Anti-glycation and anti-oxidative effects of a phenolic-enriched maple syrup extract and its protective effects on normal human colon cells.
Oxidative stress and free radical generation accelerate the formation of advanced glycation endproducts (AGEs) which are linked to several chronic diseases. Published data suggest that phenolic-rich plant foods, show promise as natural anti-AGEs agents due to their anti-oxidation capacities. A phenolic-enriched maple syrup extract (MSX) has previously been reported to show anti-inflammatory and neuroprotective effects but its anti-AGE effects remain unknown. Therefore, herein, we investigated the anti-glycation and anti-oxidation effects of MSX using biochemical and biophysical methods. MSX (500 μg mL-1) reduced the formation of AGEs by 40% in the bovine serum albumin (BSA)-fructose assay and by 30% in the BSA-methylglyoxal (MGO) assay. MSX also inhibited the formation of crosslinks typically seen in the late stage of glycation. Circular dichroism and differential scanning calorimeter analyses demonstrated that MSX maintained the structure of BSA during glycation. In the anti-oxidant assays, MSX (61.7 μg mL-1) scavenged 50% of free radicals (DPPH assay) and reduced free radical generation by 20% during the glycation process (electron paramagnetic resonance time scan). In addition, the intracellular levels of hydrogen peroxide induced reactive oxygen species were reduced by 27-58% with MSX (50-200 μg mL-1) in normal/non-tumorigenic human colon CCD-18Co cells. Moreover, in AGEs and MGO challenged CCD-18Co cells, higher cellular viabilities and rapid extracellular signal-regulated kinase (ERK) phosphorylation were observed in MSX treated cells, indicating its protective effects against AGEs-induced cytotoxicity. Overall, this study supports the biological effects of MSX, and warrants further investigation of its potential as a dietary agent against diseases mediated by oxidative stress and inflammation.


Safety profile of accelerated corneal cross-linking versus conventional cross-linking: a comparative study on ex vivo-cultured limbal epithelial cells.
To compare the effects of accelerated corneal collagen cross-linking (ACXL) and corneal collagen cross-linking (CXL) on ex vivo-cultured limbal epithelial cells (LECs). Day 14 cultured LECs were either unexposed (control) or exposed to different intensities of ultraviolet-A (UV-A) irradiance for different durations (3 mW for 30 min, 9 mW for 10 min, 18 mW for 5 min and 30 mW for 3 min) in the presence and absence of riboflavin. These cells were further processed for quantitative real-time PCR, vital staining, immunofluorescence staining and fluorescence-activated cell sorting (FACS) staining to evaluate the apoptotic status. Statistical analysis was performed using a Student t test. Vital staining showed a significantly higher (p=0.004) dead cell population with 3 mW for 30 min when compared with 30 mW for 3 min exposure (p=0.225). Quantitative PCR results revealed significantly reduced abcg2 and Δnp63 mRNA levels, while FACS analysis showed an increase in ABCG2-Annexin V positive population in cells exposed to 3 mW for 30 mins. Neither reduction of mRNA expression of abcg2 and Δnp63 nor increase in FACS-stained ABCG2-Annexin V positivity was detected in cells exposed to 30 mW for 3 min. Additionally, enhanced caspase activity was detected with fluorochrome inhibitor of caspases staining and mRNA expression of caspase 3 and 9 was upregulated in cells exposed to 3 mW for 30 min, but not at 30 mW for 3 min. The 30 mW UV-A irradiation used in ACXL appears to be safe on cultured LECs in comparison with 3 mW used in CXL.


The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation.
FAT10 is a ubiquitin-like modifier proposed to function in apoptosis induction, cell cycle control and NF-κB activation. Upon induction by pro-inflammatory cytokines, hundreds of endogenous substrates become covalently conjugated to FAT10 leading to their proteasomal degradation. Nevertheless, only three substrates have been identified so far to which FAT10 becomes covalently attached through a non-reducible isopeptide bond, and these are the FAT10-conjugating enzyme USE1 which auto-FAT10ylates itself in cis, the tumor suppressor p53 and the ubiquitin-activating enzyme UBE1 (UBA1). To identify additional FAT10 substrates and interaction partners, we used a new monoclonal FAT10-specific antibody to immunopurify endogenous FAT10 conjugates from interferon (IFN)γ-and tumor necrosis factor (TNF)α-stimulated cells for identification by mass spectrometry. In addition to two already known FAT10-interacting proteins, histone deacetylase 6 and UBA6, we identified 569 novel FAT10-interacting proteins involved in different functional pathways such as autophagy, cell cycle regulation, apoptosis and cancer. Thirty-one percent of all identified proteins were categorized as putative covalently linked substrates. One of the identified proteins, the autophagosomal receptor p62/SQSTM1, was further investigated. p62 becomes covalently mono-FAT10ylated at several lysines, and FAT10 colocalizes with p62 in p62 bodies. Strikingly, FAT10ylation of p62 leads to its proteasomal degradation, and prolonged induction of endogenous FAT10 expression by pro-inflammatory cytokines leads to a decrease of endogenous p62. The elucidation of the FAT10 degradome should enable a better understanding of why FAT10 has evolved as an additional transferable tag for proteasomal degradation.


The biobehavioral Women's Health CoOp in Pretoria, South Africa: study protocol for a cluster-randomized design.
South Africa has 6.4 million adults over the age of 15 living with HIV. Gender inequality issues continue to drive the HIV epidemic in South Africa, where Black African women bear the greatest HIV burden. Limited access to services; little capacity to negotiate sex and condom use; and other legal, social, and economic inequities make women highly vulnerable to HIV infection. Behavioral interventions have been shown to decrease risk behaviors, but they have been less successful in reducing HIV incidence. Conversely, biomedical prevention strategies have proven to be successful in reducing HIV incidence, but require behavioral interventions to increase uptake and adherence. Consequently, there is a need for integrated approaches that combine biomedical and behavioral interventions. Effective combination prevention efforts should comprise biomedical, behavioral, and structural programming proven in randomized trials that focuses on the driving forces and key populations at higher risk of HIV infection and transmission. This prospective, geographically clustered randomized field experiment is enrolling participants into two arms: a control arm that receives standard HIV testing and referral for treatment; and an intervention arm that receives an evidence-based, woman-focused behavioral intervention that emphasizes risk reduction and retention, the Women's Health CoOp. We divided the city of Pretoria into 14 mutually exclusive geographic zones and randomized these zones into either the control arm or the intervention arm. Outreach workers are recruiting drug-using women from each zone. At baseline, eligible participants complete a questionnaire and biological testing for HIV, recent drug use, and pregnancy. Follow-up interviews are completed at 6 and 12 months. The biobehavioral intervention in this study merges an efficacious behavioral HIV prevention intervention for women with biomedical prevention through HIV treatment as prevention using a Seek, Test, Treat and Retain strategy. This combination biobehavioral intervention is designed to (1) improve the quality of life and reduce HIV infectiousness among women who are HIV positive, and (2) reduce HIV risk behaviors among women regardless of their HIV status. If efficacious, this intervention could help control the HIV epidemic in South Africa. Trial registration no: NCT01497405.


Frontal infarcts and anxiety in stroke.
This study examined the association between poststroke anxiety symptoms (PSA) and frontal lobe infarcts. A cohort of 693 patients was recruited. PSA was defined as an anxiety subscale of the Hospital Anxiety and Depression Scale score of 8 or above. The presence and location of infarcts were evaluated with MRI. Compared with the non-PSA group, PSA patients were more likely to have right frontal acute infarcts. Right frontal infarcts remained independent predictors of PSA in the multivariate analysis, with an odds ratio of 4.44 (P=0.002). The results suggest that right frontal acute infarcts may play a role in the development of PSA.


Following international trends while subject to past traditions: neuropsychological test use in the Nordic countries.
Historically, the neuropsychological test traditions of the four Nordic countries have spanned from the flexible and qualitative tradition of Luria-Christensen to the quantitative large battery approach of Halstead and Kløve-Matthews. This study reports current test use and discusses whether these traditions still influence attitudes toward test use and choice of tests. The study is based on survey data from 702 Nordic neuropsychologists. The average participant used 9 tests in a standard assessment, and 25 tests overall in their practice. Test use was moderated by nationality, competence level, practice profile, and by attitude toward test selection. Participants who chose their tests flexibly used fewer tests than those adhering to the flexible battery approach, but had fewer tests from which to choose. Testing patients with psychiatric disorders was associated with using more tests. IQ, memory, attention, and executive function were the domains with the largest utilization rate, while tests of motor, visual/spatial, and language were used by few. There is a lack of academic achievement tests. Screening tests played a minor role in specialized assessments, and symptom validity tests were seldom applied on a standard basis. Most tests were of Anglo-American origin. New test methods are implemented rapidly in the Nordic countries, but test selection is also characterized by the dominating position of established and much researched tests. The Halstead-Reitan and Luria traditions are currently weak, but national differences in size of test batteries seem to be influenced by these longstanding traditions.


Blurring boundaries: professional perspectives of the emergency nurse practitioner role in a major accident and emergency department.
The role of the emergency nurse practitioner (ENP) has increasingly become part of mainstream healthcare delivery in major accident and emergency departments in the United Kingdom. Although some research data are available in this field, there has been little attempt to evaluate the impact of the implementation of the ENP role from the perspective of those healthcare professionals most closely involved at local level. This paper describes one part of a case study evaluation of the role in an accident and emergency department in the South Thames English region. Nine face-to-face semi-structured interviews were carried out with the key multidisciplinary stakeholders in the organization. Five major themes emerged from the data analysis: blurring role boundaries; managing uncertainty; individual variation; quality vs. quantity; and the organizational context. Whilst some professional consensus was evident regarding the benefits of the role, such as improved waiting times and patient satisfaction, there appeared also to be a degree of ambivalence, particularly regarding current role configuration, value for money, and the extent to which the role should be expanded in the future. These issues are discussed in terms of professional identity, changing role boundaries, and professional personhood. It is argued that the benefits and pitfalls of the ENP role need to be considered within the context of local service provision. The growing emphasis on clinical governance reinforces the need for ongoing audit of role effectiveness in order to meet the challenges and uncertainties of increasingly blurred professional boundaries.


