Isolation of a mouse cDNA encoding MTJ1, a new murine member of the DnaJ family of proteins.
We report the isolation and sequencing of MTJ1, a 1792-bp cDNA from an M27 murine lung carcinoma cell line. The largest ORF within MTJ1 encodes a 63,869-Da protein, containing a 73-amino-acid (aa) sequence (the J domain) that is conserved in proteins of the DnaJ family of chaperonins. The J domain of MTJ1 is bracketed by potential transmembrane domains in a similar configuration to the J domain of the yeast DnaJ-like protein, SEC63. Polyclonal antibodies raised against deduced aa sequences within MTJ1 recognized antigens of 62, 42 and 41 kDa that were enriched in the nuclear and heavy microsome subcellular fractions of murine tumor cells. Northern analysis detected a major 3.2-kb transcript that was present in all murine organs examined, but was relatively underexpressed in brain and heart.


Ultra Stable Molecular Sensors by Submicron Referencing and Why They Should Be Interrogated by Optical Diffraction-Part II. Experimental Demonstration.
Label-free optical biosensors are an invaluable tool for molecular interaction analysis. Over the past 30 years, refractometric biosensors and, in particular, surface plasmon resonance have matured to the de facto standard of this field despite a significant cross reactivity to environmental and experimental noise sources. In this paper, we demonstrate that sensors that apply the spatial affinity lock-in principle (part I) and perform readout by diffraction overcome the drawbacks of established refractometric biosensors. We show this with a direct comparison of the cover refractive index jump sensitivity as well as the surface mass resolution of an unstabilized diffractometric biosensor with a state-of-the-art Biacore 8k. A combined refractometric diffractometric biosensor demonstrates that a refractometric sensor requires a much higher measurement precision than the diffractometric to achieve the same resolution. In a conceptual and quantitative discussion, we elucidate the physical reasons behind and define the figure of merit of diffractometric biosensors. Because low-precision unstabilized diffractometric devices achieve the same resolution as bulky stabilized refractometric sensors, we believe that label-free optical sensors might soon move beyond the drug discovery lab as miniaturized, mass-produced environmental/medical sensors. In fact, combined with the right surface chemistry and recognition element, they might even bring the senses of smell/taste to our smart devices.


Validation of the Spanish translation of the Patient Assessment of Chronic Illness Care (PACIC) survey.
The Patient Assessment of Chronic Illness Care (PACIC) survey is a patient-centered instrument for evaluating the quality and patient-centeredness of chronic illness care received according to the Chronic Care Model paradigm. This study validates the Spanish translation of the PACIC in an urban, Spanish-speaking population. One hundred Spanish-speaking patients with diabetes completed the translated PACIC and sociodemographic and cultural questionnaires. Test-retest reliability was assessed in a subset of 20 patients who completed the questionnaire 2 to 4 months later. Internal consistency was evaluated with Cronbach alpha. PACIC score and subscale associations with sociodemographic characteristics were examined. Test-retest reliability for the overall translated PACIC scale was 0.77. Scores were not associated with patient sociodemographic characteristics, including age, country of birth, years living in the United States, or education level (P >.05). The Spanish translation of the PACIC survey demonstrated high reliability, internal consistency, and test-retest reliability. Scores showed no association with sociodemographic or cultural characteristics. The Spanish version can reliably be used to assess care delivered according to the Chronic Care Model in a heterogeneous Spanish-speaking population.


Etiological study of esophageal squamous cell carcinoma in an endemic region: a population-based case control study in Huaian, China.
Continuous exposure to various environmental carcinogens and genetic polymorphisms of xenobiotic-metabolizing enzymes (XME) are associated with many types of human cancers, including esophageal squamous cell carcinoma (ESCC). Huaian, China, is one of the endemic regions of ESCC, but fewer studies have been done in characterizing the risk factors of ESCC in this area. The aims of this study is to evaluate the etiological roles of demographic parameters, environmental and food-borne carcinogens exposure, and XME polymorphisms in formation of ESCC, and to investigate possible gene-gene and gene-environment interactions associated with ESCC in Huaian, China. A population based case-control study was conducted in 107 ESCC newly diagnosed cases and 107 residency- age-, and sex-matched controls in 5 townships of Huaian. In addition to regular epidemiological and food frequency questionnaire analyses, genetic polymorphisms of phase I enzymes CYP1A1, CYP1B1, CYP2A6, and CYP2E1, and phase II enzymes GSTM1, GSTT1, GSTP1, and microsomal epoxide hydrolase (EPHX) were assessed from genomic DNA using PCR based techniques. Consuming acrid food, fatty meat, moldy food, salted and pickled vegetables, eating fast, introverted personality, passive smoking, a family history of cancer, esophageal lesion, and infection with Helicobacter pylori were significant risk factors for ESCC (P < 0.05). Regular clean up of food storage utensils, green tea consumption, and alcohol abstinence were protective factors for ESCC (P < 0.01). The frequency of the GSTT1 null genotype was higher in cases (59.4%) compared to controls (47.2%) with an odds ratio (OR) of 1.68 and 95% confidence interval (CI) from 0.96 to 2.97 (P = 0.07), especially in males (OR = 2.78; 95% CI = 1.22-6.25; P = 0.01). No associations were found between polymorphisms of CYP1A1, CYP1B1, CYP2A6, CYP2E1, GSTM1, GSTP1, and EPHX and ESCC (P > 0.05). Our results demonstrated that dietary and environmental exposures, some demographic parameters and genetic polymorphism of GSTT1 may play important roles in the development of ESCC in Huaian area, China.


