Implementation of patients' rights legislation in the Republic of Macedonia: gaps and disparities.
Since its formation after the breakup of Yugoslavia, Macedonia has made major strides in formulating a framework for protecting patient rights through extensive legal reform. The impact of this reform had not been assessed before the work of this project. Within the context of a larger project on improving human rights in patient care, this paper provides an overview of patients' rights legislation in Macedonia and uses research, case reports, and other empirical information to highlight the gaps in the implementation of patients' rights legislation on the ground. The Law on the Protection of Patients' Rights (2008) and attendant legislation governing health care provision and other aspects of the social contract in Macedonia provide extensive protections for the rights of patients in such domains as the right to access health care, the right to information, and the right to remedy. This legislation also outlines several new procedural channels to enable patients to vindicate their rights within institutional and governmental structures on the local and national levels. Data from a number of studies and case file reviews suggest, however, that the implementation of many key provisions is lacking, both in terms of quality and presence of services or mechanisms contemplated by Macedonian law. Gaps in implementation disproportionately affect vulnerable and marginalized groups, including women, rural residents, and Roma. Although the letter of Macedonian law generally complies with international best practices in patients' rights, these rights are not fully implemented and the mechanisms implied are not fully functional. Additional investment must be made in monitoring systems, education, and incentive mechanisms to ensure effective implementation, including the formation of a mandated commission for the protection of patients' rights.


rpS6 regulates blood-testis barrier dynamics through Arp3-mediated actin microfilament organization in rat sertoli cells. An in vitro study.
In the seminiferous epithelium of rat testes, preleptotene spermatocytes residing in the basal compartment are transported across the blood-testis barrier (BTB) to enter the adluminal compartment at stage VIII of the epithelial cycle. This process involves redistribution of tight junction (TJ) proteins via reorganization of actin cytoskeleton in Sertoli cells that serves as attachment site for adhesion protein complexes. Ribosomal protein S6 (rpS6), a downstream molecule of mTORC1 (mammalian target of rapamycin complex 1), participates in this process via a yet-to-be defined mechanism. Here, we constructed an rpS6 quadruple phosphomimetic mutant by converting Ser residues at 235, 236, 240, and 244 to Glu via site-directed mutagenesis, making this mutant constitutively active. When this rpS6 mutant was overexpressed in Sertoli cells cultured in vitro with an established TJ barrier mimicking the BTB in vivo, it perturbed the TJ permeability by down-regulating and redistributing TJ proteins at the cell-cell interface. These changes are mediated by a reorganization of actin microfilaments, which was triggered by a redistribution of activated actin-related protein 3 (Arp3) as well as changes in Arp3-neuronal Wiskott-Aldrich Syndrome protein (N-WASP) interaction. This in turn induced reorganization of actin microfilaments, converting them from a "bundled" to an "unbundled/branched" configuration, concomitant with a reduced actin bundling activity, thereby destabilizing the TJ-barrier function. These changes were mediated by Akt (transforming oncogene of v-akt), because an Akt knockdown by RNA interference was able to mimic the phenotypes of rpS6 mutant overexpression at the Sertoli cell BTB. In summary, this study illustrates a mechanism by which mTORC1 signal complex regulates BTB function through rpS6 downstream by modulating actin organization via the Arp2/3 complex, which may be applicable to other tissue barriers.


Conservation in function of a SCAR/WAVE component during infection thread and root hair growth in Medicago truncatula.
Nitrogen-fixing symbioses of plants are often associated with bacterially infected nodules where nitrogen fixation occurs. The plant host facilitates bacterial infection with the formation of infection threads, unique structures associated with these symbioses, which are invaginations of the host cell with the capability of traversing cellular junctions. Here, we show that the infection thread shares mechanistic similarities to polar-growing cells, because the required for infection thread (RIT) locus of Medicago truncatula has roles in root-hair, trichome, and infection-thread growth. We show that RIT encodes the M. truncatula ortholog of NAP1, a component of the SCAR/WAVE (suppressor of cAMP receptor/WASP-family verprolin homologous protein) complex that regulates actin polymerization, through the activation of ARP2/3. NAP1 of Arabidopsis thaliana functions equivalently to the M. truncatula gene, indicating that the mode of action of NAP1 is functionally conserved across species and that legumes have not evolved a unique functionality for NAP1 during rhizobial colonization. This work highlights the surprising commonality between polar-growing cells and a polar-growing cellular intrusion and reveals important insights into the formation and maintenance of infection-thread development.


Effects of tailoring health messages on physical activity.
Computer-tailored printed education can be a promising way of promoting physical activity. The present study tested whether computer-tailored feedback on physical activity is effective and whether there are differences between respondents with low and high motivation to change. Respondents (n = 487) were randomly assigned to a tailored intervention group or a no information control group. Physical activity and determinants were measured at baseline and after 3 months. At post-test, the motivated respondents in the control group were more likely not to meet the recommendation for physical activity than to meet it, and motivated respondents in the experimental group were more likely to engage in transport-related activities and showed more improvement over time for the total activity score than respondents in the control group (beta = 0.24, P = 0.02). Both groups improved their behaviour over time. No group differences in physical activity were found for the unmotivated respondents. The results showed that the effects of the tailored feedback were restricted to respondents who had a positive motivation to change at baseline. Possible explanations could be that unmotivated respondents were unwilling to read and process the information because they felt 'no need to change'. Alternatively, one tailored feedback letter may not have been sufficient for this unmotivated group.


Determinants of condom use among HIV-positive men who have sex with men.
The main objectives of this study were to identify the determinants of condom use among HIV-positive men having anal sex with HIV-negative men or men of unknown HIV status and moderators of the intention-behaviour relationship. A cohort of 237 sexually active HIV-positive men having sex with men was followed over a period of six months. The cognitive variables measured were guided by an extended version of the theory of planned behaviour. Results indicated that past behaviour (odds ratio [OR] = 9.75; 95% confidence interval [CI]: 4.48-21.26), intention (OR = 3.13; 95% CI: 1.25-7.81), self-efficacy (OR = 3.62; 95% CI: 1.40-9.37) and use of sex drugs (OR = 0.16; 95% CI: 0.06-0.45) contributed to the prediction of 100% condom use. Self-efficacy also interacted with intention as a significant moderator of the intention-behaviour relationship (OR = 20.96; 95% CI: 2.90-151.51). Interventions promoting condom use should increase self-efficacy to use condoms and target users of sex drugs.


Detection of DNA Methylation by MeDIP and MBDCap Assays: An Overview of Techniques.
DNA methylation has been characterized as the representative example of epigenetic modifications and implicated in numerous biological processes, such as genomic imprinting and X chromosome inactivation. It primarily occurs at CpG dinucleotides in mammals and plays a critical role in transcriptional regulations. Examination of DNA methylation patterns in gene(s) or across a genome is vital to understand the role of epigenetic modulation in the progress of development and tumorigenesis. Currently, lots of approaches have been developed to investigate DNA methylation patterns for either limited regions or genome-scale studies, but some of them rely on using restriction enzymes. In this chapter, we describe two commonly used protocols to detect enrichment of methylated DNA regions, namely methylated immunoprecipitation (MeDIP) and capture of methylated DNA by methyl-CpG binding domain-based (MBD) proteins (MBDCap). They are the most economical and effective methods to study DNA methylation in either single locus or genome-wide scale.


Metabolite profiles of nodulated alfalfa plants indicate that distinct stages of nodule organogenesis are accompanied by global physiological adaptations.
An effective symbiosis between Sinorhizobium meliloti and its host plant Medicago sativa is dependent on a balanced physiological interaction enabling the microsymbiont to fix atmospheric nitrogen. Maintenance of the symbiotic interaction is regulated by still poorly understood control mechanisms. A first step toward a better understanding of nodule metabolism was the determination of characteristic metabolites for alfalfa root nodules. Furthermore, nodules arrested at different developmental stages were analyzed in order to address metabolic changes induced during the progression of nodule formation. Metabolite profiles of bacteroid-free pseudonodule extracts indicated that early nodule developmental processes are accompanied by photosynthate translocation but no massive organic acid formation. To determine metabolic adaptations induced by the presence of nonfixing bacteroids, nodules induced by mutant S. meliloti strains lacking the nitrogenase protein were analyzed. The bacteroids are unable to provide ammonium to the host plant, which is metabolically reflected by reduced levels of characteristic amino acids involved in ammonium fixation. Elevated levels of starch and sugars in Fix(-) nodules provide strong evidence that plant sanctions preventing a transformation from a symbiotic to a potentially parasitic interaction are not strictly realized via photosynthate supply. Instead, metabolic and gene expression data indicate that alfalfa plants react to nitrogen-fixation-deficient bacteroids with a decreased organic acid synthesis and an early induction of senescence. Noneffective symbiotic interactions resulting from plants nodulated by mutant rhizobia also are reflected in characteristic metabolic changes in leaves. These are typical for nitrogen deficiency, but also highlight metabolites potentially involved in sensing the N status.


Safety and pharmacokinetics of single and multiple ascending doses of avibactam alone and in combination with ceftazidime in healthy male volunteers: results of two randomized, placebo-controlled studies.
Avibactam is a novel non-β-lactam β-lactamase inhibitor effective against Ambler class A, C and some class D β-lactamases that is currently in clinical development in combination with ceftazidime for the treatment of serious Gram-negative infections. It restores the in vitro activity of a range of β-lactams, including ceftazidime, against extended-spectrum β-lactamase-producing pathogens. Two phase I studies assessed the safety and pharmacokinetics of avibactam in healthy subjects when administered alone or with ceftazidime. The first study (NXL104-1001) was a placebo-controlled, single-ascending dose study assessing avibactam 50, 100, 250, 500, 1000, 1500 or 2000 mg given as a 30-min intravenous infusion. After a 7-day washout, subjects in the 250 and 500 mg dosing groups received a second avibactam dose with concomitant ceftazidime 1000 or 2000 mg, respectively. The second study (NXL104-1002) was performed in two parts. Part 1 assessed multiple-ascending doses of avibactam. Subjects were randomized to receive avibactam 500, 750 or 1000 mg every 8 h (q8 h) over 5 days, or ceftazidime-avibactam 2000-500 mg q8 h over 10 days. Part 2 assessed bioavailability of avibactam after a single oral dose (500 mg) relative to a single 30-min intravenous infusion (500 mg). No serious or severe adverse events were reported in either study. Avibactam exposure generally increased proportionally to dose and there was no trend for accumulation after multiple doses. Almost all avibactam was excreted largely unchanged in the urine within the first 6 h. Concomitant ceftazidime did not affect avibactam's safety and pharmacokinetic profile. Avibactam exposure after oral dosing was very low at 6.2 % of that observed after intravenous infusion. Avibactam was generally well tolerated across all dosing regimens, when given alone or with ceftazidime. Avibactam exposure was dose related in both studies, and avibactam pharmacokinetics were linear and not affected by ceftazidime.