Paraneoplastic CDR2 and CDR2L antibodies affect Purkinje cell calcium homeostasis.
Paraneoplastic cerebellar degeneration (PCD) is characterized by loss of Purkinje cells (PCs) associated with progressive pancerebellar dysfunction in the presence of onconeural Yo antibodies. These antibodies recognize the cerebellar degeneration-related antigens CDR2 and CDR2L. Response to PCD therapy is disappointing due to limited understanding of the neuropathological mechanisms. Here, we report the pathological role of CDR antibodies on the calcium homeostasis in PCs. We developed an antibody-mediated PCD model based on co-incubation of cerebellar organotypic slice culture with human patient serum or rabbit CDR2 and CDR2L antibodies. The CDR antibody-induced pathology was investigated by high-resolution multiphoton imaging and biochemical analysis. Both human and rabbit CDR antibodies were rapidly internalized by PCs and led to reduced immunoreactivity of calbindin D28K (CB) and L7/Pcp-2 as well as reduced dendritic arborizations in the remaining PCs. Washout of the CDR antibodies partially recovered CB immunoreactivity, suggesting a transient structural change in CB calcium-binding site. We discovered that CDR2 and CB co-immunoprecipitate. Furthermore, the expression levels of voltage-gated calcium channel Cav2.1, protein kinase C gamma and calcium-dependent protease, calpain-2, were increased after CDR antibody internalization. Inhibition of these signaling pathways prevented or attenuated CDR antibody-induced CB and L7/Pcp-2 immunoreactivity loss, morphological changes and increased protein expression. These results signify that CDR antibody internalization causes dysregulation of cell calcium homeostasis. Hence, drugs that modulate these events may represent novel neuroprotective therapies that limit the damaging effects of CDR antibodies and prevent PC neurodegeneration.


Community to clinic navigation to improve diabetes outcomes.
Rural residents experience rates of Type 2 Diabetes Mellitus (T2DM) that are considerably higher than their urban or suburban counterparts. Two primary modifiable factors, self-management and formal clinical management, have potential to greatly improve diabetes outcomes. "Community to Clinic Navigation to Improve Diabetes Outcomes," is the first known randomized clinical trial pilot study to test a hybrid model of diabetes self-management education plus clinical navigation among rural residents with T2DM. Forty-one adults with T2DM were recruited from two federally qualified health centers in rural Appalachia from November 2014-January 2015. Community health workers provided navigation, including helping participants understand and implement a diabetes self-management program through six group sessions and, if needed, providing assistance in obtaining clinic visits (contacting providers' offices for appointments, making reminder calls, and facilitating transportation and dependent care). Pre and post-test data were collected on T2DM self-management, physical measures, demographics, psychosocial factors, and feasibility (cost, retention, and satisfaction). Although lacking statistical significance, some outcomes indicate trends in positive directions, including diet, foot care, glucose monitoring, and physical health, including decreased HbA1c and triglyceride levels. Process evaluations revealed high levels of satisfaction and feasibility. Due to the limited intervention dose, modest program expenditures (~$29,950), and a severely affected population most of whom had never received diabetes education, outcomes were not as robust as anticipated. Given high rates of satisfaction and retention, this culturally appropriate small group intervention holds promise for hard to reach rural populations. Modifications should include expanded recruitment venues, sample size, intervention dosage and longer term assessment.


A Conclusive Analysis of the Finite-Time Behavior of the Discretized Pursuit Learning Automaton.
This paper deals with the finite-time analysis (FTA) of learning automata (LA), which is a topic for which very little work has been reported in the literature. This is as opposed to the asymptotic steady-state analysis for which there are, probably, scores of papers. As clarified later, unarguably, the FTA of Markov chains, in general, and of LA, in particular, is far more complex than the asymptotic steady-state analysis. Such an FTA provides rigid bounds for the time required for the LA to attain to a given convergence accuracy. We concentrate on the FTA of the Discretized Pursuit Automaton (DPA), which is probably one of the fastest and most accurate reported LA. Although such an analysis was carried out many years ago, we record that the previous work is flawed. More specifically, in all brevity, the flaw lies in the wrongly "derived" monotonic behavior of the LA after a certain number of iterations. Rather, we claim that the property should be invoked is the submartingale property. This renders the proof to be much more involved and deep. In this paper, we rectify the flaw and reestablish the FTA based on such a submartingale phenomenon. More importantly, from the derived analysis, we are able to discover and clarify, for the first time, the underlying dilemma between the DPA's exploitation and exploration properties. We also nontrivially confirm the existence of the optimal learning rate, which yields a better comprehension of the DPA itself.


Understanding resilience in young people with complex mental health needs: A Delphi study.
Resilience is a term used to describe an individual's adaptive coping following an adverse experience; it is important for gaining insight into the development of mental health difficulties in young people and their ability to manage adversity, informing both preventive and reactive clinical practice. The Delphi method was used whereby a panel of 15 clinical psychologists rated 67 statements, generated from focus groups with young people and interviews with multi-disciplinary staff, in terms of their importance relating to resilience for young people with complex mental health needs. A consensus level of 85% across the panel was set to include/exclude statements in terms of their importance for resilience. Nineteen statements were included in the final list. These were grouped into the following four themes: (1) understanding the self, (2) agency in recovery, (3) interpersonal relationships and (4) therapeutic setting and relationships. The results highlight specific resilience factors for young people with complex mental health needs, based upon ratings by clinical psychologists. Recommendations are made which focus upon how to promote resilience within this specific population. These include offering secure therapeutic relationships and a safe environment for young people to make decisions, develop a greater understanding of themselves, and build relationships and a sense of connection with others, both within the specialist mental health service and upon discharge.


Risk factors related to ischemic and hemorrhagic cerebrovascular disease at autopsy: the Honolulu Heart Study.
This study of cerebrovascular lesions at autopsy among Hawaiian Japanese men identifies similar risks factors for cerebral infarction and hemorrhage that have been identified in a previous incidence study. Demonstrated differences were essentially the same whether subjects with these tissue changes were compared to men showing no central nervous system disease at autopsy or when they were compared with men still alive. Cerebral infarcts accompanied myocardial infarction (CHD) in 58% of autopsy cases and were associated with CHD risk factors (high serum cholesterol, hypertension, severe atherosclerosis of the coronary arteries and aorta). These associations did not persist when CHD cases were removed from the analysis, indicating there were two subsets of men with cerebral infarction. Hypertension was strongly associated with hemorrhagic disease, as were cigarette use and alcohol consumption.


Grafting of ARPE-19 and Schwann cells to the subretinal space in RCS rats.
To study the distribution of the human retinal pigment epithelium (hRPE) cell line ARPE-19 and human Schwann (hSC) cells grafted to the subretinal space of the Royal College of Surgeon (RCS) rat and the relation of graft cell distribution to photoreceptor rescue. Cell suspensions of both donor types were injected into the subretinal space of 3-week-old dystrophic RCS rats through a transscleral approach, human fibroblast and medium were used as control grafts. All animals were maintained on oral cyclosporine. At 1, 2, 4, 6, 15, 28, and 36 weeks after grafting, animals were killed. Human cell-specific markers were used to localize donor cells. Both donor cell types, as revealed by antibodies survived for a substantial time. Their distribution was very different: hRPE cells formed a large clump early on and, with time, spread along the host RPE in a layer one to two cells deep, whereas hSCs formed many smaller clumps, mainly in the subretinal space. Both cells rescued photoreceptors beyond the area of donor cell distribution. The number of surviving cells declined with time. Both hRPE and hSC grafts can survive and rescue photoreceptors for a substantial time after grafting. The number of both donor cell types declined with time, which could be an immune-related problem and/or due to other factors intrinsic to the host RCS retina. The fact that rescue occurred beyond the area of donor cell distribution suggests that diffusible factors are involved, raising the possibility that the two cell types function in a similar manner to rescue photoreceptors.


Optical mechanisms regulating emmetropisation and refractive errors: evidence from animal models.
Our current understanding of emmetropisation and myopia development has evolved from decades of work in various animal models, including chicks, non-human primates, tree shrews, guinea pigs, and mice. Extensive research on optical, biochemical, and environmental mechanisms contributing to refractive error development in animal models has provided insights into eye growth in humans. Importantly, animal models have taught us that eye growth is locally controlled within the eye, and can be influenced by the visual environment. This review will focus on information gained from animal studies regarding the role of optical mechanisms in guiding eye growth, and how these investigations have inspired studies in humans. We will first discuss how researchers came to understand that emmetropisation is guided by visual feedback, and how this can be manipulated by form-deprivation and lens-induced defocus to induce refractive errors in animal models. We will then discuss various aspects of accommodation that have been implicated in refractive error development, including accommodative microfluctuations and accommodative lag. Next, the impact of higher order aberrations and peripheral defocus will be discussed. Lastly, recent evidence suggesting that the spectral and temporal properties of light influence eye growth, and how this might be leveraged to treat myopia in children, will be presented. Taken together, these findings from animal models have significantly advanced our knowledge about the optical mechanisms contributing to eye growth in humans, and will continue to contribute to the development of novel and effective treatment options for slowing myopia progression in children.


Provoking an end-to-end continuous direct compression line with raw materials prone to segregation.
Continuous manufacturing of solid oral dosage forms is promising for increasing the efficiency and quality of pharmaceutical production and products. In this study a whole train continuous direct compression (CDC) line has been provoked using challenging formulations typically prone to segregation in batch powder processing. Industrial compositions including components with variable size, bulk density and cohesive nature were selected. An experimental design, including variables such as API/mannitol particle size, API amount, powder feed rate and mixer speed, enabled the output quality of the provoked process to be assessed. Contrary to previous studies, a broader range of finished tablet quality attributes were probed, including content, uniformity of content, tensile strength as well as release performance. Overall, the continuous direct compression line was found to be a capable and efficient manufacturing process for the challenging compositions studied and surprisingly tolerable to handle the materials susceptible to segregation in typical batch settings. As expected, and given the 'fixed' apparatus configuration used in this study, the particulate material properties were found to have the most significant impact on the finished tablet quality attributes. The results emphasize the importance for taking a holistic approach when developing the operational windows and the strategy for control, e.g. by integrating the appropriate material properties, the actual apparatus design, and the relevant formulation design. The CDC line's ability to handle cohesive materials also seem to be one of the key advantages, thus confirming the recent promising results from other continuous direct compression studies.


