Improvement of biomaterials used in tissue engineering by an ageing treatment.
Biomaterials based on crosslinked sponges of biopolymers have been extensively used as scaffolds to culture mammal cells. It is well known that single biopolymers show significant change over time due to a phenomenon called physical ageing. In this research, it was verified that scaffolds used for skin tissue engineering (based on gelatin, chitosan and hyaluronic acid) express an ageing-like phenomenon. Treatments based on ageing of scaffolds improve the behavior of skin-cells for tissue engineering purposes. Physical ageing of dry scaffolds was studied by differential scanning calorimetry and was modeled with ageing kinetic equations. In addition, the physical properties of wet scaffolds also changed with the ageing treatments. Scaffolds were aged up to 3 weeks, and then skin-cells (fibroblasts) were seeded on them. Results indicated that adhesion, migration, viability, proliferation and spreading of the skin-cells were affected by the scaffold ageing. The best performance was obtained with a 2-week aged scaffold (under cell culture conditions). The cell viability inside the scaffold was increased from 60% (scaffold without ageing treatment) to 80%. It is concluded that biopolymeric scaffolds can be modified by means of an ageing treatment, which changes the behavior of the cells seeded on them. The ageing treatment under cell culture conditions might become a bioprocess to improve the scaffolds used for tissue engineering and regenerative medicine.


Epitopes of streptococcal M proteins that evoke antibodies that cross-react with human brain.
There is evidence suggesting that Sydenham's chorea, which is a major manifestation of acute rheumatic fever, may be mediated by streptococcal antibodies that cross-react with the brain. Our studies were undertaken to determine whether streptococcal M protein, the major virulence factor of group A streptococci, evoked antibodies that cross-react with human brain. Rabbits were immunized with pepsin-extracted M protein from rheumatogenic type 6 streptococci. Immune sera were screened for the presence of antibodies that cross-reacted with human brain by indirect immunofluorescence tests and immunoblot analyses. Type 6 M protein evoked antibodies that cross-reacted with several brain proteins and antibody binding to these proteins was completely inhibited by type 6 M protein and partially inhibited by types 5 and 19 M proteins, suggesting that these heterologous M proteins contain conserved brain-cross-reactive epitopes. Using synthetic peptides from several serotypes of M proteins, the conserved brain-cross-reactive epitopes were localized to a decapeptide contained within the covalent structure of the B repeat region of type 6 M protein. These peptides also inhibited brain-cross-reactive antibodies in the serum of a patient with active Sydenham's chorea. Our data indicate that streptococcal M proteins contain brain-cross-reactive epitopes that could potentially be involved in the pathogenesis of Sydenham's chorea.


Complex archaea that bridge the gap between prokaryotes and eukaryotes.
The origin of the eukaryotic cell remains one of the most contentious puzzles in modern biology. Recent studies have provided support for the emergence of the eukaryotic host cell from within the archaeal domain of life, but the identity and nature of the putative archaeal ancestor remain a subject of debate. Here we describe the discovery of 'Lokiarchaeota', a novel candidate archaeal phylum, which forms a monophyletic group with eukaryotes in phylogenomic analyses, and whose genomes encode an expanded repertoire of eukaryotic signature proteins that are suggestive of sophisticated membrane remodelling capabilities. Our results provide strong support for hypotheses in which the eukaryotic host evolved from a bona fide archaeon, and demonstrate that many components that underpin eukaryote-specific features were already present in that ancestor. This provided the host with a rich genomic 'starter-kit' to support the increase in the cellular and genomic complexity that is characteristic of eukaryotes.


Comparison of statistical approaches for analyzing incomplete longitudinal patient-reported outcome data in randomized controlled trials.
Missing data are a potential source of bias in the results of RCTs, but are often unavoidable in clinical research, particularly in patient-reported outcome measures (PROMs). Maximum likelihood (ML), multiple imputation (MI), and inverse probability weighting (IPW) can be used to handle incomplete longitudinal data. This paper compares their performance when analyzing PROMs, using a simulation study based on an RCT data set. Realistic missing-at-random data were simulated based on patterns observed during the follow-up of the knee arthroscopy trial (ISRCTN45837371). Simulation scenarios covered different sample sizes, with missing PROM data in 10%-60% of participants. Monotone and nonmonotone missing data patterns were considered. Missing data were addressed by using ML, MI, and IPW and analyzed via multilevel mixed-effects linear regression models. Root mean square errors in the treatment effects were used as performance parameters across 1,000 simulations. Nonconvergence issues were observed for IPW at small sample sizes. The performance of all three approaches worsened with decreasing sample size and increasing proportions of missing data. MI and ML performed similarly when the MI model was restricted to baseline variables, but MI performed better when using postrandomization data in the imputation model and also in nonmonotone versus monotone missing data scenarios. IPW performed worse than ML and MI in all simulation scenarios. When additional postrandomization information is available, MI can be beneficial over ML for handling incomplete longitudinal PROM data. IPW is not recommended for handling missing PROM data in the simulated scenarios.


Risk and predictors of dementia and parkinsonism in idiopathic REM sleep behaviour disorder: a multicentre study.
Idiopathic REM sleep behaviour disorder (iRBD) is a powerful early sign of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. This provides an unprecedented opportunity to directly observe prodromal neurodegenerative states, and potentially intervene with neuroprotective therapy. For future neuroprotective trials, it is essential to accurately estimate phenoconversion rate and identify potential predictors of phenoconversion. This study assessed the neurodegenerative disease risk and predictors of neurodegeneration in a large multicentre cohort of iRBD. We combined prospective follow-up data from 24 centres of the International RBD Study Group. At baseline, patients with polysomnographically-confirmed iRBD without parkinsonism or dementia underwent sleep, motor, cognitive, autonomic and special sensory testing. Patients were then prospectively followed, during which risk of dementia and parkinsonsim were assessed. The risk of dementia and parkinsonism was estimated with Kaplan-Meier analysis. Predictors of phenoconversion were assessed with Cox proportional hazards analysis, adjusting for age, sex, and centre. Sample size estimates for disease-modifying trials were calculated using a time-to-event analysis. Overall, 1280 patients were recruited. The average age was 66.3 ± 8.4 and 82.5% were male. Average follow-up was 4.6 years (range = 1-19 years). The overall conversion rate from iRBD to an overt neurodegenerative syndrome was 6.3% per year, with 73.5% converting after 12-year follow-up. The rate of phenoconversion was significantly increased with abnormal quantitative motor testing [hazard ratio (HR) = 3.16], objective motor examination (HR = 3.03), olfactory deficit (HR = 2.62), mild cognitive impairment (HR = 1.91-2.37), erectile dysfunction (HR = 2.13), motor symptoms (HR = 2.11), an abnormal DAT scan (HR = 1.98), colour vision abnormalities (HR = 1.69), constipation (HR = 1.67), REM atonia loss (HR = 1.54), and age (HR = 1.54). There was no significant predictive value of sex, daytime somnolence, insomnia, restless legs syndrome, sleep apnoea, urinary dysfunction, orthostatic symptoms, depression, anxiety, or hyperechogenicity on substantia nigra ultrasound. Among predictive markers, only cognitive variables were different at baseline between those converting to primary dementia versus parkinsonism. Sample size estimates for definitive neuroprotective trials ranged from 142 to 366 patients per arm. This large multicentre study documents the high phenoconversion rate from iRBD to an overt neurodegenerative syndrome. Our findings provide estimates of the relative predictive value of prodromal markers, which can be used to stratify patients for neuroprotective trials.


X-linked H3K27me3 demethylase Utx is required for embryonic development in a sex-specific manner.
Embryogenesis requires the timely and coordinated activation of developmental regulators. It has been suggested that the recently discovered class of histone demethylases (UTX and JMJD3) that specifically target the repressive H3K27me3 modification play an important role in the activation of "bivalent" genes in response to specific developmental cues. To determine the requirements for UTX in pluripotency and development, we have generated Utx-null ES cells and mutant mice. The loss of UTX had a profound effect during embryogenesis. Utx-null embryos had reduced somite counts, neural tube closure defects and heart malformation that presented between E9.5 and E13.5. Unexpectedly, homozygous mutant female embryos were more severely affected than hemizygous mutant male embryos. In fact, we observed the survival of a subset of UTX-deficient males that were smaller in size and had reduced lifespan. Interestingly, these animals were fertile with normal spermatogenesis. Consistent with a midgestation lethality, UTX-null male and female ES cells gave rise to all three germ layers in teratoma assays, though sex-specific differences could be observed in the activation of developmental regulators in embryoid body assays. Lastly, ChIP-seq analysis revealed an increase in H3K27me3 in Utx-null male ES cells. In summary, our data demonstrate sex-specific requirements for this X-linked gene while suggesting a role for UTY during development.


Identification of endothelial cells and progenitor cell subsets in human peripheral blood.
An assay for circulating cell subsets in human peripheral blood by flow cytometry is used as a biomarker to determine cardiovascular disease risk and tumor responsiveness to chemotherapy since endothelial progenitor cells (EPCs) function in vasculogenesis and angiogenesis. Despite analytical advances in polychromatic flow cytometry (PFC), conventional approaches are routinely utilized to enumerate and isolate EPCs, which has led to varied results in clinical studies, potential cellular misidentification, and thus a lack of a plausible biological explanation for how purported EPCs function. Herein, a reproducible PFC protocol is provided to identify a rare circulating endothelial colony-forming cell (ECFC) with proliferative potential, along with a population of circulating progenitor cells (CPCs) in which the ratio analysis distinguishes between healthy and disease populations. In sum, a reliable PFC protocol, which can be used to investigate the roles of human hematopoietic and endothelial elements in the growth and maintenance of the vasculature, is described.