Public perspectives on protective measures during the COVID-19 pandemic in the Netherlands, Germany and Italy: A survey study.
The extent to which people implement government-issued protective measures is critical in preventing further spread of coronavirus disease 2019 (COVID-19) caused by coronavirus SARS-CoV-2. Our study aimed to describe the public belief in the effectiveness of protective measures, the reported implementation of these measures, and to identify communication channels used to acquire information on COVID-19 in European countries during the early stage of the pandemic. An online survey available in multiple languages was disseminated starting on March 19th, 2020. After five days, we computed descriptive statistics for countries with more than 500 respondents. Each day, we assessed enacted community containment measures by stage of stringency (I-IV). In total, 9,796 adults responded, of whom 8,611 resided in the Netherlands (stage III), 604 in Germany (stage III), and 581 in Italy (stage IV). To explore possible dynamics as containment strategies intensified, we also included 1,365 responses submitted during the following week. Participants indicated support for governmental measures related to avoiding social gatherings, selective closure of public places, and hand hygiene and respiratory measures (range for all measures: 95.0%-99.7%). Respondents from the Netherlands less frequently considered a complete social lockdown effective (59.2%), compared to respondents in Germany (76.6%) or Italy (87.2%). Italian residents applied enforced social distancing measures more frequently (range: 90.2%-99.3%, German and Dutch residents: 67.5%-97.0%) and self-initiated hygienic and social distancing behaviors (range: 36.3%-96.6%, German and Dutch residents: 28.3%-95.7%). Respondents reported being sufficiently informed about the outbreak and behaviors to avoid infection (range: 90.2%-91.1%). Information channels most commonly reported included television newspapers, official health websites, and social media. One week later, we observed no major differences in submitted responses. During the early stage of the COVID-19 pandemic, belief in the effectiveness of protective measures among survey respondents from three European countries was high and participants reported feeling sufficiently informed. In March 2020, implementation of measures differed between countries and were highest among respondents from Italy, who were subjected to the most stringent lockdown measures and greatest COVID-19 burden in Europe during this period.


Water balance in desert arthropods. Despite their small size, arthropods may be highly adapted for life in xeric conditions.
As judged by the number of species, or of individuals, arthropods are an extremely successful group of desert inhabitants. There is very great structural and physiological diversity within the group, and since adaptations to desert life open to one are not open to all. we should not expect to find the maximum possible development of adaptive features in any arthropod simply because it lives in a desert. Most adult insects fly; their larvae and all other arthropods do not, and their adaptations will differ accordingly. Desert beetles have very impermeable cuticles and tolerate high body temperatures, while desert cockroaches live below the sand. have more permeable cuticles, and absorb water vapor. There is probably no single respect in which all desert arthropods differ from insects of other environments. Perhaps a profitable way of viewing desert animals is to recognize that each is a whole organism with a specific collection of adaptations that must be consistent within themselves and which are associated with a specific mode of life and a specific evolutionary history. The arthropod organization is capable of producing highly efficient desert species. There is, however, a converse way of looking at the situation, Which is often neglected but which may be of general biological interest: does the evolution of adaptations to desert environments necessarily involve loss of viability in more mesic habitats? If so, then what are these disavantages- what, for example, is the disadvantage of a highly impermeable cuticle? In some cases the answer is clear: sandroaches need sand dunes to live in because they are morphologically and behaviorly specialized for this habitat. More often the answer is not obvious.


Systemic sclerosis without antinuclear antibodies or Raynaud's phenomenon: a multicentre study in the prospective EULAR Scleroderma Trials and Research (EUSTAR) database.
To assess patients with SSc who present without circulating ANAs or RP. Five thousand three hundred and ninety patients who fulfilled the ACR criteria for SSc and were enrolled in the EULAR Scleroderma Trials and Research (EUSTAR) database were screened for the absence of both RP and circulating ANA. To differentiate SSc from its mimics, additional information was gathered using a standardized questionnaire. Five thousand three hundred and seventy-eight (99.8%) of the 5390 SSc patients in the EUSTAR database had either detectable ANA or a history of RP. Twelve (0.2%) patients lacked both circulating ANA and RP. Details of the medical history could be obtained for seven patients. Three cases were compatible with ANA-negative and RP-negative SSc and were not typical of any known SSc mimic. Four patients had a malignancy: two had breast cancer, one had multiple myeloma with possible scleromyxoedema and one had bladder carcinoma. There was no temporal relationship between the onset of skin fibrosis and that of the tumour. Although no patient with confirmed nephrogenic systemic fibrosis was identified among the cases of ANA-negative and RP-negative SSc, the presentation of one patient could be compatible with that of nephrogenic systemic fibrosis other than for the absence of chronic kidney disease or of known prior gadolinium exposure. We have identified a very small subgroup of SSc patients who lack both circulating ANA and RP, none of whom fulfils the diagnostic criteria for any known SSc mimic. Prospective studies are needed to elucidate the clinical presentation, evolution and outcome of such patients.


Operant conditioning of single-unit response patterns in visual cortex.
Unit responses to photic stimuli were studied in cat visual cortex. After the baseline response pattern of a cell was determined, conditioning trials were given during which reinforcement was contingent upon increased firing during a selected segment of the poststimulus interval. Density of reinforcement increased substantially in about half the cells studied; significant increases in firing occurred within, but not outside, the criterion segment.


Elevated Serum Levels of SCUBE1, a Marker for Coagulation, in Patients with Breast Cancer.
Breast cancer (BC) is the most common cancer among women and a major cause of death. Signal Peptide-Cub-Epidermal growth factor domain-containing protein-1 (SCUBE1) is secreted under hypoxia and inflammatory conditions from platelet alpha granules. Its biological function is uncertain, although it may be a procoagulant substance in cancer patients. SCUBE1 is useful for identifying thrombotic diseases, including cancers and acute coronary syndromes. D-dimer reflects the relationship between coagulation activation and fibrinolysis; namely, thrombosis and D-dimer levels are closely linked. This is the first investigation of the potential diagnostic and prognostic value of SCUBE1 levels in patients with BC. Fifty patients and 33 age-matched and body mass index-matched healthy controls were enrolled. Blood samples were collected before chemotherapy regimens commenced. Serum SCUBE1 and D-dimer levels were measured before adjuvant chemotherapy and were compared to the healthy controls. SCUBE1 levels were determined using an enzyme-linked immunosorbent assay (ELISA) method. SCUBE1 and D-dimer levels were significantly higher in patients than in the controls (p = 0.03 and p < 0.001, respectively). A cut-off value of 1.55 ng/mL for SCUBE1 was associated with 62% sensitivity and 72.7% specificity and with positive predictive value of 77.5% and negative predictive value of 55.8%. Two patients with high SCUBE1 and D-dimer levels also developed pulmonary embolism. SCUBE1 may indicate hypercoagulability in patients with BC and thus help identify patients at greater risk of thrombosis and requiring anti-thrombosis treatment. SCUBE1 may also be used as an assistant test for identifying patients at risk of BC.


Comparing ancient DNA survival and proteome content in 69 archaeological cattle tooth and bone samples from multiple European sites.
Ancient DNA (aDNA) is the most informative biomolecule extracted from skeletal remains at archaeological sites, but its survival is unpredictable and its extraction and analysis is time consuming, expensive and often fails. Several proposed methods for better understanding aDNA survival are based upon the characterisation of some aspect of protein survival, but these are typically non-specific; proteomic analyses may offer an attractive method for understanding preservation processes. In this study, in-depth proteomic (LC-Orbitrap-MS/MS) analyses were carried out on 69 archaeological bovine bone and dentine samples from multiple European archaeological sites and compared with mitochondrial aDNA and amino acid racemisation (AAR) data. Comparisons of these data, including estimations of the relative abundances for seven selected non-collagenous proteins, indicate that the survival of aDNA in bone or dentine may correlate with the survival of some proteins, and that proteome complexity is a more useful predictor of aDNA survival than protein abundance or AAR. The lack of a strong correlation between the recovery of aDNA and the proteome abundance may indicate that the survival of aDNA is more closely linked to its ability to associate with bone hydroxyapatite crystals rather than to associate with proteins. Ancient biomolecule survival remains poorly understood, even with great advancements in 'omics' technologies, both in genomics and proteomics. This study investigates the survival of ancient DNA in relation to that of proteins, taking into account proteome complexity and the relative protein abundances to improve our understanding of survival mechanisms. The results show that although protein abundance is not necessarily directly related to aDNA survival, proteome complexity appears to be.


Fluxes of oxidised and reduced nitrogen above a mixed coniferous forest exposed to various nitrogen emission sources.
Concentrations of nitrogen gases (NH(3), NO(2), NO, HONO and HNO(3)) and particles (pNH(4) and pNO(3)) were measured over a mixed coniferous forest impacted by high nitrogen loads. Nitrogen dioxide (NO(2)) represented the main nitrogen form, followed by nitric oxide (NO) and ammonia (NH(3)). A combination of gradient method (NH(3) and NO(x)) and resistance modelling techniques (HNO(3), HONO, pNH(4) and pNO(3)) was used to calculate dry deposition of nitrogen compounds. Net flux of NH(3) amounted to -64 ng N m(-2) s(-1) over the measuring period. Net fluxes of NO(x) were upward (8.5 ng N m(-2) s(-1)) with highest emission in the morning. Fluxes of other gases or aerosols substantially contributed to dry deposition. Total nitrogen deposition was estimated at -48 kg N ha(-1) yr(-1) and consisted for almost 80% of NH(x). Comparison of throughfall nitrogen with total deposition suggested substantial uptake of reduced N (+/-15 kg N ha(-1) yr(-1)) within the canopy.