Polydesigns and causal inference.
In an increasingly common class of studies, the goal is to evaluate causal effects of treatments that are only partially controlled by the investigator. In such studies there are two conflicting features: (1) a model on the full cohort design and data can identify the causal effects of interest, but can be sensitive to extreme regions of that design's data, where model specification can have more impact; and (2) models on a reduced design (i.e., a subset of the full data), for example, conditional likelihood on matched subsets of data, can avoid such sensitivity, but do not generally identify the causal effects. We propose a framework to assess how inference is sensitive to designs by exploring combinations of both the full and reduced designs. We show that using such a "polydesign" framework generates a rich class of methods that can identify causal effects and that can also be more robust to model specification than methods using only the full design. We discuss implementation of polydesign methods, and provide an illustration in the evaluation of a needle exchange program.


Multiplexed detection of xylene and trichloroethylene in water by photonic crystal absorption spectroscopy.
We experimentally demonstrate simultaneous selective detection of xylene and trichloroethylene (TCE) using multiplexed photonic crystal waveguides (PCWs) by near-infrared optical absorption spectroscopy on a chip. Based on the slow light effect of photonic crystal structure, the sensitivity of our device is enhanced to 1 ppb (v/v) for xylene and 10 ppb (v/v) for TCE in water. Multiplexing is enabled by multimode interference power splitters and Y-combiners that integrate multiple PCWs on a silicon chip in a silicon-on-insulator platform.


Protection against lethal toxic shock by targeted disruption of the CD28 gene.
Toxic shock syndrome (TSS) is a multi system disorder resulting from superantigen-mediated cytokine production. Nearly 90% of the clinical cases of TSS arise due to an exotoxin, toxic shock syndrome toxin-1 (TSST-1), elaborated by toxigenic strains of Staphylococcus aureus. It is clearly established that besides antigen-specific signals a variety of costimulatory signals are required for full T cell activation. However, the nature and potential redundancy of costimulatory signals are incompletely understood, particularly with regards to superantigen-mediated T cell activation in vivo. Here we report that CD28-deficient mice (CD28-/-) are completely resistant to TSST-1-induced lethal TSS while CD28 (+/-) littermate mice were partially resistant to TSST-1. The mechanism for the resistance of the CD28 (-/-) mice was a complete abrogation of TNF-alpha accumulation in the serum and a nearly complete (90%) impairment of IFN-gamma secretion in response to TSST-1 injection. In contrast, the serum level of IL-2 was only moderately influenced by the variation of CD28 expression. CD28 (-/-) mice retained sensitivity to TNF-alpha as demonstrated by equivalent lethality after cytokine injection. These findings establish an essential requirement for CD28 costimulatory signals in TSST-1-induced TSS. The hierarchy of TSST-1 resistance among CD28 wild-type (CD28+/+), CD28 heterozygous (CD28+/-), and CD28-/- mice suggests a gene-dose effect, implying that the levels of T cell surface CD28 expression critically regulate superantigen-mediated costimulation. Finally, as these results demonstrate the primary and non-redundant role of CD28 receptors in the initiation of the in vivo cytokine cascade, they suggest therapeutic approaches for superantigen-mediated immunopathology.


A Century of Progress: Reading Interventions for Students in Grades 4-12, 1914-2014.
The history of research on interventions for struggling readers in Grades 4 through 12 dates back to 19th-century case studies of seemingly intelligent children who were unable to learn to read. Physicians, psychologists, educators, and others were determined to help them. In the process, they launched a century of research on a wide variety of approaches to reading intervention. As shown in this systematic narrative review, much has changed over time in the conceptualization of reading interventions and the methods used to determine their efficacy in improving outcomes for struggling readers. Building on the knowledge gathered over the past 100 years, researchers and practitioners are well-poised to continue to make progress in developing and testing reading interventions over the next 100 years.


Blood biomarkers on admission in acute traumatic brain injury: Relations to severity, CT findings and care path in the CENTER-TBI study.
Serum biomarkers may inform and improve care in traumatic brain injury (TBI). We aimed to correlate serum biomarkers with clinical severity, care path and imaging abnormalities in TBI, and explore their incremental value over clinical characteristics in predicting computed tomographic (CT) abnormalities. We analyzed six serum biomarkers (S100B, NSE, GFAP, UCH-L1, NFL and t-tau) obtained <24 h post-injury from 2867 patients with any severity of TBI in the Collaborative European NeuroTrauma Effectiveness Research (CENTER-TBI) Core Study, a prospective, multicenter, cohort study. Univariable and multivariable logistic regression analyses were performed. Discrimination was assessed by the area under the receiver operating characteristic curve (AUC) with 95% confidence intervals. All biomarkers scaled with clinical severity and care path (ER only, ward admission, or ICU), and with presence of CT abnormalities. GFAP achieved the highest discrimination for predicting CT abnormalities (AUC 0•89 [95%CI: 0•87-0•90]), with a 99% likelihood of better discriminating CT-positive patients than clinical characteristics used in contemporary decision rules. In patients with mild TBI, GFAP also showed incremental diagnostic value: discrimination increased from 0•84 [95%CI: 0•83-0•86] to 0•89 [95%CI: 0•87-0•90] when GFAP was included. Results were consistent across strata, and injury severity. Combinations of biomarkers did not improve discrimination compared to GFAP alone. Currently available biomarkers reflect injury severity, and serum GFAP, measured within 24 h after injury, outperforms clinical characteristics in predicting CT abnormalities. Our results support the further development of serum GFAP assays towards implementation in clinical practice, for which robust clinical assay platforms are required. CENTER-TBI study was supported by the European Union 7th Framework program (EC grant 602150).


Atezolizumab for the treatment of renal cell carcinoma.
The therapeutic landscape of renal cell cancer has evolved rapidly over the past 2 years with nivolumab and ipilimumab for patients with metastatic disease and an intermediate or poor prognosis, in the first line setting. More recently, data from trials combining antiangiogenic agents and immune checkpoint inhibitors demonstrated a major benefit of this treatment approach for all patients. One of three recent trials evaluated the combination of atezolizumab, an anti-programmed death ligand 1 antibody, with bevacizumab, an anti-vascular endothelial growth factor monoclonal antibody. In this manuscript, we summarize the preclinical, clinical, and safety data on atezolizumab for treatment of renal cell carcinoma and describe ongoing trials. Atezolizumab was evaluated in combination with an antiangiogenic agent. These trials were designed based on the hypothesis that selecting patients according to the expression of programmed death ligand 1 would increase the benefit of the treatment combination. Despite positive effects on the primary endpoints progression-free survival and response rate in this selected population, overall survival in the global population did not meet the criteria for significance at the time of the intermediate analysis. The major information was a proposed tumor gene expression signature. The signature was predictive of the sensitivity to anti-angiogenic and/or immune checkpoint inhibitor therapy.


Variability of respiratory rate measurements in children suspected with non-severe pneumonia in north-east Tanzania.
Measurement of respiratory rate is an important clinical sign in the diagnosis of pneumonia but suffers from interobserver variation. Here, we assess the use of video recordings as a quality assurance tool that could be useful both in research and in training of staff. Respiratory rates (RR) were recorded in children aged 2-59 months presenting with cough or difficulty breathing at two busy outpatient clinics in Tanzania. Measurements were repeated at 10-min intervals in a quiet environment with simultaneous video recordings that were independently reviewed by two paediatricians. Eight hundred and fifty-nine videos were sent to two paediatricians; 148 (17.2%) were considered unreadable by one or both. For the 711 (82.8%) videos that were readable by both paediatricians, there was perfect agreement for the presence of raised RR with a kappa value (κ) of 0.85 (P < 0.001); and in 476 (66.9%) cases, both paediatricians agreed on the RR within 2 breaths per minute (±2 bpm). A reported illness of 5 days or more was associated with unreadable video recordings (OR = 3.44, CI: 1.5-6.08; P < 0.001). The multilevel model showed that differences between observers accounted for only 13% of the variability in RR. Video recordings are reliable tools for quality assurance of RR measurements in children with suspected pneumonia. Videos with a clear view of respiratory movements may also be useful in training primary healthcare staff.


Mutations of optineurin in amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) has its onset in middle age and is a progressive disorder characterized by degeneration of motor neurons of the primary motor cortex, brainstem and spinal cord. Most cases of ALS are sporadic, but about 10% are familial. Genes known to cause classic familial ALS (FALS) are superoxide dismutase 1 (SOD1), ANG encoding angiogenin, TARDP encoding transactive response (TAR) DNA-binding protein TDP-43 (ref. 4) and fused in sarcoma/translated in liposarcoma (FUS, also known as TLS). However, these genetic defects occur in only about 20-30% of cases of FALS, and most genes causing FALS are unknown. Here we show that there are mutations in the gene encoding optineurin (OPTN), earlier reported to be a causative gene of primary open-angle glaucoma (POAG), in patients with ALS. We found three types of mutation of OPTN: a homozygous deletion of exon 5, a homozygous Q398X nonsense mutation and a heterozygous E478G missense mutation within its ubiquitin-binding domain. Analysis of cell transfection showed that the nonsense and missense mutations of OPTN abolished the inhibition of activation of nuclear factor kappa B (NF-kappaB), and the E478G mutation revealed a cytoplasmic distribution different from that of the wild type or a POAG mutation. A case with the E478G mutation showed OPTN-immunoreactive cytoplasmic inclusions. Furthermore, TDP-43- or SOD1-positive inclusions of sporadic and SOD1 cases of ALS were also noticeably immunolabelled by anti-OPTN antibodies. Our findings strongly suggest that OPTN is involved in the pathogenesis of ALS. They also indicate that NF-kappaB inhibitors could be used to treat ALS and that transgenic mice bearing various mutations of OPTN will be relevant in developing new drugs for this disorder.


