and the distance traveled by the marked (field-collected) bees into each of the alfalfa fields could be pinpointed. Honey bee self-marking devices were installed on 14.4 and 11.2% of the total hives located within the research area in 2006 and 2007, respectively. The frequency of field-collected bees possessing a distinct mark was similar, averaging 14.0% in 2006 and 12.6% in 2007. A grand total of 12,266 bees were collected from the various alfalfa fields on seven sampling dates over the course of the study. The distances traveled by marked bees ranged from a minimum of 45 m to a maximum of 5983 m. On average, marked bees were recovered ~ 800 m from their apiary of origin and the recovery rate of marked bees decreased exponentially as the distance from the apiary of origin increased. Ultimately, these data will be used to identify the extent of pollen-mediated gene flow from Roundup Ready to conventional alfalfa.


Effects of interleukin-1alpha administration on intestinal ischemia and reperfusion injury, mucosal permeability, and bacterial translocation in burn and sepsis.
To evaluate the effect of interleukin-1alpha (IL-1alpha) on the mesenteric circulation, intestinal mucosal integrity, and bacterial translocation in a burn/endotoxemia chronic porcine model. Major burn and sepsis are associated with a high mortality, ischemia/reperfusion injury to the intestine, and an increased rate of bacterial translocation. Pathologic alterations of IL-1 synthesis, degradation, and binding to receptors have been reported. Manipulation of IL-1-mediated effects might be of therapeutic utility. Twenty-one female pigs were instrumented with an ultrasonic flow probe on the superior mesenteric artery and a catheter into the superior mesenteric vein. After 5 days, all animals were anesthetized, and 14 received 40% total body surface area third-degree burn. IL-1alpha was administered intravenously at 1,000 ng/kg to seven pigs immediately after burn. Eighteen hours after burn, 100 microg/kg lipopolysaccharide (LPS) was administered intravenously. Systemic and splanchnic hemodynamics were measured and blood samples were drawn for blood gas analysis. Intestinal permeability was assessed every 6 hours by measuring the lactulose/mannitol (L/M) excretion ratio. At the end of the study (42 hours), tissue samples were harvested for bacteriologic cultures. Mesenteric blood flow was significantly decreased after burn and endotoxin. Administration of IL-1alpha significantly improved mesenteric blood flow postburn and post-LPS. Mesenteric oxygen supply and consumption showed a significant reduction after burn. In contrast, animals treated with IL-1alpha showed an increase in postburn mesenteric oxygen supply and consumption. LPS-induced mesenteric hypoxia was also ameliorated by IL-1alpha treatment. Intestinal permeability, as assessed by the L/M ratio, showed a 7- and 10-fold elevation after thermal injury and LPS, respectively. In contrast, IL-1alpha-treated animals showed an increase of only three- and fourfold in the L/M ratio, respectively. Bacterial translocation was significantly increased in the burn/endotoxin group. IL-1alpha significantly reduced the rates of bacterial translocation. IL-1alpha treatment attenuates mesenteric ischemia and reperfusion injury induced by thermal injury and endotoxemia by improving mesenteric blood flow and oxygenation. Subsequently, IL-1alpha reduces intestinal permeability and bacterial translocation after burn and sepsis.


Breast cancer risk for noncarriers of family-specific BRCA1 and BRCA2 mutations: findings from the Breast Cancer Family Registry.
Women with germline BRCA1 and BRCA2 mutations have five- to 20-fold increased risks of developing breast and ovarian cancer. A recent study claimed that women testing negative for their family-specific BRCA1 or BRCA2 mutation (noncarriers) have a five-fold increased risk of breast cancer. We estimated breast cancer risks for noncarriers by using a population-based sample of patients with breast cancer and their female first-degree relatives (FDRs). Patients were women with breast cancer and their FDRs enrolled in the population-based component of the Breast Cancer Family Registry; patients with breast cancer were tested for BRCA1 and BRCA2 mutations, as were FDRs of identified mutation carriers. We used segregation analysis to fit a model that accommodates familial correlation in breast cancer risk due to unobserved shared risk factors. We studied 3,047 families; 160 had BRCA1 and 132 had BRCA2 mutations. There was no evidence of increased breast cancer risk for noncarriers of identified mutations compared with FDRs from families without BRCA1 or BRCA2 mutations: relative risk was 0.39 (95% CI, 0.04 to 3.81). Residual breast cancer correlation within families was strong, suggesting substantial risk heterogeneity in women without BRCA1 or BRCA2 mutations, with some 3.4% of them accounting for roughly one third of breast cancer cases. These results support the practice of advising noncarriers that they do not have any increase in breast cancer risk attributable to the family-specific BRCA1 or BRCA2 mutation.