The role of the father in parental postnatal mental health.
Contrary to idealized images of parenthood, the months immediately following childbirth are characterized by enhanced rates of parental mental illness and filicide. In this paper we consider fathers and their role in the postnatal well-being of the parturient couple. The process of becoming a father is influenced by not only the man's internal and external experiences of 'father', but also the woman's, since the new infant's relationship with father is mediated by mother and her conjugal relationship with father. When a couple's prenatal relationship is based on a system of shared parental constructs involving a denigrated father and overvalued mother and a concomitant intolerance of the idea of a creative (Oedipal) couple, they will experience particular difficulties in adjusting to the new family configuration. In working with individuals or couples who have experienced post-partum psychological difficulties it is important to remember that not only maternal imagos but also paternal and conjugal ones need to be addressed.


Angiotensin II-induced protein kinase D activates the ATF/CREB family of transcription factors and promotes StAR mRNA expression.
Aldosterone synthesis is initiated upon the transport of cholesterol from the outer to the inner mitochondrial membrane, where the cholesterol is hydrolyzed to pregnenolone. This process is the rate-limiting step in acute aldosterone production and is mediated by the steroidogenic acute regulatory (StAR) protein. We have previously shown that angiotensin II (AngII) activation of the serine/threonine protein kinase D (PKD) promotes acute aldosterone production in bovine adrenal glomerulosa cells, but the mechanism remains unclear. Thus, the purpose of this study was to determine the downstream signaling effectors of AngII-stimulated PKD activity. Our results demonstrate that overexpression of the constitutively active serine-to-glutamate PKD mutant enhances, whereas the dominant-negative serine-to-alanine PKD mutant inhibits, AngII-induced StAR mRNA expression relative to the vector control. PKD has been shown to phosphorylate members of the activating transcription factor (ATF)/cAMP response element binding protein (CREB) family of leucine zipper transcription factors, which have been shown previously to bind the StAR proximal promoter and induce StAR mRNA expression. In primary glomerulosa cells, AngII induces ATF-2 and CREB phosphorylation in a time-dependent manner. Furthermore, overexpression of the constitutively active PKD mutant enhances the AngII-elicited phosphorylation of ATF-2 and CREB, and the dominant-negative mutant inhibits this response. Furthermore, the constitutively active PKD mutant increases the binding of phosphorylated CREB to the StAR promoter. Thus, these data provide insight into the previously reported role of PKD in AngII-induced acute aldosterone production, providing a mechanism by which PKD may be mediating steroidogenesis in primary bovine adrenal glomerulosa cells.


[Ertapenem and other antibacterials].
Ertapenem is a carbapenem that shares the activity of imipenem and meropenem against most species, but is less active against non-fermenters. The activity is retained against most strains with AmpC and extended-spectrum beta-lactamases, although the resistance can arise if these enzymes are combined with extreme impermeability. The resistance can also be caused by IMP, VIM, KPC and NMC carbapenemases, but again, co-requires impermeability. Although the spread of carbapenemases in the future is a concern, they are currently very rare. Given as a 1 g intravenous (i.v.) infusion once daily, ertapenem has a plasma half-life of -4 h in healthy volunteers, and a Cn,a, of 155 mg/l and 13 mg/l for total and free drug, respectively. Excretion is largely renal, divided equally between native drug and an open-ring derivative. The trials showed equivalence to piperacillin/tazobactam or ceftriaxone in (a) abdominal infections, (b) community-acquired pneumonia, (c) acute pelvic infections, (d) skin and skin structure infections and (e) complicated urinary tract infections. The USA licence grants all these five indications; the ED licence grants the first three. Further potential uses include home i.v. therapy, target therapy against Enterobacteriaceae with AmpC or extended-spectrum cephalosporinases, and tentatively, surgical prophylaxis. Widening the usage of carbapenems raises public health concerns, somewhat allayed by the continued rarity of carbapenemases after 17 years of imipenem use, and by the fact that carbapenemases occur mostly in non-fermenters outside the spectrum of ertapenem, and co-require impermeability to confer resistance in Enterobacteriaceae. Nevertheless, if ertapenem is to be used widely, its effects on the microbial resistance ecology need to be monitored carefully.