Development of a support tool for complex decision-making in the provision of rural maternity care.
Decisions in the organization of safe and effective rural maternity care are complex, difficult, value laden and fraught with uncertainty, and must often be based on imperfect information. Decision analysis offers tools for addressing these complexities in order to help decision-makers determine the best use of resources and to appreciate the downstream effects of their decisions. To develop a maternity care decision-making tool for the British Columbia Northern Health Authority (NH) for use in low birth volume settings. Based on interviews with community members, providers, recipients and decision-makers, and employing a formal decision analysis approach, we sought to clarify the influences affecting rural maternity care and develop a process to generate a set of value-focused objectives for use in designing and evaluating rural maternity care alternatives. Four low-volume communities with variable resources (with and without on-site births, with or without caesarean section capability) were chosen. Physicians (20), nurses (18), midwives and maternity support service providers (4), local business leaders, economic development officials and elected officials (12), First Nations (women [pregnant and non-pregnant], chiefs and band members) (40), social workers (3), pregnant women (2) and NH decision-makers/administrators (17). We developed a Decision Support Manual to assist with assessing community needs and values, context for decision-making, capacity of the health authority or healthcare providers, identification of key objectives for decision-making, developing alternatives for care, and a process for making trade-offs and balancing multiple objectives. The manual was deemed an effective tool for the purpose by the client, NH. Beyond assisting the decision-making process itself, the methodology provides a transparent communication tool to assist in making difficult decisions. While the manual was specifically intended to deal with rural maternity issues, the NH decision-makers feel the method can be easily adapted to assist decision-making in other contexts in medicine where there are conflicting objectives, values and opinions. Decisions on the location of new facilities or infrastructure, or enhancing or altering services such as surgical or palliative care, would be examples of complex decisions that might benefit from this methodology.


Age-related differences in walking stability.
a large proportion of falls in older people occur when walking; however the mechanisms underlying impaired balance during gait are poorly understood. to evaluate acceleration patterns at the head and pelvis in young and older subjects when walking on a level and an irregular walking surface, in order to develop an understanding of how ageing affects postural responses to challenging walking conditions. temporo-spatial gait parameters and variables derived from acceleration signals were recorded in 30 young people aged 22-39 years (mean 29.0, SD 4.3), and 30 older people with a low risk of falling aged 75-85 years (mean 79.0, SD 3.0) while walking on a level and an irregular walking surface. Subjects also underwent tests of vision, sensation, strength, reaction time and balance. older subjects exhibited a more conservative gait pattern, characterised by reduced velocity, shorter step length and increased step timing variability. These differences were particularly pronounced when walking on the irregular surface. The magnitude of accelerations at the head and pelvis were generally smaller in older subjects; however the smoothness of the acceleration signals did not differ between the two groups. Older subjects performed worse on tests of vision, peripheral sensation, strength, reaction time and balance. the adoption of a more conservative basic gait pattern by older people with a low risk of falling reduces the magnitude of accelerations experienced by the head and pelvis when walking, which is likely to be a compensatory strategy to maintain balance in the presence of age-related deficits in physiological function, particularly reduced lower limb strength.


A Review of Copy Number Variants in Inherited Neuropathies.
The rapid development in the last 10-15 years of microarray technologies, such as oligonucleotide array Comparative Genomic Hybridization (CGH) and Single Nucleotide Polymorphisms (SNP) genotyping array, has improved the identification of fine chromosomal structural variants, ranging in length from kilobases (kb) to megabases (Mb), as an important cause of genetic differences among healthy individuals and also as disease-susceptibility and/or disease-causing factors. Structural genomic variations due to unbalanced chromosomal rearrangements are known as Copy-Number Variants (CNVs) and these include variably sized deletions, duplications, triplications and translocations. CNVs can significantly contribute to human diseases and rearrangements in several dosage-sensitive genes have been identified as an important causative mechanism in the molecular aetiology of Charcot-Marie-Tooth (CMT) disease and of several CMT-related disorders, a group of inherited neuropathies with a broad range of clinical phenotypes, inheritance patterns and causative genes. Duplications or deletions of the dosage-sensitive gene PMP22 mapped to chromosome 17p12 represent the most frequent causes of CMT type 1A and Hereditary Neuropathy with liability to Pressure Palsies (HNPP), respectively. Additionally, CNVs have been identified in patients with other CMT types (e.g., CMT1X, CMT1B, CMT4D) and different hereditary poly- (e.g., giant axonal neuropathy) and focal- (e.g., hereditary neuralgic amyotrophy) neuropathies, supporting the notion of hereditary peripheral nerve diseases as possible genomic disorders and making crucial the identification of fine chromosomal rearrangements in the molecular assessment of such patients. Notably, the application of advanced computational tools in the analysis of Next-Generation Sequencing (NGS) data has emerged in recent years as a powerful technique for identifying a genome-wide scale complex structural variants (e.g., as the ones resulted from balanced rearrangements) and also smaller pathogenic (intragenic) CNVs that often remain beyond the detection limit of most conventional genomic microarray analyses; in the context of inherited neuropathies where more than 70 disease-causing genes have been identified to date, NGS and particularly Whole-Genome Sequencing (WGS) hold the potential to reduce the number of genomic assays required per patient to reach a diagnosis, analyzing with a single test all the Single Nucleotide Variants (SNVs) and CNVs in the genes possibly implicated in this heterogeneous group of disorders.


The effects of iron deficiency on the hepatocyte: a biochemical and ultrastructural study.
Effects of iron deficiency on the hepatocyte were studied quantitatively in the rat by combining ultrastructural and biochemical techniques. After 3-8 wk of an iron-deficient diet, the percentage of cytoplasm occupied by mitochondria increased progressively compared with complete diet values. The increment resulted primarily from an enlargement of individual mitochondria rather than from an increased mitochondrial number. Many mitochondria were completely divided by a double membrane, often at a point of constriction. After 2 days of iron administration, mitochondria were of heterogeneous size, shape, and electron opacity. After 5 days, essentially all mitochondria had become normal in configuration. The rate of reversal of the morphological abnormality was more rapid than would be anticipated if it coincided with known rates of renewal of mitochondrial DNA or protein. The concentrations of mitochondrial cytochromes were more rapidly depressed as a result of iron deprivation than those of microsomal cytochromes. Cytochromes c and a were decreased after 3 and 8 wk of exposure to the deficient regimen. Cytochrome P 450 was not decreased after a 3 wk exposure to the deficient diet and responded normally to phenobarbital treatment with a fourfold increase in total hepatic content; its concentration was depressed only after 8 wk of exposure to the deficient diet. There was no reduction in cytochrome b(5) concentration.


Basic life support training for single rescuers efficiently augments their willingness to make early emergency calls with no available help: a cross-over questionnaire survey.
The aim of this study was to investigate effects of basic life support (BLS) training on willingness of single rescuers to make emergency calls during out-of-hospital cardiac arrests (OHCAs) with no available help from others. A cross-over questionnaire survey was conducted with two questionnaires. Questionnaires were administered before and after two BLS courses in fire departments. One questionnaire included two scenarios which simulate OHCAs occurring in situations where help from other rescuers is available (Scenario-M) and not available (Scenario-S). The conventional BLS course was designed for multiple rescuers (Course-M), and the other was designed for single rescuers (Course-S). Of 2,312 respondents, 2,218 (95.9%) answered all questions and were included in the analysis. Although both Course-M and Course-S significantly augmented willingness to make early emergency calls not only in Scenario-M but also in Scenario-S, the willingness for Scenario-M after training course was significantly higher in respondents of Course-S than in those of Course-M (odds ratio 1.706, 95% confidential interval 1.301-2.237). Multiple logistic regression analysis for Scenario-M disclosed that post training (adjusted odds ratio 11.6, 95% confidence interval 7.84-18.0), age (0.99, 0.98-0.99), male gender (1.77, 1.39-2.24), prior BLS experience of at least three times (1.46, 1.25-2.59), and time passed since most recent training during 3 years or less (1.80, 1.25-2.59) were independently associated with willingness to make early emergency calls and that type of BLS course was not independently associated with willingness. Therefore, both Course-M and Course-S similarly augmented willingness in Scenario-M. However, in multiple logistic regression analyses for Scenario-S, Course-S was independently associated with willingness to make early emergency calls in Scenario-S (1.26, 1.00-1.57), indicating that Course-S more efficiently augmented willingness. Moreover, post training (2.30, 1.86-2.83) and male gender (1.26, 1.02-1.57) were other independent factors associated with willingness in Scenario-S. BLS courses designed for single rescuers with no help available from others are likely to augment willingness to make early emergency calls more efficiently than conventional BLS courses designed for multiple rescuers.


Impaired islet function in commonly used transgenic mouse lines due to human growth hormone minigene expression.
The human growth hormone (hGH) minigene is frequently used in the derivation of transgenic mouse lines to enhance transgene expression. Although this minigene is present in the transgenes as a secondcistron, and thus not thought to be expressed, we found that three commonly used lines, Pdx1-Cre(Late), RIP-Cre, and MIP-GFP, each expressed significant amounts of hGH in pancreatic islets. Locally secreted hGH binds to prolactin receptors on β cells, activates STAT5 signaling, and induces pregnancy-like changes in gene expression, thereby augmenting pancreatic β cell mass and insulin content. In addition, islets of Pdx1-Cre(Late) mice have lower GLUT2 expression and reduced glucose-induced insulin release and are protected against the β cell toxin streptozotocin. These findings may be important when interpreting results obtained when these and other hGH minigene-containing transgenic mice are used.


Predicting the location and spatial extent of submerged coral reef habitat in the Great Barrier Reef world heritage area, Australia.
Coral reef communities occurring in deeper waters have received little research effort compared to their shallow-water counterparts, and even such basic information as their location and extent are currently unknown throughout most of the world. Using the Great Barrier Reef as a case study, habitat suitability modelling is used to predict the distribution of deep-water coral reef communities on the Great Barrier Reef, Australia. We test the effectiveness of a range of geophysical and environmental variables for predicting the location of deep-water coral reef communities on the Great Barrier Reef. Great Barrier Reef, Australia. Maximum entropy modelling is used to identify the spatial extent of two broad communities of habitat-forming megabenthos phototrophs and heterotrophs. Models were generated using combinations of geophysical substrate properties derived from multibeam bathymetry and environmental data derived from Bio-ORACLE, combined with georeferenced occurrence records of mesophotic coral communities from autonomous underwater vehicle, remotely operated vehicle and SCUBA surveys. Model results are used to estimate the total amount of mesophotic coral reef habitat on the GBR. Our models predict extensive but previously undocumented coral communities occurring both along the continental shelf-edge of the Great Barrier Reef and also on submerged reefs inside the lagoon. Habitat suitability for phototrophs is highest on submerged reefs along the outer-shelf and the deeper flanks of emergent reefs inside the GBR lagoon, while suitability for heterotrophs is highest in the deep waters along the shelf-edge. Models using only geophysical variables consistently outperformed models incorporating environmental data for both phototrophs and heterotrophs. Extensive submerged coral reef communities that are currently undocumented are likely to occur throughout the Great Barrier Reef. High-quality bathymetry data can be used to identify these reefs, which may play an important role in resilience of the GBR ecosystem to climate change.