The genetic history of indigenous populations of the Peruvian and Bolivian Altiplano: the legacy of the Uros.
The Altiplano region of the South American Andes is marked by an inhospitable climate to which the autochthonous human populations adapted and then developed great ancient civilizations, such as the Tiwanaku culture and the Inca Empire. Since pre-Columbian times, different rulers established themselves around the Titicaca and Poopo Lakes. By the time of the arrival of Spaniards, Aymara and Quechua languages were predominant on the Altiplano under the rule of the Incas, although the occurrence of other spoken languages, such as Puquina and Uruquilla, suggests the existence of different ethnic groups in this region. In this study, we focused on the pre-Columbian history of the autochthonous Altiplano populations, particularly the Uros ethnic group, which claims to directly descend from the first settlers of the Andes, and some linguists suggest they might otherwise be related to Arawak speaking groups from the Amazon. Using phylogeographic, population structure and spatial genetic analyses of Y-chromosome and mtDNA data, we inferred the genetic relationships among Uros populations (Los Uros from Peru, Uru-Chipaya and Uru-Poopo from Bolivia), and compared their haplotype profiles with eight Aymara, nine Quechua and two Arawak (Machiguenga and Yanesha) speaking populations from Peru and Bolivia. Our results indicated that Uros populations stand out among the Altiplano populations, while appearing more closely related to the Aymara and Quechua from Lake Titicaca and surrounding regions than to the Amazon Arawaks. Moreover, the Uros populations from Peru and Bolivia are genetically differentiated from each other, indicating a high heterogeneity in this ethnic group. Finally, our results support the distinctive ancestry for the Uros populations of Peru and Bolivia, which are likely derived from ancient Andean lineages that were partially replaced during more recent farming expansion events and the establishment of complex civilizations in the Andes.


[Nerve growth factor strategy and preparation of animal model for Alzheimer-type senile dementia].
Nerve growth factor (NGF) plays an important role in the survival and maintenance of cholinergic neurons in the central nervous system. In senile dementia of the Alzheimer type (SDAT), learning and memory are impaired by the loss of neurons in the magnocellular cholinergic neuronal system. It is, therefore, of interest to investigate the role of NGF in this degenerative disorder. Since NGF does not cross the blood-brain barrier and is easily metabolized by peptidases when administered peripherally, it can be used for medical treatment only when directly injected into the brain. We tried to develop drugs which could be taken orally and stimulate the NGF synthesis in the brain. In addition, we attempted to develop a SDAT animal model using osmotic minipump to infuse beta-amyloid protein into cerebral ventricle, since there are no SDAT model animals accompanied with various pathophysiological changes. We demonstrate here that the oral administration of propentofylline, idebenone and trimethylquinone derivative, potent in vitro NGF synthesis stimulators, induced the increase in NGF protein and mRNA, and in choline acetyltransferase activity, in basal forebrain-lesioned and aged rats, but not in intact young rats. These drugs also ameliorated the behavioral deficits in habituation, water maze, and passive avoidance tasks in these animals. These results suggest that these drugs stimulated NGF synthesis in vivo and ameliorated the behavioral deficits which were accompanied with the reduced choline acetyltransferase activity in the basal forebrain-lesioned and aged rats. In terms of the SDAT animal model, the performance of the water maze and passive avoidance tasks was impaired and choline acetyltransferase activity significantly decreased in beta-amyloid protein-treated rats. Histochemical results showed the deposition of beta-amyloid protein in the cortex and hippocampus and atrophy and loss of hippocampal neurons. These results suggest that the deposition of beta-amyloid protein in the brain is related to the impairment of learning and cholinergic neuronal degeneration, and that beta-amyloid protein-treated rats could be an animal model for SDAT and used for the screening of drugs for SDAT.