The Case of the Cognitive (Opti)miser: Electrophysiological Correlates of Working Memory Maintenance Predict Demand Avoidance.
People are often considered cognitive misers. When given a free choice between two tasks, people tend to choose tasks requiring less cognitive effort. Such demand avoidance (DA) is associated with cognitive control, but it is still not clear to what extent individual differences in cognitive control can account for variations in DA. We sought to elucidate the relation between cognitive control and cognitive effort preferences by investigating the extent to which sustained neural activity in a task requiring cognitive control is correlated with DA. We hypothesized that neural measures of efficient filtering will predict individual variations in demand preferences. To test this hypothesis, we had participants perform a delayed-match-to-sample paradigm with their ERPs recorded, as well as a separate behavioral demand-selection task. We focused on the ERP correlates of cognitive filtering efficiency (CFE)-the ability to ignore task-irrelevant distractors during working memory maintenance-as it manifests in a modulation of the contralateral delay activity, an ERP correlate of cognitive control. As predicted, we found a significant positive correlation between CFE and DA. Individuals with high CFE tended to be significantly more demand avoidant than their low-CFE counterparts. Low-CFE individuals, in comparison, did not form distinct cognitive effort preferences. Overall, our results suggest that cognitive control over the contents of visual working memory contribute to individual differences in the expression of cognitive effort preferences. This further implies that these observed preferences are the product of sensitivity to cognitive task demands.


The role of lifestyle risk factors on mortality from colorectal cancer in populations of the Asia-Pacific region.
Although colorectal cancer is one of the leading malignancies worldwide, there are few data on aetiological relationships from the Asia-Pacific region. Therefore, a collaborative study was conducted involving over half a million subjects from 33 cohort studies in the region. Age-adjusted death rates from colorectal cancer, over an average of 6.8 years follow-up, were 12 and 14 per 100,000 person-years among Asian women and men, respectively; corresponding values in Australasia were 31 and 41. Height was strongly associated with death from colorectal cancer: an extra 5 cm of height was associated with 10% (95%confidence interval, 3% - 18% additional risk, after adjustment for other factors. Smoking increased risk by 43% (9% - 88%), although no significant dose-response relationship was discerned (p>0.05). Other significant (p <0.05) risk factors were body mass index and lack of physical activity. There was no significant effect on colorectal cancer mortality for alcohol consumption, waist circumference, fasting blood glucose or diabetes, although the latter conferred a notable 26% additional risk. Height may be a biomarker for some currently unknown genetic, or environmental, risk factors that are related both to skeletal growth and mutanogenesis. Understanding such mechanisms could provide opportunities for novel preventive and therapeutic intervention.


Impaired lymphatic function accelerates cancer growth.
Increased lymphangiogenesis is a common feature of cancer development and progression, yet the influence of impaired lymphangiogenesis on tumor growth is elusive. C3HBA breast cancer and KHT-1 sarcoma cell lines were implanted orthotopically in Chy mice, harboring a heterozygous inactivating mutation of vascular endothelial growth factor receptor-3, resulting in impaired dermal lymphangiogenesis. Accelerated tumor growth was observed in both cancer models in Chy mice, coinciding with reduced peritumoral lymphangiogenesis. An impaired lymphatic washout was observed from the peritumoral area in Chy mice with C3HBA tumors, and the number of macrophages was significantly reduced. While fewer macrophages were detected, the fraction of CD163+ M2 macrophages remained constant, causing a shift towards a higher M2/M1 ratio in Chy mice. No difference in adaptive immune cells was observed between wt and Chy mice. Interestingly, levels of pro- and anti-inflammatory macrophage-associated cytokines were reduced in C3HBA tumors, pointing to an impaired innate immune response. However, IL-6 was profoundly elevated in the C3HBA tumor interstitial fluid, and treatment with the anti-IL-6 receptor antibody tocilizumab inhibited breast cancer growth. Collectively, our data indicate that impaired lymphangiogenesis weakens anti-tumor immunity and favors tumor growth at an early stage of cancer development.


Inhibition of peptidoglycan biosynthesis in vancomycin-susceptible and -resistant bacteria by a semisynthetic glycopeptide antibiotic.
LY191145 is a p-chlorobenzyl derivative of LY264826 (A82846B) with activity against both vancomycin-susceptible and -resistant enterococci. Incorporation of L-[14C]lysine into peptidoglycan of intact vancomycin-susceptible and -resistant Enterococcus faecium was inhibited by LY191145 (50% inhibitory concentrations of 1 and 5 microgram/ml, respectively). Inhibition was accompanied by accumulation of UDP-muramyl-peptide precursors in the cytoplasm. This agent inhibited late-stage steps in peptidoglycan biosynthesis in permeabilized E. faecium when either the UDP-muramyl-pentapeptide precursor from vancomycin-susceptible E. faecium or the UDP-muramyl-pentadepsipeptide precursor from vancomycin-resistant E. faecium was used as a substrate. Inhibition of late-stage steps led to accumulation of an N-acetyl-[14C]glucosamine-labeled lipid intermediate indicative of inhibition of the transglycosylation step. Inhibition of peptidoglycan polymerization without affecting cross-linking in a particulate membrane-plus-wall-fragment assay from Aerococcus viridans was consistent with this explanation. The fact that inhibition of peptidoglycan biosynthesis by LY191145 was not readily antagonized by an excess of free acyl-D-alanyl-D-alanine or acyl-D-alanyl-D-lactate ligands indicates that the manner in which this compound inhibits transglycosylation may not be identical to that of vancomycin.


A discriminative learning framework with pairwise constraints for video object classification.
To deal with the problem of insufficient labeled data in video object classification, one solution is to utilize additional pairwise constraints that indicate the relationship between two examples, i.e., whether these examples belong to the same class or not. In this paper, we propose a discriminative learning approach which can incorporate pairwise constraints into a conventional margin-based learning framework. Different from previous work that usually attempts to learn better distance metrics or estimate the underlying data distribution, the proposed approach can directly model the decision boundary and, thus, require fewer model assumptions. Moreover, the proposed approach can handle both labeled data and pairwise constraints in a unified framework. In this work, we investigate two families of pairwise loss functions, namely, convex and nonconvex pairwise loss functions, and then derive three pairwise learning algorithms by plugging in the hinge loss and the logistic loss functions. The proposed learning algorithms were evaluated using a people identification task on two surveillance video data sets. The experiments demonstrated that the proposed pairwise learning algorithms considerably outperform the baseline classifiers using only labeled data and two other pairwise learning algorithms with the same amount of pairwise constraints.


Prospective study of catheter-related central vein thrombosis in home parenteral nutrition patients with benign disease using serial venous Doppler ultrasound.
Catheter-related central vein thrombosis (CRVT) is a severe complication of home parenteral nutrition (HPN) that may be clinically manifest or subclinical. The aims of the study were to prospectively investigate the incidence of CRVT in patients on HPN with benign disease and determine the influence of different variables on this complication. A prospective, multicentre, observational study in the Home Artificial Nutrition-Chronic Intestinal Failure ESPEN group was performed. Patients with benign disease starting HPN or already on HPN after the insertion of a new catheter, were recruited and followed up with Color Doppler Duplex Sonography (CDDS) evaluations at baseline, 1 week, 3, 6 and 12 months after catheter insertion. Fisher's exact test was used to calculate the association of different variables (related to the patient, type of catheter, vascular access, insertion method, catheter care and anticoagulant treatment) with CRVT events. Sixty-two patients (31 males, 31 females) aged 50 ± 19 (19-83) years were included and followed for a median 363 days, with an Inter Quartile Range of 180-365 days, and a total of 16,186 catheter-days. Six patients had previous CRVT and 16 had history of thromboembolic disease (pulmonary and mesenteric). Forty one patients were receiving anticoagulant treatment. Fifty two patients had tunneled catheters and 10 implanted ports. Two patients had symptomatic thrombosis at 3 and 12 months of follow-up (2 and 3 weeks after normal routine CDDS evaluation). The incidence of CRVT was 0.045/catheter/year. CRVT was not significantly associated with any of the variables analyzed. The incidence of CRVT in patients on HPN for benign disease followed by CDDS is low in the first year of catheterization. We did not observe any case of asymptomatic CRVT. Based on our data, CDDS seems to have low effectiveness as a screening tool for CRVT in asymptomatic patients on HPN with benign disease.


Immunohistochemical localization of mdm-2, p27Kip1 and bcl-2 in Warthin's tumor of the parotid gland.
Warthin's tumor is a benign monomorphic adenoma with unclear origin that almost occur exclusively in the parotid gland. Etiology of Warthin's tumor as well as its malignant potential are still unclear. Therefore immunohistochemical assessment of Warthin's tumor may be useful to detect its origin or its malignant transformation potential. The present study aims to investigate the immunohistochemical expression of murine double minute-2 (mdm-2), p27Kip1 and B cell lymphoma-2 (bcl-2) in Warthin's tumor of parotid gland and also to clarify the role of these proteins in the behavior of that tumor. Twenty paraffin blocks of cases previously diagnosed as Warthin's tumor were collected for immunohistochemical staining with primary antibodies against mdm-2, p27Kip1 and bcl-2 using streptavidin-biotin immunoperoxidase staining system. All cases showed immunopositivity for mdm-2 and p27Kip1 while 18/20 showed bcl-2 immunopositivity. Both layers of the neoplastic epithelial cells that line the cystic spaces showed immunopositivity with all antibodies used. Goblet cells were mdm-2 immunonegative while myoepithelial cells were p27Kip-1 immunonegative. Areas of epithelial proliferation that formed buds were p27Kip-1 and bcl-2 immunopositive. Mdm-2 played a tumor-suppressor role that might be implicated with the benign behavior of Warthin's tumor. The mutual expression of both p27Kip1 and bcl-2 suggested a protective role of these slowly proliferating cells from apoptosis to maintain their survival and elevated bcl-2 expression offers a significant protection against p27Kip1-mediated apoptosis.