Evolutionary advantages of polyploidy in halophilic archaea.
Several species of haloarchaea have been shown to be polyploid and thus this trait might be typical for and widespread in haloarchaea. In the present paper, nine different possible evolutionary advantages of polyploidy for haloarchaea are discussed, including low mutation rate, radiation/desiccation resistance, gene redundancy and survival over geological times and at extraterrestrial sites. Experimental indications exist for all but one of these evolutionary advantages. Several of the advantages require gene conversion, which has been shown to be present and active in haloarchaea.


Assessing the influence of the rhizosphere on soil hydraulic properties using X-ray computed tomography and numerical modelling.
Understanding the dynamics of water distribution in soil is crucial for enhancing our knowledge of managing soil and water resources. The application of X-ray computed tomography (CT) to the plant and soil sciences is now well established. However, few studies have utilized the technique for visualizing water in soil pore spaces. Here this method is utilized to visualize the water in soil in situ and in three-dimensions at successive reductive matric potentials in bulk and rhizosphere soil. The measurements are combined with numerical modelling to determine the unsaturated hydraulic conductivity, providing a complete picture of the hydraulic properties of the soil. The technique was performed on soil cores that were sampled adjacent to established roots (rhizosphere soil) and from soil that had not been influenced by roots (bulk soil). A water release curve was obtained for the different soil types using measurements of their pore geometries derived from CT imaging and verified using conventional methods, such as pressure plates. The water, soil, and air phases from the images were segmented and quantified using image analysis. The water release characteristics obtained for the contrasting soils showed clear differences in hydraulic properties between rhizosphere and bulk soil, especially in clay soil. The data suggest that soils influenced by roots (rhizosphere soil) are less porous due to increased aggregation when compared with bulk soil. The information and insights obtained on the hydraulic properties of rhizosphere and bulk soil will enhance our understanding of rhizosphere biophysics and improve current water uptake models.


Sequential triangle strip generator based on Hopfield networks.
The important task of generating the minimum number of sequential triangle strips (tristrips) for a given triangulated surface model is motivated by applications in computer graphics. This hard combinatorial optimization problem is reduced to the minimum energy problem in Hopfield nets by a linear-size construction. In particular, the classes of equivalent optimal stripifications are mapped one to one to the minimum energy states reached by a Hopfield network during sequential computation starting at the zero initial state. Thus, the underlying Hopfield network powered by simulated annealing (i.e., Boltzmann machine), which is implemented in the program HTGEN, can be used for computing the semioptimal stripifications. Practical experiments confirm that one can obtain much better results using HTGEN than by a leading conventional stripification program FTSG (a reference stripification method not based on neural nets), although the running time of simulated annealing grows rapidly near the global optimum. Nevertheless, HTGEN exhibits empirical linear time complexity when the parameters of simulated annealing (i.e., the initial temperature and the stopping criterion) are fixed and thus provides the semioptimal offline solutions, even for huge models of hundreds of thousands of triangles, within a reasonable time.


Attention-deficit/hyperactivity disorder after early exposure to procedures requiring general anesthesia.
To study the association between exposure to procedures performed under general anesthesia before age 2 years and development of attention-deficit/hyperactivity disorder (ADHD). Study patients included all children born between January 1, 1976, and December 31, 1982, in Rochester, MN, who remained in Rochester after age 5. Cases of ADHD diagnosed before age 19 years were identified by applying stringent research criteria. Cox proportional hazards regression assessed exposure to procedures requiring general anesthesia (none, 1, 2 or more) as a predictor of ADHD using a stratified analysis with strata based on a propensity score including comorbid health conditions. Among the 5357 children analyzed, 341 ADHD cases were identified (estimated cumulative incidence, 7.6%; 95% confidence interval [CI], 6.8%-8.4%). For children with no postnatal exposure to procedures requiring anesthesia before the age of 2 years, the cumulative incidence of ADHD at age 19 years was 7.3% (95% CI, 6.5%-8.1%). For single and 2 or more exposures, the estimates were 10.7% ( 95% CI, 6.8%-14.4%) and 17.9% ( 95% CI, 7.2%-27.4%), respectively. After adjusting for gestational age, sex, birth weight, and comorbid health conditions, exposure to multiple (hazard ratio, 1.95; 95% CI, 1.03-3.71), but not single (hazard ratio,1.18; 95% CI, 0.79-1.77), procedures requiring general anesthesia was associated with an increased risk for ADHD. Children repeatedly exposed to procedures requiring general anesthesia before age 2 years are at increased risk for the later development of ADHD even after adjusting for comorbidities.


Ase1p phosphorylation by cyclin-dependent kinase promotes correct spindle assembly in S. cerevisiae.
Spindle morphogenesis and dynamics follow an orderly sequence of events coupled to the oscillatory activation of cyclin-dependent kinase (CDK). Using S. cerevisiae, we have addressed the requirement of CDK for phosphorylation of the spindle midzone component Ase1p and its significance to spindle assembly. Ase1p is related to human PRC1, a protein negatively regulated by CDK until late mitosis, when it is required for central spindle organization and cytokinesis. By contrast, we show here that Ase1p phosphorylation by CDK promotes correct spindle assembly. Indeed, Ase1p phosphorylation coincident with spindle assembly requires Clb5p, Clb3p and Clb4p. Moreover, in clb5Δ cells, Ase1p recruitment and the kinetics of spindle formation were perturbed. These phenotypes were enhanced in a cdc28-4 clb5Δ mutant to the extent that midzone disruption resulted in transient breaks of the short spindle. By contrast, clb3Δ clb4Δ cells delayed spindle assembly downstream to Ase1p recruitment. Expression of Ase1(7D) p that mimics the phosphorylated state restored timely recruitment in clb5Δ cells and fully rescued the corresponding spindle phenotypes. Finally, Ase1(7D) p partially suppressed the spindle assembly delay in clb3Δ clb4Δ cells. Thus, Ase1p phosphorylation by CDK promotes the assembly and stability of the mitotic spindle. It follows that CDK may differentially alter the functionality of members of the Ase1p/PRC1 family to place their distinct roles in their respective stage-specific contexts, a further factor of complexity in the organization of pathways promoting spindle assembly and dynamics.


What works in falls prevention after stroke?: a systematic review and meta-analysis.
Falls are common after stroke. Despite evidence that single and multifactorial interventions can reduce falls in older people, this issue remains relatively underexplored in stroke survivors. Effective fall prevention in this population has the potential to prevent injury, improve quality of life, and decrease the likelihood of subsequent fear of falling and activity restriction. The aim of this article was to review and integrate the research evidence relating to interventions that reduce falls after stroke. Published studies evaluating interventions to reduce falls in stroke survivors were retrieved and screened according to predetermined criteria. Included studies were independently assessed. Quality of trials was assessed using the Physiotherapy Evidence Database score. Pooling of results was undertaken for similar interventions with comparable outcomes using the inverse variance method. Thirteen studies met the inclusion criteria, with pooling of results possible for only 2 types of intervention. Methodological quality of the included studies was variable with the main bias because of lack of blinding of participants and those administering the intervention. Variability in falls data reporting was seen across the studies. The only intervention shown to be effective in reducing falls was vitamin D for female stroke survivors in an institutional setting. Other interventions were no more effective than usual care. Fall risk is high in stroke survivors; however, the only intervention shown to be effective in reducing falls in this review was vitamin D supplementation. Consistency in outcome measurement would enable comparisons across studies. Additionally, further research evaluating a range of single and multifactorial interventions for fall prevention in the stroke population is required.


Prevalence and Patterns of Marijuana Use in Young Adults With Inflammatory Bowel Disease.
Recent studies in adults report symptom relief with marijuana use in patients with inflammatory bowel disease (IBD). We assessed the prevalence, pattern, effects, and adverse effects of marijuana use in young adults with IBD. We conducted a prospective questionnaire survey study at a pediatric IBD clinic. All patients (18-21 years of age) answered anonymous questionnaires about demographics, IBD, medications, and marijuana use. Fifty-three patients (mean age 18.7 years, 32 boys) were enrolled. Thirty-seven patients (70%) reported using marijuana currently or in the past. There was no statistically significant difference between the users and nonusers of marijuana regarding demographics, disease activity, or medications. Despite prolonged use of marijuana, 70% of patients did not discuss it with their gastroenterologists. Twenty-four patients used marijuana medicinally for IBD symptoms in addition to medical therapy. Although majority found marijuana to be moderately/very helpful, complete relief of symptoms such as abdominal pain, poor appetite, nausea, and diarrhea was seen in 29%, 37%, 14%, and 10% of patients, respectively. Only half of patients reported knowledge of possible adverse effects of marijuana and 19% of patients reported mild neuropsychiatric adverse effects. Overall, 98% of patients supported legalization of marijuana and 85% were interested in using medical marijuana if it became legally available. We found a high rate of marijuana use in our cohort of young adults with IBD. Majority of users report symptom improvement but do not inform physicians. Future well-controlled studies are necessary to assess role of marijuana in IBD therapy.