Aortic endothelial and smooth muscle histamine metabolism in experimental diabetes.
We studied histamine metabolism, i.e., histidine decarboxylase (HD)-mediated synthesis and histaminase-mediated catabolism, in relation to intracellular histamine content in both aortic endothelial and subjacent smooth muscle cells of control and diabetic rats. Diabetes was induced by a single jugular vein injection of streptozotocin (55 mg/kg in acidified saline, pH 4.5), and animals were held for either 2 or 4 weeks following overt manifestation of diabetes. An additional 4-week diabetic group received insulin (Iletin NPH, 10 U per 24 hour) during the last week. With respect to control values, the histamine content of aortic endothelial cells increased 138%, HD activity increased 250%, and histaminase activity decreased 50% over the 4-week period. In subjacent smooth muscle cells, the histamine content increased in excess of 150%, HD activity increased more than 300%, and histaminase activity decreased in excess of 30%. Insulin treatment for the last week resulted in complete reversal of all these changes. These results support the concept that a large vessel response similar to the microcirculatory prolonged phase of inflammation occurs in experimental diabetes, a change similar to that occurring in experimental atherosclerosis. They also indicate that both synthetic and catabolic changes occur in histamine metabolism under these conditions, changes that alter arterial wall histamine pools, and suggest that insulin administration under conditions of experimental diabetes may modulate aortic histamine metabolism and the resultant intraaortic histamine pools.


Text-based phenotypic profiles incorporating biochemical phenotypes of inborn errors of metabolism improve phenomics-based diagnosis.
Phenomics is the comprehensive study of phenotypes at every level of biology: from metabolites to organisms. With high throughput technologies increasing the scope of biological discoveries, the field of phenomics has been developing rapid and precise methods to collect, catalog, and analyze phenotypes. Such methods have allowed phenotypic data to be widely used in medical applications, from assisting clinical diagnoses to prioritizing genomic diagnoses. To channel the benefits of phenomics into the field of inborn errors of metabolism (IEM), we have recently launched IEMbase, an expert-curated knowledgebase of IEM and their disease-characterizing phenotypes. While our efforts with IEMbase have realized benefits, taking full advantage of phenomics requires a comprehensive curation of IEM phenotypes in core phenomics projects, which is dependent upon contributions from the IEM clinical and research community. Here, we assess the inclusion of IEM biochemical phenotypes in a core phenomics project, the Human Phenotype Ontology. We then demonstrate the utility of biochemical phenotypes using a text-based phenomics method to predict gene-disease relationships, showing that the prediction of IEM genes is significantly better using biochemical rather than clinical profiles. The findings herein provide a motivating goal for the IEM community to expand the computationally accessible descriptions of biochemical phenotypes associated with IEM in phenomics resources.


Collagen fragment biomarkers as serological biomarkers of lean body mass - a biomarker pilot study from the DAHANCA25B cohort and matched controls.
Loss of muscle mass and function is an important complication to ageing and a range of pathologies, including, but not restricted to, cancer, organ failures, and sepsis. A number of interventions have been proposed ranging from exercise to anabolic pharmacological therapy, with varying success. Easily applicable serological biomarkers of lean and/or muscle mass and change therein would benefit monitoring of muscle mass during muscle atrophy as well as during recovery. We set out to validate if novel peptide biomarkers derived from Collagen III and VI were markers of lean body mass (LBM) or change therein in head and neck cancer patients in the Danish Head and Neck Cancer Group(DAHANCA) 25B cohort subjected to resistance training as well as in an age-matched and gender-matched control group. Blood samples and dual X-ray absorptiometry data were measured at baseline, after 12 and 24 weeks in 41 HNSCC subjects of the DAHANCA 25B cohort of subjects recovering from neck and head cancer (stages provided in Table 1), and at baseline only in 21 healthy age-matched and gender-matched controls. Serum from blood was analyzed for the ProC3, IC6, and C6M peptide biomarkers and LBM were derived from the dual X-ray absorptiometry scans. We were not able to show any correlation between biomarkers and LBM or C6M and anabolic response to exercise in recovering head and neck cancer patients. However, we did find that the biomarkers IC6, IC6/C6M, and ProC3 are biomarkers of LBM in the control group subjects (R (2)/P of 0.249/0.035, 0.416/0.007 and 0.178 and P = 0.057, respectively). In conclusion, the IC6, ProC3, and IC6/C6M biomarkers are indeed biomarkers of LBM in healthy individuals of both genders, but not in HNSCC patients.