Replication of hepatitis B virus in first-degree relatives of patients with hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) may occur in family clusters. No genetic mechanism has been identified as responsible for this familial tendency. We suspected that a longer hepatitis B virus (HBV) replication phase might be the reason for a higher risk of HCC in families with this disease. We performed liver biochemical tests, test for viral hepatitis markers and hepatitis B e antigen (HBeAg), and liver ultrasonography in relatives of patients with HCC. A total of 1,885 first-degree relatives from 688 families participated in this study. Seven hundred fifty-two relatives were found to be carriers of hepatitis B surface antigen (HBsAg) and 675 of them were tested for HBeAg. The prevalence of HBeAg was 27.4% in relatives of those with HCC and 20% in asymptomatic HBsAg carriers. The HBeAg prevalence rate was higher in relatives of those with HCC > or = 40 years old than in asymptomatic HBsAg carriers. Moreover, HBeAg was more likely to persist in men than in women > or = 40 years old. We conclude that families with HCC showed a prolonged HBV replication phase that may be one of the cofactors for a familial tendency for HCC.


A signal amplification strategy and sensing application using single gold nanoelectrodes.
In this work, a label-free electrochemical apta-nanosensor was fabricated on a single gold nanodisk electrode (AuNDE) for thrombin sensing with high sensitivity via a novel signal amplification strategy. This recognition platform was fabricated via self-assembly of helper DNA (HP-DNA), thrombin-binding aptamer (TBA) and gold nanoparticle (AuNP)-DNA complexes to form a sandwich structure on the AuNDE surface. A novel signal amplification strategy via designed AuNP-DNA complexes was introduced using Ru(NH3)63+ as the signal reporter based on the electrostatic interaction. In the presence of thrombin, the strong interaction between the TBA and target led to the dissociation of sandwich DNA complexes from the AuNDE, which resulted in the reduction current of Ru(NH3)63+. This proposed sensing platform showed a wide detection range of 0.1 pM-5 nM and a low detection limit of 0.02 pM. Considering the small overall dimensions and high sensitivity, this nanosensor can be potentially applied for bioanalysis in living biosystems.


Opposing wall mechanics are significantly influenced by longitudinal cardiac rotation in the assessment of ventricular dyssynchrony.
This study sought to assess whether longitudinal rotation (LR) affects myocardial systolic velocity profiles and to compare velocity-based measures of dyssynchrony with LR for predicting cardiac resynchronization therapy (CRT) response. Longitudinal rotation, a rocking motion often seen when the dilated left ventricle (LV) is imaged in its horizontal long-axis plane, is a recently recognized phenomenon and a new predictor of response to CRT. One hundred patients with CRT implants and suitable baseline echocardiograms were identified. Longitudinal rotation was assessed in the apical 4-chamber view by speckle-tracking techniques and myocardial systolic velocities for basal septum, and lateral LV were analyzed from tissue Doppler images. The quartiles of LR distribution were analyzed for differences in their systolic velocities. Correlation between measurements and reduction in LV end-systolic volume (ESV) at follow-up was performed. Quartile 1 had a mean LR of -6.8 +/- 2.3 degrees ; quartile 4 showed a mean LR of 2.3 +/- 1.6 degrees . A depressed peak velocity of lateral wall, when compared with the septum, was found for quartile 1 (p = 0.01), whereas the converse was noted in quartile 4 (p = 0.0001). The difference in amplitude of peak velocity between septal and lateral walls was found to correlate with the pattern of LR and with percentage reduction in LV ESV at follow-up in nonischemic patients. Septal-lateral delay was not correlated with the presence of LR, nor was it predictive of reduction in LV ESV. Patients with prominent clockwise LR have depressed long-axis systolic velocities of the lateral wall, whereas the patients with counterclockwise LR have depressed septal wall velocities. The difference in peak amplitude of basal septal and lateral systolic velocities is predictive of LR, and in the nonischemic subgroup correlates with quantitative LV reverse remodeling at follow-up. Velocity time-based measures, including septal-lateral delay were not predictive of CRT response.


Escherichia coli lac repressor-lac operator interaction and the influence of allosteric effectors.
The wild type E. coli lac operator is embedded in a 35 base-pair DNA sequence containing extensive 2-fold symmetry, suggesting a symmetric repressor operator complex. However, deviations from strict 2-fold symmetry occur at the central base-pair and at three additional base-pairs. Using an operator fragment binding analysis we have determined: (a) a relative contribution each pair provides to the lac repressor-lac operator DNA complex, (b) the operator DNA length necessary for maximum binding to lac repressor; and (c) the contribution of the several non-symmetric base in the wild-type operator to the binding affinity. Since lac repressor-lac operator DNA interaction is reduced upon binding of the gratuitous inducer, isopropyl-beta-D-galactoside (IPTG), the same DNA fragment binding analysis was performed with the low affinity form of lac repressor. In the presence of inducer, the affinity for the left half site of the wild-type lac operator is reduced without significant reduction on the right half of the operator. Conversely, the anti-inducer orthonitrophenylfucoside (ONPF) which stabilizes the lac repressor-lac operator complex increases the binding affinity, particularly to the right half of the operator.


