Comparison of Ensemble and Single Molecule Methods for Particle Characterization and Binding Analysis of a PEGylated Single-Domain Antibody.
Domain antibodies (dAbs) are single immunoglobulin domains that form the smallest functional unit of an antibody. This study investigates the behavior of these small proteins when covalently attached to the polyethylene glycol (PEG) moiety that is necessary for extending the half-life of a dAb. The effect of the 40 kDa PEG on hydrodynamic properties, particle behavior, and receptor binding of the dAb has been compared by both ensemble solution and surface methods [light scattering, isothermal titration calorimetry (ITC), surface Plasmon resonance (SPR)] and single-molecule atomic force microscopy (AFM) methods (topography, recognition imaging, and force microscopy). The large PEG dominates the properties of the dAb-PEG conjugate such as a hydrodynamic radius that corresponds to a globular protein over four times its size and a much reduced association rate. We have used AFM single-molecule studies to determine the mechanism of PEG-dependent reductions in the effectiveness of the dAb observed by SPR kinetic studies. Recognition imaging showed that all of the PEGylated dAb molecules are active, suggesting that some may transiently become inactive if PEG sterically blocks binding. This helps explain the disconnect between the SPR, determined kinetically, and the force microscopy and ITC results that demonstrated that PEG does not change the binding energy.


Writing and probing light-induced waveguides thanks to an endlessly single-mode photonic crystal fiber.
We demonstrate writing and probing of light-induced waveguides in photorefractive bulk LiNbO3 crystal using an endlessly single-mode photonic crystal fiber. The optical waveguides are written at visible wavelengths by slightly raising the ferroelectric crystal temperature to benefit from the pyroelectric-driven photorefractive effect and the guiding properties are investigated at telecom wavelengths using the same photonic crystal fiber. End butt coupling with this photonic crystal fiber enables writing and probing of optical waveguides due to the self-alignment properties of spatial solitons.


Both information and social cohesion determine collective decisions in animal groups.
During consensus decision making, individuals in groups balance personal information (based on their own past experiences) with social information (based on the behavior of other individuals), allowing the group to reach a single collective choice. Previous studies of consensus decision making processes have focused on the informational aspects of behavioral choice, assuming that individuals make choices based solely on their likelihood of being beneficial (e.g., rewarded). However, decisions by both humans and nonhuman animals systematically violate such expectations. Furthermore, the typical experimental paradigm of assessing binary decisions, those between two mutually exclusive options, confounds two aspects common to most group decisions: minimizing uncertainty (through the use of personal and social information) and maintaining group cohesion (for example, to reduce predation risk). Here we experimentally disassociate cohesion-based decisions from information-based decisions using a three-choice paradigm and demonstrate that both factors are crucial to understanding the collective decision making of schooling fish. In addition, we demonstrate how multiple informational dimensions (here color and stripe orientation) are integrated within groups to achieve consensus, even though no individual is explicitly aware of, or has a unique preference for, the consensus option. Balancing of personal information and social cues by individuals in key frontal positions in the group is shown to be essential for such group-level capabilities. Our results demonstrate the importance of integrating informational with other social considerations when explaining the collective capabilities of group-living animals.


Serum testing of genetically modified soybeans with special emphasis on potential allergenicity of the heterologous protein CP4 EPSPS.
Roundup Ready soy contains the CP4-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein. Serum IgE from two distinct populations of soy-allergic patients were recruited to determine their IgE-binding specificity. One population consisted of 10 adult patients from Europe, whose primary diagnosis was soy food allergy with some also having mite allergy. In addition, 6 primarily mite-allergic, 6 food-allergic (celery, carrot, milk, shrimp, walnut, and apple), and 5 non-allergic patients were tested. Another population consisted of 13 children from Korea, whose primary diagnosis was atopic dermatitis and secondarily soy and egg sensitization. In addition, 11 non-allergic patients were tested. Each patient population was extensively characterized with respect to clinical symptoms, specific IgE (CAP) scores, and total IgE. Immunoblots and ELISA assays were developed using serum IgE from these patients and soy extracts, CP4 EPSPS, rice extract, ovalbumin, rubisco, purified major peanut allergen Ara h 2, the putative soy allergen Gly m Bd 30k and mite allergen Der f 2 proteins as the intended targets. Immunoblot results indicated that soy-allergic patients bound soy extracts but did not specifically bind rubisco or CP4 EPSPS. ELISA results were in general agreement with the immunoblot results except that rubisco bound significant quantities of serum IgE from some patients. These results indicate that the CP4 EPSPS protein does not bind significant quantities of IgE from two geographically distinct sensitive populations and there is no evidence for an increased allergenic potential of this biotech protein.


Estrogen metabolism and mammographic density in postmenopausal women: a cross-sectional study.
Prospective studies have consistently found that postmenopausal breast cancer risk increases with circulating estrogens; however, findings from studies of estrogens and mammographic density (MD), an intermediate marker of breast cancer risk, have been inconsistent. We investigated the cross-sectional associations of urinary estrogens, and their 2-, 4-, and 16-hydroxylated metabolites with MD. Postmenopausal women without breast cancer (n = 194), ages 48 to 82 years, and reporting no current menopausal hormone therapy use were enrolled at a clinic in Western NY in 2005. Urinary estrogens and estrogen metabolites were measured using mass spectrometry. Percent MD and dense area (cm(2)) were measured using computer-assisted analyses of digitized films. Linear regression models were used to estimate associations of log-transformed estrogen measures with MD while adjusting for age, body mass index (BMI), parity, and past hormone therapy use. Urinary concentrations of most individual estrogens and metabolites were not associated with MD; however, across the interdecile range of the ratio of parent estrogens (estrone and estradiol) to their metabolites, MD increased by 6.8 percentage points (P = 0.02) and dense area increased by 10.3 cm(2) (P = 0.03). Across the interdecile ranges of the ratios of 2-, 4-, and 16-hydroxylation pathways to the parent estrogens, MD declined by 6.2 (P = 0.03), 6.4 (P = 0.04), and 5.7 (P = 0.05) percentage points, respectively. All associations remained apparent in models without adjustment for BMI. In this study of postmenopausal women, less extensive hydroxylation of parent estrogens was associated with higher MD. Hydroxylation of estrogens may modulate postmenopausal breast cancer risk through a pathway involving MD.


AGSE: A Novel Grape Seed Extract Enriched for PP2A Activating Flavonoids That Combats Oxidative Stress and Promotes Skin Health.
Environmental stimuli attack the skin daily resulting in the generation of reactive oxygen species (ROS) and inflammation. One pathway that regulates oxidative stress in skin involves Protein Phosphatase 2A (PP2A), a phosphatase which has been previously linked to Alzheimer's Disease and aging. Oxidative stress decreases PP2A methylation in normal human dermal fibroblasts (NHDFs). Thus, we hypothesize agents that increase PP2A methylation and activity will promote skin health and combat aging. To discover novel inhibitors of PP2A demethylation activity, we screened a library of 32 natural botanical extracts. We discovered Grape Seed Extract (GSE), which has previously been reported to have several benefits for skin, to be the most potent PP2A demethylating extract. Via several fractionation and extraction steps we developed a novel grape seed extract called Activated Grape Seed Extract (AGSE), which is enriched for PP2A activating flavonoids that increase potency in preventing PP2A demethylation when compared to commercial GSE. We then determined that 1% AGSE and 1% commercial GSE exhibit distinct gene expression profiles when topically applied to a 3D human skin model. To begin to characterize AGSE's activity, we investigated its antioxidant potential and demonstrate it reduces ROS levels in NHDFs and cell-free assays equal to or better than Vitamin C and E. Moreover, AGSE shows anti-inflammatory properties, dose-dependently inhibiting UVA, UVB and chemical-induced inflammation. These results demonstrate AGSE is a novel, multi-functional extract that modulates methylation levels of PP2A and supports the hypothesis of PP2A as a master regulator for oxidative stress signaling and aging in skin.


Cannabinoid receptor type-1: breaking the dogmas.
The endocannabinoid system (ECS) is abundantly expressed in the brain. This system regulates a plethora of physiological functions and is composed of cannabinoid receptors, their endogenous ligands (endocannabinoids), and the enzymes involved in the metabolism of endocannabinoids. In this review, we highlight the new advances in cannabinoid signaling, focusing on a key component of the ECS, the type-1 cannabinoid receptor (CB 1). In recent years, the development of new imaging and molecular tools has demonstrated that this receptor can be distributed in many cell types (e.g., neuronal or glial cells) and intracellular compartments (e.g., mitochondria). Interestingly, cellular and molecular effects are differentially mediated by CB 1 receptors according to their specific localization (e.g., glutamatergic or GABAergic neurons). Moreover, this receptor is expressed in the periphery, where it can modulate periphery-brain connections. Finally, the better understanding of the CB 1 receptor structure led researchers to propose interesting and new allosteric modulators. Thus, the advances and the new directions of the CB 1 receptor field will provide new insights and better approaches to profit from its interesting therapeutic profile.


Pathologic alterations in the diabetic neuropathies of humans: a review.
Pathologic study of nerve tissue is especially useful for the recognition of interstitial events such as inflammation or vascular alterations that cannot be inferred from clinical or electrophysiologic findings and may provide insight into an underlying mechanism or cause. Considerable variation in the natural history and pathologic alterations among diabetic neuropathies suggests that they are heterogeneous. For diabetic polyneuropathy, two mechanisms need to be considered. The first assumes that hyperglycemia induces metabolic derangements that directly affect Schwann cells (or myelin), nodes of Ranvier, or axons. The second assumes that hyperglycemia and metabolic derangement affect the structure and function of endoneurial microvessels, which then induce fiber changes by altering the blood-nerve barrier, inducing hypoxia or ischemia, or by unknown mechanisms. In proximal diabetic neuropathy, there is increasing evidence that the characteristic lesion is an inflammatory (immune) vasculitis that induces ischemic nerve fiber degeneration. Truncal radiculopathy may be due to an inflammatory polyganglionopathy. In cranial nerve III neuropathy, the monophasic course and the pathologic alterations of ischemia suggest that localized vasculitis should be excluded in future cases. Many upper limb mononeuropathies in diabetes mellitus are from carpal tunnel syndrome. Repetitive shear forces, anatomic factors, and excessive stiffness of connective tissues may cause these mononeuropathies.