Interaction of the molecular chaperone DNAJB6 with growing amyloid-beta 42 (Aβ42) aggregates leads to sub-stoichiometric inhibition of amyloid formation.
The human molecular chaperone protein DNAJB6 was recently found to inhibit the formation of amyloid fibrils from polyglutamine peptides associated with neurodegenerative disorders such as Huntington disease. We show in the present study that DNAJB6 also inhibits amyloid formation by an even more aggregation-prone peptide (the amyloid-beta peptide, Aβ42, implicated in Alzheimer disease) in a highly efficient manner. By monitoring fibril formation using Thioflavin T fluorescence and far-UV CD spectroscopy, we have found that the aggregation of Aβ42 is retarded by DNAJB6 in a concentration-dependent manner, extending to very low sub-stoichiometric molar ratios of chaperone to peptide. Quantitative kinetic analysis and immunochemistry studies suggest that the high inhibitory efficiency is due to the interactions of the chaperone with aggregated forms of Aβ42 rather than the monomeric form of the peptide. This interaction prevents the growth of such species to longer fibrils and inhibits the formation of new amyloid fibrils through both primary and secondary nucleation. A low dissociation rate of DNAJB6 from Aβ42 aggregates leads to its incorporation into growing fibrils and hence to its gradual depletion from solution with time. When DNAJB6 is eventually depleted, fibril proliferation takes place, but the inhibitory activity can be prolonged by introducing DNAJB6 at regular intervals during the aggregation reaction. These results reveal the highly efficacious mode of action of this molecular chaperone against protein aggregation, and demonstrate that the role of molecular chaperones can involve interactions with multiple aggregated species leading to the inhibition of both principal nucleation pathways through which aggregates are able to form.


Endovascular embolization of cerebral arteriovenous malformations: results of the Japanese Registry of Neuroendovascular Therapy (JR-NET) 1 and 2.
This retrospective study constitutes a part of the Japanese Registry of Neuroendovascular Therapy (JR-NET) 1 and 2. Its purpose is to evaluate the feasibility, safety, and outcome of endovascular embolization for cerebral arteriovenous malformations (AVMs) in Japan. Nine hundred and eighty-seven embolization procedures were registered with JR-NET 1 and 2 (424 procedures in 122 institutions with JRNET 1 and 563 procedures in 150 institutions with JRNET 2). In total, 790 patients (80.1%) had favourable clinical outcomes defined as modified Rankin Scale (mRS) scores 0-2 at 30 days after embolization. Complete AVM obliteration by embolization alone was achieved in 90 procedures (9.1%). The procedural morbidity and mortality rate was 2.5% and 0.3% per procedure, respectively. In the multivariate logistic regression models, deep venous drainage and embolization of four or more feeding pedicles per session were significantly associated with any treatment-related complications (P = 0.02 and P = 0.003, respectively). About 6 cm or more in maximum nidus diameter had a negative correlation with complications (P = 0.003). Our study shows that embolization of cerebral AVMs was performed with a high degree of safety and a low rate of symptomatic complications in Japan.


The molecular diversity of adaptive convergence.
To estimate the number and diversity of beneficial mutations, we experimentally evolved 115 populations of Escherichia coli to 42.2°C for 2000 generations and sequenced one genome from each population. We identified 1331 total mutations, affecting more than 600 different sites. Few mutations were shared among replicates, but a strong pattern of convergence emerged at the level of genes, operons, and functional complexes. Our experiment uncovered a set of primary functional targets of high temperature, but we estimate that many other beneficial mutations could contribute to similar adaptive outcomes. We inferred the pervasive presence of epistasis among beneficial mutations, which shaped adaptive trajectories into at least two distinct pathways involving mutations either in the RNA polymerase complex or the termination factor rho.


miR-128 regulates non-myocyte hyperplasia, deposition of extracellular matrix and Islet1 expression during newt cardiac regeneration.
Cardiovascular disease is a global scourge to society, with novel therapeutic approaches required in order to alleviate the suffering caused by sustained cardiac damage. MicroRNAs (miRNAs) are being touted as one such approach in the fight against heart disease, acting as possible post-transcriptional molecular triggers responsible for invoking cardiac regeneration. To further ones understanding of miRNAs and cardiac regeneration, it is prudent to learn from organisms that can intrinsically regenerate their hearts following injury. Using the red-spotted newt, an adult chordate capable of cardiac regeneration, we decided to delve deeper into the role miRNAs play during this process. RNA isolated from regenerating newt heart samples, was used in a microarray screen, to identify significantly expressed candidate miRNAs during newt cardiac regeneration. We performed quantitative qPCR analysis on several conserved miRNAs and found one in particular, miR-128, to be significantly elevated when cardiac hyperplasia is at its peak following injury. In-situ hybridisation techniques revealed a localised expression pattern for miR-128 in the cardiomyocytes and non-cardiomyocytes in close proximity to the regeneration zone and in vivo knockdown studies revealed a regulatory role for miR-128 in proliferating non-cardiomyocyte populations and extracellular matrix deposition. Finally, 3'UTR reporter assays revealed Islet1 as a biological target for miR-128, which was confirmed further through in vivo Islet1 transcriptional and translational expression analysis in regenerating newt hearts. From these studies we conclude that miR-128 regulates both cardiac hyperplasia and Islet1 expression during newt heart regeneration and that this information could be translated into future mammalian cardiac studies.