Phenazine-Based Covalent Organic Framework Cathode Materials with High Energy and Power Densities.
Redox-active covalent organic frameworks (COFs) are promising materials for energy storage devices because of their high density of redox sites, permanent and controlled porosity, high surface areas, and tunable structures. However, the low electrochemical accessibility of their redox-active sites has limited COF-based devices either to thin films (<250 nm) grown on conductive substrates or to thicker films (1 μm) when a conductive polymer is introduced into the COF pores. Electrical energy storage devices constructed from bulk microcrystalline COF powders, eliminating the need for both thin-film formation and conductive polymer guests, would offer both improved capacity and potentially scalable fabrication processes. Here we report on the synthesis and electrochemical evaluation of a new phenazine-based 2D COF (DAPH-TFP COF), as well as its composite with poly(3,4-ethylenedioxythiophene) (PEDOT). Both the COF and its PEDOT composite were evaluated as powders that were solution-cast onto bulk electrodes serving as current collectors. The unmodified DAPH-TFP COF exhibited excellent electrical access to its redox sites, even without PEDOT functionalization, and outperformed the PEDOT composite of our previously reported anthraquinone-based system. Devices containing DAPH-TFP COF were able to deliver both high-energy and high-power densities, validating the promise of unmodified redox-active COFs that are easily incorporated into electrical energy storage devices.


A double-blind, randomized, and active-controlled phase III study of Herbiron drink in the treatment of iron-deficiency anemia in premenopausal females in Taiwan.
About 468 million non-pregnant women are estimated to suffer from iron-deficiency anemia (IDA) worldwide. The highest prevalence of IDA occurs in the Taiwanese population. To evaluate the effectiveness of Herbiron to increase iron absorption in women with IDA. Phase III double-blind, randomized, active-controlled, and parallel comparative study enrolled 124 patients with IDA and consisted of a 2-week run-in period, randomization, 12 weeks of supplementation, and 4 weeks of follow-up. The treatment group received Herbiron drink 50 mL p.o., b.i.d., before meals (daily iron intake: 21 mg/day) plus placebo tablets. The control group received a ferrous sulfate tablet, t.i.d., plus placebo 50-mL drink before meals (daily iron intake: 195 mg/day). Both treatments significantly improved hemoglobin and all secondary efficacy endpoints. Most IDA patients treated with Herbiron or ferrous sulfate finished the study in the normal range. Ferrous sulfate treatment induced a rapid rate of hemoglobin synthesis, which plateaued by week 8, whereas Herbiron treatment increased the rate of hemoglobin synthesis more slowly, likely due to its nine-fold lower iron content. Gastrointestinal adverse events (diarrhea, abdominal pain, dyspepsia, and nausea) but not infectious adverse events were significantly more common in the ferrous sulfate group (n=11, 18.3%) than those in the Herbiron group (n=1, 1.6%) (p=0.004). Twelve weeks of Herbiron treatment delivering 21mg of iron or ferrous sulfate treatment delivering 195 mg of iron induced normal hemoglobin levels in 62 or 91% of non-pregnant women with IDA in Taiwan, respectively, suggesting dose-dependent and bioavailability effects.


How do vegetable oils (hazelnut and canola) affect the reproductive system in male rats?
Vegetable oils have an important place in our daily diet. This study starts from this point to investigate the effects of canola oil and hazelnut oil in the male reproductive system in rats. 30 male rats were used in this 16-week study. The animals were divided into three groups: the animals in group I served as the control group, while the animals in group II and group III were fed with hazelnut and canola oil, respectively. The testes of all rats were excised for histopathologic evaluation and immunohistochemical (IHC) evaluation with a standard method. Blood samples were obtained for determination of serum hormone levels. No significant differences were noted with respect to behavior or weight among the three groups. Rats in the canola oil group (group III) had higher luteinizing hormone (LH) and higher testosterone levels than rats in the control group. Rats who received hazelnut oil (group II) exhibited similar findings, with these levels being higher than they were in the control group. No statistical differences were shown for histopathology or IHC testosterone antibody levels across all treatment groups. Conclussion: Canola oil was shown to have a greater effect on serum LH and testosterone compared to the control group and the group fed with hazelnut oil. Further investigation is required into how these oils affect serum hormone and sperm activity.


Transgenic monkeys produced by retroviral gene transfer into mature oocytes.
Transgenic rhesus monkeys carrying the green fluorescent protein (GFP) gene were produced by injecting pseudotyped replication-defective retroviral vector into the perivitelline space of 224 mature rhesus oocytes, later fertilized by intracytoplasmic sperm injection. Of the three males born from 20 embryo transfers, one was transgenic when accessible tissues were assayed for transgene DNA and messenger RNA. All tissues that were studied from a fraternal set of twins, miscarried at 73 days, carried the transgene, as confirmed by Southern analyses, and the GFP transgene reporter was detected by both direct and indirect fluorescence imaging.


Chromosome evolution in Eulipotyphla.
We integrated chromosome painting information on 5 core-insectivora species available in the literature with new Zoo-FISH data for Iberian shrew (Sorex granarius) and Altai mole (Talpa altaica). Our analysis of these 7 species allowed us to determine the chromosomal features of Eulipotyphla genomes and to update the previously proposed ancestral karyotype for 2 main groups of the Sorex genus. The chromosome painting evidence with human painting probes (HSA) reveals the presence of the 2 unique associations HSA4/5 and 1/10p/12/22b, which support Eulipotyphla. There are a series of synapomorphies both for Erinaceidae (HSA3/1/5, 3/17, 11/15 and 10/20) and for Soricinae (HSA5/9, 6/7/16, 8/3/21 and 11/12/22). We found associations that link Talpidae/Erinaceidae (HSA7/8, 1/5 and 1/19p), Talpidae/Soricidae (HSA1/8/4) and Erinaceidae/Soricidae (HSA4/20 and 2/13). Genome conservation in Eulipotyphla was estimated on the basis of the number of evolutionary breaks in the ancestral mammalian chromosomes. In total, 7 chromosomes of the boreo-eutherian ancestor (BEA8 or 10, 9, 17, 18, 20-22) were retained in all eulipotyphlans studied; among them moles show the highest level of chromosome conservation. The integration of sequence data into the chromosome painting information allowed us to further examine the chromosomal syntenies within a phylogenetic perspective. Based on our analysis we offer the most parsimonious reconstruction of phylogenetic relationships in Eulipotyphla. The cytogenetic reconstructions based on these data do not conflict with molecular phylogenies supporting basal position of Talpidae in the order.


CSF biomarkers and clinical progression of Parkinson disease.
To investigate whether certain CSF biomarkers at baseline can predict future progression of motor symptoms and cognitive decline in patients with Parkinson disease (PD). Patients and controls were recruited from hospitals in southern Sweden as part of the prospective and longitudinal Swedish BioFinder Study. In the present study, we included 42 patients with PD and 69 controls who had clinical assessment and lumbar puncture at baseline. Baseline CSF samples were analyzed for α-synuclein (αSyn), β-amyloid 1-42 (Aβ42), tau, phosphorylated tau, and neurofilament light. Associations between CSF markers at baseline and change in clinical characteristics after 2 years of follow-up were investigated using multivariate models adjusting for age, sex, disease duration, and levodopa-equivalent daily dose. Higher levels of αSyn within the PD group were associated with progression of motor symptoms and cognitive decline over 2 years, indicated by significant relationships between αSyn and change in Hoehn and Yahr (β = 0.394, p = 0.043), Unified Parkinson's Disease Rating Scale, Part III (UPDRS-III) (β = 0.449, p = 0.013), Timed Up and Go (β = 0.406, p = 0.023), and A Quick Test of Cognitive Speed (β = 0.423, p = 0.018). Lower levels of Aβ42 were associated with worsening of performance on delayed memory recall (F = 5.834, p = 0.022). Finally, high levels of phosphorylated tau were associated with worsening in motor symptoms (UPDRS-III, β = 0.350, p = 0.045; Hoehn and Yahr, β = 0.366, p = 0.038). We found evidence of a link between higher levels of αSyn at baseline and worsening of motor symptoms and cognitive speed over 2 years in PD. Increased αSyn might be a marker of more intense synaptic degeneration in PD. The results indicate that cortical amyloid pathology (low CSF Aβ42) is associated with memory decline.


Treatment Trends in Dry Eye Disease and Factors Associated with Ophthalmic Follow-up Discontinuation in Japan.
Despite the importance of dry eye disease (DED) treatment, the rate of DED treatment discontinuation, especially discontinuation of ophthalmic follow-up, remains unknown. This study aimed to assess the prevalence and risk factors of ophthalmic follow-up discontinuation for DED. A cross-sectional survey of 1030 participants was conducted using a self-administered web-survey instrument. We collected lifestyle information, history of DED diagnosis, types of treatment, frequency of eye-drop usage, symptoms, and the reasons for discontinuing treatment. Statistical analyses including logistic regression were used to evaluate the risk factors of discontinuing ophthalmic follow-up for DED. A past history of clinical DED diagnosis was reported by 155 (15.0%) subjects. Of those, 130 had persistent DED, and 88 (67.7%) of the subjects reported discontinuation of ophthalmic follow-up for DED. The most prevalent reasons for ophthalmic follow-up discontinuation were time restrictions, followed by dissatisfaction with the DED treatment. Duration after DED diagnosis was the only significant risk factor for discontinuing ophthalmic follow-up after adjusting for age and sex (odds ratio = 1.09, 95% confidence interval = 1.02-1.17, p = 0.009). In conclusion, longer DED duration after diagnosis was a significant risk factor for discontinuing ophthalmic follow-up for DED. This study showed that DED ophthalmic follow-up discontinuation involves both medical and non-medical reasons. Clinicians need to be aware of them, and preventative effort is needed to avoid discontinuation.