Maternity records in Edinburgh and Aberdeen in 1936: a comparison.
Historians have long used maternity records to understand the evolution of maternity services. More recently, epidemiologists have become interested in obstetric hospital records as a source of data (e.g. birth weight, social class), to study the influence of early life on future health and disease: life course epidemiology. Edinburgh and Aberdeen are unusual in holding detailed records from several maternity institutions. The records of 1936 are of particular interest because all children born in this year and at school in Scotland at age 11 sat a cognitive ability test, the Scottish Mental Survey 1947. This study aims to describe the maternity services in Edinburgh and Aberdeen in 1936, between the First and Second World Wars. Understanding the richness of data in birth records, the manner in which they were recorded, and the context of the institutions in their community is essential for interpreting life course epidemiology studies.


Role of the Sln1-phosphorelay pathway in the response to hyperosmotic stress in the yeast Kluyveromyces lactis.
The Kluyveromyces lactis SLN1 phosphorelay system includes the osmosensor histidine kinase Sln1, the phosphotransfer protein Ypd1 and the response regulator Ssk1. Here we show that K. lactis has a functional phosphorelay system. In vitro assays, using a heterologous histidine kinase, show that the phosphate group is accepted by KlYpd1 and transferred to KlSsk1. Upon hyperosmotic stress the phosphorelay is inactivated, KlYpd1 is dephosphorylated in a KlSln1 dependent manner, and only the version of KlSsk1 that lacks the phosphate group interacts with the MAPKKK KlSsk2. Interestingly, inactivation of the KlPtp2 phosphatase in a ΔKlsln1 mutant did not lead to KlHog1 constitutive phosphorylation. KlHog1 can replace ScHog1p and activate the hyperosmotic response in Saccharomyces cerevisiae, and when ScSln1 is inactivated, KlHog1 becomes phosphorylated and induces cell lethality. All these observations indicate that the phosphorelay negatively regulates KlHog1. Nevertheless, in the absence of KlSln1 or KlYpd1, no constitutive phosphorylation is detected and cells are viable, suggesting that a strong negative feedback that is independent of KlPtp2 operates in K. lactis. Compared with S. cerevisiae, K. lactis has only a moderate accumulation of glycerol and fails to produce trehalose under hyperosmotic stress, indicating that regulation of osmolyte production is different in K. lactis.


Population-based pediatric reference intervals for hematology, iron and transferrin.
Reference intervals are crucial decision-making tools aiding clinicians in differentiating between healthy and diseased populations. However, for children such values often are lacking or incomplete. Blood samples were obtained from 689 healthy children, aged 6 months to 18 years, recruited in day care centers and schools. Hematology and anemia analytes were measured on the Siemens Advia 2120 and Abbott Architect ci8200 platforms (hemoglobin, erythrocyte volume fraction [EVF], erythrocytes, mean corpuscular volume [MCV], mean corpuscular hemoglobin [MCH], mean corpuscular hemoglobin concentration [MCHC], reticulocytes, leukocytes, lymphocytes, monocytes, neutrophils, eosinophils, basophils, platelets, iron, transferrin, transferrin saturation). Age- and gender-specific pediatric reference intervals were defined by calculating 2.5th and 97.5th percentiles. The data generated is primarily applicable to a Caucasian population, but could be used by any laboratory if verified for the local patient population.


Milk volatile organic compounds and fatty acid profile in cows fed timothy as hay, pasture, or silage.
Nutrient composition and organoleptic properties of milk can be influenced by cow diets. The objective of this study was to evaluate the forage type effects on volatile organic compounds, fatty acid (FA) profile, and organoleptic properties of milk. Timothy grass was fed as hay, pasture, or silage during a period of 27 d to a group of 21 cows in a complete block design based on days in milk. Each cow also received 7.2 kg/d of a concentrate mix to meet their nutrient requirements. Forage dry matter intake averaged 13.9 kg/d and was not different among treatments. Milk yield was higher for cows fed pasture, intermediate for cows fed silage, and lowest for cows fed hay. However, milk fat content was higher for cows fed hay and silage, compared with cows fed pasture. As a result, fat-corrected milk and fat yield were not different among treatments. Increasing the supply of dietary cis-9,cis-12 18:2 (linoleic acid) and cis-9,cis-12,cis-15 18:3 (α-linolenic acid) when feeding pasture enhanced the concentration of these 2 essential FA in milk fat compared with feeding hay or silage. Moreover, the ratio of 16:0 (palmitic acid) to cis-9 18:1 (oleic acid), which is closely related to the melting properties of milk fat, was lower in milk from cows on pasture than in milk from cows fed hay or silage. Cows fed hay produced milk with higher levels of several free FA and γ-lactones, but less pentanal and 1-pentanol. More dimethyl sulfone and toluene were found in milk of cows on pasture. Cows fed silage produced milk with higher levels of acetone, 2-butanone, and α-pinene. Results from a sensory evaluation showed that panelists could not detect a difference in flavor between milk from cows fed hay compared with silage. However, a significant number of assessors perceived a difference between milk from cows fed hay compared with milk from cows fed pasture. In a sensory ranking test, the percentage of assessors ranking for the intensity of total (raw milk, fresh milk, and farm milk), sweet (empyreumatic, vanilla, caramel, and sugar), and grassy (grass, leafy vegetable, and plant) flavors was higher for milk from cows fed pasture compared with hay and silage. Using timothy hay, pasture, or silage harvested at a similar stage of development, the current study shows that the taste of milk is affected by the forage type fed to cows. More research is, however, needed to establish a link between the sensory attributes of milk and the observed changes in volatile organic compounds and FA profile.


Acute panmyelosis with myelofibrosis - a rare subtype of acute myeloid leukemia.
One case of acute panmyelosis with myelofibrosis (APMF) is here reported. A 45 year old male presented with abrupt onset of rapidly progressing low backache, weakness and pancytopenia. On examination there was no organomegaly. Peripheral blood examination revealed normocytic normochromic red blood cells with 10% circulating blasts. Flowcytometric examination of peripheral blood revealed blasts which were positive for CD34, HLA- DR and myeloid associated antigens (i.e. CD13 and CD33). Blasts were negative for anti MPO. Bone marrow aspirate resulted in a dry tap. Bone marrow biopsy revealed panmyeloid proliferation with scattered blasts which were CD34 positive on imunohistochemistry and negative for anti MPO. Reticulin stain showed grade III myelofibrosis (WHO). Differential diagnosis considered included AML-M7, MDS-RAEB II and AML with myelodysplasia. He was started on chemotherapy [idarubicin and cytarabine; 3+7 induction regimen followed by three cycles of HIDAC (High dose cytosine arabinoside)] after which patient was in complete morphological remission with markedly reduced bone marrow fibrosis. He is now being worked up for allogeneic stem cell transplantation. Patient is asymptomatic at eight months of diagnosis. In conclusion these patients should be managed aggressively with AML therapy and this case report reaffirms the fact that APMF is subtype of AML.


Risk of Injection-Site Abscess among Infants Receiving a Preservative-Free, Two-Dose Vial Formulation of Pneumococcal Conjugate Vaccine in Kenya.
There is a theoretical risk of adverse events following immunization with a preservative-free, 2-dose vial formulation of 10-valent-pneumococcal conjugate vaccine (PCV10). We set out to measure this risk. Four population-based surveillance sites in Kenya (total annual birth cohort of 11,500 infants) were used to conduct a 2-year post-introduction vaccine safety study of PCV10. Injection-site abscesses occurring within 7 days following vaccine administration were clinically diagnosed in all study sites (passive facility-based surveillance) and, also, detected by caregiver-reported symptoms of swelling plus discharge in two sites (active household-based surveillance). Abscess risk was expressed as the number of abscesses per 100,000 injections and was compared for the second vs first vial dose of PCV10 and for PCV10 vs pentavalent vaccine (comparator). A total of 58,288 PCV10 injections were recorded, including 24,054 and 19,702 identified as first and second vial doses, respectively (14,532 unknown vial dose). The risk ratio for abscess following injection with the second (41 per 100,000) vs first (33 per 100,000) vial dose of PCV10 was 1.22 (95% confidence interval [CI] 0.37-4.06). The comparator vaccine was changed from a 2-dose to 10-dose presentation midway through the study. The matched odds ratios for abscess following PCV10 were 1.00 (95% CI 0.12-8.56) and 0.27 (95% CI 0.14-0.54) when compared to the 2-dose and 10-dose pentavalent vaccine presentations, respectively. In Kenya immunization with PCV10 was not associated with an increased risk of injection site abscess, providing confidence that the vaccine may be safely used in Africa. The relatively higher risk of abscess following the 10-dose presentation of pentavalent vaccine merits further study.


Are women ready for the new cervical screening protocol in England? A systematic review and qualitative synthesis of views about human papillomavirus testing.
A new protocol for human papillomavirus (HPV) testing within the UK cervical screening programme commenced in April 2011, creating new patient experiences. This is the first review to synthesise a substantial body of international evidence of women's information needs, views and preferences regarding HPV testing. We aimed to inform the development of educational materials to promote informed choice, reduce anxiety and improve disease control. We searched 12 bibliographic databases. Two reviewers independently screened papers and assessed study quality; disagreements were resolved by discussion. Results were extracted verbatim and authors' findings treated as primary data. Studies were synthesised collaboratively using framework methods. We synthesised findings from 17 studies. Women had overwhelmingly negative concerns; an HPV diagnosis was daunting, had associated problems of disclosure of a sexually transmitted infection (STI), impacted on relationships and provoked fear of stigmatisation. Nevertheless, many thought HPV testing could be a preferable alternative to repeat cytology. Knowledge was poor; women struggled to interpret limited information in the context of existing knowledge about STIs and cervical cancer. Women are likely to be poorly informed, have limited understanding and many unanswered questions. This could increase anxiety and reduce ability to make informed choices, presenting a substantial challenge for those who design and provide information.