Association of opioid misuse with anxiety and depression: A systematic review of the literature.
The opioid epidemic is a public health problem associated with a host of negative outcomes. Although clinicians recognize covariation between opioid misuse with anxiety and depressive symptoms and disorders, research on this topic has only recently accumulated. Progress in this domain is impeded by the lack of systematic and integrative research to better understand and treat these co-occurring problems. This paper represents the first attempt to systematically review the empirical literature examining relations between opioid use and misuse, and anxiety and depression. In the first section, we define key terms and describe the article selection strategy. In the second section, we review the prevalence of anxiety and depressive symptoms among individuals who use and misuse prescription and illicit opioids. In the third section, we review the magnitude of associations between anxiety and depressive symptoms and disorders with opioid misuse, as well as highlight studies examining the longitudinal and temporal sequence of the relations between these variables. In the fourth section, we focus on experimental therapeutics, reviewing what is known about individual difference and transdiagnostic vulnerability factors for anxiety and depression that might contribute to opioid misuse and its symptoms. Finally, we discuss current knowledge gaps and present a heuristic model to guide future research.


Metabotyping: a new approach to investigate rapeseed (Brassica napus L.) genetic diversity in the metabolic response to clubroot infection.
Clubroot disease affects all Brassicaceae spp. and is caused by the obligate biotroph pathogen Plasmodiophora brassicae. The development of galls on the root system is associated with the establishment of a new carbon metabolic sink. Here, we aimed to deepen our knowledge of the involvement of primary metabolism in the Brassica napus response to clubroot infection. We studied the dynamics and the diversity of the metabolic responses to the infection. Root system metabotyping was carried out for 18 rapeseed genotypes displaying different degrees of symptom severity, under inoculated and noninoculated conditions at 42 days postinoculation (dpi). Clubroot susceptibility was positively correlated with clubroot-induced accumulation of several amino acids. Although glucose and fructose accumulated in some genotypes with minor symptoms, their levels were negatively correlated to the disease index across the whole set of genotypes. The dynamics of the metabolic response were studied for the susceptible genotype 'Yudal,' which allowed an "early" metabolic response (established from 14 to 28 dpi) to be differentiated from a "late" response (from 35 dpi). We discuss the early accumulation of amino acids in the context of the establishment of a nitrogen metabolic sink and the hypothetical biological role of the accumulation of glutathione and S-methylcysteine.


A network-based gene expression signature informs prognosis and treatment for colorectal cancer patients.
Several studies have reported gene expression signatures that predict recurrence risk in stage II and III colorectal cancer (CRC) patients with minimal gene membership overlap and undefined biological relevance. The goal of this study was to investigate biological themes underlying these signatures, to infer genes of potential mechanistic importance to the CRC recurrence phenotype and to test whether accurate prognostic models can be developed using mechanistically important genes. We investigated eight published CRC gene expression signatures and found no functional convergence in Gene Ontology enrichment analysis. Using a random walk-based approach, we integrated these signatures and publicly available somatic mutation data on a protein-protein interaction network and inferred 487 genes that were plausible candidate molecular underpinnings for the CRC recurrence phenotype. We named the list of 487 genes a NEM signature because it integrated information from Network, Expression, and Mutation. The signature showed significant enrichment in four biological processes closely related to cancer pathophysiology and provided good coverage of known oncogenes, tumor suppressors, and CRC-related signaling pathways. A NEM signature-based Survival Support Vector Machine prognostic model was trained using a microarray gene expression dataset and tested on an independent dataset. The model-based scores showed a 75.7% concordance with the real survival data and separated patients into two groups with significantly different relapse-free survival (p = 0.002). Similar results were obtained with reversed training and testing datasets (p = 0.007). Furthermore, adjuvant chemotherapy was significantly associated with prolonged survival of the high-risk patients (p = 0.006), but not beneficial to the low-risk patients (p = 0.491). The NEM signature not only reflects CRC biology but also informs patient prognosis and treatment response. Thus, the network-based data integration method provides a convergence between biological relevance and clinical usefulness in gene signature development.