Evidence that phosphorylated ubiquitin signaling is involved in the etiology of Parkinson's disease.
The ubiquitin (Ub) kinase PINK1 and the E3 Ub ligase Parkin, two gene products associated with young-onset Parkinson's disease (PD), participate in mitochondrial quality control. The phosphorylation of mitochondrial polyUb by PINK1, which is activated in a mitochondrial membrane potential (ΔΨm)-dependent manner, facilitates the mitochondrial translocation and concomitant enzymatic activation of Parkin, leading to the clearance of phospho-polyUb-tagged mitochondria via mitophagy. Thus, Ub phosphorylation is a key event in PINK1-Parkin-mediated mitophagy. Here, we examined the role of phospho-Ub signaling in the pathogenesis of PD using fly PD models, human brain tissue and dopaminergic neurons derived from induced pluripotent stem cells (iPSCs) containing Parkin or PINK1 mutations, as well as normal controls. We report that phospho-Ub signaling is highly conserved between humans and Drosophila, and that phospho-Ub signaling and the relocation of axonal mitochondria upon ΔΨm reduction are indeed compromised in human dopaminergic neurons containing Parkin or PINK1 mutations. Moreover, phospho-Ub signaling is prominent in tyrosine hydroxylase-positive neurons compared with tyrosine hydroxylase-negative neurons, suggesting that PINK1-Parkin signaling is more required for dopaminergic neurons. These results shed light on the particular vulnerability of dopaminergic neurons to mitochondrial stress.


Transformations to additivity in measurement error models.
In many problems, one wants to model the relationship between a response Y and a covariate X. Sometimes it is difficult, expensive, or even impossible to observe X directly, but one can instead observe a substitute variable W that is easier to obtain. By far, the most common model for the relationship between the actual covariate of interest X and the substitute W is W = X + U, where the variable U represents measurement error. This assumption of additive measurement error may be unreasonable for certain data sets. We propose a new model, namely h(W) = h(X) + U, where h(.) is a monotone transformation function selected from some family H of monotone functions. The idea of the new model is that, in the correct scale, measurement error is additive. We propose two possible transformation families H. One is based on selecting a transformation that makes the within-sample mean and standard deviation of replicated W's uncorrelated. The second is based on selecting the transformation so that the errors (U's) fit a prespecified distribution. Transformation families used are the parametric power transformations and a cubic spline family. Several data examples are presented to illustrate the methods.


Rat Genome Database: a unique resource for rat, human, and mouse quantitative trait locus data.
The rat has been widely used as a disease model in a laboratory setting, resulting in an abundance of genetic and phenotype data from a wide variety of studies. These data can be found at the Rat Genome Database (RGD, http://rgd.mcw.edu/), which provides a platform for researchers interested in linking genomic variations to phenotypes. Quantitative trait loci (QTLs) form one of the earliest and core datasets, allowing researchers to identify loci harboring genes associated with disease. These QTLs are not only important for those using the rat to identify genes and regions associated with disease, but also for cross-organism analyses of syntenic regions on the mouse and the human genomes to identify potential regions for study in these organisms. Currently, RGD has data on >1,900 rat QTLs that include details about the methods and animals used to determine the respective QTL along with the genomic positions and markers that define the region. RGD also curates human QTLs (>1,900) and houses>4,000 mouse QTLs (imported from Mouse Genome Informatics). Multiple ontologies are used to standardize traits, phenotypes, diseases, and experimental methods to facilitate queries, analyses, and cross-organism comparisons. QTLs are visualized in tools such as GBrowse and GViewer, with additional tools for analysis of gene sets within QTL regions. The QTL data at RGD provide valuable information for the study of mapped phenotypes and identification of candidate genes for disease associations.


Long-term occupational exposures on disability-free survival and mortality in older adults.
The impact of long-term occupational exposures on health in older adults is increasingly relevant as populations age. To date, no studies have reported their impact on survival free of disability in older adults. We aimed to investigate the association between long-term occupational exposure and disability-free survival (DFS), all-cause mortality and cause-specific mortality in initially healthy older adults. We analysed data from 12 215 healthy participants in the ASPirin in Reducing Events in the Elderly (ASPREE) study whose mean age was 75 years. Their work history was collated with the 'ALOHA-plus JEM' (Job Exposure Matrix) to assign occupational exposures. The primary endpoint, DFS, was a composite measure of death, dementia or persistent physical disability. The secondary endpoint, mortality, was classified according to the underlying cause. Cox proportional hazard models were used to calculate hazard ratios and 95% confidence intervals, adjusted for confounders. A total of 1835 individuals reached the DFS endpoint during the median 4.7 years follow-up period. Both ever-high and cumulative exposure to all dusts and all pesticides during a person's working years were associated with reduced DFS. Compared to no exposure, men with high exposure to dusts and pesticides had a reduced DFS. Neither of these exposures were significantly associated with all-cause mortality. Men with high occupational exposure to solvents and women exposed to dusts experienced higher all-cause and cancer-related mortality. Long-term occupational exposure to all dusts and pesticides was associated with a reduced DFS and increased mortality in community-dwelling healthy older adults.


Contribution of CYP2E1 polymorphism to aging in the mechanical workshop workers.
Aging is attributed to both genetic and environmental factors. Occupational exposure is one of the environmental factors with potential genotoxic effects. Researchers try to determine factors involved in genetic damages at hazards exposure that could accelerate aging. Cytochrome P450 2E1 (CYP2E1) gene contributes in activation and detoxification of the environmental hazards. This polymorphism plays an important role in susceptibility of inter-individuals to DNA damage at the occupational exposure. The current study evaluated the possible influence of this gene polymorphism in aging by genomic damages through the biomarkers alterations of micronuclei (MN), comet tail length and telomere length shortening at the exposure. In this study, buccal cells were collected from the oral cavity of exposed workers and non-exposed controls. The CYP2E1 genotypes were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The wild genotype significantly affected MN frequency (p = 0.007) and relative telomere length (p = 0.047) in the older group of workers. It was concluded that the interaction of gene polymorphism and exposure enhances DNA damage and accelerates aging consequently.


Metronomic PDT and cell death pathways.
The term "metronomic" was recently introduced to describe continuous low-dose administration of chemotherapeutics following the discovery that this causes minimal side effects (Hanahan et al. 2000, J Clin Invest, 105(8), 1045-1047; Bisland et al. 2004, Photochem Photobiol, 80, 22-30). Metronomic dosing in PDT is proposed by analogy and the rationale is as a means to improve the tumor-specific response through cell death by apoptosis. We investigated the molecular mechanisms associated with apoptosis following ALA-PDT treatment in two brain glioma cell lines, namely U87 (human) and CNS-1 (rat) cells. We used the high energy of light at a short time (acute PDT) and the low energy of light at a long time of exposure (metronomic PDT) to treat both cell lines. To identify potential cell death pathways associated with metronomic PDT, microarray analysis of gene expression was conducted on RNA from glioblastoma cells with metronomic ALA-PDT. The apoptosis mechanism for metronomic ALA-PDT occurred via the inhibition of LTbetaR and the transcription factor NF-kappaB. This inhibition was ALA concentration dependent.


High-sensitivity C-reactive protein in heart failure with preserved ejection fraction.
Microvascular inflammation may contribute to the pathogenesis of both heart failure with preserved ejection fraction (HFpEF) and pulmonary hypertension (PH). We investigated whether the inflammation biomarker C-reactive protein (CRP) was associated with clinical characteristics, disease severity or PH in HFpEF. Patients in the Phosphodiesterase-5 Inhibition to Improve Clinical Status and Exercise Capacity in Diastolic Heart failure (RELAX) trial had baseline high-sensitivity CRP levels measured (n = 214). Clinical characteristics, exercise performance, echocardiographic variables and biomarkers of neurohumoral activation, fibrosis and myocardial necrosis were assessed. Patients with normal (≤3mg/L) versus high (>3mg/L) CRP levels were compared. The median CRP level was 3.69mg/L. CRP was elevated in 57% of patients. High CRP levels were associated with younger age, higher body mass index (BMI), chronic obstructive pulmonary disease (COPD), lower peak oxygen consumption and higher endothelin-1 and aldosterone levels. CRP increased progressively with the number of comorbidities (0.7mg/L per increment in comorbidity number, P = 0.02). Adjusting for age, BMI and statin use, high CRP levels were additionally associated with atrial fibrillation, right ventricular dysfunction, and higher N-terminal pro-B-type natriuretic peptide levels (P<0.05 for all). CRP was not associated with PH or left ventricular function. CRP did not identify responders to sildenafil(P-value for interaction 0.13). In HFpEF, high CRP is associated with greater comorbidity burden and some markers of disease severity but CRP was normal in 40% of patients. These findings support the presence of comorbidity-driven systemic inflammation in HFpEF but also the need to study other biomarkers which may better reflect the presence of systemic inflammation.


Pharmacological reduction of NEFA restores the efficacy of incretin-based therapies through GLP-1 receptor signalling in the beta cell in mouse models of diabetes.
Type 2 diabetes mellitus is associated with reduced incretin effects. Although previous studies have shown that hyperglycaemia contributes to impaired incretin responses in beta cells, it is largely unknown how hyperlipidaemia, another feature of type 2 diabetes, contributes to impaired glucagon-like peptide 1 (GLP-1) response. Here, we investigated the effects of NEFA on incretin receptor signalling and examined the glucose-lowering efficacy of incretin-based drugs in combination with the lipid-lowering agent bezafibrate. We used db/db mice to examine the in vivo efficacy of the treatment. Beta cell lines and mouse islets were used to examine GLP-1 and glucose-dependent insulinotropic peptide receptor signalling. Palmitate treatment decreased Glp1r expression in rodent insulinoma cell lines and isolated islets. This was associated with impairment of the following: GLP-1-stimulated cAMP production, phosphorylation of cAMP-responsive elements binding protein (CREB) and insulin secretion. In insulinoma cell lines, the expression of exogenous Glp1r restored cAMP production and the phosphorylation of CREB. Treatment with bezafibrate in combination with des-fluoro-sitagliptin or exendin-4 led to more robust glycaemic control, associated with improved islet morphology and beta cell mass in db/db mice. Elevated NEFA contributes to impaired responsiveness to GLP-1, partially through downregulation of GLP-1 receptor signalling. Improvements in lipid control in mouse models of obesity and diabetes increase the efficacy of incretin-based therapy.