Next Generation Sequencing Based Multiplex Long-Range PCR for Routine Genotyping of Autoinflammatory Disorders.
During the last decade, remarkable progress with massive sequencing has been made in the identification of disease-associated genes for AIDs using next-generation sequencing technologies (NGS). An international group of experts described the ideal genetic screening method which should give information about SNVs, InDels, Copy Number Variations (CNVs), GC rich regions. We aimed to develop and validate a molecular diagnostic method in conjunction with the NGS platform as an inexpensive, extended and uniform coverage and fast screening tool which consists of nine genes known to be associated with various AIDs. For the validation of basic and expanded panels, long-range multiplex models were setup on healthy samples without any known variations for MEFV, MVK, TNFRSF1A, NLRP3, PSTPIP1, IL1RN, NOD2, NLRP12 and LPIN2 genes. Patients with AIDs who had already known causative variants in these genes were sequenced for analytical validation. As a last step, multiplex models were validated on patients with pre-diagnosis of AIDs. All sequencing steps were performed on the Illumina NGS platform. Validity steps included the selection of related candidate genes, primer design, development of screening methods, validation and verification of the product. The GDPE (Gentera) bioinformatics pipeline was followed. Although there was no nonsynonymous variation in 21 healthy samples, 107 synonymous variant alleles and some intronic and UTR variants were detected. In 10 patients who underwent analytical validation, besides the 11 known nonsynonymous variant alleles, 11 additional nonsynonymous variant alleles and a total of 81 synonymous variants were found. In the clinical validation phase, 46 patients sequenced with multiplex panels, genetic and clinical findings were combined for diagnosis. In this study, we describe the development and validation of an NGS-based multiplex array enabling the "long-amplicon" approach for targeted sequencing of nine genes associated with common AIDs. This screening tool is less expensive and more comprehensive compared to other methods and more informative than traditional sequencing. The proposed panel offers advantages to WES or hybridization probe equivalents in terms of CNV analysis, high sensitivity and uniformity, GC-rich region sequencing, InDel detection and intron covering.


Exercise intensity and its effects on thyroid hormones.
Physical activity influences energy metabolism in human subjects by increasing activity-induced energy expenditure and resting metabolic rate for several hours after exercise. Effects of exercise on circulating thyroid hormone values remain controversial. We have investigated the effect of acute aerobic exercise on thyroid hormone values. The effect of different intensity levels of acute aerobic exercise on thyroid hormones was investigated in 60 male well-trained athletes by performing bicycle ergometer at 45% (low intensity), 70% (moderate intensity), and 90% (high intensity). These intensities were selected according to their maximum heart rate (MHR). At each intensity level, heart rate, blood lactic acid, serum total thyroxine (T4), free thyroxine (fT4), total triiodothyronine (T3), free triiodothyronine (fT3) and thyroid stimulating hormone (TSH) values were measured. The results of this study show that exercise performed at the anaerobic threshold (70% of maximum heart rate, lactate level 4.59 +/- 1.75 mmol/l) caused the most prominent changes in the amount of any hormone values. While the rate of T4, fT4, and TSH continued to rise at 90% of maximum heart rate, the rate of T3 and fT3 started to fall. Maximal aerobic exercise greatly affects the level of circulating thyroid hormones.


US state- and county-level social capital in relation to obesity and physical inactivity: a multilevel, multivariable analysis.
Although social capital has been linked to a variety of health outcomes, its association with obesity has yet to be elucidated. This study explored the relations between social capital measured at the US state and county levels and individual obesity and leisure-time physical inactivity. Individual-level data were drawn from the 2001 Behavioral Risk Factor Surveillance System survey, while data from other surveys and administrative sources were used to construct contextual measures. Two state-level social capital scales were derived from 10 indicators and two county-level scales from five indicators. In 2-level analyses of over 167,000 adults nested within 48 states plus the District of Columbia, residence in a state above the median on one or both state social capital scales (vs. neither scale) was associated with lower relative odds of obesity and physical inactivity, controlling for individual-level covariables and state-level estimates of mean household income, the Gini coefficient, and the percentage of Black residents. In 3-level analyses, the adjusted odds ratio (OR) for physical inactivity associated with residence in a county above the median on one or both county social capital scales was significantly below 1, while the association with obesity was not significant. Significantly weaker inverse ORs for the relations between state- and county-level social capital and obesity were observed among American Indians/Alaska Natives compared to Whites. Meanwhile, little support was found for mediation by social capital of the associations of urban sprawl and income inequality with obesity or physical inactivity. Overall, this study provides some evidence for the promotion of social capital as a potential strategy for addressing the burgeoning obesity epidemic.


Monoallelic expression of the interleukin-2 locus.
The lymphokine interleukin-2 (IL-2) is responsible for autocrine cell cycle progression and regulation of immune responses. Uncontrolled secretion of IL-2 results in adverse reactions ranging from anergy, to aberrant T cell activation, to autoimmunity. With the use of fluorescent in situ hybridization and single-cell polymerase chain reaction in cells with different IL-2 alleles, IL-2 expression in mature thymocytes and T cells was found to be tightly controlled by monoallelic expression. Because IL-2 is encoded at a nonimprinted autosomal locus, this result represents an unusual regulatory mode for controlling the precise expression of a single gene.


An engineered solution to the maladministration of spinal injections.
An overview is provided of the progress made in a Department of Health (DH) initiative to implement an engineered solution to the maladministration of spinal injections. In an effort to eliminate the possibility of misconnection errors at the spinal route, the DH is investigating the potential for dedicated spinal equipment that will be incompatible with standard Luer syringes, needles and associated devices. Background information on the problem of misconnection errors is provided and a systematic approach to their eradication is detailed. Research to date has entailed extensive bench-testing of prototype non-Luer connectors, a prospective hazard analysis of spinal procedures in haematology and anaesthesia and usability evaluation of prototype non-Luer devices in simulated environments. The prospective hazard analysis identified two potential risks which will need to be managed as part of a successful implementation programme. Usability testing of two prototype connection systems concluded that one design was inadequate, as the non-Luer element was provided as a separable adapter. The second connection system was modified following the first round of testing, and achieved improved satisfaction ratings from clinicians in round two. This system was selected to proceed to a pre-implementation evaluation and the research team are currently evaluating its acceptability in clinical use.


Ketone Bodies and Exercise Performance: The Next Magic Bullet or Merely Hype?
Elite athletes and coaches are in a constant search for training methods and nutritional strategies to support training and recovery efforts that may ultimately maximize athletes' performance. Recently, there has been a re-emerging interest in the role of ketone bodies in exercise metabolism, with considerable media speculation about ketone body supplements being routinely used by professional cyclists. Ketone bodies can serve as an important energy substrate under certain conditions, such as starvation, and can modulate carbohydrate and lipid metabolism. Dietary strategies to increase endogenous ketone body availability (i.e., a ketogenic diet) require a diet high in lipids and low in carbohydrates for ~4 days to induce nutritional ketosis. However, a high fat, low carbohydrate ketogenic diet may impair exercise performance via reducing the capacity to utilize carbohydrate, which forms a key fuel source for skeletal muscle during intense endurance-type exercise. Recently, ketone body supplements (ketone salts and esters) have emerged and may be used to rapidly increase ketone body availability, without the need to first adapt to a ketogenic diet. However, the extent to which ketone bodies regulate skeletal muscle bioenergetics and substrate metabolism during prolonged endurance-type exercise of varying intensity and duration remains unknown. Therefore, at present there are no data available to suggest that ingestion of ketone bodies during exercise improves athletes' performance under conditions where evidence-based nutritional strategies are applied appropriately.


Annexin A6 interacts with p65 and stimulates NF-κB activity and catabolic events in articular chondrocytes.
ANXA6, the gene for annexin A6, is highly expressed in osteoarthritic (OA) articular chondrocytes but not in healthy articular chondrocytes. This study was undertaken to determine whether annexin A6 affects catabolic events in these cells. Articular chondrocytes were isolated from Anxa6-knockout mice, wild-type (WT) mice, and human articular cartilage in which ANXA6 was overexpressed. Cells were treated with interleukin-1β (IL-1β) or tumor necrosis factor α (TNFα), and expression of catabolic genes and activation of NF-κB were determined by real-time polymerase chain reaction and luciferase reporter assay. Anxa6(-/-) and WT mouse knee joints were injected with IL-1β or the medial collateral ligament was transected and partial resection of the medial meniscus was performed to determine the role of Anxa6 in IL-1β-mediated cartilage destruction and OA progression. The mechanism by which Anxa6 stimulates NF-κB activity was determined by coimmunoprecipitation and immunoblot analysis of nuclear and cytoplasmic fractions of IL-1β-treated Anxa6(-/-) and WT mouse chondrocytes for p65 and Anxa6. Loss of Anxa6 resulted in decreased NF-κB activation and catabolic marker messenger RNA (mRNA) levels in IL-1β- or TNFα-treated articular chondrocytes, whereas overexpression of ANXA6 resulted in increased NF-κB activity and catabolic marker mRNA levels. Annexin A6 interacted with p65, and loss of Anxa6 caused decreased nuclear translocation and retention of the active p50/p65 NF-κB complex. Cartilage destruction in Anxa6(-/-) mouse knee joints after IL-1β injection or partial medial meniscectomy was reduced as compared to that in WT mouse joints. Our data define a role of annexin A6 in the modulation of NF-κB activity and in the stimulation of catabolic events in articular chondrocytes.


Possibility of middle meningeal artery-to-petrous internal carotid artery bypass: an anatomic study.
The possibility of creating a middle meningeal artery (MMA)-to-petrous internal carotid artery (ICA) bypass was investigated in six cadavers (bilaterally). Such a procedure could be used to treat patients with high cervical vascular lesions and those with tumors of the infratemporal fossa invading the high cervical ICA. After a frontotemporal craniotomy, the foramen spinosum and foramen ovale were exposed extradurally. Immediately posterior to the foramen ovale and medial to the foramen spinosum, the petrous portion of the ICA was exposed with a diamond-tipped drill. The MMA was lifted from its groove, and a sufficient length was transected to perform a bypass with the petrous ICA medially. The mean width of the MMA at the site of anastomosis was 2.3 +/- 0.35 mm. The mean length of MMA from the foramen spinosum to the site of the anastomosis was 9.6 +/- 1.7 mm. Based on these measurements, width and length of MMA appear to be sufficient for a bypass with petrous ICA.


IRF6 mutation screening in non-syndromic orofacial clefting: analysis of 1521 families.
Van der Woude syndrome (VWS) is an autosomal dominant malformation syndrome characterized by orofacial clefting (OFC) and lower lip pits. The clinical presentation of VWS is variable and can present as an isolated OFC, making it difficult to distinguish VWS cases from individuals with non-syndromic OFCs. About 70% of causal VWS mutations occur in IRF6, a gene that is also associated with non-syndromic OFCs. Screening for IRF6 mutations in apparently non-syndromic cases has been performed in several modestly sized cohorts with mixed results. In this study, we screened 1521 trios with presumed non-syndromic OFCs to determine the frequency of causal IRF6 mutations. We identified seven likely causal IRF6 mutations, although a posteriori review identified two misdiagnosed VWS families based on the presence of lip pits. We found no evidence for association between rare IRF6 polymorphisms and non-syndromic OFCs. We combined our results with other similar studies (totaling 2472 families) and conclude that causal IRF6 mutations are found in 0.24-0.44% of apparently non-syndromic OFC families. We suggest that clinical mutation screening for IRF6 be considered for certain family patterns such as families with mixed types of OFCs and/or autosomal dominant transmission.