Myelodysplastic syndromes: revisiting the role of the bone marrow microenvironment in disease pathogenesis.
Myelodysplastic syndromes are a heterogeneous group of diseases characterized by ineffective hematopoiesis and the propensity to leukemic transformation. Their pathogenesis is complex and likely depends on interplay between aberrant hematopoietic cells and their microenvironment. How niche cells play a role in disease evolution is poorly defined, but the delineation of the hematopoietic stem cell niche and the ability to interrogate its role in hematopoietic disease in animal models have furthered our insights in recent years. The data support a view in which the microenvironment can play an active role in the evolution of myelodysplasia and myeloproliferative disorders, thus providing further rationale to explore therapeutic targeting of mesenchymal-hematopoietic interactions in these diseases.


Decoding key nodes in the metabolism of cancer cells: sugar & spice and all things nice.
In the past 5 years, a convergence of studies has resulted in a broad appreciation in the cancer research community that reprogramming of cellular metabolism may be more central to cancer than appreciated in the past 30 years. The re-emergence of cancer metabolism stems in part from discoveries that a number of common oncogenes and tumor suppressor genes more directly control cell metabolism than previously thought. In addition, a number of what would previously have been called "card-carrying" metabolic enzymes have been identified as human tumor suppressors or oncogenes, causally mutated in a variety of human cancers. This growing appreciation of the role of altered cell metabolism has led to further investigation into the rate-limiting proteins involved in different aspects of the unique metabolism of tumor cells. Targeting cancer metabolism with drugs requires a therapeutic window in which tumor cells, compared to normal tissues, have a greater dependence on specific metabolic enzymes. Themes that have emerged in the past decade of developing oncogene-targeted cancer therapeutics suggest that tumors with distinct oncogenic lesions are likely to require drugs that target distinct metabolic pathways. Ultimately, the hope is that detailed knowledge of oncogene and tumor suppressor gene functions and their effects on metabolism will lead to drug combinations that will be far more effective in treating cancers.


Genome-wide association analyses of chronotype in 697,828 individuals provides insights into circadian rhythms.
Being a morning person is a behavioural indicator of a person's underlying circadian rhythm. Using genome-wide data from 697,828 UK Biobank and 23andMe participants we increase the number of genetic loci associated with being a morning person from 24 to 351. Using data from 85,760 individuals with activity-monitor derived measures of sleep timing we find that the chronotype loci associate with sleep timing: the mean sleep timing of the 5% of individuals carrying the most morningness alleles is 25 min earlier than the 5% carrying the fewest. The loci are enriched for genes involved in circadian regulation, cAMP, glutamate and insulin signalling pathways, and those expressed in the retina, hindbrain, hypothalamus, and pituitary. Using Mendelian Randomisation, we show that being a morning person is causally associated with better mental health but does not affect BMI or risk of Type 2 diabetes. This study offers insights into circadian biology and its links to disease in humans.