Melatonin alleviates postinfarction cardiac remodeling and dysfunction by inhibiting Mst1.
Melatonin reportedly protects against several cardiovascular diseases including ischemia/reperfusion (I/R), atherosclerosis, and hypertension. The present study investigated the effects and mechanisms of melatonin on cardiomyocyte autophagy, apoptosis, and mitochondrial injury in the context of myocardial infarction (MI). We demonstrated that melatonin significantly alleviated cardiac dysfunction after MI. Four weeks after MI, echocardiography and Masson staining indicated that melatonin notably mitigated adverse left ventricle remodeling. The mechanism may be associated with increased autophagy, reduced apoptosis, and alleviated mitochondrial dysfunction. Furthermore, melatonin significantly inhibited Mst1 phosphorylation while promoting Sirt1 expression after MI, which indicates that Mst1/Sirt1 signaling may serve as the downstream target of melatonin. We thus constructed a MI model using Mst1 transgenic (Mst1 Tg) and Mst1 knockout (Mst1-/- ) mice. The absence of Mst1 abolished the favorable effects of melatonin on cardiac injury after MI. Consistently, melatonin administration did not further increase autophagy, decrease apoptosis, or alleviate mitochondrial integrity and biogenesis in Mst1 knockout mice subjected to MI injury. These results suggest that melatonin alleviates postinfarction cardiac remodeling and dysfunction by upregulating autophagy, decreasing apoptosis, and modulating mitochondrial integrity and biogenesis. The attributed mechanism involved, at least in part, Mst1/Sirt1 signaling.


Smoking initiation among youth: the role of cigarette excise taxes and prices by race/ethnicity and gender.
Existing evidence for the role of cigarette excise taxes and prices as significant determinants of youth smoking initiation is mixed. A few studies have considered the possibility that the impact of cigarette taxes and prices might differ by gender or race/ethnicity. In this paper, we address the role of cigarette taxes and prices on youth smoking initiation using the National Longitudinal Survey of Youth 1997 cohort and discrete-time survival methods. We present results overall and by gender, race/ethnicity, and gender by race/ethnicity. We examine initiation over the age range during which youth are most at risk of initiation and over a period in which substantial changes have occurred in tax and price. The result for cigarette excise taxes is small and mixed across alternative specifications, with the effect strongest for black youth. Cigarette prices are more consistently a significant determinant of youth smoking initiation, especially for black youth.


In vitro amplification of BVDV field strains isolated in Argentina: effect of cell line and culture conditions.
The aim of this work was to study the in vitro amplification of BVDV (Pestivirus, Flaviridae) field isolates from Argentina in MDBK, BoTur and BHK-21 continuous cell lines. Field isolates 99/134 (mucosal disease), 00/693 (mucosal disease), 04P7016 (respiratory disease) and 04/89 (mucosal disease), genotype 1b, were used and compared with the Singer and NADL reference strains, genotype 1a. Additionally, cell lines derived from explants of bovine testis (RD-420), bovine uterus (NCL-1) and porcine kidney (PKZ) were tested as alternative substrates for BVDV propagation in vitro. The effect of cell line, harvest time and infection protocol was evaluated. The viral titers observed depended on the virus and harvest time but not on the infection protocol. We found that MDBK and BoTur cell lines were susceptible to the infection whereas BHK-21 and PKZ were not. NADL viral titers, 00/693 and 04/89, increased from 24 to 48 h p.i. in BoTur cells and then reached a plateau, whereas those of 99/134 and 04P7016 remained constant between 24 and 72 h p.i. BVDV Singer, on the other hand, presented a maximum titer at 24 h p.i. and then decreased. BVDV-NADL titers increased in MDBK and NCL-1 but not in RD-420 between 24 and 48 h p.i., and then decreased at 72 h p.i. These facts lead us to conclude that neither the subgenotypes (1a, 1b) nor the clinical symptoms of the animal from the virus had been isolated seem to affect the virus cell line kinetics of viral replication in vitro. On the other hand, the most homogenous behavior, the most similar replication curves, and highest titers observed in MDBK and NCL-1 seem to indicate that these lines are generally more susceptible to BVDV replication.


U.S. Ebola Treatment Center Clinical Laboratory Support.
Fifty-five hospitals in the United States have been designated Ebola treatment centers (ETCs) by their state and local health authorities. Designated ETCs must have appropriate plans to manage a patient with confirmed Ebola virus disease (EVD) for the full duration of illness and must have these plans assessed through a CDC site visit conducted by an interdisciplinary team of subject matter experts. This study determined the clinical laboratory capabilities of these ETCs. ETCs were electronically surveyed on clinical laboratory characteristics. Survey responses were returned from 47 ETCs (85%). Forty-one (87%) of the ETCs planned to provide some laboratory support (e.g., point-of-care [POC] testing) within the room of the isolated patient. Forty-four (94%) ETCs indicated that their hospital would also provide clinical laboratory support for patient care. Twenty-two (50%) of these ETC clinical laboratories had biosafety level 3 (BSL-3) containment. Of all respondents, 34 (72%) were supported by their jurisdictional public health laboratory (PHL), all of which had available BSL-3 laboratories. Overall, 40 of 44 (91%) ETCs reported BSL-3 laboratory support via their clinical laboratory and/or PHL. This survey provided a snapshot of the laboratory support for designated U.S. ETCs. ETCs have approached high-level isolation critical care with laboratory support in close proximity to the patient room and by distributing laboratory support among laboratory resources. Experts might review safety considerations for these laboratory testing/diagnostic activities that are novel in the context of biocontainment care.


The effect of various cold-water immersion protocols on exercise-induced inflammatory response and functional recovery from high-intensity sprint exercise.
The purpose of this study was to investigate the effects of different cold-water immersion (CWI) protocols on the inflammatory response to and functional recovery from high-intensity exercise. Eight healthy recreationally active males completed five trials of a high-intensity intermittent sprint protocol followed by a randomly assigned recovery condition: 1 of 4 CWI protocols (CWI-10 min × 20 °C, CWI-30 min × 20 °C, CWI-10 min × 10 °C, or CWI-30 min × 10 °C) versus passive rest. Circulating mediators of the inflammatory response were measured from EDTA plasma taken pre-exercise (baseline), immediately post-exercise, and at 2, 24, and 48 h post-exercise. Ratings of perceived soreness and impairment were noted on a 10-pt Likert scale, and squat jump and drop jump were performed at these time points. IL-6, IL-8, and MPO increased significantly from baseline immediately post-exercise in all conditions. IL-6 remained elevated from baseline at 2 h in the CWI-30 min × 20 °C, CWI-10 min × 10 °C, and CWI-30 min × 10 °C conditions, while further increases were observed for IL-8 and MPO in the CWI-30 min × 20 °C and CWI-30 min × 10 °C conditions. Squat jump and drop jump height were significantly lower in all conditions immediately post-exercise and at 2 h. Drop jump remained below baseline at 24 and 48 h in the CON and CWI-10 min × 20 °C conditions only, while squat jump height returned to baseline in all conditions. Cold-water immersion appears to facilitate restoration of muscle performance in a stretch-shortening cycle, but not concentric power. These changes do not appear to be related to inflammatory modulation. CWI protocols of excessive duration may actually exacerbate the concentration of cytokines in circulation post-exercise; however, the origin of the circulating cytokines is not necessarily skeletal muscle.


Measurement of telomere length: a new assay using QuantiGene chemistry on a Luminex platform.
Telomeres are tandem repeats of sequences present at the end of the chromosomes that maintain chromosomal integrity. After repeated cell division, telomeres shorten to a critical level, triggering replicative senescence or apoptosis, which is a key determinant of cellular aging. Short telomeres also contribute to genome instability and are a hallmark of many cancers. There are several methods for estimating telomere length (TL) from extracted DNA samples. Southern blot is accurate but requires a large quantity of DNA and is expensive. qPCR is cost-effective and requires a small quantity of DNA and is therefore widely used for large-scale epidemiologic studies; however, it typically requires triplicates. We describe a novel multiplexed probe-based non-PCR method for TL measurement. A small amount of DNA (∼50 ng) is hybridized to telomere repeat sequence-specific probes (T) and a reference single gene probes (R). T and R signals are detected from a single reaction well containing the same input DNA. Branching DNA technology is used to amplify the signal, which is detected by Luminex technology. The intra- and interassay CV (∼3% and ∼5%, respectively) shows the precision of the new assay and the measurements from single well correlated well with traditional single-plex qPCR run in triplicate (r = 0.7 to 0.8). The assay was also validated in an independent set of samples using Southern blot (r = 0.74). We describe a novel assay for TL assessment using the Luminex platform. This may offer an alternative cost-efficient way to study TL in extracted DNA samples. See all the articles in this CEBP Focus section, "Biomarkers, Biospecimens, and New Technologies in Molecular Epidemiology."


Association of Cardiovascular Health With Subclinical Disease and Incident Events: The Multi-Ethnic Study of Atherosclerosis.
Few adults have ideal cardiovascular health (CVH). We studied associations of an overall CVH score with subclinical cardiovascular disease and events. We assessed whether associations varied by race/ethnicity. Among 5961 participants in the Multi-Ethnic Study of Atherosclerosis, components of CVH were measured at baseline, 2000-2002: systolic blood pressure, total cholesterol, fasting glucose, smoking, physical activity, diet, and body mass index. Levels were classified as ideal (2 points), intermediate (1 point), and poor (0 points) according to American Heart Association definitions. Points were summed to produce a CVH score (0-7 low, 8-11 moderate, 12-14 high). Coronary artery calcium, carotid intima-media thickness, and left ventricular mass were measured at baseline. Cardiovascular disease was defined as myocardial infarction, coronary heart disease death, resuscitated cardiac arrest, stroke, heart failure, or peripheral artery disease. Follow-up was 10.3 years. Regression models were used to examine associations of the CVH score with subclinical disease and events, adjusting for age, sex, and education. Analyses were stratified by race/ethnicity. Adults with high or moderate CVH scores had significantly lower odds of coronary artery calcium and lower carotid intima-media thickness and left ventricular mass than adults with low CVH scores. Adults with high or moderate CVH scores were 67% (95%CI 41% to 82%) and 37% (95%CI 22% to 49%) less likely, respectively, to experience a cardiovascular disease event than adults with low scores. There was no interaction with race/ethnicity. There is a graded inverse association between CVH scores and measures of subclinical and overt cardiovascular disease that is similar across race/ethnic groups.