Public health nurses and the prevention of and intervention in family violence.
This article seeks to describe the current efforts of public health nurses involved in family violence prevention while on home visits, participating in community groups, working on special projects, and developing innovative policies. Despite a dearth of outcomes research and data regarding public health nursing practice in this area, both strengths and weakness in public health nursing practice are evident. Improvements in nursing education related to family violence, an increase in multidisciplinary collaboration by public health nurses, and improvements in the utilization of public health nurses in the public health infrastructure and in primary prevention efforts would remove many barriers to effective family violence prevention strategies.


Effect of modified physiology laboratory sessions on the awareness and knowledge of obesity of health-related university students.
Obesity is a global health problem of immense magnitude. Increased awareness and knowledge about obesity in health-care providers are essential to addressing this global issue. The aim of the present study was to assess the baseline knowledge on obesity of university students in health-related programs and evaluate the effectiveness of modified physiology laboratory (MPL) sessions in enhancing the awareness of obesity of these students by repeated exposure to physiological differences between normal weight (NW) and overweight or obese (OW/OB) individuals. It was a quasi-experimental study in which three regular physiology laboratory exercises were modified (MPL) to address different aspects of obesity. A total of 319 students (181 male and 138 female) from health-related colleges taking the physiology course during academic session 2011-2012 were included in the study. A questionnaire on obesity served as a measure of prior knowledge and improvement of evolution of student knowledge. The questionnaire was administered thrice: Beginning of the first MPL session (pretest), at the end of the second session of the first MPL (posttest-1), and the end of second session of the final MPL (posttest-2). Correct knowledge (CK), perceived knowledge (PK), and accuracy of knowledge (KA) were evaluated. Students gathered and analyzed physiological data in these sessions to explore the differences between NW and OW/OB subjects. The students' level of PK on obesity was high, but KA was low in pretest. The three knowledge indices improved significantly (P <0.001) from pretest (PK = 77.2%; KA = 52.8%; CK = 40.5%) to posttest 1 (PK = 93.5%; KA = 70.1%; CK = 65.9%). However, from posttest 1 to posttest 2, only small but significant increments were observed in CK (10.5%) and KA (8.7%). The awareness and knowledge on obesity of health-related undergraduate university students at baseline were low, but significantly improved after MPLs. The students' knowledge did not only increase but also improved in accuracy. This increase in awareness and knowledge of obesity is expected to directly impact on the students' lifestyles and boost their confidence to counsel others on obesity.


Griffithsin carrageenan fast dissolving inserts prevent SHIV HSV-2 and HPV infections in vivo.
Human immunodeficiency virus (HIV) pre-exposure prophylaxis (PrEP) strategies with proven in vivo efficacy rely on antiretroviral drugs, creating the potential for drug resistance and complicated treatment options in individuals who become infected. Moreover, on-demand products are currently missing from the PrEP development portfolio. Griffithsin (GRFT) is a non-antiretroviral HIV entry inhibitor derived from red algae with an excellent safety profile and potent activity in vitro. When combined with carrageenan (CG), GRFT has strong activity against herpes simplex virus-2 (HSV-2) and human papillomavirus (HPV) in vitro and in vivo. Here, we report that GRFT/CG in a freeze-dried fast dissolving insert (FDI) formulation for on-demand use protects rhesus macaques from a high dose vaginal SHIV SF162P3 challenge 4 h after FDI insertion. Furthermore, the GRFT/CG FDI also protects mice vaginally against HSV-2 and HPV pseudovirus. As a safe, potent, broad-spectrum, on-demand non-antiretroviral product, the GRFT/CG FDI warrants clinical development.


Failure of ventral closure and axial rotation in embryos lacking the proprotein convertase Furin.
We have examined the role of Furin in postimplantation-stage mouse embryos by analyzing both the expression pattern of fur mRNA and the developmental consequences of a loss-of-function mutation at the fur locus. At early stages (day 7.5), fur mRNA is abundant in extraembryonic endoderm and mesoderm, anterior visceral endoderm, and in precardiac mesoderm. 1 day later fur is expressed throughout the heart tube and in the lateral plate mesoderm, notochordal plate and definitive gut endoderm. Embryos lacking Furin die between days 10.5 and 11.5, presumably due to hemodynamic insufficiency associated with severe ventral closure defects and the failure of the heart tube to fuse and undergo looping morphogenesis. Morphogenesis of the yolk sac vasculature is also abnormal, although blood islands and endothelial precursors form. Analysis of cardiac and endodermal marker genes shows that while both myocardial precursors and definitive endoderm cells are specified, their numbers and migratory properties are compromised. Notably, mutant embryos fail to undergo axial rotation, even though Nodal and eHand, two molecular markers of left-right asymmetry, are appropriately expressed. Overall, the present data identify Furin as an important activator of signals responsible for ventral closure and embryonic turning.


Influence of pH, light cycle, and temperature on ecotoxicity of four sulfonylurea herbicides towards Lemna gibba.
In chemical regulation, e.g. the EU Water Framework Directive, REACH, or the Pesticide Directive, standardized ecotoxicological tests are applied to evaluate and rank the hazard of compounds and for deriving environmental quality standards (EQS). Standardized test methods prescribe fixed testing conditions e.g. specific temperature, pH, light intensity etc. However, environmental conditions under which the organisms live are rarely identical to the standard conditions. Thus, the ecotoxicity of compounds found in standard test is not only a function of the compounds inherent physico-chemical properties but is also affected by test conditions. It is therefore important to study the effect of changes in test conditions in order to get reliable input ecotoxicity data for assessing the potential risk posed by a compound. The objective of this study was to investigate the implications of changing test conditions on the toxicity of four sulfonylurea herbicides (SUs). The toxicity of the four SUs towards Lemna gibba was investigated at three pH levels (6, 7.5 and 9), at two temperatures (15 and 24 °C) and two light regimes (continuous and 12:12 h light:dark cycle) The EC50 increased twofold to tenfold for the four SUs when pH was increased from 6 to 9. Decreasing the temperature from 24 to 15 °C or introducing a dark:light cycle did not cause any trends in changes in toxicity. The results show that test conditions can have an effect on the toxicity and this should be considered when the standard test results are used for derivation of EQS.


Unraveling the multiscale structural organization and connectivity of the human brain: the role of diffusion MRI.
The structural architecture and the anatomical connectivity of the human brain show different organizational principles at distinct spatial scales. Histological staining and light microscopy techniques have been widely used in classical neuroanatomical studies to unravel brain organization. Using such techniques is a laborious task performed on 2-dimensional histological sections by skilled anatomists possibly aided by semi-automated algorithms. With the recent advent of modern magnetic resonance imaging (MRI) contrast mechanisms, cortical layers and columns can now be reliably identified and their structural properties quantified post-mortem. These developments are allowing the investigation of neuroanatomical features of the brain at a spatial resolution that could be interfaced with that of histology. Diffusion MRI and tractography techniques, in particular, have been used to probe the architecture of both white and gray matter in three dimensions. Combined with mathematical network analysis, these techniques are increasingly influential in the investigation of the macro-, meso-, and microscopic organization of brain connectivity and anatomy, both in vivo and ex vivo. Diffusion MRI-based techniques in combination with histology approaches can therefore support the endeavor of creating multimodal atlases that take into account the different spatial scales or levels on which the brain is organized. The aim of this review is to illustrate and discuss the structural architecture and the anatomical connectivity of the human brain at different spatial scales and how recently developed diffusion MRI techniques can help investigate these.


Sustained therapeutic effects of percutaneous tibial nerve stimulation: 24-month results of the STEP study.
To evaluate the safety, sustained effectiveness, and treatment interval for percutaneous tibial nerve stimulation (PTNS) for overactive bladder (OAB) therapy through 24 months. A prospective study following treatment success after 12 weekly PTNS treatments, subjects were prescribed a 14-week tapering protocol, followed by ongoing therapy with a Personal Treatment Plan determined by the investigator and subject to sustain subject OAB symptom improvement. Questionnaires were completed every 3 months, voiding diaries every 6 months; adverse events were reported throughout. Of 50 subjects enrolled, 35 remained in the study at 24 months. During the 24 months following initial treatment success and a 14-week tapering protocol, mean treatments per month was 1.3. Voiding diary and OAB-q data demonstrate sustained improvement reported at 13 weeks through 24 months. Improvements in frequency, urge incontinence episodes, night-time voids and moderate-to-severe urgency episodes from voiding diaries at 6, 12, 18, and 24 months were statistically significant compared to baseline (prior to initial 12 weekly treatments). Compared to baseline, OAB-q symptom severity scores and health related quality of life scores were statistically significant for improvement at each tested time point. Five mild adverse events of unknown relation to treatment were reported. Sustained safety and efficacy of PTNS were demonstrated over 24 months with initial success after 12 weekly treatments, followed by a 14-week prescribed tapering protocol and a Personalized Treatment Plan. With an average of 1.3 treatments per month, PTNS therapy is a safe, durable, and valuable long-term OAB treatment option to sustain clinically significant OAB symptom control.


Game movement demands and physical profiles of junior, senior and elite male and female rugby sevens players.
To inform recruitment, selection, training and testing of male and female rugby sevens players game running movement patterns and physical characteristics were quantified across junior, senior, and elite playing levels. Anthropometric and physical testing (40 m sprint, vertical jump, Yo-Yo IR1) occurred prior to players' national championships or international tournaments (n = 110 players), while game movements were obtained via GPS (n = 499 game files). The game movements of male players were similar across playing levels except for number of impacts >10 g which were 2 to 4-fold higher in elite (25.0 ± 11.2 impacts · game-1; mean ± SD), than junior (6.3 ± 3.5) and senior (11.8 ± 6.6) players. In men, there were fewer substantial correlations between on- and off-field measures which may reflect similar physical attributes across playing levels, and that other (strength, technical or tactical) factors may better differentiate these players. In females, elite players had more favourable on- and off-field performance measures than juniors and seniors, with moderate to strong correlations between on- and off-field variables. Female players should benefit from additional fitness training, while male players need to balance fitness with other technical and tactical factors.