The 2000 Canadian recommendations for the management of hypertension: Part one--therapy.
To provide updated, evidence-based recommendations for the therapy of hypertension in adults. For patients with hypertension, there are a number of lifestyle manoeuvres and antihypertensive agents that may control blood pressure. Randomized trials evaluating first- line therapy with thiazides, beta-adrenergic antagonists, angiotensin-converting enzyme inhibitors, calcium channel blockers, alpha-blockers, centrally acting agents or angiotensin II receptor antagonists were reviewed. The health outcomes considered were changes in blood pressure, cardiovascular morbidity, and cardiovascular and/or all-cause mortality rates. Economic outcomes were not considered due to insufficient evidence. Medline searches were conducted from the period of the last revision of the Canadian Recommendations for the Management of Hypertension (May 1998 to October 2000). Reference lists were scanned, experts were polled, and the personal files of the subgroup members and authors were used to identify other studies. All relevant articles were reviewed and appraised, using prespecified levels of evidence, by content experts and methodological experts. A high value was placed on the avoidance of cardiovascular morbidity and mortality. Various lifestyle manoeuvres and antihypertensive agents reduce the blood pressure of patients with sustained hypertension. In certain settings, and for specific classes of drugs, blood pressure lowering has been associated with reduced cardiovascular morbidity and/or mortality. The present document contains detailed recommendations pertaining to all aspects of the therapy of patients with hypertension, including lifestyle modifications proven to lower blood pressure, treatment thresholds, target blood pressures, choice of agents in various settings and strategies to enhance adherence. Lower thresholds for blood pressure treatment are advocated for people with other cardiovascular risk factors or established hypertensive target organ damage. Implicit in the recommendations for therapy is the principle that treatment should be individualized for each patient and the choice of agent should be dictated by coexistent conditions. For the treatment of uncomplicated essential hypertension, thiazides, beta-adrenergic antagonists, angiotensin-converting enzyme inhibitors or calcium channel blockers may be appropriate, depending on individual circumstances. All recommendations were graded according to strength of the evidence and voted on by the Canadian Hypertension Recommendations Working Group. Only those recommendations achieving high levels of consensus are reported here. These guidelines will be updated annually.


Extra corporeal membrane oxygenation support: ethical dilemmas.
The vast expansion of patients treated with of extra corporeal membrane oxygenation (ECMO) emerge novel ethical questions about the use of this new technology. In regard the indications, duration of support and timing of withdrawal of support, these questions sometimes create disagreement among surrogates, between health care team and surrogates, and even disagreement among health care team, these disagreements occurs because of the extreme emergency of support initiation, the ambiguity of the outcome as well as lack of clarity on the intended treatment direction, whether it is ineffective, bridge to recovery or bridge to lifetime mechanical support or transplant. In this article we discuss these questions through patients' scenarios.


Geochemical behavior of heavy metals in differents environments in Rodrigo de Freitas lagoon - RJ/Brazil.
The accelerated urbanisation without a planning, brought several environmental problems to Rio de Janeiro coastal zone, especially in areas such as Rodrigo de Freitas lagoon, which receives a great amount of untreated sewage every day. To assess the nature, potentially sources and extent of heavy metal pollution in the lagoon, sediments from the surrounding streets, from the entrance of the main canal that drains to the lagoon and from the bottom of the lagoon were collected and analysed by a modified selective extraction procedure in order to study the geochemical partitioning and bioavailability of Zn, Cu, Cr, Ni and Pb in these three compartments. The present study verified an increase in the Cu, Pb and Zn concentrations in the north of the Rodrigo de Freitas lagoon. Despite the different levels of oxidation between the sediments accumulated in the streets and in the bottom of the lagoon, the geochemical partitioning of the heavy metals did not show any pattern of variation for the metals, except for the element Cu. No concentrations were found in the soluble phase of samples collected in the surfacial sediments of the lagoon, suggesting no bioavailability of heavy metals.


Mechanism responsible for the induction of I-J restriction on TS3 suppressor cells.
The mechanisms responsible for the induction of I-J restrictions on third-order suppressor T cells (TS3) were analyzed. The I-J phenotype of the antigen-coupled cells used for priming restricted the specificity of the TS3 population. Thus, TS3 cells were only generated after priming with antigen-coupled I-J homologous cells. Identity at the I-JM (and I-E) subregions was sufficient for TS3 induction. Furthermore, priming of H-2 heterozygous mice with antigen-coupled parental cells generated TS3 that were restricted to the parental haplotype used for priming. The splenic cell population responsible for antigen presentation and induction of TS3 cells was fractionated. The cells involved in antigen presentation were found in the splenic adherent population and were absent in the fraction containing splenic nonadherent T and B cells. The subsequent activation and interaction of TS3 cells is also restricted by genes in the H-2 complex. The results are discussed in terms of a general mechanism responsible for the induction of restrictions in T helper and TS3 cells.


Imaging of meniscal cyst of the knee in three cases.
Three cases of solitary meniscal cyst in the knee have recently been diagnosed in our departments using ultrasound and nuclear magnetic resonance (MRI). Two cysts involved the lateral and one the medial meniscus. The appearance of these lesions on ultrasound and MR images is shown. All three cysts had low intensity on T1-weighted images and high intensity on T2-weighted images and clearly communicated with a large horizontal tear in the meniscus. Ultrasonography showed a relatively hypoechoic lesion and, in the two cases of lateral meniscal cyst, focal areas of relatively increased echoes which are probably due to meniscal debris. Ultrasound showed the abnormal meniscus in both of these cases. MRI is the best way of showing the cyst and an accompanying meniscal tear but, where MRI is not available, ultrasound may be an easy and inexpensive way of making a diagnosis whether or not it is used in conjunction with positive contrast arthrography.


Priority-setting and rationing in German health care.
While the framework for the German statutory health insurance system is determined at the national level and the states have a significant influence on the provision of hospital care and public health services, most decisions on the actual delivery of curative health care services are made through joint negotiations between the associations of the physicians and the sickness funds at various levels. Decisions with effects on priority-setting include the determination of the benefit package and financial regulations. The actual provision of care is further regulated through guidelines, e. g. about the prescription of pharmaceuticals. Financial regulations include both the setting of budget limits and the determination of relative point values which affect remuneration and serve as incentives to provide certain services (or not). A survey of 2000 citizens reveals, however, that the majority of the public favours unlimited funding for health services and that treatment decisions should be made by doctors. Limiting the benefits catalogue to a core of essential services is rejected as well as priorities based on age. Separately interviewed experts expect further limitations of therapeutic freedom, however. Germany seems to be just at the beginning of a public debate on priority-setting and rationing in health care.


Magnesium repletion therapy improved lipid metabolism in hypomagnesemic renal transplant recipients: a pilot study.
Hypomagnesemia has been associated with hypertension, abnormal glucose and lipid metabolism, and accelerated atherosclerosis in nontransplant patients. In this prospective short-term pilot study, 14 hypomagnesemic renal transplant recipients with stable renal function were evaluated monthly over a 6-month interval. The first 3 months was the baseline observation period. During the second 3 months, MgO2 was administered to normalize the serum Mg level. Glucose tolerance, lipid levels, blood pressure, weight, and routine chemistries were assessed before and after Mg replacement. All others medications were held constant during the 6-month study. Serum Mg levels increased to normal range after MgO2 therapy, which was well tolerated. There were significant decreases in total cholesterol, low density lipoprotein, and total cholesterol/high density lipoprotein ratio after 3 months of MgO2 therapy. Only three patients had abnormal baseline glucose tolerance tests. All three patients showed improved glucose tolerance after MgO2, but this was not statistically significant. Mg repletion may be an important ancillary therapy in hypomagnesemic renal transplant patients with hyperlipidemia.


Intravenous hydrochloric acid in patients with metabolic alkalosis and hypercapnia.
During a seven-year period, 15 patients admitted to an intensive care unit with a metabolic alkalosis resistant to saline and potassium chloride infusions and with an arterial pH of greater than 7.44 and arterial carbon dioxide pressure (PaCO2) of greater than 50 mm Hg, while breathing spontaneously, were given hydrochloric acid at 200 mmol/24 h through a central venous line until the arterial pH had decreased to less than 7.36 or PaCO2 had decreased to less than 40 mm Hg. Five patients without respiratory failure had a significant decrease in PaCO2, pH, bicarbonate Ion (HCO3-), and base excess (BE). Four patients with acute respiratory failure had a significant decrease in PaCO2, HCO3-, and BE and an increase in arterial oxygen pressure, indicating that in both of these patient groups alveolar ventilation had improved. Six patients with chronic respiratory failure had a significant decrease in pH, HCO3-, and BE, although there was no significant change in PaCO2, indicating that in this patient group alveolar ventilation could not be consistently increased in response to a reduction in arterial pH.


What iconic gesture fragments reveal about gesture-speech integration: when synchrony is lost, memory can help.
The present series of experiments explores several issues related to gesture-speech integration and synchrony during sentence processing. To be able to more precisely manipulate gesture-speech synchrony, we used gesture fragments instead of complete gestures, thereby avoiding the usual long temporal overlap of gestures with their coexpressive speech. In a pretest, the minimal duration of an iconic gesture fragment needed to disambiguate a homonym (i.e., disambiguation point) was therefore identified. In three subsequent ERP experiments, we then investigated whether the gesture information available at the disambiguation point has immediate as well as delayed consequences on the processing of a temporarily ambiguous spoken sentence, and whether these gesture-speech integration processes are susceptible to temporal synchrony. Experiment 1, which used asynchronous stimuli as well as an explicit task, showed clear N400 effects at the homonym as well as at the target word presented further downstream, suggesting that asynchrony does not prevent integration under explicit task conditions. No such effects were found when asynchronous stimuli were presented using a more shallow task (Experiment 2). Finally, when gesture fragment and homonym were synchronous, similar results as in Experiment 1 were found, even under shallow task conditions (Experiment 3). We conclude that when iconic gesture fragments and speech are in synchrony, their interaction is more or less automatic. When they are not, more controlled, active memory processes are necessary to be able to combine the gesture fragment and speech context in such a way that the homonym is disambiguated correctly.


Emotions in cognitive conflicts are not aversive but are task specific.
It has been suggested that cognitive conflicts require effortful processing and, therefore, are aversive (Botvinick, 2007). In the present study, we compared conflicts emerging from the inhibition of a predominant response tendency in a go/no-go task with those between incompatible response activations in a Simon task in a within-subjects design, using the same type of stimuli. Whereas no-go trials elicited reduced skin conductance and pupillometric responses, but prolonged corrugator muscle activity, as compared with go trials, incompatible and compatible Simon trials were indistinguishable with respect to these parameters. Furthermore, the conflict-sensitive N2 components of the event-related brain potential were similar in amplitude, but showed significantly different scalp distributions, indicating dissociable neural generator systems. The present findings suggest the involvement of different emotional and cognitive processes in both types of cognitive conflicts-none being aversive, however. In addition, the N2 findings call into question claims of common monitoring systems for all kinds of cognitive conflicts.