Driving evaluation methods for able-bodied persons and individuals with lower extremity disabilities: a review of assessment modalities.
Assessing the driving abilities of individuals with disabilities is often a very challenging task because each medical condition is accompanied by physical impairments and because relative individual functional performance may vary depending on personal characteristics.We identified existing driving evaluation modalities for able-bodied and lower extremity-impaired subjects (spinal cord injury patients and amputees) and evaluated the potential relationships between driving performance and the motor component of driving.An extensive scoping review of the literature was conducted to identify driving assessment tools that are currently used for able-bodied individuals and for those with spinal cord injury or lower extremity amputation. The literature search focused on the assessment of the motor component of driving. References were electronically obtained via Medline from the PubMed, Ovid, Web of Science and Google Scholar databases.This article compares the current assessments of driving performance for those with lower extremity impairments with the assessments used for able-bodied persons. Very few articles were found concerning "Lower Extremity Disabilities," thus confirming the need for further studies that can provide evidence and guidance for such assessments in the future. Little is known about the motor component of driving and its association with the other driving domains, such as vision and cognition. The available research demonstrates the need for a more evidenced-based understanding of how to best evaluate persons with lower extremity impairment.


Prediction of prolonged pain experiences during orthodontic treatment.
In this study, we investigated prolonged pain reactions in teenage orthodontic patients during a common orthodontic treatment. The aim was to examine factors predicting pain at the end of a follow-up week after placement of elastic separators. Fifty-five patients (ages, 12-18 years) were included. Baseline assessments were made of perceived intensity of general and dental pain experiences, motivation for treatment, dental anxiety, and personality factors (self-esteem and temperament). Pain intensity was assessed on a visual analog scale, and pain medications were recorded. The patients were separated into pain and no-pain groups according to pain experiences at day 7. The pain group (mainly girls) had significantly higher ratings of treatment pain than in the non-pain group at all times measured except for the treatment day. Bivariate and multiple logistic regressions showed significant predictive power from motivation, dental anxiety, activity temperament, and vaccination pain. In this adolescent patient sample, low motivation for orthodontic treatment, high ratings of vaccination pain, elevated dental anxiety level, and low activity temperament characterized patients reporting pain 1 week after the elastic separators were placed.


Weekly versus daily changes of in-line suction catheters: impact on rates of ventilator-associated pneumonia and associated costs.
An earlier randomized, controlled trial showed that weekly or as-needed (as opposed to daily) changes of in-line suction catheters were associated with substantial cost savings, without a higher rate of ventilator-associated pneumonia (VAP). To examine the impact of decreasing the frequency of in-line suction catheter changes in our medical intensive care unit, we conducted an observational study, comparing the catheter costs and frequency of VAP during (1) a control period, during which in-line suction catheters were changed daily, and (2) a treatment period, during which the catheters were changed every 7 days or sooner if needed, for mechanical failure or soilage. All adult patients admitted to our 18-bed medical intensive care unit were evaluated for the 3-month interval 1 year prior to the practice change (May through July 1998) and for the 3 months after implementing the new policy (May through July 1999). To avoid bias related to usual seasonal variation in VAP frequency, we also determined (via medical records) the VAP rate during May through July 1997. The occurrence of VAP was ascertained by an infection control practitioner, using criteria established by the Centers for Disease Control and applied in a standard fashion. The VAP rate was calculated as the mean number of VAPs per 100 ventilator-days for each 3-month interval. Use of ventilators, humidifiers, and non-heated-wire, disposable circuits was uniform during the study, as were policies regarding humidity, temperature settings, and frequency of routine ventilator circuit changes. During the control period 146 patients accounted for 1,075 ventilator-days and there were 2 VAPs (0.19 VAPs per 100 ventilator-days). During the treatment period 143 patients accounted for 1,167 ventilator-days and there were no VAPs. The mean +/- SD duration of in-line suction catheter use during the treatment period was 3.8 +/- 0.8 days, and 51% of the patients had the same catheter in place for > 3 days (range 4-9 days). The actual cost of catheters used during the treatment period was lower than during the control period ($1,330 vs $6,026), predicting annual catheter cost savings of $18,782. We conclude that (1) a policy of weekly (vs daily) change of in-line suction catheter is associated with substantial cost savings, with no significant increase in the frequency of VAP, and (2) to the extent that these findings confirm the results of prior studies they support a policy of changing in-line suction catheters weekly rather than daily.


Sexual risk behavior associated with transition to injection among young non-injecting heroin users.
The aim of this study was to investigate the effects of transitioning from non-injection heroin use to injection drug use on sexual risk behavior. Non-injecting heroin users age 16-30 were enrolled from 2002 to 2005, and were re-interviewed at 6-month intervals for up to three years; 561 participants completed at least one follow-up interview. The majority of participants were non-Hispanic (NH) Black (54 %), 23 % were Hispanic, and 21 % were NH white. During follow-up, 154 participants (27.5 %) transitioned to injecting drugs. Logistic regression analyses were conducted using generalized estimating equations (GEE) to estimate the effect of transition to injection drug use on changes in sexual risk behavior during follow-up. Transition to injection drug use during follow-up was associated with increased likelihood of sexual risk behavior, especially for men. Harm reduction efforts that focus on preventing initiation or return to injection among non-injecting drug users may also ameliorate HIV sexual risk behaviors.


Comparison of weather station and climate reanalysis data for modelling temperature-related mortality.
Epidemiological analyses of health risks associated with non-optimal temperature are traditionally based on ground observations from weather stations that offer limited spatial and temporal coverage. Climate reanalysis represents an alternative option that provide complete spatio-temporal exposure coverage, and yet are to be systematically explored for their suitability in assessing temperature-related health risks at a global scale. Here we provide the first comprehensive analysis over multiple regions to assess the suitability of the most recent generation of reanalysis datasets for health impact assessments and evaluate their comparative performance against traditional station-based data. Our findings show that reanalysis temperature from the last ERA5 products generally compare well to station observations, with similar non-optimal temperature-related risk estimates. However, the analysis offers some indication of lower performance in tropical regions, with a likely underestimation of heat-related excess mortality. Reanalysis data represent a valid alternative source of exposure variables in epidemiological analyses of temperature-related risk.


Ace/Fgf8 is required for forebrain commissure formation and patterning of the telencephalon.
Fibroblast growth factors (Fgfs) form a large family of secreted signalling proteins that have a wide variety of roles during embryonic development. Within the central nervous system (CNS) Fgf8 is implicated in patterning neural tissue adjacent to the midbrain-hindbrain boundary. However, the roles of Fgfs in CNS tissue rostral to the midbrain are less clear. Here we examine the patterning of the forebrain in zebrafish embryos that lack functional Fgf8/Ace. We find that Ace is required for the development of midline structures in the forebrain. In the absence of Ace activity, midline cells fail to adopt their normal morphology and exhibit altered patterns of gene expression. This disruption to midline tissue leads to severe commissural axon pathway defects, including misprojections from the eye to ectopic ipsilateral and contralateral targets. Ace is also required for the differentiation of the basal telencephalon and several populations of putative telencephalic neurons but not for overall regional patterning of forebrain derivatives. Finally, we show that ace expression co-localises with anterior neural plate cells that have previously been shown to have forebrain patterning activity. Removal of these cells leads to a failure in induction of ace expression indicating that loss of Ace activity may contribute to the phenotypes observed when anterior neural plate cells are ablated. However, as ace mutant neural plate cells still retain at least some inductive activity, then other signals must also be produced by the anterior margin of the neural plate.


Pharmacodynamics of dose-escalated 'front-loading' polymyxin B regimens against polymyxin-resistant mcr-1-harbouring Escherichia coli.
Gram-negative bacteria harbouring the mcr-1 plasmid are resistant to the 'last-line' polymyxins and have been reported worldwide. Our objective was to define the impact of increasing the initial polymyxin B dose intensity against an mcr-1 -harbouring strain to delineate the impact of plasmid-mediated polymyxin resistance on the dynamics of bacterial killing and resistance. A hollow fibre infection model (HFIM) was used to simulate polymyxin B regimens against an mcr-1 -harbouring Escherichia coli (MIC 8 mg/L) over 10 days. Four escalating polymyxin B 'front-loading' regimens (3.33, 6.66, 13.3 or 26.6 mg/kg for one dose followed by 1.43 mg/kg every 12 h starting 12 h later) simulating human pharmacokinetics were utilized in the HFIM. A mechanism-based, mathematical model was developed using S-ADAPT to characterize bacterial killing. The 3.33 mg/kg 'front-loading' regimen resulted in regrowth mirroring the growth control. The 6.66, 13.3 and 26.6 mg/kg 'front-loading' regimens resulted in maximal bacterial reductions of 1.91, 3.79 and 6.14 log 10 cfu/mL, respectively. Irrespective of the early polymyxin B exposure (24 h AUC), population analysis profiles showed similar growth of polymyxin B-resistant subpopulations. The HFIM data were well described by the mechanism-based model integrating three subpopulations (susceptible, intermediate and resistant). Compared with the susceptible subpopulation of mcr-1 -harbouring E. coli , the resistant subpopulation had an approximately 10-fold lower rate of killing due to polymyxin B treatment. Manipulating initial dose intensity of polymyxin B was not able to overcome plasmid-mediated resistance due to mcr-1 in E. coli . This reinforces the need to develop new combinatorial strategies to combat these highly resistant Gram-negative bacteria.


Atlantoaxial rotatory subluxation in patients with Marfan syndrome. A report of three cases.
This is a retrospective case series of three patients, ages 9 1/2, 13, and 20 years old, with Marfan syndrome treated for atlantoaxial rotatory subluxation. In the first two cases, acute torticollis was noted postoperatively, following pectus excavatum repair. The diagnosis was made in the third patient after she presented to the emergency room with a week-long history of unresolved neck pain following minor trauma. To report and discuss the courses and clinical sequelae of atlantoaxial subluxation in patients with Marfan syndrome. Radiographic analysis of patients with Marfan syndrome has shown that increased atlantoaxial translation, larger odontoid height, and basilar impression are more prevalent in this population compared to age-matched controls. Despite these findings, there are sparse data on injuries secondary to cervical spine instability or abnormalities in this population. To the authors' best knowledge, no report of atlantoaxial rotatory subluxation in patients with Marfan syndrome exists in the literature. Case records of rotatory instability of the atlanto-axial level were reviewed and are presented in the following report. The first two patients described in this report were noted to have "cock robin" posturing of their necks following pectus excavatum repairs. The first patient's subluxation was partially reduced with halo traction, and he subsequently underwent posterior spinal fusion of C1-C2 with internal fixation. The patient was well aligned postoperatively, and had no neurologic deficits. The second patient's subluxation reduced after 20 days of halter and traction; he was immobilized in a collar following discharge and reduction was maintained. The third patient's subluxation failed to reduce with halo traction; further imaging studies revealed odontoid prominence in the foramen magnum. She underwent posterior spinal fusion, occiput to C3, with satisfactory result. The cervical bony and ligamentous abnormalities seen in patients with Marfan syndrome may slightly increase their risk for atlantoaxial rotatory instability. Special attention to intubation and positioning, both intraoperatively and postoperatively, may be necessary in patients with Marfan syndrome. Additionally, rotatory subluxation should be included in the differential diagnosis for Marfan patients with neck pain after injury.