Banking of biological fluids for studies of disease-associated protein biomarkers.
With the increasing demand of providing personalized medicine the need for biobanking of biological material from individual patients has increased. Such samples are essential for molecular research aimed at characterizing diseases at several levels ranging from epidemiology and diagnostic and prognostic classification to prediction of response to therapy. Clinically validated biomarkers may provide information to be used for diagnosis, screening, evaluation of risk/predisposition, assessment of prognosis, monitoring (recurrence of disease), and prediction of response to treatment and as a surrogate response marker. Many types of biological fluids or tissues can be collected and stored in biorepositories. Samples of blood can be further processed into plasma and serum, and tissue pieces can be either frozen or fixed in formalin and then embedded into paraffin. The present review focuses on biological fluids, especially serum and plasma, intended for study of protein biomarkers. In biomarker studies the process from the decision to take a sample from an individual to the moment the sample is safely placed in the biobank consists of several phases including collection of samples, transport of the samples, and handling and storage of samples. Critical points in each step important for high quality biomarker studies are described in this review. Failure to develop and adhere to robust standardized protocols may have significant consequences as the quality of the material stored in the biobank as well as conclusions and clinical recommendations based on analysis of such material may be severely affected.


Many college freshmen drink at levels far beyond the binge threshold.
Binge drinking is a dichotomous variable that allows researchers to sort students into categories based upon a specific threshold of consumption, commonly 4 (females) or 5 (males) drinks. Crossing the binge threshold increases the risk of negative alcohol-related consequences. The use of such thresholds has played a vital role in the study of college drinking. While extremely valuable, the dichotomous nature of binge drinking variables removes information about how heavily students actually drink, leaving the characterization of college drinking incomplete. The present study examined patterns of alcohol use beyond the binge threshold. The data set consisted of self-reported 2-week drinking histories from 10,424 first-semester freshmen at 14 schools across the United States during the fall of 2003. The number of students who reached the 4+/5+ binge-drinking threshold was calculated, as was the number who reached 2 times (8+/10+ drinks) or 3 times (12+/15+ drinks) the binge threshold. Logistic regression analyses were used to explore gender differences and to assess whether frequent binge drinkers (3+ binges per 2 weeks) were more likely than infrequent binge drinkers (1-2 binges per 2 weeks) to reach high peak levels of consumption. Roughly 1 of 5 males consumed 10+ drinks and 1 of 10 females consumed 8+ drinks, twice the binge threshold, at least once in the previous 2 weeks. Gender differences were observed at every drinking level and were particularly large at higher peak levels. Frequent binge drinkers were more likely than infrequent binge drinkers to consume 2 or 3 times the binge threshold. A surprisingly large percentage of students, particularly males, drink at peak levels well beyond the binge threshold. Such findings suggest that schools might make additional progress in the battle against alcohol misuse by focusing on extreme drinking practices in addition to binge drinking per se.


Racial differences in blood pressure control: potential explanatory factors.
The objective of the study was to identify potential explanatory factors for racial differences in blood pressure (BP) control. The design of the study was a cross-sectional study The study included 608 patients with hypertension who were either African American (50%) or white (50%) and who received primary care in Durham, NC. Baseline data were obtained from the Take Control of Your Blood pressure study and included clinical, demographic, and psychosocial variables potentially related to clinic BP measures. African Americans were more likely than whites to have inadequate baseline clinic BP control as defined as greater than or equal to 140/90 mmHg (49% versus 34%; unadjusted odds ratio [OR] 1.8; 95% confidence interval [CI] 1.3-2.5). Among factors that may explain this disparity, being older, reporting hypertension medication nonadherence, reporting a hypertension diagnosis for more than 5 years, reporting high levels of stress, being worried about hypertension, and reporting an increased number of medication side effects were related to inadequate BP control. In adjusted analyses, African Americans continue to have poor BP control relative to whites; the magnitude of the association was reduced (OR = 1.5; 95% CI 1.0-2.1). Medication nonadherence, worries about hypertension, and older age (>70) continued to be related to poor BP control. In this sample of hypertensive patients, there were a number of factors associated with poor BP control that partially explained the observed racial disparity in hypertension control including age, medication nonadherence, and worry about BP. Medication nonadherence is of particular interest because it is a potentially modifiable factor that might be used to reduce the racial disparity in BP control.


A rapid procedure for selectively isolating the major glycoprotein from purified rat brain myelin.
Treatment of the chloroform/methanol-insoluble residue of rat brain myelin with lithium 3,5-di-iodosalicylate solubilized the major myelin-associated glycoprotein along with most other proteins and glycoproteins. Equilibration of the extract with phenol resulted in the selective partitioning of the major glycoprotein into the aqueous phase, whereas nearly all of the other proteins and glycoproteins went into the phenol phase.


Morphological Modification of TiO₂ Thin Films as Highly Efficient Photoanodes for Photoelectrochemical Water Splitting.
TiO2 films with modified morphology have been successfully synthesized via a facile spray-pyrolysis method in the presence of poly(ethylene glycol) (PEG) as a templating agent. The effects of the PEG concentration on the relevant properties of TiO2 films were investigated by means of scanning electron microscopy, X-ray diffraction, and UV-vis absorbance spectroscopy, while their photocatalytic properties were assessed by photoelectrochemical (PEC) water-splitting measurements. The introduction of 10 g·L(-1) PEG into the precursor solution leads to surface roughening with an exceptional improvement in PEC responses, revealing a photoconversion efficiency of 1.15% at -0.50 V vs HgO|Hg (in a 1 M NaOH electrolyte under broad-spectrum illumination), which is nearly triple that of the unmodified film (0.45% at -0.38 V vs HgO|Hg). Although the efficiency of the rough-surface photoanodes deteriorates upon increasing the PEG content, their PEC responses are still superior to those of smooth-surface films. Possible phenomena that might be responsible for the experimental observations are suggested and discussed accordingly.


Drug susceptibility testing and mortality in patients treated for tuberculosis in high-burden countries: a multicentre cohort study.
Drug resistance is a challenge for the global control of tuberculosis. We examined mortality in patients with tuberculosis from high-burden countries, according to concordance or discordance of results from drug susceptibility testing done locally and in a reference laboratory. This multicentre cohort study was done in Côte d'Ivoire, Democratic Republic of the Congo, Kenya, Nigeria, South Africa, Peru, and Thailand. We collected Mycobacterium tuberculosis isolates and clinical data from adult patients aged 16 years or older. Patients were stratified by HIV status and tuberculosis drug resistance. Molecular or phenotypic drug susceptibility testing was done locally and at the Swiss National Center for Mycobacteria, Zurich, Switzerland. We examined mortality during treatment according to drug susceptibility test results and treatment adequacy in multivariable logistic regression models adjusting for sex, age, sputum microscopy, and HIV status. We obtained M tuberculosis isolates from 871 patients diagnosed between 2013 and 2016. After exclusion of 237 patients, 634 patients with tuberculosis were included in this analysis; the median age was 33·2 years (IQR 26·9-42·5), 239 (38%) were women, 272 (43%) were HIV-positive, and 69 (11%) patients died. Based on the reference laboratory drug susceptibility test, 394 (62%) strains were pan-susceptible, 45 (7%) monoresistant, 163 (26%) multidrug-resistant (MDR), and 30 (5%) had pre-extensively or extensively drug resistant (pre-XDR or XDR) tuberculosis. Results of reference and local laboratories were concordant for 513 (81%) of 634 patients and discordant for 121 (19%) of 634. Overall, sensitivity to detect any resistance was 90·8% (95% CI 86·5-94·2) and specificity 84·3% (80·3-87·7). Mortality ranged from 6% (20 of 336) in patients with pan-susceptible tuberculosis treated according to WHO guidelines to 57% (eight of 14) in patients with resistant strains who were under-treated. In logistic regression models, compared with concordant drug susceptibility test results, the adjusted odds ratio of death was 7·33 (95% CI 2·70-19·95) for patients with discordant results potentially leading to under-treatment. Inaccurate drug susceptibility testing by comparison with a reference standard leads to under-treatment of drug-resistant tuberculosis and increased mortality. Rapid molecular drug susceptibility test of first-line and second-line drugs at diagnosis is required to improve outcomes in patients with MDR tuberculosis and pre-XDR or XDR tuberculosis. National Institutes of Allergy and Infectious Diseases, Swiss National Science Foundation, Swiss National Center for Mycobacteria.


Inhibition of Hedgehog signaling sensitizes NSCLC cells to standard therapies through modulation of EMT-regulating miRNAs.
Epidermal growth factor receptor- tyrosine kinase inhibitors (EGFR-TKIs) benefit Non-small cell lung cancer (NSCLC) patients, and an EGFR-TKIi erlotinib, is approved for patients with recurrent NSCLC. However, resistance to erlotinib is a major clinical problem. Earlier we have demonstrated the role of Hedgehog (Hh) signaling in Epithelial-to-Mesenchymal transition (EMT) of NSCLC cells, leading to increased proliferation and invasion. Here, we investigated the role of Hh signaling in erlotinib resistance of TGF-β1-induced NSCLC cells that are reminiscent of EMT cells. Hh signaling was inhibited by specific siRNA and by GDC-0449, a small molecule antagonist of G protein coupled receptor smoothened in the Hh pathway. Not all NSCLC patients are likely to benefit from EGFR-TKIs and, therefore, cisplatin was used to further demonstrate a role of inhibition of Hh signaling in sensitization of resistant EMT cells. Specific pre- and anti-miRNA preparations were used to study the mechanistic involvement of miRNAs in drug resistance mechanism. siRNA-mediated inhibition as well as pharmacological inhibition of Hh signaling abrogated resistance of NSCLC cells to erlotinib and cisplatin. It also resulted in re-sensitization of TGF-β1-induced A549 (A549M) cells as well the mesenchymal phenotypic H1299 cells to erlotinib and cisplatin treatment with concomitant up-regulation of cancer stem cell (CSC) markers (Sox2, Nanog and EpCAM) and down-regulation of miR-200 and let-7 family miRNAs. Ectopic up-regulation of miRNAs, especially miR-200b and let-7c, significantly diminished the erlotinib resistance of A549M cells. Inhibition of Hh signaling by GDC-0449 in EMT cells resulted in the attenuation of CSC markers and up-regulation of miR-200b and let-7c, leading to sensitization of EMT cells to drug treatment, thus, confirming a connection between Hh signaling, miRNAs and drug resistance. We demonstrate that Hh pathway, through EMT-induction, leads to reduced sensitivity to EGFR-TKIs in NSCLCs. Therefore, targeting Hh pathway may lead to the reversal of EMT phenotype and improve the therapeutic efficacy of EGFR-TKIs in NSCLC patients.