Evaluation of diffusion models in breast cancer.
The purpose of this study is to investigate whether the microvascular pseudodiffusion effects resulting with non-monoexponential behavior are present in breast cancer, taking into account tumor spatial heterogeneity. Additionally, methodological factors affecting the signal in low and high diffusion-sensitizing gradient ranges were explored in phantom studies. The effect of eddy currents and accuracy of b-value determination using a multiple b-value diffusion-weighted MR imaging sequence were investigated in test objects. Diffusion model selection and noise were then investigated in volunteers (n = 5) and breast tumor patients (n = 21) using the Bayesian information criterion. 54.3% of lesion voxels were best fitted by a monoexponential, 26.2% by a stretched-exponential, and 19.5% by a biexponential intravoxel incoherent motion (IVIM) model. High correlation (0.92) was observed between diffusion coefficients calculated using mono- and stretched-exponential models and moderate (0.59) between monoexponential and IVIM (medians: 0.96/0.84/0.72 × 10(-3) mm(2)/s, respectively). Distortion due to eddy currents depended on the direction of the diffusion gradient and displacement varied between 1 and 6 mm for high b-value images. Shift in the apparent diffusion coefficient due to intrinsic field gradients was compensated for by averaging diffusion data obtained from opposite directions. Pseudodiffusion and intravoxel heterogeneity effects were not observed in approximately half of breast cancer and normal tissue voxels. This result indicates that stretched and IVIM models should be utilized in regional analysis rather than global tumor assessment. Cross terms between diffusion-sensitization gradients and other imaging or susceptibility-related gradients are relevant in clinical protocols, supporting the use of geometric averaging of diffusion-weighted images acquired with diffusion-sensitization gradients in opposite directions.


Bi-stable depth ordering of superimposed moving gratings.
Ambiguous stimuli with two distinct interpretations give rise to perceptual alternations between them. During prolonged viewing of transparently moving gratings, observers report periods of perceiving one grating in front of the other, alternating with periods of the reverse depth ordering. We measured the percepts' dominance times to study the effect of depth cues (wavelength, duty cycle, and speed) on the perceived depth ordering. The grating with shorter wavelength, lower duty cycle, or higher speed was perceived as being behind the other for a fraction of time larger than one half. The fraction of time spent perceiving each grating as behind changed gradually as a function of the parameters. The fraction of dominance depended on the ratio between the gratings' wavelengths, not on their absolute sizes. The wavelength ratio had a stronger effect on perceived depth than that of duty cycle or speed and could override stereoscopic disparity cues. Similar results were obtained with superimposed moving surfaces of random dots. The findings are interpreted in terms of their relation to statistical properties of natural surfaces and provide evidence that the fraction of dominance of each percept represents the likelihood that it corresponds to the true interpretation of the underlying scene.


Death as a stimulus to improving the African environment for more healthful living.
In D. Leviton's (1991) conception of horrendous death, a poorly managed environment can contribute to widespread mortality, and the deaths so caused can further create an environment for subsequent death. The African environment in particular is characterized by a number of "deathogenic" factors that must be understood and confronted by health educators committed to minimizing or eliminating the impact of horrendous death on the African continent. In this comment, the author argues that attention to characteristic but preventable forms of death in the African context can lead to greater public advocacy among African health educators, contributing to the physical and psychological well-being of the populations they serve.


Environmental impact of estrogens on human, animal and plant life: A critical review.
Since the inception of global industrialization, steroidal estrogens have become an emerging and serious concern. Worldwide, steroid estrogens including estrone, estradiol and estriol, pose serious threats to soil, plants, water resources and humans. Indeed, estrogens have gained notable attention in recent years, due to their rapidly increasing concentrations in soil and water all over the world. Concern has been expressed regarding the entry of estrogens into the human food chain which in turn relates to how plants take up and metabolism estrogens. In this review we explore the environmental fate of estrogens highlighting their release through effluent sources, their uptake, partitioning and physiological effects in the ecological system. We draw attention to the potential risk of intensive modern agriculture and waste disposal systems on estrogen release and their effects on human health. We also highlight their uptake and metabolism in plants. We use MEDLINE and other search data bases for estrogens in the environment from 2005 to the present, with the majority of our sources spanning the past five years. Published acceptable daily intake of estrogens (μg/L) and predicted no effect concentrations (μg/L) are listed from published sources and used as thresholds to discuss reported levels of estrogens in the aquatic and terrestrial environments. Global levels of estrogens from river sources and from Waste Water Treatment Facilities have been mapped, together with transport pathways of estrogens in plants. Estrogens at polluting levels have been detected at sites close to waste water treatment facilities and in groundwater at various sites globally. Estrogens at pollutant levels have been linked with breast cancer in women and prostate cancer in men. Estrogens also perturb fish physiology and can affect reproductive development in both domestic and wild animals. Treatment of plants with steroid estrogen hormones or their precursors can affect root and shoot development, flowering and germination. However, estrogens can ameliorate the effects of other environmental stresses on the plant. There is published evidence to establish a causal relationship between estrogens in the environment and breast cancer. However, there are serious gaps in our knowledge about estrogen levels in the environment and a call is required for a world wide effort to provide more data on many more samples sites. Of the data available, the synthetic estrogen, ethinyl estradiol, is more persistent in the environment than natural estrogens and may be a greater cause for environmental concern. Finally, we believe that there is an urgent requirement for inter-disciplinary studies of estrogens in order to better understand their ecological and environmental impact.