Pax6a and Pax6b are required at different points in neuronal progenitor cell proliferation during zebrafish photoreceptor regeneration.
The light-damaged zebrafish retina results in the death of photoreceptor cells and the subsequent regeneration of the missing rod and cone cells. Photoreceptor regeneration initiates with asymmetric Müller glial cell division to produce neuronal progenitor cells, which amplify, migrate to the outer nuclear layer (ONL), and differentiate into both classes of photoreceptor cells. In this study, we examined the role of the Pax6 protein in regeneration. In zebrafish, there are two Pax6 proteins, one encoded by the pax6a gene and the other encoded by the pax6b gene. We intravitreally injected and electroporated morpholinos that were complementary to either the pax6a or pax6b mRNA to knockdown the translation of the corresponding protein. Loss of Pax6b expression did not affect Müller glial cell division, but blocked the subsequent first cell division of the neuronal progenitors. In contrast, the paralogous Pax6a protein was required for later neuronal progenitor cell divisions, which maximized the number of neuronal progenitors. Without neuronal progenitor cell amplification, proliferation of resident ONL rod precursor cells, which can only regenerate rods, increased inversely proportional to the number of INL neuronal progenitor cells. This confirmed that Müller glial-derived neuronal progenitor cells are necessary to regenerate cones and that distinct mechanisms selectively regenerate rod and cone photoreceptors. This work also defines distinct roles for Pax6a and Pax6b in regulating neuronal progenitor cell proliferation in the adult zebrafish retina and increases our understanding of the molecular pathways required for photoreceptor cell regeneration.


A high-throughput and quantitative hierarchical oligonucleotide primer extension (HOPE)-based approach to identify sources of faecal contamination in water bodies.
As faecal contamination of recreational and drinking water impairs the water quality and threatens public health, water bodies are routinely monitored for faecal coliforms to detect contamination. However, faecal coliforms are facultative anaerobes that survive and reproduce in ambient waters, and their presence does not depict the origin of contamination. Therefore, the use of Bacteroides-Prevotella 16S rRNA gene to perform faecal source tracking has been proposed and applied. Here, we demonstrate the use of a new molecular method termed hierarchical oligonucleotide primer extension (HOPE) to simultaneously detect human-associated Bacteroides spp. and three clusters of cow-, pig- and dog-specific uncultivated Bacteroidales. The method correctly identifies the origin of faecal contamination when tested against human, cow, pig and dog faeces (n = 17, 17, 16 and 13 respectively), and in waters contaminated with faeces of known origins. Subsequent tests with a total of 21 blind samples show that HOPE is able to accurately indicate single or multiple sources of faecal contamination originating from pigs, cows and humans in 81% of the blind samples. HOPE can further correctly detect and identify faecal contamination in five sampling sites located along a canal in southern Taiwan, and the results are validated against conventional faecal coliform tests and quantitative PCR. Overall, this study demonstrates HOPE as a quantitative and high-throughput method that can identify sources of faecal contamination.


[Is the general practice laboratory medically and economically valuable?].
Between 1998 and 2000 we evaluated the office-based laboratory activities of general practitioners. The aim was to clarify whether there is a medical and economic benefit of these activities. The study was performed in four parts: I. A cross-sectional study with a random sample of general practitioners of the German and French speaking part of Switzerland. II. A prospective evaluation of the office-based laboratory activities of 56 GP's. III. A cross-sectional study of the preference of 837 patients in 52 of GP's offices. IV. A consensus panel with nine experts using the RAND method. 1999 there were 55.4 Million laboratory tests ordered by GP's (excl. pediaters) of which 78.9% were analysed in the office-based laboratory. The probability of a second visit is reduced by 60%, if all of the tests could be performed in the office-based laboratory. 85% of the patients appreciate the possibility to discuss the test results within the same consulation. In the consensus panel, 43 tests were proposed of which only bicarbonate, chloride and urea were assessed as not useful for the office based laboratory. The office-based laboratory is a well embodied institution in Switzerland. It's predominant advantage is the possibility of point of care testing. It allows a quick management of the patient and avoids unnecessary second consultations.


Machine learning-aided quantification of antibody-based cancer immunotherapy by natural killer cells in microfluidic droplets.
Natural killer (NK) cells have emerged as an effective alternative option to T cell-based immunotherapies, particularly against liquid (hematologic) tumors. However, the effectiveness of NK cell therapy has been less than optimal for solid tumors, partly due to the heterogeneity in target interaction leading to variable anti-tumor cytotoxicity. This paper describes a microfluidic droplet-based cytotoxicity assay for quantitative comparison of immunotherapeutic NK-92 cell interaction with various types of target cells. Machine learning algorithms were developed to assess the dynamics of individual effector-target cell pair conjugation and target death in droplets in a semi-automated manner. Our results showed that while short contacts were sufficient to induce potent killing of hematological cancer cells, long-lasting stable conjugation with NK-92 cells was unable to kill HER2+ solid tumor cells (SKOV3, SKBR3) significantly. NK-92 cells that were engineered to express FcγRIII (CD16) mediated antibody-dependent cellular cytotoxicity (ADCC) selectively against HER2+ cells upon addition of Herceptin (trastuzumab). The requirement of CD16, Herceptin and specific pre-incubation temperature served as three inputs to generate a molecular logic function with HER2+ cell death as the output. Mass proteomic analysis of the two effector cell lines suggested differential changes in adhesion, exocytosis, metabolism, transport and activation of upstream regulators and cytotoxicity mediators, which can be utilized to regulate specific functionalities of NK-92 cells in future. These results suggest that this semi-automated single cell assay can reveal the variability and functional potency of NK cells and may be used to optimize immunotherapeutic efficacy for preclinical analyses.


Anatomical study in the Western world before the Middle Ages: historical evidence.
Although modern anatomy is commonly retained to begin in the XVI century, the roots of anatomical study in the Western world may be identified beforehand. An anatomical practice was present in the Western world well before the Middle Ages, starting in ancient Greece. Hippocrates of Cos (V-IV centuries B.C.) provided descriptions of the heart and vessels, and the so-called "Hippocratic Corpus" largely deals with anatomy. Aristotle of Stagira (IV century B.C.) was one of the first well-known scholars of the past to perform dissections of animals. The anatomical interest of Aristotle contained a "physiological" background too, since he was convinced that all parts of human organisms had one or more specific functions. Galen of Pergamum (II century A.D.) was the performer of hundreds of dissections of animals, and he described a great number of anatomical parts of apes, dogs, goats and pigs. The anatomical system of Galen became a gold standard for medicine for more than a thousand years, and in the Middle Ages (V-XV centuries A.D.) the human anatomy that was taught and acquired in European universities remained based on Galenic anatomy. In conclusion, Greek-speaking scholars between the IV century B.C. and the II century A.D. set the basis for the systematic dissection of animals and the comparative investigation of animal anatomical findings. These scholars also began to study the structures of the human body, interestingly taking into account the relationship between the macroscopical morphology of observed structures and their more evident functions. (www.actabiomedica.it).


Predictors of early acute lung injury at a combat support hospital: a prospective observational study.
Acute lung injury (ALI) is a syndrome consisting of noncardiogenic acute hypoxemic respiratory failure with the presence of bilateral pulmonary infiltrates and occurs in up to 33% of critically ill trauma patients. Retrospective and observational studies have suggested that a blood component resuscitation strategy using equal ratios of packed red blood cells (PRBCs) and fresh frozen plasma (FFP) may have a survival benefit in combat casualties. The purpose of this study was to determine whether this strategy is associated with an increased incidence of ALI. We performed a prospective observational study of all injured patients admitted to an intensive care unit (ICU) at a combat support hospital who required >5 units of blood transfusion within the first 24 hours of admission. Baseline demographic data along with Injury Severity Score (ISS), pulmonary injury, presence of long bone fracture, blood products transfused, mechanical ventilation data, and arterial blood gas analysis were collected. The primary endpoint of the study was the development of ALI at 48 hours after injury. Those who did not survive to ICU admission were excluded from analysis. Follow-up (including mortality) longer than 48 hours was unavailable secondary to rapid transfer out of our facility. A multivariate logistic regression was performed to determine the independent effects of variables on the incidence of early ALI. During a 12-month period (from January 2008 to December 2008), 87 subjects were studied; of these, 66 patients met inclusion criteria, and 22 patients developed ALI at 48 hours (33%). Overall, the ratio of FFP to PRBC was 1:1.1. Those who developed ALI had a higher ISS (32 +/- 15 vs. 26 +/- 11; p = 0.04) and received more units of FFP (22 +/- 15 vs. 12 +/- 7; p < 0.001), PRBCs (22 +/- 16 vs. 13 +/- 7; p = 0.008), and platelets (5 +/- 11 vs. 1 +/- 2; p = 0.004) compared with those who did not develop ALI. Multivariate logistic regression analysis revealed that presence of pulmonary injury (odds ratio, 5.4; 95% confidence interval, 1.3-21.9) and volume of FFP transfused (odds ratio, 1.2; 95% confidence interval, 1.1-1.3) had independent effects on ALI at 48 hours. On the basis of this small, prospective, descriptive study of severely injured patients admitted to the ICU, we determined that the presence of pulmonary injury had the greatest impact on the incidence of early ALI. There was also an independent relationship between the amount of FFP transfused and the incidence of early ALI. Further studies are required to determine the effects of the development of early ALI from FFP transfusion on short- and long-term survival.