Non-smoking policies, tobacco education, and smoking cessation programmes in facilities serving the elderly in Michigan, United States.
To determine the extent of and impetus for smoke-free policies in facilities serving Michigan's elderly, and the extent of tobacco education and smoking cessation programmes for elders and staff of these facilities. Telephone survey in February 1997 of three types of facilities serving Michigan's elderly population. Area Agencies on Aging (n = 12), Councils and Commissions on Aging (n = 31), and senior centres (n = 98) located in Michigan, USA. Prevalence of smoke-free policies, tobacco education, and smoking cessation programmes in facilities serving the elderly. 99% (95% confidence interval (CI) = 97% to 100%) of 141 facilities surveyed have an indoor smoke-free policy. Eighty-five per cent (95% CI = 79% to 91%) of these policies prohibit all smoking inside the facility. Forty-five per cent (95% CI = 37% to 54%) cited a law as requiring the smoke-free policy, whereas 38% (95% CI = 30% to 46%) indicated the policy was adopted voluntarily for health reasons. Forty-two per cent (95% CI = 34% to 50%) of the facilities provided some education on the dangers of tobacco, while 11% (95% CI = 6% to 16%) arranged smoking cessation programmes for staff or elders. In Michigan, a very high percentage of non-institutional facilities serving the elderly have smoke-free policies, which appear to increase participation at these facilities. Tobacco education programmes are provided in less than half the facilities, and very few arrange smoking cessation programmes for elders or staff.


Helping policy-makers address rural health access problems.
This paper provides a comprehensive review of the key dimensions of access and their significance for the provision of primary health care and a framework that assists policy-makers to evaluate how well policy targets the dimensions of access. Access to health care can be conceptualised as the potential ease with which consumers can obtain health care at times of need. Disaggregation of the concept of access into the dimensions of availability, geography, affordability, accommodation, timeliness, acceptability and awareness allows policy-makers to identify key questions which must be addressed to ensure reasonable primary health care access for rural and remote Australians. Evaluating how well national primary health care policies target these dimensions of access helps identify policy gaps and potential inequities in ensuring access to primary health care. Effective policies must incorporate the multiple dimensions of access if they are to comprehensively and effectively address unacceptable inequities in health status and access to basic health services experienced by rural and remote Australians.


Congruency effects between auditory and tactile motion: extending the phenomenon of cross-modal dynamic capture.
Behavioral studies of multisensory integration in motion perception have focused on the particular case of visual and auditory signals. Here, we addressed a new case: audition and touch. In Experiment 1, we tested the effects of an apparent motion stream presented in an irrelevant modality (audition or touch) on the perception of apparent motion streams in the other modality (touch or audition, respectively). We found significant congruency effects (lower performance when the direction of motion in the irrelevant modality was incongruent with the direction of the target) for the two possible modality combinations. This congruency effect was asymmetrical, with tactile motion distractors having a stronger influence on auditory motion perception than vice versa In Experiment 2, we used auditory motion targets and tactile motion distractors while participants adopted one of two possible postures: arms uncrossed or arms crossed. The effects of tactile motion on auditory motion judgments were replicated in the arms-uncrossed posture, but they dissipated in the arms-crossed posture. The implications of these results are discussed in light of current findings regarding the representation of tactile and auditory space.


Assignment of the ferriheme resonances of the low-spin complexes of nitrophorins 1 and 4 by (1)H and (13)C NMR spectroscopy: comparison to structural data obtained from X-ray crystallography.
In this work we report the assignment of the majority of the ferriheme resonances of low-spin nitrophorins (NP) 1 and 4 and compare them to those of NP2, published previously. It is found that the structure of the ferriheme complexes of NP1 and NP4, in terms of the orientation of the ligand(s), can be determined with good accuracy by NMR techniques in the low-spin forms and that angle plots proposed previously (Shokhirev, N. V.; Walker, F. A. J. Biol. Inorg. Chem. 1998, 3, 581-594) describe the angle of the effective nodal plane of the axial ligands in solution. The effective nodal plane of low-spin NP1, NP4, and NP2 complexes is in all cases of imidazole and histamine complexes quite similar to the average of the His-59 or -57 and the exogenous ligand angles seen in the X-ray crystal structures. For the cyanide complexes of the nitrophorins, however, the effective nodal plane of the axial ligand does not coincide with the actual histidine-imidazole plane orientation. This appears to be a result of the contribution of an additional source of asymmetry, the orientation of one of the zero-ruffling lines of the heme. Probably this effect exists for the imidazole and histamine complexes as well, but because the effect of asymmetry that occurs from planar exogenous axial ligands is much larger than the effect of heme ruffling the effect of the zero-ruffling line can only be detected for the cyanide complexes, where the only ligand plane is that of the proximal histidine. The three-dimensional structures of the three NP-CN complexes, including that of NP2-CN reported herein, confirm the high degree of ruffling of these complexes. There is an equilibrium between the two heme orientations (A and B) that depends on the heme cavity shape and changes somewhat with exogenous axial ligand. The A:B ratio can be much more accurately measured by NMR spectroscopy than by X-ray crystallography.


Characterization and modeling of the Haemophilus influenzae core and supragenomes based on the complete genomic sequences of Rd and 12 clinical nontypeable strains.
The distributed genome hypothesis (DGH) posits that chronic bacterial pathogens utilize polyclonal infection and reassortment of genic characters to ensure persistence in the face of adaptive host defenses. Studies based on random sequencing of multiple strain libraries suggested that free-living bacterial species possess a supragenome that is much larger than the genome of any single bacterium. We derived high depth genomic coverage of nine nontypeable Haemophilus influenzae (NTHi) clinical isolates, bringing to 13 the number of sequenced NTHi genomes. Clustering identified 2,786 genes, of which 1,461 were common to all strains, with each of the remaining 1,328 found in a subset of strains; the number of clusters ranged from 1,686 to 1,878 per strain. Genic differences of between 96 and 585 were identified per strain pair. Comparisons of each of the NTHi strains with the Rd strain revealed between 107 and 158 insertions and 100 and 213 deletions per genome. The mean insertion and deletion sizes were 1,356 and 1,020 base-pairs, respectively, with mean maximum insertions and deletions of 26,977 and 37,299 base-pairs. This relatively large number of small rearrangements among strains is in keeping with what is known about the transformation mechanisms in this naturally competent pathogen. A finite supragenome model was developed to explain the distribution of genes among strains. The model predicts that the NTHi supragenome contains between 4,425 and 6,052 genes with most uncertainty regarding the number of rare genes, those that have a frequency of <0.1 among strains; collectively, these results support the DGH.


Physiological effects of subthalamic nucleus deep brain stimulation surgery in cervical dystonia.
Subthalamic nucleus deep brain stimulation (STN DBS) surgery is clinically effective for treatment of cervical dystonia; however, the underlying physiology has not been examined. We used transcranial magnetic stimulation (TMS) to examine the effects of STN DBS on sensorimotor integration, sensorimotor plasticity and motor cortex excitability, which are identified as the key pathophysiological features underlying dystonia. TMS paradigms of short latency afferent inhibition (SAI) and long latency afferent inhibition (LAI) were used to examine the sensorimotor integration. Sensorimotor plasticity was measured with paired associative stimulation paradigm, and motor cortex excitability was examined with short interval intracortical inhibition and intracortical facilitation. DBS was turned off and on to record these measures. STN DBS modulated SAI and LAI, which correlated well with the acute clinical improvement. While there were no changes seen in the motor cortex excitability, DBS was found to normalise the sensorimotor plasticity; however, there was no clinical correlation. Modulation of sensorimotor integration is a key contributor to clinical improvement with acute stimulation of STN. Since the motor cortex excitability did not change and the change in sensorimotor plasticity did not correlate with clinical improvement, STN DBS demonstrates restricted effects on the underlying physiology. NCT01671527.


On the Discretization of the Power-Law Hemolysis Model.
Flow-induced hemolysis remains a concern for blood-contacting devices, and computer-based prediction of hemolysis could facilitate faster and more economical refinement of such devices. While evaluation of convergence of velocity fields obtained by computational fluid dynamics (CFD) simulations has become conventional, convergence of hemolysis calculations is also essential. In this paper, convergence of the power-law hemolysis model is compared for simple flows, including pathlines with exponentially increasing and decreasing stress, in gradually expanding and contracting Couette flows, in a sudden radial expansion and in the Food and Drug Administration (FDA) channel. In the exponential cases, convergence along a pathline required from one to tens of thousands of timesteps, depending on the exponent. Greater timesteps were required for rapidly increasing (large exponent) stress and for rapidly decreasing (small exponent) stress. Example pathlines in the Couette flows could be fit with exponential curves, and convergence behavior followed the trends identified from the exponential cases. More complex flows, such as in the radial expansion and the FDA channel, increase the likelihood of encountering problematic pathlines. For the exponential cases, comparison of converged hemolysis values with analytical solutions demonstrated that the error of the converged solution may exceed 10% for both rapidly decreasing and rapidly increasing stress.


Neural injury after use of vasopressin and adrenaline during porcine cardiopulmonary resuscitation.
Our aim was to investigate cerebral and cardiac tissue injury subsequent to use of vasopressin and adrenaline in combination compared with vasopressin alone during cardiopulmonary resuscitation (CPR). In a randomized, prospective, laboratory animal study 28 anesthetized piglets were subject to a 12-min untreated cardiac arrest and subsequent CPR. After 1 min of CPR, 10 of the piglets received 0.4 U/kg of arg(8)-vasopressin (V group), and 10 piglets received 0.4 U/kg of arg(8)-vasopressin, 1 min later followed by 20 µg/kg body weight of adrenaline, and another 1 min later continuous administration (10 µg/kg/min) of adrenaline (VA group). After 8 min of CPR, the piglets were defibrillated and monitored for another 3 h. Then they were killed and the brain immediately removed pending histological analysis. During CPR, the VA group had higher mean blood pressure and cerebral cortical blood flow (CCBF) but similar coronary perfusion pressure. After restoration of spontaneous circulation there was no difference in the pressure variables, but CCBF tended to be (36% ± 16%) higher in the V group. Neuronal injury and signs of a disrupted blood-brain barrier (BBB) were greater, 20% ± 4% and 21% ± 4%, respectively, in the VA group. In a background study of repeated single doses of adrenaline every third minute after 5 min arrest but otherwise the same protocol, histological measurements showed even worse neural injury and disruption of the BBB. Combined use of vasopressin and adrenaline caused greater signs of cerebral and cardiac injury than use of vasopressin alone during experimental cardiopulmonary resuscitation.