First Year Course-Based Undergraduate Research Experience (CURE) Using the CRISPR/Cas9 Genome Engineering Technology in Zebrafish.
Genetic analysis in model systems can provide a rich context for conceptual understanding of gene structure, regulation, and function. With an intent to create a rich learning experience in molecular genetics, we developed a semester-long course-based undergraduate research experience (CURE) using the CRISPR-Cas9 gene editing system to disrupt specific genes in the zebrafish. The course was offered to freshman students; nine students worked in four groups (two to three members per group) to design, synthesize, and test the nuclease activity of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/sgRNAs for targeted disruption of specific genes in the zebrafish. Each group worked with a gene with an already known mutant phenotype that can be visually scored and a gene that had not been studied in zebrafish previously. Embedded in the course were a series of workshop-styled units or tutorials, including tours to core facilities. The focus was on introducing and developing skills that could be accommodated within the span of a semester. Each group successfully cloned at least one plasmid-encoding CRISPR/sgRNA template, visually analyzed injected embryos, and performed genotyping assays to detect CRISPR-Cas9 activity. In-class discussions, a final end-of-semester written test, and group oral presentations were assessed for an understanding of the CRISPR-Cas9 system, application of the CRISPR-Cas9 system as a gene manipulation tool, and experimental methods used to create plasmid vectors and synthesize sgRNA. In addition, poster presentations were evaluated by faculty, graduate students, and senior undergraduate students at a University research exposition. Self-reflections in the form of group conversations were video recorded. All students (9/9) distinctly showed learning gains after completing the activity, but the extent of the gains was variable, as seen from results of a written test and poster presentation assessment. Qualitative analysis of evaluations and self-reporting data indicated several gains, suggesting that all students found many aspects of the CURE valuable and gained project-specific (conceptual) and transferrable skills (science process and science identity).


Oceanobacillus locisalsi sp. nov., isolated from a marine solar saltern.
A Gram-stain-variable, motile, moderately halophilic bacterial strain, CHL-21(T), was isolated from a marine solar saltern and its taxonomic position was investigated using a polyphasic approach. Optimal growth of strain CHL-21(T) occurred at 30-37 °C, at pH 7.0-7.5 and in the presence of 5-10 % (w/v) NaCl. In phylogenetic trees based on 16S rRNA gene sequences, strain CHL-21(T) fell within the cluster comprising members of the genera Oceanobacillus, Ornithinibacillus and Paucisalibacillus. Strain CHL-21(T) exhibited 97.1-97.2 % 16S rRNA gene sequence similarity to the type strains of the two subspecies of Oceanobacillus oncorhynchi and 92.0-94.7 % 16S rRNA gene sequence similarity to the type strains of other members of the genus Oceanobacillus and members of the genera Ornithinibacillus and Paucisalibacillus. Mean DNA-DNA reassociation values between strain CHL-21(T) and the type strains of the two subspecies of Oceanobacillus oncorhynchi were 19-21 %. The cell-wall peptidoglycan of strain CHL-21(T) was based on meso-diaminopimelic acid, MK-7 was the predominant menaquinone, and anteiso-C(15 : 0) and anteiso-C(17 : 0) were the major fatty acids. The DNA G+C content was 39.8 mol%. Differential phenotypic properties, including facultatively anaerobic growth and acid production from substrates, together with its phylogenetic and genetic distinctiveness, demonstrated that strain CHL-21(T) is distinguishable from recognized Oceanobacillus species. On the basis of data presented, strain CHL-21(T) represents a novel species within the genus Oceanobacillus, for which the name Oceanobacillus locisalsi sp. nov. is proposed; the type strain is CHL-21(T) (=KCTC 13253(T) =CCUG 56608(T)).


Full vectorial BPM modeling of Index-Guiding Photonic Crystal Fibers and Couplers.
A 3D full-vectorial Beam Propagation Method is successfully applied to compute both the propagation constants and the modal profiles in high-contrast silica-air index-guiding Photonic Crystal Fibers. The approach is intrinsically suited to investigate longitudinally varying structures or propagation and polarization effects, which are of practical interest for advanced optical applications. As an example we model a dual-core coupler, showing that efficient polarization preserving coupling can be expected.