Secular trend in age at menarche for South Korean women born between 1920 and 1986: the Ansan Study.
There is strong evidence of a downward secular trend in age at menarche in Europe and the USA during the last century and in Japan and China during the past few decades. However, no study on this trend in age at menarche has been reported in South Korea. To measure the trend in age at menarche in South Korea during the past few decades and the association of height with this trend. A total of 1061 South Korean women born between 1920 and 1986 were randomly recruited from Ansan Cohort Study samples and separate school girl samples, and subjected to this analysis. The data on age at menarche were collected by the retrospective method. Height was measured at time studied and assumed to be relatively constant since age at menarche. Women were grouped with respect to decade of birth and mean age at menarche was determined. The secular trends in annual age at menarche and in height were analysed by the 3-year moving average. Mean menarcheal age decreased from 16.8 to 12.7 years during the past 67 years, corresponding to -0.64 years per decade. Height increased from 149.23 to 161.75 cm during the same period, showing an inverse relationship in the change of trend between height and mean age at menarche. Our data suggest that the downward secular trend in age at menarche may reflect the secular change in physical growth in South Korean women during the past 67 years.


Role for GABA and Glu plasma membrane transporters in the interplay of inhibitory and excitatory neurotransmission.
Neurotransmitter plasma membrane transporters do have much more to perform than simply terminating synaptic transmission and replenishing neurotransmitter pools. Findings in the past decade have evidenced their function in maintaining physiological synaptic excitability, and their actions in critical or pathological conditions, also. Conclusively these findings indicated a previously unrecognized role for neurotransmitter plasma membrane transporters in both, synaptic and nonsynaptic signaling. Major inhibitory and excitatory neurotransmitters within the brain, GABA and Glu, have long been considered to operate through independent systems (GABAergic or Gluergic), each of them characterized by its own localization, function and dedicated GABAergic or Gluergic cell phenotypes. Recent advances, however, have challenged this long-standing paradigm. Localization of GABA in Gluergic terminals and Glu in GABAergic cells were reported. Specific plasma membrane transporters for GABA and Glu are also co-localized in different brain areas. Although, their role in regulating each other's signal is still far from being understood, emerging lines of evidence on interplaying GABAergic and Gluergic processes through plasma membrane transporters opens up a new avenue in the field of more specific therapeutic intervention.


Darkfield-confocal microscopy detection of nanoscale particle internalization by human lung cells.
Concerns over the health effects of nanomaterials in the environment have created a need for microscopy methods capable of examining the biological interactions of nanoparticles (NP). Unfortunately, NP are beyond the diffraction limit of resolution for conventional light microscopy (~200 nm). Fluorescence and electron microscopy techniques commonly used to examine NP interactions with biological substrates have drawbacks that limit their usefulness in toxicological investigation of NP. EM is labor intensive and slow, while fluorescence carries the risk of photobleaching the sample and has size resolution limits. In addition, many relevant particles lack intrinsic fluorescence and therefore can not be detected in this manner. To surmount these limitations, we evaluated the potential of a novel combination of darkfield and confocal laser scanning microscopy (DF-CLSM) for the efficient 3D detection of NP in human lung cells. The DF-CLSM approach utilizes the contrast enhancements of darkfield microscopy to detect objects below the diffraction limit of 200 nm based on their light scattering properties and interfaces it with the power of confocal microscopy to resolve objects in the z-plane. Validation of the DF-CLSM method using fluorescent polystyrene beads demonstrated spatial colocalization of particle fluorescence (Confocal) and scattered transmitted light (Darkfield) along the X, Y, and Z axes. DF-CLSM imaging was able to detect and provide reasonable spatial locations of 27 nm TiO2 particles in relation to the stained nuclei of exposed BEAS 2B cells. Statistical analysis of particle proximity to cellular nuclei determined a significant difference between 5 min and 2 hr particle exposures suggesting a time-dependent internalization process. DF-CLSM microscopy is an alternative to current conventional light and electron microscopy methods that does not rely on particle fluorescence or contrast in electron density. DF-CLSM is especially well suited to the task of establishing the spatial localization of nanoparticles within cells, a critical topic in nanotoxicology. This technique has advantages to 2D darkfield microscopy as it visualizes nanoparticles in 3D using confocal microscopy. Use of this technique should aid toxicological studies related to observation of NP interactions with biological endpoints at cellular and subcellular levels.


[Lifestyle intervention in the treatment of severe obesity].
The prevalence of overweight and obesity is escalating globally, and in Denmark more than 10% of the population are now severely overweight. The aim of this study was to estimate the short-term health effects of 15 weeks of intensive lifestyle intervention composed of physical activity, dietary changes and personal development in severely obese subjects of both sexes. The 27 subjects were weighed weekly. Fat percentage, abdominal circumference, maximum oxygen uptake, heart rate, oral glucose tolerance and blood lipids were measured at baseline and in week 15. The intensive lifestyle intervention was completed under supervision at a stay at Ebeltoft Kurcenter, and the goal was a weight loss of approximately 1% per week. At baseline, the participants' average body mass index (BMI: kg/m2) was 44 +- 1; the BMI was reduced by 11% after the stay (p<0.001). Body weight was reduced by 14 +- 4 kg (p<0.001), corresponding to 76% of the desired weight loss. The subjects' fat mass was reduced by 4% (p<0.001), and their maximal oxygen uptake was enhanced by 25% (p<0.001). Concerning the blood lipids, total cholesterol was reduced by 8% (p = 0.03); there was no significant change in LDL level and a reduction of 15% (p<0.05) in HDL level. After the intervention, oral glucose tolerance was significantly improved (p<0.001). After 15 weeks of intensive lifestyle intervention, there were significant improvements in aerobic fitness and metabolic risk parameters, and the observed weight loss was equivalent to that obtained by surgical treatment. Decisive in the choice of obesity treatment will continue to be the extent of success in permanent weight loss.


Exploring oxidative modifications of tyrosine: an update on mechanisms of formation, advances in analysis and biological consequences.
Protein oxidation is increasingly recognised as an important modulator of biochemical pathways controlling both physiological and pathological processes. While much attention has focused on cysteine modifications in reversible redox signalling, there is increasing evidence that other protein residues are oxidised in vivo with impact on cellular homeostasis and redox signalling pathways. A notable example is tyrosine, which can undergo a number of oxidative post-translational modifications to form 3-hydroxy-tyrosine, tyrosine crosslinks, 3-nitrotyrosine and halogenated tyrosine, with different effects on cellular functions. Tyrosine oxidation has been studied extensively in vitro, and this has generated detailed information about the molecular mechanisms that may occur in vivo. An important aspect of studying tyrosine oxidation both in vitro and in biological systems is the ability to monitor the formation of oxidised derivatives, which depends on a variety of analytical techniques. While antibody-dependent techniques such as ELISAs are commonly used, these have limitations, and more specific assays based on spectroscopic or spectrometric techniques are required to provide information on the exact residues modified and the nature of the modification. These approaches have helped understanding of the consequences of tyrosine oxidation in biological systems, especially its effects on cell signalling and cell dysfunction, linking to roles in disease. There is mounting evidence that tyrosine oxidation processes are important in vivo and can contribute to cellular pathology.


Perspectives about adult sibling relationships: A dyadic analysis of siblings with and without intellectual and developmental disabilities.
Most siblings of individuals with intellectual and developmental disabilities (IDD) report positive sibling relationships. However, extant research often only examines the perspective of the nondisabled sibling; it is unclear whether siblings with IDD report close sibling relationships. Thus, the aim of this study was to understand adult sibling relationships from the perspectives of both siblings with and without IDD. Using dyadic interviews, we examined the perspectives of eight adult sibling dyads. The study was conducted in the United States. Data were analyzed using constant comparative analysis and cross-case analysis to identify themes within and across dyads. Overall, siblings with and without IDD reported enjoying spending time with one another. However, siblings with and without Down syndrome (versus autism spectrum disorder) reported more reciprocal sibling relationships, more frequent contact, and a greater range of shared activities. Implications for future research and practice are discussed.


Management of central venous catheters at the intensive care units in Yemen. Survey of practices.
To describe the intensive care units (ICU) current infection control practices regarding the management of central venous catheters (CVCs) in Yemeni hospitals and compare the current practices with the evidence-based guidelines. This study was carried out in ICUs of Sana'a hospitals, Republic of Yemen, in July 2010. We gathered the data regarding the infection control practices associated with CVC management in 25 ICUs of 14 hospitals. A self-administered questionnaire was distributed to ICUs' nurse managers in Sana'a city. The results were analyzed and tabulated using the Statistical Package for Social Sciences software version 11, and compared with the evidence-based guidelines. Only 44% of units had written policies for CVC management. The 2 most commonly used practices that comply with the guidelines were: wearing of gloves and dressing material. None of the units used 2% chlorhexidine solutions. More than half of the units were adherent to the recommended practice for hand hygiene (before and after insertion, accessing, dressing or replacing/repairing of CVC), preferred insertion site, antimicrobial-coated catheters, aseptic technique during catheter insertion and site care, disinfection of intravenous access ports, capping stopcocks and infusion set tips while they are not in use, and CVC replacement/removal. In all other sections, only the minority were adherent to the recommended practices. There is a diversity of current practices and lack of consistent adherence to the evidence-based guidelines for the prevention of intravascular catheter-related infections.


Quantum-to-continuum prediction of ductility loss in aluminium-magnesium alloys due to dynamic strain aging.
Negative strain-rate sensitivity due to dynamic strain aging in Aluminium-5XXX alloys leads to reduced ductility and plastic instabilities at room temperature, inhibiting application of these alloys in many forming processes. Here a hierarchical multiscale model is presented that uses (i) quantum and atomic information on solute energies and motion around a dislocation core, (ii) dislocation models to predict the effects of solutes on dislocation motion through a dislocation forest, (iii) a thermo-kinetic constitutive model that faithfully includes the atomistic and dislocation scale mechanisms and (iv) a finite-element implementation, to predict the ductility as a function of temperature and strain rate in AA5182. The model, which contains no significant adjustable parameters, predicts the observed steep drop in ductility at room temperature, which can be directly attributed to the atomistic aging mechanism. On the basis of quantum inputs, this multiscale theory can be used in the future to design new alloys with higher ductility.