Mental illness and well-being: the central importance of positive psychology and recovery approaches.
A new evidence base is emerging, which focuses on well-being. This makes it possible for health services to orientate around promoting well-being as well as treating illness, and so to make a reality of the long-standing rhetoric that health is more than the absence of illness. The aim of this paper is to support the re-orientation of health services around promoting well-being. Mental health services are used as an example to illustrate the new knowledge skills which will be needed by health professionals. New forms of evidence give a triangulated understanding about the promotion of well-being in mental health services. The academic discipline of positive psychology is developing evidence-based interventions to improve well-being. This complements the results emerging from synthesising narratives about recovery from mental illness, which provide ecologically valid insights into the processes by which people experiencing mental illness can develop a purposeful and meaningful life. The implications for health professionals are explored. In relation to working with individuals, more emphasis on the person's own goals and strengths will be needed, with integration of interventions which promote well-being into routine clinical practice. In addition, a more societally-focussed role for professionals is envisaged, in which a central part of the job is to influence local and national policies and practices that impact on well-being. If health services are to give primacy to increasing well-being, rather than to treating illness, then health workers need new approaches to working with individuals. For mental health services, this will involve the incorporation of emerging knowledge from recovery and from positive psychology into education and training for all mental health professionals, and changes to some long-established working practices.


Reconsidering the effects of seat belt laws and their enforcement status.
The debate over the benefits of mandatory seat belt laws and their enforcement status has focused on a controversial empirical enigma: why have these policies, which appear to have increased belt use sharply, had a relatively small impact on traffic fatalities? In this paper, I offer new insights into this question by examining panel data on observed belt use from the National Highway Traffic Safety Administration and self-reported data on belt use from pooled cross-sections of the Centers for Disease Control and Prevention's 1985-1993 Behavioral Risk Factor Surveillance System. By exploiting the panel nature of these data, I demonstrate that prior estimates, which have not conditioned on the unobserved time-varying determinants of belt use, have dramatically overestimated the impact of seat belt laws and their enforcement status on belt use. The true effects are more consistent with the modest impact these policies have had on traffic fatalities without having to appeal to the possibility of risk compensation by drivers. However, I find strong evidence in support of the selective recruitment hypothesis. Belt use among those most likely to be involved in traffic accidents (e.g. males, drinkers of alcohol, the young) has been significantly less responsive to seat belt laws and their enforcement status.


Identification and evolution of a plant cell wall specific glycoprotein glycosyl transferase, ExAD.
Extensins are plant cell wall glycoproteins that act as scaffolds for the deposition of the main wall carbohydrate polymers, which are interlocked into the supramolecular wall structure through intra- and inter-molecular iso-di-tyrosine crosslinks within the extensin backbone. In the conserved canonical extensin repeat, Ser-Hyp4, serine and the consecutive C4-hydroxyprolines (Hyps) are substituted with an α-galactose and 1-5 β- or α-linked arabinofuranoses (Arafs), respectively. These modifications are required for correct extended structure and function of the extensin network. Here, we identified a single Arabidopsis thaliana gene, At3g57630, in clade E of the inverting Glycosyltransferase family GT47 as a candidate for the transfer of Araf to Hyp-arabinofuranotriose (Hyp-β1,4Araf-β1,2Araf-β1,2Araf) side chains in an α-linkage, to yield Hyp-Araf4 which is exclusively found in extensins. T-DNA knock-out mutants of At3g57630 showed a truncated root hair phenotype, as seen for mutants of all hitherto characterized extensin glycosylation enzymes; both root hair and glycan phenotypes were restored upon reintroduction of At3g57630. At3g57630 was named Extensin Arabinose Deficient transferase, ExAD, accordingly. The occurrence of ExAD orthologs within the Viridiplantae along with its' product, Hyp-Araf4, point to ExAD being an evolutionary hallmark of terrestrial plants and charophyte green algae.


Endothelial cell tetrahydrobiopterin deficiency attenuates LPS-induced vascular dysfunction and hypotension.
Overproduction of nitric oxide (NO) is thought to be a key mediator of the vascular dysfunction and severe hypotension in patients with endotoxaemia and septic shock. The contribution of NO produced directly in the vasculature by endothelial cells to the hypotension seen in these conditions, vs. the broader systemic increase in NO, is unclear. To determine the specific role of endothelium derived NO in lipopolysaccharide (LPS)-induced vascular dysfunction we administered LPS to mice deficient in endothelial cell tetrahydrobiopterin (BH4), the essential co-factor for NO production by NOS enzymes. Mice deficient in endothelial BH4 production, through loss of the essential biosynthesis enzyme Gch1 (Gch1(fl/fl)Tie2cre mice) received a 24hour challenge with LPS or saline control. In vivo LPS treatment increased vascular GTP cyclohydrolase and BH4 levels in aortas, lungs and hearts, but this increase was significantly attenuated in Gch1(fl/fl)Tie2cre mice, which were also partially protected from the LPS-induced hypotension. In isometric tension studies, in vivo LPS treatment reduced the vasoconstriction response and impaired endothelium-dependent and independent vasodilatations in mesenteric arteries from wild-type mice, but not in Gch1(fl/fl)Tie2cre mesenteric arteries. Ex vivo LPS treatment decreased vasoconstriction response to phenylephrine in aortic rings from wild-type and not in Gch1(fl/fl)Tie2cre mice, even in the context of significant eNOS and iNOS upregulation. These data provide direct evidence that endothelial cell NO has a significant contribution to LPS-induced vascular dysfunction and hypotension and may provide a novel therapeutic target for the treatment of systemic inflammation and patients with septic shock.


Vascular endothelial growth factor C-induced lymphangiogenesis decreases tumor interstitial fluid pressure and tumor.
Characteristically, most solid tumors exhibit an increased tumor interstitial fluid pressure (TIFP) that directly contributes to the lowered uptake of macromolecular therapeutics into the tumor interstitium. Abnormalities in the tumor-associated lymph vessels are a central brick in the development and prolonged sustaining of an increased TIFP. In the current study, vascular endothelial growth factor C (VEGF-C) was used to enhance tumor-associated lymphangiogenesis as a new mechanism to actively reduce the TIFP by increased lymphatic drainage of the tumor tissue. Human A431 epidermoid vulva carcinoma cells were inoculated in NMRI nu/nu mice to generate a xenograft mouse model. Seven days after tumor cell injection, VEGF-C was peritumorally injected to induce lymphangiogenesis. Tumor growth and TIFP was lowered significantly over time in VEGF-C-treated tumors in comparison to control or VEGF-A-treated animals. These data demonstrate for the first time that actively induced lymphangiogenesis can lower the TIFP in a xenograft tumor model and apparently reduce tumor growth. This model represents a novel approach to modulate biomechanical properties of the tumor interstitium enabling a lowering of TIFP in vivo.


Human epidermal growth factor receptor 2 expression in gastric carcinoma and its association with histopathological parameters in Indian population.
Gastric carcinoma is a leading cause of death worldwide with a five year survival of 10-15% even after curative resection. Trastuzumab has emerged as a potential targeted therapy in treatment of Her 2 positive gastric cancer. Her2 positivity ranges from 7-34% in studies across the world. There is a paucity of Indian studies hence a need for determination of Her2 expression in Indian population for better patient management. This study was carried out to determine the frequency of Her 2 expression in gastric carcinoma by immunohistochemistry (IHC) technique and to evaluate its association with histopathological parameters. A total of 110 cases of gastric adenocarcinoma diagnosed on histopathological examination from July 2013 to June 2015 were included. Of these, 40 cases were resection specimens and 70 were biopsies. Histological typing of gastric carcinoma was done on the basis of Lauren classification. IHC for Her2 was done in all 110 cases. Her 2 expression was correlated with various histopathological parameters. Positive Her 2 expression (IHC 3+) was seen in 24.5% cases of gastric carcinoma. Patients in older age group(> 60 years) showed higher Her 2 positivity rate as compared to middle age (40-60 years) and younger population (<40 years). Higher percentage of Her 2 positivity was noted in male patients as compared to female patients. Her 2 positivity was seen more commonly in intestinal type of tumor as compared to diffuse and mixed types. Her 2 positivity was seen more in well differentiated carcinoma and higher stage tumors (pT3 and pT4). However, out of all, a statistically significant association of Her2 expression was found only with (intestinal) type of tumors (p= 0.005) and no significant association was seen with age, gender, site of tumor, tumor stage, lymph node status, lymphovascular, perineural and perinodal invasion or survival. Inspired by promising results of Trastuzumab therapy in Her 2 positive gastric carcinoma worldwide, it is recommended to routinely test all cases of gastric carcinoma for Her 2 expression and to use the targeted therapy with curative intentions in Indian population.


Blockage of cell-to-cell communication within pancreatic acini is associated with increased basal release of amylase.
To assess whether junctional coupling is involved in the secretory activity of pancreatic acinar cells, dispersed rat acini were incubated for 30 min in the presence of either heptanol (3.5 mM) or octanol (1.0 mM). Exposure to either alkanol caused a marked uncoupling of the acinar cells which, in control acini, were extensively coupled. Uncoupling was associated with an increased basal release of amylase that was at least twice that of controls. By contrast, carbamylcholine (10(-5) M)-induced maximal amylase secretion, cytosolic pH, and free Ca2+, as well as the structure of gap junctions joining the acinar cells, were unaffected. Both uncoupling and the alteration of basal secretion were already observed after only 5 min of exposure to heptanol, they both persisted throughout the 30-min exposure to the alkanols, and were reversible after removal of either heptanol or octanol. Since neither of the two uncouplers appeared to alter unspecifically the secretory machinery and the nonjunctional membrane of acinar cells, the data are consistent with the view that junctional coupling participates in the control of the basal secretion of acinar cells.