Sweet's syndrome in a patient with Wegener's granulomatosis and ESRD.
A female patient in her late 60s with end stage renal disease secondary to Wegener's granulomatosis presented with a 2-day history of fever, nausea, somnolence and purple coloured exudative plaques on her forehead and index finger. Herpes zoster was the initial diagnosis and acyclovir was prescribed. Twenty-four hours later, her level of consciousness (LOC) had declined to a Glascow coma scale of 10/15 and she started exhibiting myoclonic jerks. The lesions progressed to both sides of the face and acyclovir was discontinued as it was felt to be contributing to the myoclonic jerks. CT scan of the brain and lumbar puncture revealed no abnormalities to explain diminished LOC. Septic screen was negative, and an EEG showed no ictal activity. Skin biopsy revealed neutrophilic dermal infiltrates, leading to the diagnosis of Sweet's syndrome and prednisone (1 mg/kg) was initiated. The patient's condition improved over 12 h and she was discharged 5 days later. She died 6 weeks later from her cardiac disease.


Potential implication of new torque teno mini viruses in parapneumonic empyema in children.
An unexplained increase in the incidence of parapneumonic empyema (PPE) in pneumonia cases has been reported in recent years. The present study investigated the genetic and biological specifications of new isolates of torque teno mini virus (TTMV) detected in pleural effusion samples from children hospitalised for severe pneumonia with PPE. A pathogen discovery protocol was applied in undiagnosed pleural effusion samples and led to the identification of three new isolates of TTMV (TTMV-LY). Isolated TTMV-LY genomes were transfected into A549 and human embryonic kidney 293T cells and viral replication was assessed by quantitative real-time PCR and full-length genome amplification. A549 cells were further infected with released TTMV-LY virions and the induced-innate immune response was measured by multiplex immunoassays. Genetic analyses of the three TTMV-LY genomes revealed a classic genomic organisation but a weak identity (<64%) with known sequences. We demonstrated the in vitro replication of TTMV-LY in alveolar epithelial cells and the effective release of infectious viral particles. We also showed a selective production of inflammatory mediators in response to TTMV infection. This study reports the description of replicative TTMV-LY isolated from parapneumonic effusions of children hospitalised with PPE, suggesting a potential role of the virus in the pathogenesis of pneumonia.


Clinical and Epidemiological Implications of 24-Hour Ambulatory Blood Pressure Monitoring for the Diagnosis of Hypertension in Kenyan Adults: A Population-Based Study.
The clinical and epidemiological implications of using ambulatory blood pressure monitoring (ABPM) for the diagnosis of hypertension have not been studied at a population level in sub-Saharan Africa. We examined the impact of ABPM use among Kenyan adults. We performed a nested case-control study of diagnostic accuracy. We selected an age-stratified random sample of 1248 adults from the list of residents of the Kilifi Health and Demographic Surveillance System in Kenya. All participants underwent a screening blood pressure (BP) measurement. All those with screening BP ≥140/90 mm Hg and a random subset of those with screening BP <140/90 mm Hg were invited to undergo ABPM. Based on the 2 tests, participants were categorized as sustained hypertensive, masked hypertensive, "white coat" hypertensive, or normotensive. Analyses were weighted by the probability of undergoing ABPM. Screening BP ≥140/90 mm Hg was present in 359 of 986 participants, translating to a crude population prevalence of 23.1% (95% CI 16.5-31.5%). Age standardized prevalence of screening BP ≥140/90 mm Hg was 26.5% (95% CI 19.3-35.6%). On ABPM, 186 of 415 participants were confirmed to be hypertensive, with crude prevalence of 15.6% (95% CI 9.4-23.1%) and age-standardized prevalence of 17.1% (95% CI 11.0-24.4%). Age-standardized prevalence of masked and white coat hypertension were 7.6% (95% CI 2.8-13.7%) and 3.8% (95% CI 1.7-6.1%), respectively. The sensitivity and specificity of screening BP measurements were 80% (95% CI 73-86%) and 84% (95% CI 79-88%), respectively. BP indices and validity measures showed strong age-related trends. Screening BP measurement significantly overestimated hypertension prevalence while failing to identify ≈50% of true hypertension diagnosed by ABPM. Our findings suggest significant clinical and epidemiological benefits of ABPM use for diagnosing hypertension in Kenyan adults.


Concentration of trichloroethylene in breast milk and household water from Nogales, Arizona.
The United States Environmental Protection Agency has identified quantification of trichloroethylene (TCE), an industrial solvent, in breast milk as a high priority need for risk assessment. Water and milk samples were collected from 20 households by a lactation consultant in Nogales, Arizona. Separate water samples (including tap, bottled, and vending machine) were collected for all household uses: drinking, bathing, cooking, and laundry. A risk factor questionnaire was administered. Liquid-liquid extraction with diethyl ether was followed by GC-MS for TCE quantification in water. Breast milk underwent homogenization, lipid hydrolysis, and centrifugation prior to extraction. The limit of detection was 1.5 ng/mL. TCE was detected in 7 of 20 mothers' breast milk samples. The maximum concentration was 6 ng/mL. TCE concentration in breast milk was significantly correlated with the concentration in water used for bathing (ρ = 0.59, p = 0.008). Detection of TCE in breast milk was more likely if the infant had a body mass index <14 (RR = 5.2, p = 0.02). Based on average breast milk consumption, TCE intake for 5% of the infants may exceed the proposed U.S. EPA Reference Dose. Results of this exploratory study warrant more in depth studies to understand risk of TCE exposures from breast milk intake.


Ovarian Dermoid Cyst Causing Distal Ureteral Obstruction.
A case of a 45-year old woman with an ovarian dermoid cyst causing ureteric colic secondary to distal ureteral obstruction is reported. The dermoid cyst was observed on computed tomography to be adjacent to and compressing the distal left ureter and this was confirmed at surgical exploration. Following oophorectomy, the patient's symptoms completely resolved and the excised ovarian cyst was confirmed on pathological evaluation to be a dermoid cyst. This appears to be the first reported case of ureteral obstruction caused by an ovarian dermoid cyst in the English medical literature.


Analysis of local perspectives on sustainable forest management: an Indonesian case studyThis study was funded through a collaborative project between the University of Illinois and the Center for International Forestry Research (CIFOR). The ideas contained herein are solely of the authors and do not necessarily reflect official views of CIFOR.
Despite widespread acceptance of sustainability as the ultimate goal of forest management, perspectives about its meaning, significance, and relevant indicators may still differ. This paper examines local perspectives on sustainability, and evaluates their similarities and differences. A systematic procedure based on criteria of proximity, pre-existing rights, dependency, knowledge of forest management, forestry spirit, daily activity, and legal rights was used to identify a small group of relevant stakeholders representing different groups, institutions, and organizations. Using participatory action research (PAR), stakeholders were asked to identify relevant indicators of sustainable forest management. The indicators identified by each stakeholder were then compared to a consolidated list assembled by field facilitators with respect to whether relevant indicators are present or not. Based on the resulting presence/absence matrix, a statistical tool called the simple matching coefficient was used to estimate the similarity measures among the stakeholders' perspectives. In addition, cluster analysis was used to classify groups of stakeholders depending on their similarities to each other. Finally, hypotheses related to the 'closeness' of perspectives among local communities, non-governmental organizations, a timber company, and government organizations, as revealed by their selection of indicators, were tested. Results show that: (a) local communities have different perceptions in terms of what they consider to be important indicators compared to the NGOs, (b) there are significantly different perceptions between the government and the timber companies, and (c) there are also different perceptions between urban and field-based personnel of the same organization.


Actinomycosis of the Pancreas: A Case Report and Review.
Actinomyces is a normal commensal of the upper aerodigestive tract, colon and female reproductive tract. It can give rise to invasive disease in case of any breach in mucosal integrity, as well as, in patients with immunosuppression. Rarely, actinomycosis can involve the pancreas especially after episodes of pancreatitis or in post operative patients. We observed a case of actinomycosis affecting recurrent intraductal papillary mucinous neoplasm (IPMN) of pancreatic remnant, 5 years after a Whipple's procedure. Our patient, a 66 years old male with a history of Whipple's procedure for IPMN of pancreatic uncinate process, presented with repeated episodes of acute pancreatitis. Repeated radiological investigations (CT, MRI and EUS) revealed resolving pancreatitis with recurrent IPMN of the pancreatic tail. The patient underwent laparobotic assisted resection of the remnant pancreas and spleen 3 months later. Intraoperatively, in addition to the recurrent IPMN of pancreatic tail, we found a dense peripancreatic desmoplastic reaction with areas of thick yellow pus pockets in the remnant pancreatic body. Bacteriology and histopathology revealed it as a recurrent IPMN associated with actinomycosis of pancreas with chronic xanthogranulomatous changes. We conclude that actinomycosis of the pancreas is a rare entity with only 5 cases reported in English literature to the best of our knowledge. If diagnosed preoperatively, early institution of antibiotics can improve the surgical outcome. Fortunately, after diagnosis, we were able to start antibiotics in early postoperative period with successful outcome.


Simultaneous all-optical demodulation and format conversion for multi-channel (CS)RZ-DPSK signals.
We proposed and demonstrate an all-optical demodulation and format conversion scheme for multi-channel (carrier suppressed) return-to-zero differential phase shift keying ((CS)RZ-DPSK) signals. By utilizing a single delay interferometer (DI) with half bit delay, multi-channel (CS)RZ-DPSK signals can be demodulated simultaneously at the destructive port of the DI, with the corresponding converted nonreturn-to-zero differential phase shift keying (NRZ-DPSK) signals obtained at the constructive port. The proposed multi-channel operation has been demonstrated for 6*20 Gb/s RZ-DPSK and 6*40 Gb/s CSRZ-DPSK signals, with ~0.8 and 1.2 dB average power penalties for the format conversions respectively.