Active gaming in Dutch adolescents: a descriptive study.
Adequate levels of physical activity are part of a healthy lifestyle and in this way linked to better health outcomes. For children and adolescents, the physical activity guideline recommends at least 60 minutes of moderate-to-vigorous physical activity every day. However, many adolescents are not physically active enough and they spend a lot of their time on sedentary activities (such as video games). A new generation of video games that require body movements to play them, so-called "active games", could serve to increase physical activity in adolescents. The activity level while playing these games is comparable to light-to-moderate intensity physical activity. The current study aims to increase our understanding of 1) the demographic characteristics of adolescents who play active games regularly (≥ 1 hour per week) and non-regularly (< 1 hour per week), 2) time spent on active games, 3) the contribution of active games to daily physical activity and 4) the type and amount of activities being replaced by active gaming. A cross-sectional survey was conducted in a Dutch internet panel, questioning adolescents in conjunction with one of their parents. A random sample of 320 households (with stratification on gender of the parent and the adolescent, the age of the adolescent and the region of the household) was selected that owned a console or application for active video games and that had a child aged 12 through 16 years. 201 child-parent couples (63% response) completed an internet survey with questions about demographics, physical activity and sedentary behaviour, and gaming behaviour. The questionnaire also contained questions designed to assess whether and how active gaming replaces other activities. Besides descriptive analyses, independent t-test, Pearson's chi-square and Mann-Whitney test (when data were not normally distributed) were used for comparisons between regular and non-regular active gamers. Eleven percent of the adolescents with an active game in their household never used the game. There were no significant differences in gender, education level (of adolescent and parent), ethnicity and sedentary behaviour between regular (n = 65) and non-regular active gamers (n = 114). Adolescents' (regular and non-regular active gamers) meantime spent on active gaming was 80 (± 136) minutes a week; this potentially amounts to 11% of total physical activity. When time spent on active gaming was included in the calculation of the percentage of adolescents that met the physical activity guideline, the percentage increased significantly (p < 0.05) from 67 to 73%. According to the adolescents, active gaming mainly replaces sedentary screen time such as TV viewing, internet and non-active gaming. Parental opinions concurred with this appraisal. The results of this study confirm the idea that active gaming may contribute to an active lifestyle in adolescents, primarily because it potentially contributes substantially to time spent on physical activity. Secondly, active gamers indicate that they spent time on active games which they would have spent otherwise on less active activities.


Psychological Treatment of Depression in People Aged 65 Years and Over: A Systematic Review of Efficacy, Safety, and Cost-Effectiveness.
Depression in elderly people is a major public health concern. As response to antidepressants is often unsatisfactory in this age group, there is a need for evidence-based non-pharmacological treatment options. Our objectives were twofold: firstly, to synthesize published trials evaluating efficacy, safety and cost-effectiveness of psychological treatment of depression in the elderly and secondly, to assess the quality of evidence. The electronic databases PubMed, EMBASE, Cochrane Library, CINAL, Scopus, and PsycINFO were searched up to 23 May 2016 for randomized controlled trials (RCTs) of psychological treatment for depressive disorders or depressive symptoms in people aged 65 years and over. Two reviewers independently assessed relevant studies for risk of bias. Where appropriate, the results were synthesized in meta-analyses. The quality of the evidence was graded according to GRADE (Grading of Recommendations Assessment, Development and Evaluation). Twenty-two relevant RCTs were identified, eight of which were excluded from the synthesis due to a high risk of bias. Of the remaining trials, six evaluated problem-solving therapy (PST), five evaluated other forms of cognitive behavioural therapy (CBT), and three evaluated life review/reminiscence therapy. In frail elderly with depressive symptoms, the evidence supported the efficacy of PST, with large but heterogeneous effect sizes compared with treatment as usual. The results for life-review/reminiscence therapy and CBT were also promising, but because of the limited number of trials the quality of evidence was rated as very low. Safety data were not reported in any included trial. The only identified cost-effectiveness study estimated an incremental cost per additional point reduction in Beck Depression Inventory II score for CBT compared with talking control and treatment as usual. Psychological treatment is a feasible option for frail elderly with depressive symptoms. However, important questions about efficacy, generalizability, safety and cost-effectiveness remain.


A classifier ensemble approach for the missing feature problem.
Many classification problems must deal with data that contains missing values. In such cases data imputation is critical. This paper evaluates the performance of several statistical and machine learning imputation methods, including our novel multiple imputation ensemble approach, using different datasets. Several state-of-the-art approaches are compared using different datasets. Some state-of-the-art classifiers (including support vector machines and input decimated ensembles) are tested with several imputation methods. The novel approach proposed in this work is a multiple imputation method based on random subspace, where each missing value is calculated considering a different cluster of the data. We have used a fuzzy clustering approach for the clustering algorithm. Our experiments have shown that the proposed multiple imputation approach based on clustering and a random subspace classifier outperforms several other state-of-the-art approaches. Using the Wilcoxon signed-rank test (reject the null hypothesis, level of significance 0.05) we have shown that the proposed best approach is outperformed by the classifier trained using the original data (i.e., without missing values) only when >20% of the data are missed. Moreover, we have shown that coupling an imputation method with our cluster based imputation we outperform the base method (level of significance ∼0.05). Starting from the assumptions that the feature set must be partially redundant and that the redundancy is distributed randomly over the feature set, we have proposed a method that works quite well even when a large percentage of the features is missing (≥30%). Our best approach is available (MATLAB code) at bias.csr.unibo.it/nanni/MI.rar.