Effects of purified eicosapentaenoic and docosahexaenoic acids in nonalcoholic fatty liver disease: results from the Welcome* study.
There is no licensed treatment for non-alcoholic fatty liver disease (NAFLD), a condition that increases risk of chronic liver disease, type 2 diabetes and cardiovascular disease. We tested whether 15-18 months treatment with docosahexaenoic acid (DHA) plus eicosapentaenoic acid (EPA) (Omacor/Lovaza) (4 g/day) decreased liver fat and improved two histologically-validated liver fibrosis biomarker scores (primary outcomes). Patients with NAFLD were randomised in a double blind placebo-controlled trial [DHA+EPA(n=51), placebo(n=52)]. We quantified liver fat percentage (%) by magnetic resonance spectroscopy in three liver zones. We measured liver fibrosis using two validated scores. We tested adherence to the intervention (Omacor group) and contamination (with DHA and EPA) (placebo group) by measuring erythrocyte percentage DHA and EPA enrichment (gas chromatography). We undertook multivariable linear regression to test effects of: a) DHA+EPA treatment (ITT analyses) and b) erythrocyte DHA and EPA enrichment (secondary analysis). Median (IQR) baseline and end of study liver fat% were 21.7 (19.3) and 19.7 (18.0) (placebo), and 23.0 (36.2) and 16.3 (22.0), (DHA+EPA). In the fully adjusted regression model there was a trend towards improvement in liver fat% with DHA+EPA treatment (β=-3.64 (95%CI -8.0,0.8); p=0.1) but there was evidence of contamination in the placebo group and variable adherence to the intervention in the Omacor group. Further regression analysis showed that DHA enrichment was independently associated with a decrease in liver fat% (for each 1% enrichment, β=-1.70 (95%CI -2.9,-0.5); p=0.007). No improvement in the fibrosis scores occurred. Conclusion. Erythrocyte DHA enrichment with DHA+EPA treatment is linearly associated with decreased liver fat%. Substantial decreases in liver fat% can be achieved with high percentage erythrocyte DHA enrichment in NAFLD. (Hepatology 2014;).


Lattice-induced frequency shifts in Sr optical lattice clocks at the 10(-17) level.
We present a comprehensive study of the frequency shifts associated with the lattice potential in a Sr lattice clock by comparing two such clocks with a frequency stability reaching 5×10(-17) after a 1 h integration time. We put the first experimental upper bound on the multipolar M1 and E2 interactions, significantly smaller than the recently predicted theoretical upper limit, and give a 30-fold improved upper limit on the effect of hyperpolarizability. Finally, we report on the first observation of the vector and tensor shifts in a Sr lattice clock. Combining these measurements, we show that all known lattice related perturbations will not affect the clock accuracy down to the 10(-17) level, even for lattices as deep as 150 recoil energies.


Clinical evaluation of posterior composite restorations: the 10-year report.
To summarize all clinically relevant data of a 10-year prospective evaluation of posterior glass-ionomer cement/composite restorations (Ketac Bond/Visio-Molar radiopaque). For this study 194 hybrid composite fillings (115 Class I, 79 Class II) were applied in 73 adult patients. The exposed dentin was covered with glass-ionomer cement. The clinical evaluation was carried out at baseline, after 6 months and at 1-year intervals up to 10 years. The USPHS-compatible CPM Index was used with the C and P criteria presented here for anatomic form, color match, surface quality, wear, marginal integrity, marginal ledge, marginal discoloration, secondary caries, and clinical acceptability. Thermal and electrical sensitivity testing were recorded annually. At baseline, 194 restorations were rated. The drop-out of patients comprised 108 fillings after 10 years. During this time, 24 restorations could not be followed up because of new carious lesions at different sites and/or prosthodontic treatment (drop-out of teeth). Forty-six composite restorations were longitudinally controlled over 10 years, and 16 more fillings failed before the end of the study (3 filling fractures, 7 partial filling losses, 1 total filling loss, 5 cases of secondary caries). Only Class II composite restorations exhibited secondary caries (4 fillings after 7 years and 1 filling after 9 years). After 10 years, the correct anatomic form was preserved in 44 fillings, but all restorations showed rough surface and wear. The majority of restorations (26 fillings) was free of marginal discoloration, and the color match showed only 3 fillings being too dark. The marginal integrity was optimal at baseline and during the first year. During the second year a continuing degradation started and lasted until the end of the study. Thirteen restorations showed optimal marginal integrity after 10 years. The USPHS-compatible CPM Index represents a sensitive rating system for long-term observation of posterior composite restorations. The early risk of failure is attributed to bulk fractures and partial loss of filling material. The longevity over 10 years is a maximum of 74.2%, and the very low secondary caries rate and the high percentage of correct anatomical form confirm the clinical safety of posterior composite restorations.


Statistical false positive or true disease pathway?
Three very recent reports provide convincing statistical evidence (P < 10(-8)), at a genome-wide level, of the association of common polymorphisms with three different common diseases: systemic lupus erythematosus (IRF5), prostate cancer and type 1 diabetes (IFIH1 region). This adds to the trickle--soon to be a flood--of disease association results that are highly unlikely to be false positives. There are other convincing examples in the last 12 months: age-related macular degeneration (CFH), type 1 diabetes (IL2RA, also known as CD25) and type 2 diabetes (TCF7L2). Given 20 years of a literature full of irreproducible results, what has changed?


Modification of the existing maximum residue levels for penthiopyrad in Florence fennels and celeries.
In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Corteva submitted a request to the competent national authority in Italy to modify the existing maximum residue levels (MRLs) for the active substance penthiopyrad in Florence fennels and celeries. The data submitted in support of the request were found to be sufficient to derive MRL proposals for Florence fennels and celeries. Adequate analytical methods for enforcement are available to control the residues of penthiopyrad in the commodities under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the short-term and long-term intake of penthiopyrad residues and of its metabolite PAM, resulting from the use of penthiopyrad according to the reported agricultural practice is unlikely to present a risk to consumer health. The reliable end points, appropriate for use in regulatory risk assessment, are presented.


The Ro/SSA autoantigen as an immunogen. Some anti-Ro/SSA antibody binds IgG.
The rheumatic disease autoantigen, Ro/SSA, was immunogenic to a rabbit host. The heteroimmune rabbit serum bound the Ro/SSA particle in immunoblots and in an ELISA. Both the rabbit anti-Ro/SSA and a human prototype anti-Ro/SSA serum also bound IgG; and moreover, IgG inhibited both rabbit and human anti-Ro/SSA activity. Anti-IgG activity of the rabbit and human anti-Ro/SSA sera bound Ro/SSA by Western blot and solid-phase assays. In addition, purified Ro/SSA inhibited the anti-IgG activity of the anti-Ro/SSA sera from rabbit and man. Affinity purification of the IgG- and Ro/SSA-binding fractions of the rabbit anti-Ro/SSA demonstrated that both the anti-Ro/SSA and anti-IgG activities were concentrated in these fractions. These data show that Ro/SSA and IgG share epitopes that are bound by anti-Ro/SSA antibody. Inhibition experiments suggest that this antibody is found in most human anti-Ro/SSA autoimmune sera and that the epitope(s) are found in the F(ab')2 fragment of IgG.


Predicted for greatness: 1994 molecule of the year--the DNA repair enzyme.
Lung cancer mortality is the highest of any cancer. Primary prevention has stalled, however, new lung cancer screening trials of low-dose computerized tomography (LDCT) have shown that the mortality from lung cancer can be reduced by up to 20% among current and former smokers. There are potential harms that must be taken into account when evaluating any screening program. With LDCT, there is a 90% rate of false positives and the potential for high doses of radiation from subsequent workup of benign lesions. The development of biomarkers that might refine the ability of screening to identify individuals at high risk for developing and dying from lung cancer is a ripe area for investigation. Sevilya and colleagues have developed a highly promising set of biomarkers of DNA repair capacity that may satisfy that goal. The large estimate of risk, the thoughtful combination of functional assays of DNA repair capacity, and the population-based design of the study make it reasonable to test these biomarkers in a larger study.


Phospholipase A2 activation as a therapeutic approach for cognitive enhancement in early-stage Alzheimer disease.
Alzheimer disease (AD) is the leading cause of dementia in the elderly and has no known cure. Evidence suggests that reduced activity of specific subtypes of intracellular phospholipases A2 (cPLA2 and iPLA2) is an early event in AD and may contribute to memory impairment and neuropathology in the disease. The objective of this study was to review the literature focusing on the therapeutic role of PLA2 stimulation by cognitive training and positive modulators, or of supplementation with arachidonic acid (PLA2 product) in facilitating memory function and synaptic transmission and plasticity in either research animals or human subjects. MEDLINE database was searched (no date restrictions) for published articles using the keywords Alzheimer disease (mild, moderate, severe), mild cognitive impairment, healthy elderly, rats, mice, phospholipase A(2), phospholipid metabolism, phosphatidylcholine, arachidonic acid, cognitive training, learning, memory, long-term potentiation, protein kinases, dietary lipid compounds, cell proliferation, neurogenesis, and neuritogenesis. Reference lists of the identified articles were checked to select additional studies of interest. Overall, the data suggest that PLA2 activation is induced in the healthy brain during learning and memory. Furthermore, learning seems to regulate endogenous neurogenesis, which has been observed in AD brains. Finally, PLA2 appears to be implicated in homeostatic processes related to neurite outgrowth and differentiation in both neurodevelopmental processes and response to neuronal injury. The use of positive modulators of PLA2 (especially of cPLA2 and iPLA2) or supplementation with dietary lipid compounds (e.g., arachidonic acid) in combination with cognitive training could be a valuable therapeutic strategy for cognitive enhancement in early-stage AD.


Morphological divergence between three Arctic charr morphs - the significance of the deep-water environment.
Morphological divergence was evident among three sympatric morphs of Arctic charr (Salvelinus alpinus (L.)) that are ecologically diverged along the shallow-, deep-water resource axis in a subarctic postglacial lake (Norway). The two deep-water (profundal) spawning morphs, a benthivore (PB-morph) and a piscivore (PP-morph), have evolved under identical abiotic conditions with constant low light and temperature levels in their deep-water habitat, and were morphologically most similar. However, they differed in important head traits (e.g., eye and mouth size) related to their different diet specializations. The small-sized PB-morph had a paedomorphic appearance with a blunt head shape, large eyes, and a deep body shape adapted to their profundal lifestyle feeding on submerged benthos from soft, deep-water sediments. The PP-morph had a robust head, large mouth with numerous teeth, and an elongated body shape strongly related to their piscivorous behavior. The littoral spawning omnivore morph (LO-morph) predominantly utilizes the shallow benthic-pelagic habitat and food resources. Compared to the deep-water morphs, the LO-morph had smaller head relative to body size. The LO-morph exhibited traits typical for both shallow-water benthic feeding (e.g., large body depths and small eyes) and planktivorous feeding in the pelagic habitat (e.g., streamlined body shape and small mouth). The development of morphological differences within the same deep-water habitat for the PB- and PP-morphs highlights the potential of biotic factors and ecological interactions to promote further divergence in the evolution of polymorphism in a tentative incipient speciation process. The diversity of deep-water charr in this study represents a novelty in the Arctic charr polymorphism as a truly deep-water piscivore morph has to our knowledge not been described elsewhere.