What is the best candidate allograft for ACL reconstruction? An in vitro mechanical and histologic study in a canine model.
The knee joint is generally characterized by very low friction and high wear resistance. Several previous studies have compared ACL with the commonly used allografts from tensile properties perspective. No study has reported about the graft tendons from a frictional perspective, which is an important parameter for ACL functional performance. Twenty hind legs were used to harvest FDP tendon, ACL, ACH, and patellar tendon. Samples were evaluated with surface friction testing, indentation testing for tendon compressive moduli, lubricin immunohistochemistry, and histologic analysis. Frictional force of FDP tendon and ACL was significantly less than that of patellar tendon and ACH at first and fifth cycles. At the tenth cycle, the FDP tendon, ACL, and ACH showed significantly less frictional force than patellar tendon; after 100 cycles, the FDP tendon and ACL showed significantly less frictional force than patellar tendon. The compressive moduli of the FDP tendon, ACL, and ACH were significantly greater than that of patellar tendon. Histologic results showed that FDP tendon and ACL had a smooth surface with a thin layer of epitenon cells; patellar tendon and ACH had a rough surface and a layer of paratenon. Lubricin was found on the surface and extracellular matrix of FDP tendon and ACL. There was only limited lubricin expression on the surface and extracellular matrix of the ACH and patellar tendon. The FDP tendon has friction force and lubricin expression similar to those of native ACL. However, patellar tendon and ACH show higher friction force and less lubricin expression than ACL.


NEMO, a Transcriptional Target of Estrogen and Progesterone, Is Linked to Tumor Suppressor PML in Breast Cancer.
The beneficial versus detrimental roles of estrogen plus progesterone (E+P) in breast cancer remains controversial. Here we report a beneficial mechanism of E+P treatment in breast cancer cells driven by transcriptional upregulation of the NFκB modulator NEMO, which in turn promotes expression of the tumor suppressor protein promyelocytic leukemia (PML). E+P treatment of patient-derived epithelial cells derived from ductal carcinoma in situ (DCIS) increased secretion of the proinflammatory cytokine IL6. Mechanistic investigations indicated that IL6 upregulation occurred as a result of transcriptional upregulation of NEMO, the gene that harbored estrogen receptor (ER) binding sites within its promoter. Accordingly, E+P treatment of breast cancer cells increased ER binding to the NEMO promoter, thereby increasing NEMO expression, NFκB activation, and IL6 secretion. In two mouse xenograft models of DCIS, we found that RNAi-mediated silencing of NEMO increased tumor invasion and progression. This seemingly paradoxical result was linked to NEMO-mediated regulation of NFκB and IL6 secretion, increased phosphorylation of STAT3 on Ser727, and increased expression of PML, a STAT3 transcriptional target. In identifying NEMO as a pivotal transcriptional target of E+P signaling in breast cancer cells, our work offers a mechanistic explanation for the paradoxical antitumorigenic roles of E+P in breast cancer by showing how it upregulates the tumor suppressor protein PML. Cancer Res; 77(14); 3802-13. ©2017 AACR.


The development of a survey to measure completeness of response to migraine therapy.
To assess the psychometric properties of a new patient-reported migraine instrument, the Completeness of Response Survey (CORS), which measures a comprehensive set of factors important to patients' decisions regarding the initiation and continuation of treatment. Traditionally, migraine treatments and the instruments used to demonstrate their efficacy have focused on the relief of headache pain. As new treatments emerge with the potential for more complete and consistent migraine relief, more comprehensive tools are needed to demonstrate these benefits. The CORS includes 2 modules, the static CORS, which comprehensively evaluates one treatment at one time point, and the comparative CORS, which provides a more global comparison between 2 treatments at one time point. Together, the 2 modules can measure unmet treatment needs and improvements over the course of a clinical study. Data from an 8-site study comparing 147 patients' recent experiences with their current triptan therapy and 2 months of study treatment with a single-tablet formulation of sumatriptan/naproxen sodium were used to conduct a preliminary psychometric evaluation of the CORS. The study included both modules of the CORS, the Headache Impact Test, the revised Patient Perception of Migraine Questionnaire, and a migraine diary. The CORS response categories in both the static and comparative modules demonstrated limited floor or ceiling effects and few missing values (<3%). Inter-item correlations, principal components analysis (component loading range: 0.62 to 0.95), and high estimates of internal consistency (alpha range: 0.88 to 0.94) for each composite score supported the structure and proposed scoring algorithm for the static module. The pattern of correlations between the CORS static and comparative items and composites with the revised Patient Perception of Migraine Questionnaire items and subscales, as well as the relationships between responses to selected static CORS items and the migraine diary, supported the construct validity of the CORS. The CORS is capable of demonstrating advantages of more comprehensive migraine therapies over traditional therapies, which are primarily focused on the resolution of headache pain, by addressing the frequency and speed with which the most common migraine symptoms are resolved and patients' return to normal functioning. This research shows evidence for the value and utility for the CORS static and comparative items and components, and further evaluation is underway.


Regeneration of the active zone at the frog neuromuscular junction.
The active zone is a unique specialization of the presynaptic membrane and is believed to be the site of transmitter release. The formation of the active zone and the relationship of this process to transmitter release were studied at reinnervated neuromuscular junctions in the frog. At different times after a nerve crush, the cutaneous pectoris muscles were examined with intracellular recording recording and freeze-fracture electron microscopy. The P face of a normal active zone typically consists of two double rows of particles lined up in a continuous segment located opposite a junctional fold. In the initial stage of reinnervation, clusters of large intramembrane particles surrounding membrane elevations appeared on the P face of nerve terminals. Like normal active zones, these clusters were aligned with junctional folds. Vesicle openings, which indicate transmitter release, were seen at these primitive active zones, even though intramembrane particles were not yet organized into the normal pattern of two double rows. The length of active zones at this stage was only approximately 15% of normal. During the secondary stage, every junction was reinnervated and most active zones had begun to organize into the normal pattern with normal orientation. Unlike normal, there were often two or more discontinuous short segments of active zone aligned with the same junctional fold. The total length of active zone per junctional fold increased to one-third of normal, mainly because of the greater number of segments. In the third stage, the number of active zone segments per junctional fold showed almost no change when compared with the secondary stage. However, individual segments elongated and increased the total length of all active zone segments per junctional fold to about two-thirds of the normal length. The dynamic process culminated in the final stage, during which elongating active zones appeared to join together and the number of active zone segments per junctional fold decreased to normal. Thus, in most regions, regeneration of the active zones was complete. These results suggest that the normal organization of two double rows is not necessary for the active zone to be functional. Furthermore, localization of regenerating active zones is related to junctional folds and/or their associated structures.


EggLib: processing, analysis and simulation tools for population genetics and genomics.
With the considerable growth of available nucleotide sequence data over the last decade, integrated and flexible analytical tools have become a necessity. In particular, in the field of population genetics, there is a strong need for automated and reliable procedures to conduct repeatable and rapid polymorphism analyses, coalescent simulations, data manipulation and estimation of demographic parameters under a variety of scenarios. In this context, we present EggLib (Evolutionary Genetics and Genomics Library), a flexible and powerful C++/Python software package providing efficient and easy to use computational tools for sequence data management and extensive population genetic analyses on nucleotide sequence data. EggLib is a multifaceted project involving several integrated modules: an underlying computationally efficient C++ library (which can be used independently in pure C++ applications); two C++ programs; a Python package providing, among other features, a high level Python interface to the C++ library; and the egglib script which provides direct access to pre-programmed Python applications. EggLib has been designed aiming to be both efficient and easy to use. A wide array of methods are implemented, including file format conversion, sequence alignment edition, coalescent simulations, neutrality tests and estimation of demographic parameters by Approximate Bayesian Computation (ABC). Classes implementing different demographic scenarios for ABC analyses can easily be developed by the user and included to the package. EggLib source code is distributed freely under the GNU General Public License (GPL) from its website http://egglib.sourceforge.net/ where a full documentation and a manual can also be found and downloaded.


An integrin-contactin complex regulates CNS myelination by differential Fyn phosphorylation.
The understanding of how adhesion molecules mediate the axon-glial interactions in the CNS that ensure target-dependent survival of oligodendrocytes and initiate myelination remains incomplete. Here, we investigate how signals from adhesion molecules can be integrated to regulate these initial steps of myelination. We first demonstrate that the Ig superfamily molecule contactin is associated in oligodendrocytes with integrins, extracellular matrix receptors that regulate target-dependent survival by amplification of growth factor signaling. This amplification is inhibited by small interfering RNA-mediated knockdown of contactin in oligodendrocytes. In contrast, the presence of L1-Fc, the extracellular portion of a contactin ligand expressed on axons, enhanced survival and additionally promoted myelination in cocultures of neurons and oligodendrocytes. We further demonstrate that the signals from contactin and integrin are integrated by differential phosphorylation of the Src family kinase Fyn. Integrin induced dephosphorylation of the inhibitory Tyr-531, whereas contactin increased phosphorylation of both Tyr-531 and the activating Tyr-420. The combined effect is an enhanced activity of Fyn and also a dynamic regulation of the phosphorylation/dephosphorylation balance of Fyn, as required for normal cell adhesion and spreading. We conclude, therefore, that a novel integrin/contactin complex coordinates signals from extracellular matrix and the axonal surface to regulate both oligodendrocyte survival and myelination by controlling Fyn activity.


Gene therapy for the treatment of X-linked retinitis pigmentosa.
X-linked retinitis pigmentosa caused by mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene is the most common form of recessive RP. The phenotype is characterised by its severity and rapid disease progression. Gene therapy using adeno-associated viral vectors is currently the most promising therapeutic approach. However, the construction of a stable vector encoding the full-length RPGR transcript has previously proven to be a limiting step towards gene therapy clinical trials. Recently however, a codon optimised version of RPGR has been shown to increase the stability and fidelity of the sequence, conferring a therapeutic effect in murine and canine animal models. This manuscript reviews the natural history of X-linked retinitis pigmentosa and the research performed from the discovery of the causative gene, RPGR, to the preclinical testing of potential therapies that have led to the initiation of three clinical trials. X-linked retinitis pigmentosa is an amenable disease to be treated by gene therapy. Codon optimisation has overcome the challenge of designing an RPGR vector without mutations, and with a therapeutic effect in different animal models. With the RPGR gene therapy clinical trials still in the early stages, the confirmation of the safety, tolerability and potency of the therapy is still ongoing.