Quantitation of antigen-specific CD8+ T-cell responses.
Enumeration of antigen-specific CD8+ T-cells has traditionally been largely dependent on the use of techniques that require a period of expansion in vitro. However the recent emergence of methods that can be used directly ex vivo to quantify antigen-specific CD8+ T cells has prompted a re-evaluation of the magnitude of viral-specific cellular immune responses. The purpose of this review is to describe the commonly used methods currently available to quantify and characterise antigen-specific T-cells and then to focus in more detail on the use of tetrameric complexes of HLA/peptide to study the cellular immune response.


PREVALENCE OF ANTI-CYCLIC CITRULLINATED PEPTIDE ANTIBODIES IN PATIENTS CLASSIFIED AS RHEUMATOID AND UNDIFFERENTIATED ARTHRITIS AT KENYATTA NATIONAL HOSPITAL.
Rheumatoid arthritis(RA) is a debilitating condition.Early diagnosis of RA can be difficult as the disease may initially be indistinguishable from Undifferentiated arthritis(UA). American College of Rheumatology criteria(ACR)is not suitable for early diagnosis as its characteristics are fulfilled when bone damage has already taken place.Anti-cyclic citrullinated antibodies(Anti-CCP) are highly specific for RA and have been used to confirm early diagnosis. To determine the prevalence and clinical utility of Anti-CCP antibodies in patients with rheumatoid and undifferentiated arthritis at presentation to KNH medical clinics. Design: A cross-sectional descriptive study. Kenyatta National Hospital Medical Outpatient Clinics (MOPCs) between the month of October 2008 to February 2009. A total of 95 patients were recruited.The mean age of the patients studied in the RA and UA was 44.7 and 41.2(p=0.356) respectively. Sixty four patients(64) satisfied ACR criteria.The overall prevalence of Anti-ccp antibodies in the population studied was 47.4%.The prevalence of Anti-ccp antibodies in patients who satisfied the ACR criteria was 62.5%.The prevalence of Rheumatoid Factor (RF) in patients who satisfied the ACR criteria was 50% compared to 9.7% for those who did not(p=0.000). The male to female ratio of subjects studied was 1:11. Anti-ccp antibodies are more prevalent in this cohort of patients with rheumatoid and undifferentiated arthritis than RF It was also concluded that ACR characteristics correlated well with Anti-ccp and RF. A greater percentage of patients who were RF negative were Anti-ccp positive.


[Community acquired pneumonia: from intravenous to oral cephalosporin sequential therapy].
Many hospitalized patients with community acquired pneumoniae can be switched early in the course of therapy from intravenous to oral antibiotics, when there are subjective and objective indicators of improvement. This modality of treatment is called "switch therapy". To compare sequential therapy using an oral third generation cephalosporin, with conventional therapy using intravenous ceftriaxone in community acquired pneumonia. Forty patients admitted due to community acquired pneumonia, initially treated with ceftriaxone 1 g/day i.v. and that showed clinical improvement after three days of therapy, were studied. They were randomly assigned to continue intravenous therapy with ceftriaxone for a total of 10 days or switched to ceftibuten 400 mg od for seven days. Twenty one patients continued i.v. treatment and 19 were switched to ceftibuten. There were no differences between both groups in terms of clinical cure, radiological improvement or normalisation of white blood cell count. Patients with community acquired pneumonia that have a good initial response to intravenous antimicrobials, can be safely switched to oral therapy. This therapy will shorten hospital stay and thereby treatment costs.


Deaf adults' attitudes toward career choices for deaf and hearing people in India.
This study investigated the expressed attitudes of deaf people in India toward career choices for deaf and hearing people. Deaf adults from Pune, India rated the suitability of 12 professions for deaf and hearing people and gave written comments on the suitability of any other professions they could list. The results, in general, were consistent with those of other studies in the United States, England, Italy, South Africa, and India with hearing teachers and parents of deaf children, which indicated that the hearing status of imagined deaf and hearing advisees selectively influenced attitudes toward the suitability of certain professions. Some differences in profession preferences also emerged indicating that the deaf respondents' criteria for career choice appeared to be primarily based on the use of hearing, speech, and visual skills required for a particular career. There was some suggestion that culturally specific factors played a role in shaping attitudes. These findings underscore the importance of understanding the attitudes of deaf people.


Protective Effects of Melatonin on Retinal Inflammation and Oxidative Stress in Experimental Diabetic Retinopathy.
Oxidative stress and inflammation are important pathogenic factors contributing to the etiology of diabetic retinopathy (DR). Melatonin is an endogenous hormone that exhibits a variety of biological effects including antioxidant and anti-inflammatory functions. The goals of this study were to determine whether melatonin could ameliorate retinal injury and to explore the potential mechanisms. Diabetes was induced by a single intraperitoneal (i.p.) injection of STZ (60 mg/kg) in Sprague-Dawley rats. Melatonin (10 mg kg(-1) daily, i.p.) was administered from the induction of diabetes and continued for up to 12 weeks, after which the animals were sacrificed and retinal samples were collected. The retina of diabetic rats showed depletion of glutathione and downregulation of glutamate cysteine ligase (GCL). Melatonin significantly upregulated GCL by retaining Nrf2 in the nucleus and stimulating Akt phosphorylation. The production of proinflammatory cytokines and proteins, including interleukin 1β, TNF-α, and inducible nitric oxide synthase (iNOS), was inhibited by melatonin through the NF-κB pathway. At 12 weeks, melatonin prevented the significant decrease in the ERG a- and b-wave amplitudes under the diabetic condition. Our results suggest potent protective functions of melatonin in diabetic retinopathy. In addition to being a direct antioxidant, melatonin can exert receptor-mediated signaling effects to attenuate inflammation and oxidative stress of the retina.


Multiple responses to EGF receptor activation and their abrogation by a specific EGF receptor tyrosine kinase inhibitor.
Epidermal growth factor receptor (EGF-R) autophosphorylation is essential for its intracellular mitogenic signaling via the MAPK pathway and for interaction in other cellular processes. Inhibition of this activity in tumor cells that predominantly utilise EGF-R therefore offers an alternative approach to therapy. The ability of a specific inhibitor of EGF-R tyrosine kinase, ZM 252868, (TKI) to alter various parameters related to growth in DU145 and PC3 cell lines was investigated, by immunocytochemistry, Northern blotting, Western blotting and invasion assays. In DU145 cultures, the total cell population and number of cells in cell cycle decreased in the presence of TKI whilst the apoptotic rate was significantly increased. Reduction in autophosphorylation of the EGF-R, membrane expression of EGF-R, activation of the MAPK, p38, and JNK enzymes and the invasive capacity of DU145 cells was observed in the TKI treated cells. Under the same conditions, PC3 cell growth and EGF-R expression and MAPK activation were not affected. The use of inhibitors of intracellular signaling indicated that the DU145 cells, in contrast to PC3 cells, predominantly utilize EGF-R activation of the MAPK signaling pathway for growth. In prostatic cancer patients, in whom androgen resistance has developed and whose tumors have upregulated EGF-R for growth, specific TKI's may offer an important therapy option.


Reducing intrapartum-related deaths and disability: can the health system deliver?
Each year 1.02 million intrapartum stillbirths and 904,000 intrapartum-related neonatal deaths (formerly called "birth asphyxia") occur, closely linked to 536,000 maternal deaths, an estimated 42% of which are intrapartum-related. To summarize the results of a systematic evidence review, and synthesize actions required to strengthen healthcare delivery systems and home care to reduce intrapartum-related deaths. For this series, systematic searches were undertaken, data synthesized, and meta-analyses carried out for various aspects of intrapartum care, including: obstetric care, neonatal resuscitation, strategies to link communities with facility-based care, care within communities for 60 million non-facility births, and perinatal audit. We used the Lives Saved Tool (LiST) to estimate neonatal deaths prevented with relevant interventions under 2 scenarios: (1) to address missed opportunities for facility and home births; and (2) assuming full coverage of comprehensive emergency obstetric care and emergency newborn care. Countries were first grouped into 5 Categories according to level of neonatal mortality rate and examined, and then priorities were suggested to reduce intrapartum-related deaths for each Category based on health performance and possible lives saved. There is moderate GRADE evidence of effectiveness for the reduction of intrapartum-related mortality through facility-based neonatal resuscitation, perinatal audit, integrated community health worker packages, and community mobilization. The quality of evidence for obstetric care is low, requiring further evaluation for effect on perinatal outcomes, but is expected to be high impact. Over three-quarters of intrapartum-related deaths occur in settings with weak health systems marked by low coverage of skilled birth attendance (<50%), low density of skilled human resources (<0.9 per 1000 population) and low per capita spending on health (<US $20 per year). By providing comprehensive emergency obstetric care and emergency newborn care for births already occurring in facilities, 327,200 intrapartum-related neonatal deaths could be averted globally, and with full (90%) coverage, 613,000 intrapartum-related neonatal deaths could be saved, primarily in high mortality settings. Even in high-performance settings, there is scope to improve intrapartum care and especially reduce impairment and disability. Addressing missed opportunities for births already occurring in facilities could avert 36% of intrapartum-related deaths. Improved quality of care through drills and audit are promising strategies. However, the majority of deaths occur in poorly performing health systems requiring urgent strategic planning and investment to scale up effective care at birth, neonatal resuscitation, and community mobilization as well as to develop, adapt, and introduce tools, technologies, and task shifting to reach the poorest.