Age-related differences in signaling efficiency of human lens cells underpin differential wound healing response rates following cataract surgery.
Cataract surgery is blighted by posterior capsule opacification (PCO), which is more severe and frequent in the young than the elderly (>60 years). Our aim was to understand the biological basis for these age-related differences in PCO/wound healing rates. Human capsular bags were prepared by cataract surgery on donor lenses (young [<40 years] and elderly [>60 years] groups) and maintained in serum-free Eagle's minimum essential medium. Cell growth was determined using the MTS assay. Fibroblast growth factor (FGF) and hepatocyte growth factor (HGF) levels were determined using ELISA. Protein synthesis rates were elucidated by 35S-methionine incorporation. U0126, SB203580, and SP600125 were used to disrupt ERK-, p38-, and JNK-mediated signaling, respectively. Level of total and phospho-ERK, -c-jun, -P38, and -JNK plus cytokines were detected using a BIOPLEX array system. Following a 2-day culture period, significant decreases in IL-1β and IL-6, and increases in IL-10, IL-12, IL-13, and VEGF in the >60 years group were observed compared with their younger counterparts. Capsular bags (cells and capsule) from aged donors contained greater than or equal levels of HGF and FGF than younger counterparts and had greater rates of protein synthesis. Inhibition of ERK, p38, and JNK signaling significantly suppressed cell coverage on the posterior capsule. pERK, p-c-jun, p-p38, and pJNK were consistently lower in aged cell populations; total signaling protein expression was unaffected by age. Serum stimulation increased pERK, p-c-jun, and pJNK levels in cells of all ages; p-p38 was significantly increased in the >60 years group only. Ligand availability to cells is not a limiting factor as we age, but the ability to convert this resource into signaling activity is. We therefore propose that overall signaling efficiency is reduced as a function of age, which consequently limits wound-healing response rates after injury.


Injury biomechanics of the human cervical column.
In this study, the authors have developed a technique to replicate clinically relevant traumatic cervical spine injuries and determined the injury biomechanics. Because of the importance of compressive forces in neck injuries, this research was conducted using compression as the primary load vector. Six fresh human cadaveric head-neck complexes were prepared by fixing the distal end in methylmethacrylate. Tests were done with varying loading rates to include quasistatic and dynamic conditions. For quasistatic experiments, the proximal end was fixed to the piston of the testing device. In dynamic tests, the cranium was unconstrained, and to maintain stability, the effects of the spinal musculature were simulated by means of pulleys, deadweights, and springs in the anterior and posterior parts of the head-neck complex. Quasistatic tests conducted at a rate of 2.0 mm/sec produced cervical spine trauma at forces ranging from 1.7 to 2.3 kN, with deformations ranging from 2.2 to 3.7 cm. The specimens were deep-frozen at the level of injury, preserving the local deformation of the tissues to enable a detailed evaluation immediately after the injury. Dynamic tests conducted at velocities of 3.2 to 5.7 m/sec resulted in impact injuries at one level of the head-neck complex. The applied forces at the vertex were considerably higher than those recorded at the distal end. The failure deformations for both the quasistatic (2.2-3.7 cm) and dynamic (1.7-3.2 cm) tests, however, were found to be similar, suggesting that the human head-neck complex is a deformation-sensitive structure.


Obesity and type-2 diabetes as inducers of premature cellular senescence and ageing.
Cellular senescence is now considered as a major mechanism in the development and progression of various diseases and this may include metabolic diseases such as obesity and type-2 diabetes. The presence of obesity and diabetes is a major risk factor in the development of additional health conditions, such as cardiovascular disease, kidney disease and cancer. Since senescent cells can drive disease development, obesity and diabetes can potentially create an environment that accelerates cell senescence within other tissues of the body. This can consequently manifest as age-related biological impairments and secondary diseases. Cell senescence in cell types linked with obesity and diabetes, namely adipocytes and pancreatic beta cells will be explored, followed by a discussion on the role of obesity and diabetes in accelerating ageing through induction of premature cell senescence mediated by high glucose levels and oxidised low-density lipoproteins. Particular emphasis will be placed on accelerated cell senescence in endothelial progenitor cells, endothelial cells and vascular smooth muscle cells with relation to cardiovascular disease and proximal tubular cells with relation to kidney disease. A summary of the potential strategies for therapeutically targeting senescent cells for improving health is also presented.


Supplemental Perioperative Oxygen to Reduce Surgical Site Infection After High-Energy Fracture Surgery (OXYGEN Study).
Supplemental perioperative oxygen (SPO) therapy has been proposed as one approach for reducing the risk of surgical site infection (SSI). Current data are mixed regarding efficacy in decreasing SSI rates and hospital inpatient stays in general and few data exist for orthopaedic trauma patients. This study is a phase III, double-blind, prospective randomized clinical trial with a primary goal of assessing the efficacy of 2 different concentrations of perioperative oxygen in the prevention of SSIs in adults with tibial plateau, pilon (tibial plafond), or calcaneus fractures at higher risk of infection and definitively treated with plate and screw fixation. Patients are block randomized (within center) in a 1:1 ratio to either treatment group (FiO2 80%) or control group (FiO2 30%) and stratified by each study injury location. Secondary objectives of the study are to compare species and antibacterial sensitivities of the bacteria in patients who develop SSIs, to validate a previously developed risk prediction model for the development of SSI after fracture surgery, and to measure and compare resource utilization and cost associated with SSI in the 2 study groups. SPO is a low cost and readily available resource that could be easily disseminated to trauma centers across the country and the world if proved to be effective.


Physical activity and sedentary behaviour typologies of 10-11 year olds.
Targeted interventions may be more effective at increasing children's physical activity. The aim of this study was to identify clusters of children based on physical activity and sedentary patterns across the week. Participants were 761, 10-11 year old children. Participant's self-reported time spent in eight physical activity and sedentary contexts and wore an accelerometer. Cluster analysis was conducted on the time spent in the self-reported physical activity and sedentary contexts. Mean minutes of accelerometer derived of moderate to vigorous physical activity (MVPA) and sedentary time were derived for the entire week, weekdays only, weekend days and four different time periods across each type (weekend or weekday) of days. Differences in the physical activity patterns of the groups derived from the cluster analysis were assessed for overall physical activity as well as for the four time periods on weekdays and weekend days. Three clusters emerged: 1) High active/Low sedentary; 2) Low active/Moderate sedentary; and 3) High Active/High sedentary. Patterns of activity differed across the week for each group and the High Active/High sedentary obtained the most minutes of MVPA. Patterns of physical activity and sedentary time differed across the week for each cluster. Interventions could be targeted to the key periods when each group is inactive.


Global gene expression profiles during acute pathogen-induced pulmonary inflammation reveal divergent roles for Th1 and Th2 responses in tissue repair.
T helper 1 responses are typically proinflammatory, while Th2 responses have been considered regulatory. Interestingly, Th2 responses characterize a number of pulmonary diseases, many of which terminate in tissue remodeling and fibrosis. We developed a mouse model using Schistosoma mansoni eggs and cytokine-deficient mice to induce highly polarized Th1- or Th2-type inflammation in the lung. In this study, we examined the pathology and cytokine profiles in Th1- and Th2-polarized environments and used oligonucleotide microarray analysis to decipher the genes responsible for these effects. We further elaborated on the results using IL-10- and IL-13-deficient mice because these cytokines are believed to be the central regulators of Th2-associated pathology. We found that the Th1-polarized mice developed small granulomas with less fibrosis while expressing genes characteristic of tissue damage. Th2-polarized mice, in contrast, formed large granulomas with massive collagen deposition and up-regulated genes associated with wound healing, specifically, arginase, collagens, matrix metalloproteinases (MMPs), and tissue inhibitors of MMP. In addition, several members of the chitinase-like family were up-regulated in the lung following egg challenge. We also developed a method of defining the net collagen deposition using the expression profiles of several collagen, MMP, and tissue inhibitors of MMP genes. We found that Th1-polarized mice did not elaborate collagens or MMPs and therefore did not have a significant capacity for repair in this model. Thus, Th1-mediated inflammation is characterized by tissue damage, while Th2 directs wound healing and fibrosis.


Implantation of a novel telemetric intraocular pressure sensor in patients with glaucoma (ARGOS study): 1-year results.
We investigated the safety of a telemetric IOP sensor and the accuracy of its IOP measurements in six patients with open-angle glaucoma and cataract. The study design was a prospective, single-center clinical trial. Here we present 1-year follow-up data. A ring-shaped telemetric IOP sensor was implanted in the ciliary sulcus after implantation of the intracapsular lens, during planned cataract surgery. The sensor is encapsulated in silicone rubber and consists of a miniature device with eight pressure-sensitive capacitors and a circular microcoil antenna. IOP measurements are performed with a reader unit held in front of the eye. IOP is calculated as the differences between the absolute pressure inside the eye (pressure sensor) and that outside the eye (reader unit). The sensor was successfully implanted in all patients. Four patients developed sterile anterior chamber inflammation that resolved completely within 9 days after surgery with anti-inflammatory treatment. All patients showed mild to moderate pupillary distortion and pigment dispersion after surgery. Telemetric IOP measurement was performed in all patients at all visits, and the patients successfully performed self-tonometry at home after receiving instructions. Telemetric IOP values showed similar profiles compared to those of Goldmann applanation tonometry (GAT). Three patients showed a relevant IOP step during follow-up, and in one patient, negative values were obtained throughout the study. Despite early postoperative anterior chamber inflammation, the IOP sensor was well tolerated by all patients. We describe the first prospective clinical study of a noncontact IOP sensor that potentially enables continuous IOP monitoring in patients with glaucoma. The sensor shape and size needs to be adapted to avoid pupillary distortion and to confirm that IOP measurements are accurately recorded in comparison to those of GAT. ( www.germanctr.de; number DRKS00003335.).