Contrasting effects of nitrogen addition on soil respiration in two Mediterranean ecosystems.
Increased atmospheric nitrogen (N) deposition is known to alter ecosystem carbon source-sink dynamics through changes in soil CO2 fluxes. However, a limited number of experiments have been conducted to assess the effects of realistic N deposition in the Mediterranean Basin, and none of them have explored the effects of N addition on soil respiration (R s ). To fill this gap, we assessed the effects of N supply on R s dynamics in the following two Mediterranean sites: Capo Caccia (Italy), where 30 kg ha-1 year-1 was supplied for 3 years, and El Regajal (Spain), where plots were treated with 10, 20, or 50 kg N ha-1 year-1 for 8 years. Results show a complex, non-linear response of soil respiration (R s ) to N additions with R s overall increasing at Capo Caccia and decreasing at El Regajal. This suggests that the response of R s to N addition depends on dose and duration of N supply, and the existence of a threshold above which the N introduced in the ecosystem can affect the ecosystem's functioning. Soil cover and seasonality of precipitations also play a key role in determining the effects of N on R s as shown by the different responses observed across seasons and in bare soil vs. the soil under canopy of the dominant species. These results show how increasing rates of N addition may influence soil C dynamics in semiarid ecosystems in the Mediterranean Basin and represent a valuable contribution for the understanding and the protection of Mediterranean ecosystems.


Effects of antigen and genetic adjuvants on immune responses to human immunodeficiency virus DNA vaccines in mice.
The effects of genetic adjuvants on humoral and cell-mediated immunity to two human immunodeficiency virus antigens, Env and Nef, have been examined in mice. Despite similar levels of gene expression and the same gene delivery vector, the immune responses to these two gene products differed following DNA immunization. Intramuscular immunization with a Nef expression vector plasmid generated a humoral response and antigen-specific gamma interferon (IFN-gamma) production but little cytotoxic-T-lymphocyte (CTL) immunity. In contrast, immunization with an Env vector stimulated CTL activity but did not induce a high-titer antibody response. The ability to modify these antigen-specific immune responses was investigated by coinjection of DNA plasmids encoding cytokine and/or hematopoietic growth factors, interleukin-2 (IL-2), IL-12, IL-15, Flt3 ligand (FL), and granulocyte-macrophage colony-stimulating factor (GM-CSF). Coadministration of these genes largely altered the immune responses quantitatively but not qualitatively. IL-12 induced the greatest increase in IFN-gamma and immunoglobulin G responses to Nef, and GM-CSF induced the strongest IFN-gamma and CTL responses to Env. A dual approach of expanding innate immunity by administering the FL gene, together with a cytokine that enhances adaptive immune responses, IL-2, IL-12, or IL-15, generated the most potent immune response at the lowest doses of Nef antigen. These findings suggest that intrinsic properties of the antigen determine the character of immune reactivity for this method of immunization and that specific combination of innate and adaptive immune cytokine genes can increase the magnitude of the response to DNA vaccines.


ADAM9 Mediates Triple-Negative Breast Cancer Progression via AKT/NF-κB Pathway.
Upregulation of a disintegrin and metalloprotease 9 (ADAM9) is correlated with progression of cancers, such as prostate, bladder, and pancreatic cancers. However, its role in triple-negative breast cancer (TNBC) is still unclear. Our study aimed to investigate whether ADAM9 is upregulated and promoted the aggressiveness in TNBC. Breast cancer cell lines and patient specimens were used to evaluate the ADAM9 expression by western blotting and immunohistochemistry staining, respectively. Compared with the non-TNBC, ADAM9 expression was significantly increased in TNBC cells and TNBC patient specimens. Based on the data acquired from public databases, the correlation between ADAM9 expression and breast cancer patient survival was analyzed by Kaplan-Meier method. It was shown that ADAM9 overexpression was significantly correlated with poorer survival in patients with TNBC. Furthermore, ADAM9 in TNBC cells was knocked down by small interference RNA and then studied by the MTT/colony formation assay, wound healing assay and transwell invasion assay on the cell proliferation, migration, and invasion, respectively. We found that inhibiting ADAM9 expression suppressed TNBC cell proliferation, migration, and invasion by lowering the activation of AKT/NF-κB pathway. Our results demonstrated that ADAM9 is an important molecule in mediating TNBC aggressiveness and may be a potential useful therapeutic target in TNBC treatment.


Large bowel mucosal dysplasia and carcinoma in ulcerative colitis.
The clinicopathological details of eight cases of ulcerative colitis complicated by carcinoma of the colon are described. There was a total of 14 primary colonic cancers, six of which were not detected before pathological examination of the resection specimens. The reason for this may be related to atypical tumour growth patterns. Three occurred in flat mucosa, one in a mucosal plaque lesion, and another in polypoidal mucosa. The occurrence, distribution, and morphology of mucosal dysplasia were noted in both resection specimens and biopsies taken at varying stages before resection. Tumour was associated with normal and adjacent dysplastic mucosa of varying grades. The extent and grade of dysplasia were not reliable indicators of tumour differentiation or subsequent clinical outcome. Only two cancers were poorly differentiated. In five cases a total of 23 mucosal biopsies were taken, all less than 12 months before resection. Three rectal biopsies were graded positive for dysplasia and three colonic biopsies indefinite for dysplasia. The subsequent resection specimens showed both dysplastic and carcinomatous changes. Three rectal and 14 colonic biopsies were graded negative for dysplasia despite positive findings in the subsequent resection specimens. This anomaly is partly attributed to the patchy nature of dysplasia in colitic mucosa. Two cases illustrate the possibility of dysplasia pursuing a rapidly progressive course. The mucosal changes of ulcerative colitis were assessed using a recently introduced and standardised international classification.


Coeliac disease: a potentially treatable health problem of Saharawi refugee children.
To characterize the clinical and nutritional impact of coeliac disease (gluten-sensitive enteropathy) among Saharawi children living as refugees in Algeria. A total of 65 Saharawi children with coeliac disease were compared with 71 age-matched non-coeliac controls. For each participant, the clinical history was taken and a clinical examination, non-quantitative 24-hour dietary recall, anthropometric and skinfold measurements, bioelectric impedance analysis (BIA) of body composition, and venous blood sampling for haemoglobin determination were performed. Gluten-containing food, especially bread, was the staple diet of Saharawi children. Abdominal pain and distension were significantly commoner among children with coeliac disease than in controls (P < 0.05). The mean height-for-age was significantly lower in such children than in controls (-2.5 +/- 1.4 units vs -1.8 +/- 1.3 units, respectively, P < 0.01). No significant differences were found for either skinfold or BIA measurements. Haemoglobin values tended to be lower in children with coeliac disease than in controls. Coeliac disease has a negative effect on the health status of Saharawi refugee children. Because of the high prevalence of the condition in the Saharawi, a specific programme for treating all affected individuals should be established. Further studies are required to quantify the impact of coeliac disease in other areas of the developing world.


An educational intervention to update health workers about HIV and infant feeding.
Clinical guidelines are used to translate research findings into evidence-based clinical practice but are frequently not comprehensively adopted by health workers (HWs). HIV and infant feeding guidelines were revised by the World Health Organization to align feeding advice for HIV-exposed and unexposed infants, and these were adopted in South Africa in 2017. We describe an innovative, team-based, mentoring programme developed to update HWs on these guidelines. The intervention was underpinned by strong theoretical frameworks and aimed to improve HWs' attitudes, knowledge, confidence, and skills about breastfeeding in the context of HIV. On-site workshops and clinical mentoring used interactive participatory methods and a simple low-tech approach, guided by participants' self-reported knowledge gaps. Workshops were conducted at 24 participating clinics over three sessions, each lasting 1-2 hr. Evaluation data were collected using a self-administered questionnaire. Of 303 participating HWs, 249/303 (82.2%) attended all workshops. Achieving high workshop attendance was challenging and "catch-up" sessions were required to achieve good coverage. Common knowledge gaps identified included antiretroviral therapy adherence monitoring during breastfeeding and management of viral load results (173 participants), management of breast conditions (79), and advice about expressing and storing breastmilk (64). Most participants reported all their knowledge gaps were addressed and anticipated that their practice would change. We describe a feasible, sustainable approach to updating HWs on HIV and infant feeding guidelines and improving skills in breastfeeding counselling in resource-constrained settings. This approach could be adapted to other topics and, with further evaluation, implemented at scale using existing resources.


Cost of acute coronary syndrome in Switzerland in 2008.
To perform a cost-of-illness study of acute coronary syndrome (ACS) in Switzerland from a societal perspective, evaluating direct costs, production losses and intangible costs in terms of quality adjusted life years (QALYs) lost. A bottom-up incidence-based approach was used. Data concerning patients with one or more ACS events were extracted from a national hospital database and from mortality statistics. Inpatient costs included acute care and rehabilitation. Outpatient costs included costs for ambulance, visits to GP and cardiologist, outpatient diagnostics, medication and rehabilitation. Production losses included absenteeism, permanent disability and premature death. Intangible costs were calculated on previously published QALY weights. Cost data were derived from official price lists, literature and experts. Future costs and QALYs lost were discounted. In 2008 14,955 patients experienced a total of 16,815 ACS events; 2,752 died as a consequence of these. The resulting 19,064 hospital stays had an average across-hospital length of stay of 9.1 days per patient. Total direct costs of ACS amounted to 630 Mio Swiss Francs (CHF) for society and CHF 462 Mio for health insurers. Total direct costs were dominated by costs of myocardial infarction: ST-elevation 45.8%, non-ST-elevation 35.8%. Production losses were CHF 519 Mio and intangible costs resulted in 49,878 QALYs lost. ACS causes considerable costs in terms of direct medical expenditures, lost production, suffering and premature death, even without taking into account costs for its chronic consequences such as congestive heart failure.