Slowed saccades and increased square wave jerks in essential tremor.
Eye movements in essential tremor (ET) are poorly described and may present useful information on the underlying pathophysiology of the disorder. Sixty patients with ET, including 15 de novo untreated patients, and 60 age-matched controls constitute the study population. A video-based eye tracker was used to assess binocular eye position. Oculomotor function was assessed while subjects followed random horizontally and vertically step-displaced targets. For all reflexive saccades, latencies were increased in ET subjects by a mean of 16.3% (p<0.01). Saccades showed reduced peak velocities with a lengthy, wavering velocity plateau, followed by slowed decelerations. For larger 30°+ saccades, peak velocities were decreased by a mean of 25.2% (p<0.01) and durations increased by 31.8% (p<0.01). The frequency of square wave jerks (SWJs) in patients was more than triple that of controls (p<0.0001). Despite frequent interruptions by SWJs, fixations were otherwise stable and indistinguishable from controls (root mean square [RMS] velocity, p = 0.324). The abnormal eye movement parameters were independent of disease duration, tremor severity, and medication therapy. In contrast to normally swift onset and efficient acceleration/deceleration movements, saccades in ET are characterized by abnormally prolonged latencies and slowed velocity profiles. Although ET subjects maintain highly stable fixations, they are interrupted by increased numbers of SWJs. This study reveals novel oculomotor deficits in ET, which are distinct from the eye movement dysfunction of other movement disorders, supporting a role for eye tracking to assist in the differential diagnoses of not only atypical, but also more common movement disorders.


The "other" respiratory effect of opioids: suppression of spontaneous augmented ("sigh") breaths.
The purpose of this study was to examine the effects of a clinically relevant opioid on the production of augmented breaths (ABs) in unanesthetized animals breathing normal room air, using a dosage which does not depress breathing. To do this we monitored breathing noninvasively, in unrestrained animals before and after subcutaneous injection of either morphine, or a saline control. The effect of ketamine/xylazine was also studied to determine the potential effect of an alternative sedative agent. Last, the effect of naloxone was studied to determine the potential influence of endogenous opioids in regulating the normal incidence of ABs. Morphine (5 mg/kg) had no depressive effect on breathing, but completely eliminated ABs in all animals in room air (P = 0.027). However, when animals breathed hypoxic air (10% O(2)), animals did express ABs, although their incidence was still reduced by morphine (P < 0.001). This was not a result of sedation per se, as ABs continued at their normal rate in room air during sedation with ketamine. Naloxone had no effect on breathing or AB production, and so endogenous opioids are not likely involved in regulating their rate of production under normal conditions. Our results show that in unanesthetized animals breathing normal room air, a clinically relevant opioid eliminates ABs, even at a dose that does not cause respiratory depression. Despite this, hypoxia-induced stimulation of breathing can facilitate the production of ABs even with the systemic opioid present, indicating that peripheral chemoreceptor stimulation provides a potential means of overcoming the opioid-induced suppression of these respiratory events.


A Surprise in the Lacrimal Sac.
To present a case with recurrent dacryocystitis as an unusual complication of medial orbital wall fracture repair with cartilage tissue graft. A 20-year-old male had facial trauma and underwent surgery to reconstruct right medial orbital wall fracture. During follow-up, he presented with continuous epiphora, mucopurulent discharge from the right eye. A thorough history taking indicated that medial orbital fracture was reconstructed with postauricular cartilage. We planned a standard external dacryocystorhinostomy (DCR). During the creation of lacrimal sac flaps, hard tissue was noted in the lacrimal sac. This tissue was excised and sent for pathological examination. The pathological examination revealed cartilage tissue. There were no further ipsilateral symptoms or complications after DCR. In patients with lacrimal system injury related to orbital wall fracture, iatrogenic foreign bodies in the lacrimal sac should be considered in patients with recurrent dacryocystitis who had reconstructive surgery for facial and orbital trauma.


Predictors of atherosclerotic events in patients on haemodialysis: post hoc analyses from the AURORA study.
Patients on haemodialysis (HD) are at high risk for cardiovascular events, but heart failure and sudden death are more common than atherosclerotic events. The A Study to Evaluate the Use of Rosuvastatinin in Subjects on Regular Hemodialysis: An Assessment of Survival and Cardiovascular Events (AURORA) trial was designed to assess the effect of rosuvastatin on myocardial infarction and death from any cardiac cause in 2773 HD patients. We studied predictors of the atherosclerotic cardiovascular events in AURORA. We readjudicated all deaths and presumed myocardial infarctions according to the criteria used in the Study of Heart and Renal Protection (SHARP); these were specifically developed to separate atherosclerotic from non-atherosclerotic cardiovascular events. The readjudicated atherosclerotic end point included the first event of the following: non-fatal myocardial infarction, fatal coronary heart disease, non-fatal and fatal non-haemorrhagic stroke, coronary revascularization procedures and death from ischaemic limb disease. Stepwise Cox regression analysis was used to identify the predictors of such events. During a mean follow-up of 3.2 years, 506 patients experienced the new composite atherosclerotic outcome. Age, male sex, prevalent diabetes, prior cardiovascular disease, weekly dialysis duration, baseline albumin [hazard ratio (HR) 0.96; 95% confidence interval (CI) 0.94-0.99 per g/L increase], high-sensitivity C-reactive protein (HR 1.13; 95% CI 1.04-1.22 per mg/L increase) and oxidized low-density lipoprotein (LDL) cholesterol (HR 1.09; 95% CI 1.03-1.17 per 10 U/L increase) were selected as significant predictors in the model. Neither LDL cholesterol nor allocation to placebo/rosuvastatin therapy predicted the outcome. Even with the use of strict criteria for end point definition, non-traditional risk factors, but not lipid disturbances, predicted atherosclerotic events in HD patients.


Successful treatment of an MTBE-impacted aquifer using a bioreactor self-colonized by native aquifer bacteria.
A field-scale fixed bed bioreactor was used to successfully treat an MTBE-contaminated aquifer in North Hollywood, CA without requiring inoculation with introduced bacteria. Native bacteria from the MTBE-impacted aquifer rapidly colonized the bioreactor, entering the bioreactor in the contaminated groundwater pumped from the site, and biodegraded MTBE with greater than 99 % removal efficiency. DNA sequencing of the 16S rRNA gene identified MTBE-degrading bacteria Methylibium petroleiphilum in the bioreactor. Quantitative PCR showed M. petroleiphilum enriched by three orders of magnitude in the bioreactor above densities pre-existing in the groundwater. Because treatment was carried out by indigenous rather than introduced organisms, regulatory approval was obtained for implementation of a full-scale bioreactor to continue treatment of the aquifer. In addition, after confirmation of MTBE removal in the bioreactor to below maximum contaminant limit levels (MCL; MTBE = 5 μg L(-1)), treated water was approved for reinjection back into the aquifer rather than requiring discharge to a water treatment system. This is the first treatment system in California to be approved for reinjection of biologically treated effluent into a drinking water aquifer. This study demonstrated the potential for using native microbial communities already present in the aquifer as an inoculum for ex-situ bioreactors, circumventing the need to establish non-native, non-acclimated and potentially costly inoculants. Understanding and harnessing the metabolic potential of native organisms circumvents some of the issues associated with introducing non-native organisms into drinking water aquifers, and can provide a low-cost and efficient remediation technology that can streamline future bioremediation approval processes.


Antifibrotic drugs as treatment of nonidiopathic pulmonary fibrosis interstitial pneumonias: the time is now (?).
Therapeutic advances in the management of idiopathic pulmonary fibrosis (IPF) has led to improved outcomes with the use of the antifibrotic agents pirfenidone and nintedanib, with a number of randomized studies demonstrating benefits in slowing disease progression in IPF. However, treatment of other fibrosing interstitial lung diseases (ILD) remains challenging. Observational and uncontrolled studies investigating pirfenidone and nintedanib in non-IPF ILDs suggest potential benefits, although the data is weak. A number of randomized controlled trials evaluating pirfenidone and nintedanib in a variety of fibrosing ILDs are currently enrolling or planned. The review will discuss the rationale for use of established antifibrotic drugs approved for IPF for use in non-IPF ILD, describe supportive data from observational studies and ongoing clinical trials.


Detection of respiratory viruses by PCR assay of nasopharyngeal swabs stored in skim milk-tryptone-glucose-glycerol transport medium.
We analyzed 129 paired nasopharyngeal aspirates (stored in viral transport medium [VTM]) and nasopharyngeal swabs (stored in skim milk-tryptone-glucose-glycerol [STGG] bacterial transport and storage medium) using PCRs to detect adenoviruses, influenza virus A or B, and respiratory syncytial virus (RSV). Overall, swabs stored in STGG medium without antimicrobials were found to be an acceptable alternative to aspirates stored in antimicrobial-containing VTM, with PCR agreement of 90.2% (kappa of 0.8).


Self-folding nanostructures with imprint patterned surfaces (SNIPS).
A significant need in nanotechnology is the development of methods to mass-produce three-dimensional (3D) nanostructures and their ordered assemblies with patterns of functional materials such as metals, ceramics, device grade semiconductors, and polymers. While top-down lithography approaches can enable heterogeneous integration, tunability, and significant material versatility, these methods enable inherently two-dimensional (2D) patterning. Bottom-up approaches enable mass-production of 3D nanostructures and their assemblies but with limited precision, and tunability in surface patterning. Here, we demonstrate a methodology to create Self-folding Nanostructures with Imprint Patterned Surfaces (SNIPS). By a variety of examples, we illustrate that SNIPS, either individually or in ordered arrays, are mass-producible and have significant tunability, material heterogeneity, and patterning precision.


Dementia care and quality of life in assisted living and nursing homes.
There are few empirical studies relating components of long-term care to quality of life for residents with dementia. This study relates elements of dementia care in residential care/assisted living (RC/AL) facilities and nursing homes to resident quality of life and considers the guidance this information provides for practice and policy. We used a variety of report and observational measures of the structure and process of care and 11 standardized measures of quality of life to evaluate the care for and quality of life of 421 residents with dementia in 35 RC/AL facilities and 10 nursing homes in four states. Data were collected cross sectionally on-site, and we conducted a 6-month follow-up by telephone. Change in quality of life was better in facilities that used a specialized worker approach, trained more staff in more domains central to dementia care, and encouraged activity participation. Residents perceived their quality of life as better when staff was more involved in care planning and when staff attitudes were more favorable. Better resident-staff communication was related to higher quality of life as observed and reported by care providers. Also, more stable resident-staff assignment was related to care providers' lower quality-of-life ratings. Improvement in resident quality of life may be achieved by improved training and deployment of staff.


A good death: is euthanasia the answer?
Euthanasia--particularly active voluntary euthanasia--and assisted suicide are subjects of continuing controversy. Historical attitudes, current concerns, the situation in the Netherlands, and the positions of various medical associations are reviewed. Major arguments for and against active euthanasia are presented, with special consideration to the role that health care providers might be asked to perform should active euthanasia and assisted suicide be given societal sanction. The authors conclude that better pain management and A willingness to provide care within already established ethical and legal guidelines, not the legalization of active euthanasia and assisted suicide, are the appropriate responses to current proposals for assistance in dying.