Cytotoxic T lymphocyte epitopes of HIV-1 Nef: Generation of multiple definitive major histocompatibility complex class I ligands by proteasomes.
Although a pivotal role of proteasomes in the proteolytic generation of epitopes for major histocompatibility complex (MHC) class I presentation is undisputed, their precise function is currently the subject of an active debate: do proteasomes generate many epitopes in definitive form, or do they merely generate the COOH termini, whereas the definitive NH(2) termini are cleaved by aminopeptidases? We determined five naturally processed MHC class I ligands derived from HIV-1 Nef. Unexpectedly, the five ligands correspond to only three cytotoxic T lymphocyte (CTL) epitopes, two of which occur in two COOH-terminal length variants. Parallel analyses of proteasomal digests of a Nef fragment encompassing the epitopes revealed that all five ligands are direct products of proteasomes. Moreover, in four of the five ligands, the NH(2) termini correspond to major proteasome cleavage sites, and putative NH(2)-terminally extended precursor fragments were detected for only one of the five ligands. All ligands are transported by the transporter associated with antigen processing (TAP). The combined results from these five ligands provide strong evidence that many definitive MHC class I ligands are precisely cleaved at both ends by proteasomes. Additional evidence supporting this conclusion is discussed, along with contrasting results of others who propose a strong role for NH(2)-terminal trimming with direct proteasomal epitope generation being a rare event.


Dual chain extension effect and antibacterial properties of biomolecules interleaved within LDH dispersed into PBS by in situ polymerization.
Nanocomposites based on poly(butylene succinate) (PBS) and hydrotalcite-type anionic clays (HTs) organo-modified with biomolecules characterized by antibacterial and/or antioxidant activities, such as l-ascorbic acid (ASA), phloretic acid (HPP), l-tyrosine (TYR) and l-tryptophan (TRP), have been prepared by in situ polymerization. From XRD analysis and rheology experiments in a molten polymer state, intercalated HT hybrid platelets acting here as a hybrid filler are found to be well dispersed into polymers while providing a chain extension effect on PBS. Moreover, the molecules, when hosted within a HT interlayer gap, do preserve their pristine antibacterial activity, both in HT and in the resulting PBS composites. In particular, under the experimental conditions tested, HT/ASA and HT/TYR present the best combination of both properties (chain extension effect and antibacterial), especially versus E. coli as high as 90 and 97% of inhibition, respectively, using 2.5 wt% hybrid filler only. These findings open future applications for PBS associated with the hybrid HT filler as multifunctional materials in active packaging applications.


Apoptotic susceptibility of cancer cells selected for camptothecin resistance: gene expression profiling, functional analysis, and molecular interaction mapping.
To study the molecular mechanisms by which drug resistance develops, we compared DU145 humanprostate cancer cells with a subline selected for resistance to camptothecin. Differences in gene expression level were assessed by hybridizing the two cell types against each other using quadruplicate "Oncochip" cDNA microarrays that included 1648 cancer-related genes. Expression levels differing by a factor of >1.5 were detected for 181 of the genes. These differences were judged statistically reliable on the basis of a stratum-adjusted Kruskal-Wallis test, after taking into account a dye-dependent variable. The 181 expression-altered genes included a larger than expected number of the "apoptosis-related" genes (P = 0.04). To assess whether this observation reflected a generalized resistance of RCO.1 to apoptosis, we exposed the cells to a range of stresses (cisplatin, staurosporine, UV, ionizing radiation, and serum starvation) and found greatly reduced apoptotic responses for RC0.1 (relative to DU145) using flow cytometric Annexin V and terminal deoxynucleotidyl transferase-mediated nick end labeling assays. We next examined the apoptosis-related genes in the context of a molecular interaction map and found expression differences in the direction "expected" on the basis of the apoptosis-resistance of RC0.1 for BAD, caspase-6, and genes that signal via the Akt pathway. Exposure of the cells to wortmannin, an inhibitor of the Akt effector phosphatidylinositol 3-kinase, provided functional support for involvement of the Akt pathway. However, closer examination of the molecular interaction map revealed a paradox: many of the expression differences observed for apoptosis-related genes were in the direction "contrary" to that expected given the resistance of RC0.1. The map indicated that most of these unexpected expression differences were associated with genes involved in the nuclear factor kappa B and transforming growth factor beta pathways. Overall, the patterns that emerged suggested a two-step model for the selection process that led to resistance in RC0.1 cells. The first hypothesized step would involve a decrease in apoptotic susceptibility through changes in the apoptosis-control machinery associated with the Bcl-2 and caspase gene families, and also in antiapoptotic pathways operating through Akt/PKB. The second step would involve changes in multifunctional upstream genes (including some genes in the nuclear factor kappa B and transforming growth factor beta pathways) that can facilitate apoptosis but that would also tend to contribute to cell proliferation in the presence of drug. Thus, we propose that a downstream blockade of apoptosis was "permissive" for the selection of upstream pathway changes that would otherwise have induced apoptosis. This model is analogous to one suggested previously for the relationship between oncogene function and apoptosis in carcinogenesis.


Interference of the linguistic variant in the repair strategies used during the phonological acquisition process.
To investigate and compare the use of repair strategies in the acquisition of /R/ in simple onset, produced by children with typical phonological acquisition. Speech data containing the /R/ from 120 children with typical phonological acquisition (60 male and 60 female) from Santa Maria (RS) and Crissiumal (RS), Brazil, aged between 1 year and 6 months and 4 years were used. To analyze the repair strategies, the following dependent variables were considered: omission, semivocalization, and liquid substitution; as well as the following independent variables: gender, age, precedent and following context, grammatical class, tonicity, number of syllables, and position in the word. The VARBRUL program was used for statistical analysis. The statistical program selected as significant for omission in Santa Maria the variables tonicity and gender, and in Crissiumal, tonicity and age. For semivocalization in Santa Maria, the program selected the variable gender, and in Crissiumal, tonicity. For lateral liquid substitution in Santa Maria the statistical program did not select any variable. However, in Crissiumal, the variables position in the word, gender, and age were selected. It was possible to observe that the repair strategies can diverge according to the dialect being used. Hence, it is important to consider the dialectal variation to make the phonological therapy more effective.


Strong effects of genetic and lifestyle factors on biomarker variation and use of personalized cutoffs.
Ideal biomarkers used for disease diagnosis should display deviating levels in affected individuals only and be robust to factors unrelated to the disease. Here we show the impact of genetic, clinical and lifestyle factors on circulating levels of 92 protein biomarkers for cancer and inflammation, using a population-based cohort of 1,005 individuals. For 75% of the biomarkers, the levels are significantly heritable and genome-wide association studies identifies 16 novel loci and replicate 2 previously known loci with strong effects on one or several of the biomarkers with P-values down to 4.4 × 10(-58). Integrative analysis attributes as much as 56.3% of the observed variance to non-disease factors. We propose that information on the biomarker-specific profile of major genetic, clinical and lifestyle factors should be used to establish personalized clinical cutoffs, and that this would increase the sensitivity of using biomarkers for prediction of clinical end points.


Astrocyte-specific deletion of the transcription factor Yin Yang 1 in murine substantia nigra mitigates manganese-induced dopaminergic neurotoxicity.
Manganese (Mn)-induced neurotoxicity resembles Parkinson's disease (PD), but the mechanisms underpinning its effects remain unknown. Mn dysregulates astrocytic glutamate transporters, GLT-1 and GLAST, and dopaminergic function, including tyrosine hydroxylase (TH). Our previous in vitro studies have shown that Mn repressed GLAST and GLT-1 via activation of transcription factor Yin Yang 1 (YY1). Here, we investigated if in vivo astrocytic YY1 deletion mitigates Mn-induced dopaminergic neurotoxicity, attenuating Mn-induced reduction in GLAST/GLT-1 expression in murine substantia nigra (SN). AAV5-GFAP-Cre-GFP particles were infused into the SN of 8-week-old YY1 flox/flox mice to generate a region-specific astrocytic YY1 conditional knockout (cKO) mouse model. 3 weeks after adeno-associated viral (AAV) infusion, mice were exposed to 330 μg of Mn (MnCl2 30 mg/kg, intranasal instillation, daily) for 3 weeks. After Mn exposure, motor functions were determined in open-field and rotarod tests, followed by Western blotting, quantitative PCR, and immunohistochemistry to assess YY1, TH, GLAST, and GLT-1 levels. Infusion of AAV5-GFAP-Cre-GFP vectors into the SN resulted in region-specific astrocytic YY1 deletion and attenuation of Mn-induced impairment of motor functions, reduction of TH-expressing cells in SN, and TH mRNA/protein levels in midbrain/striatum. Astrocytic YY1 deletion also attenuated the Mn-induced decrease in GLAST/GLT-1 mRNA/protein levels in midbrain. Moreover, YY1 deletion abrogated its interaction with histone deacetylases in astrocytes. These results indicate that astrocytic YY1 plays a critical role in Mn-induced neurotoxicity in vivo, at least in part, by reducing astrocytic GLAST/GLT-1. Thus, YY1 might be a potential target for treatment of Mn toxicity and other neurological disorders associated with dysregulation of GLAST/GLT-1.


The effectiveness of evidence summaries on health policymakers and health system managers use of evidence from systematic reviews: a systematic review.
Systematic reviews are important for decision makers. They offer many potential benefits but are often written in technical language, are too long, and do not contain contextual details which make them hard to use for decision-making. There are many organizations that develop and disseminate derivative products, such as evidence summaries, from systematic reviews for different populations or subsets of decision makers. This systematic review aimed to (1) assess the effectiveness of evidence summaries on policymakers' use of the evidence and (2) identify the most effective summary components for increasing policymakers' use of the evidence. We present an overview of the available evidence on systematic review derivative products. We included studies of policymakers at all levels as well as health system managers. We included studies examining any type of "evidence summary," "policy brief," or other products derived from systematic reviews that presented evidence in a summarized form. The primary outcomes were the (1) use of systematic review summaries in decision-making (e.g., self-reported use of the evidence in policymaking and decision-making) and (2) policymakers' understanding, knowledge, and/or beliefs (e.g., changes in knowledge scores about the topic included in the summary). We also assessed perceived relevance, credibility, usefulness, understandability, and desirability (e.g., format) of the summaries. Our database search combined with our gray literature search yielded 10,113 references after removal of duplicates. From these, 54 were reviewed in full text, and we included six studies (reported in seven papers) as well as protocols from two ongoing studies. Two studies assessed the use of evidence summaries in decision-making and found little to no difference in effect. There was also little to no difference in effect for knowledge, understanding or beliefs (four studies), and perceived usefulness or usability (three studies). Summary of findings tables and graded entry summaries were perceived as slightly easier to understand compared to complete systematic reviews. Two studies assessed formatting changes and found that for summary of findings tables, certain elements, such as reporting study event rates and absolute differences, were preferred as well as avoiding the use of footnotes. Evidence summaries are likely easier to understand than complete systematic reviews. However, their ability to increase the use of systematic review evidence in policymaking is unclear. The protocol was published in the journal Systematic Reviews (2015;4:122).


Risk factors for Plasmodium falciparum gametocyte positivity in a longitudinal cohort.
Malaria transmission intensity is highly heterogeneous even at a very small scale. Implementing targeted intervention in malaria transmission hotspots offers the potential to reduce the burden of disease both locally and in adjacent areas. Transmission of malaria parasites from man to mosquito requires the production of gametocyte stage parasites. Cluster analysis of a 19-year long cohort study for gametocyte carriage revealed spatially defined gametocyte hotspots that occurred during the time when chloroquine was the drug used for clinical case treatment. In addition to known risk factors for gametocyte carriage, notably young age (<15 years old) and associated with a clinical episode, blood groups B and O increased risk compared to groups A and AB. A hotspot of clinical P. falciparum clinical episodes that overlapped the gametocyte hotspots was also identified. Gametocyte positivity was found to be increased in individuals who had been treated with chloroquine, as opposed to other drug treatment regimens, for a clinical P. falciparum episode up to 30 days previously. It seems likely the hotspots were generated by a vicious circle of ineffective treatment of clinical cases and concomitant gametocyte production in a sub-population characterized by an increased prevalence of all the identified risk factors. While rapid access to treatment with an effective anti-malarial can reduce the duration of gametocyte carriage and onward parasite transmission, localised hotspots represent a challenge to malaria control and eventual eradication.


Base composition bias might result from competition for metabolic resources.
The GC content of bacterial genomes varies from 25 to 75%, but the reason for this variation is unclear. Here, we show that genomes of bacteria that rely on their host for survival (obligatory pathogens or symbionts) tend to be AT rich. Furthermore, we have analysed bacterial phages, plasmids and insertion sequences, which might also be regarded as 'intracellular pathogens', and show that they too are significantly richer in AT than their hosts. We suggest that the higher energy cost and limited availability of G and C over A and T/U could be a basis for the understanding of these differences.


Socio-economic position and subjective health and well-being among older people in Europe: a systematic narrative review.
Previous studies of older European populations have established that disability and morbidity vary with indicators of socio-economic position (SEP). We undertook a systematic narrative review of the literature to ascertain to what extent there is evidence of similar inequalities in the subjective health and well-being of older people in Europe. Relevant original research articles were searched for using Medline, Global Health, Embase, Social Policy and Practice, Cinahl, Web of Science and International Bibliography of the Social Sciences (IBSS). We included studies of SEP and indicators of subjective health and well-being (self-rated health; life satisfaction; quality of life) conducted since 1991 using population-based samples of older people in Europe and published 1995-2013. A total of 71 studies were identified. Poorer SEP was associated with poorer subjective health and well-being. Associations varied somewhat depending on the SEP measure and subjective health and well-being outcome used. Associations were weaker when social support and health-related behaviours were adjusted for suggesting that these factors mediate the relationship between SEP and subjective health and well-being. Associations tended to be weaker in the oldest age groups. The patterns of associations by gender were not consistent and tended to diminish after adjusting for indicators of health and life circumstances. The results of this systematic narrative review of the literature demonstrate the importance of social influences on later life subjective health and well-being and indicate areas which need further investigation, such as more studies from Eastern Europe, more longitudinal studies and more research on the role of mediating factors.


The determinants of dentists' productivity and the measurement of output.
Improving the productivity of the healthcare system, for example by taking advantage of scale economies or encouraging substitution of expensive specialist personnel with less expensive workers, is often seen as an attractive way to meet increasing demand within a constrained budget. Using data on 558 dentists participating in the Longitudinal Study of Dentists' Practice Activity (LSDPA) survey between 1993 and 2003 linked to patient data and average fee schedules, we estimate production functions for private dental services in Australia to quantify the contribution of different capital and labour inputs and identify economies of scale in the production of dental care. Given the challenges in measuring output in the healthcare setting, we discuss three different output measures (raw activity, time-, and price-weighted activity) and test the sensitivity of results to the choice of measure. Our results suggest that expansion of the scale of dental services is unlikely to be constrained by decreasing returns to scale. We note that conclusions about the contribution of individual input factors and the estimated returns to scale are sensitive to the choice of output measure employed.


Associations between conspiracism and the rejection of scientific innovations.
Public opinion regarding scientific developments such as genetically modified food can be mixed. We suggest such science-based technological innovations are rejected by some because they are perceived to be advanced as part of a conspiracy. In nationally representative samples (Australia n = 1011; New Zealand n = 754), we report the associations between five conspiracism facets and anti-science attitudes. Results indicate broad public opposition to genetically modified food and use of nuclear power, but more acceptance of renewable power, potable recycled water, 5G networks, and childhood vaccinations. There were small to moderate associations between the rejection of scientific innovations and conspiracism. Multivariate models estimating unique associations of conspiracism facets with anti-science attitudes suggested several novel and important relationships, particularly for childhood vaccination, genetically modified food, and 5G networks. We discuss the importance of examining factors such as conspiracism in understanding what may motivate and sustain rejection of scientific evidence-based claims about socially contentious technological innovations.


Analysis of Long Non-Coding RNA in Cryptosporidium parvum Reveals Significant Stage-Specific Antisense Transcription.
Cryptosporidium is a protist parasite that has been identified as the second leading cause of moderate to severe diarrhea in children younger than two and a significant cause of mortality worldwide. Cryptosporidium has a complex, obligate, intracellular but extra cytoplasmic lifecycle in a single host. How genes are regulated in this parasite remains largely unknown. Long non-coding RNAs (lncRNAs) play critical regulatory roles, including gene expression across a broad range of organisms. Cryptosporidium lncRNAs have been reported to enter the host cell nucleus and affect the host response. However, no systematic study of lncRNAs in Cryptosporidium has been conducted to identify additional lncRNAs. In this study, we analyzed a C. parvum in vitro strand-specific RNA-seq developmental time series covering both asexual and sexual stages to identify lncRNAs associated with parasite development. In total, we identified 396 novel lncRNAs, mostly antisense, with 86% being differentially expressed. Surprisingly, nearly 10% of annotated mRNAs have an antisense transcript. lncRNAs occur most often at the 3' end of their corresponding sense mRNA. Putative lncRNA regulatory regions were identified and many appear to encode bidirectional promoters. A positive correlation between lncRNA and upstream mRNA expression was observed. Evolutionary conservation and expression of lncRNA candidates was observed between C. parvum, C. hominis and C. baileyi. Ten C. parvum protein-encoding genes with antisense transcripts have P. falciparum orthologs that also have antisense transcripts. Three C. parvum lncRNAs with exceptional properties (e.g., intron splicing) were experimentally validated using RT-PCR and RT-qPCR. This initial characterization of the C. parvum non-coding transcriptome facilitates further investigations into the roles of lncRNAs in parasite development and host-pathogen interactions.


Uncontrolled, open-label, pilot study of tea tree (Melaleuca alternifolia) oil solution in the decolonisation of methicillin-resistant Staphylococcus aureus positive wounds and its influence on wound healing.
Many complementary and alternative products are used to treat wounds. The essential oil of Melaleuca alternifolia, tea tree oil, has proven antimicrobial and anti-inflammatory properties, may be useful in methicillin-resistant Staphylococcus aureus (MRSA) decolonisation regimens and is reputed to have 'wound-healing' properties, but more data are required to support these indications. The primary aim of this uncontrolled case series was to assess whether a tea tree oil solution used in a wound cleansing procedure could decolonise MRSA from acute and chronic wounds of mixed aetiology. The secondary aim was to determine if the tea tree oil solution influenced wound healing outcomes. Nineteen participants with wounds suspected of being colonised with MRSA were enrolled in a pilot study. Seven were subsequently shown not to have MRSA and were withdrawn from the study. As many as 11 of the remaining 12 participants were treated with a water-miscible tea tree oil (3·3%) solution applied as part of the wound cleansing regimen at each dressing change. Dressing changes were three times per week or daily as deemed necessary by the study nurse following assessment. One participant withdrew from the study before treatment. No participants were MRSA negative after treatment. After treatment had been implemented, 8 of the 11 treated wounds had begun to heal and reduced in size as measured by computer planimetry. Although this formulation and mode of delivery did not achieve the primary aim of the study, tea tree oil did not appear to inhibit healing and the majority of wounds reduced in size after treatment.


Contingencies and compounded rare perturbations dictate sudden distributional shifts during periods of gradual climate change.
Ecological responses to climate change may occur gradually with changing conditions, or they may occur rapidly once some threshold or "tipping point" has been reached. Here, we use a high-resolution, 30-year data set on the upper vertical limit of a high intertidal alga to demonstrate that distributional shifts in this species do not keep pace with gradual trends in air temperature or sea level, but rather occur in sudden, discrete steps. These steps occur when unusually warm air temperatures are associated with unusually calm seas and are contingent in the sense that neither atmospheric nor sea conditions by themselves were sufficient to generate the underlying physiological challenge. Shifts in the upper limit did not correlate with large environmental perturbations such as El Niños; rather, they appeared to be associated with stochastic departures from otherwise gradual environmental trends. Our results exemplify the view that multiple environmental factors should be considered when attempting to understand ecological responses to climate change. Furthermore, punctuated responses such as those we have identified urge caution when attempting to infer causal mechanisms and project future distributional shifts using data of limited temporal resolution or scope.