Growth characteristics of selected fungi on polyvinyl chloride film.
The objective of this study was to determine if plasticized polyvinyl chloride film would support the growth of any of nine species of fungi. The film was suspended in distilled water with no nutrients or with glucose or ammonium sulfate. Spores of each of the test species were inoculated into the suspension medium, and the mixture was incubated at 30 degrees C for up to 18 weeks. Most species were found to be capable of utilizing the film for carbon or nitrogen when the other nutrient was supplied. Only two species, Aspergillus fischeri and Paecilomyces sp., were found to be capable of utilizing components of the film without added nutrients. Components of the polyvinyl chloride film were then incorporated into mineral salts medium to determine if these components could serve as carbon sources in the presence of ammonium nitrate. The only component found to be utilized by all the fungi as a carbon source was epoxidized oil, a plasticizer-stabilizer. Calcium-zinc stearate was an available carbon source for all except the Penicillium and Verticillium strains. The only other component utilized was a stearamide, which was metabolized solely by the Aspergillus sp. Only the stearamide contained enough nitrogen to serve as a primary source in the film. The compound, however, did not support growth of fungi in the presence of glucose. It was theorized that either the nitrogen of the stearamide was more readily available to the fungi in the whole film due to the presence of trace nutrients or the nitrogen was supplied by exogenous sources.


Quantifying the effects of diuretics and β-adrenoceptor blockers on glycaemic control in diabetes mellitus - a systematic review and meta-analysis.
Although there are reports that β-adrenoceptor antagonists (beta-blockers) and diuretics can affect glycaemic control in people with diabetes mellitus, there is no clear information on how blood glucose concentrations may change and by how much. We report results from a systematic review to quantify the effects of these antihypertensive drugs on glycaemic control in adults with established diabetes. We systematically reviewed the literature to identify randomized controlled trials in which glycaemic control was studied in adults with diabetes taking either beta-blockers or diuretics. We combined data on HbA1c and fasting blood glucose using fixed effects meta-analysis. From 3864 papers retrieved, we found 10 studies of beta-blockers and 12 studies of diuretics to include in the meta-analysis. One study included both comparisons, totalling 21 included reports. Beta-blockers increased fasting blood glucose concentrations by 0.64 mmol l(-1) (95% CI 0.24, 1.03) and diuretics by 0.77 mmol l(-1) (95% CI 0.14, 1.39) compared with placebo. Effect sizes were largest in trials of non-selective beta-blockers (1.33, 95% CI 0.72, 1.95) and thiazide diuretics (1.69, 95% CI 0.60, 2.69). Beta-blockers increased HbA1c concentrations by 0.75% (95% CI 0.30, 1.20) and diuretics by 0.24% (95% CI -0.17, 0.65) compared with placebo. There was no significant difference in the number of hypoglycaemic events between beta-blockers and placebo in three trials. Randomized trials suggest that thiazide diuretics and non-selective beta-blockers increase fasting blood glucose and HbA1c concentrations in patients with diabetes by moderate amounts. These data will inform prescribing and monitoring of beta-blockers and diuretics in patients with diabetes.


Removal of monocular interactions equates rivalry behavior for monocular, binocular, and stimulus rivalries.
When the two eyes are presented with conflicting stimuli, perception starts to fluctuate over time (i.e., binocular rivalry). A similar fluctuation occurs when two patterns are presented to a single eye (i.e., monocular rivalry), or when they are swapped rapidly and repeatedly between the eyes (i.e., stimulus rivalry). Although all these cases lead to rivalry, in quantitative terms these modes of rivalry are generally found to differ significantly. We studied these different modes of rivalry with identical intermittently shown stimuli while varying the temporal layout of stimulation. We show that the quantitative differences between the modes of rivalry are caused by the presence of monocular interactions between the rivaling patterns; the introduction of a blank period just before a stimulus swap changed the number of rivalry reports to the extent that monocular and stimulus rivalries were inducible over ranges of spatial frequency content and contrast values that were nearly identical to binocular rivalry. Moreover when monocular interactions did not occur the perceptual dynamics of monocular, binocular, and stimulus rivalries were statistically indistinguishable. This range of identical behavior exhibited a monocular (approximately 50 ms) and a binocular (approximately 350 ms) limit. We argue that a common binocular, or pattern-based, mechanism determines the temporal constraints for these modes of rivalry.


Humphry Fortescue Osmond (1917-2004), a radical and conventional psychiatrist: The transcendent years.
This article describes the life and work of the psychiatrist Humphry Osmond who pursued a radical path as a psychiatrist while he remained within the establishment. To the public mind however, he is best known as the man who introduced Aldous Huxley to mescaline and coined the iconic word psychedelic. From an early stage of his career, Henry Osmond embraced new ideas to break the nexus in psychiatry at a time when neither biological nor psychoanalytic treatments were shown to have much benefit. To do this, he joined the radical social experiment in health in the Canadian province of Saskatchewan where he initiated a range of innovations that attracted international attention, as well as controversy over his espousal of the use of hallucinogens better to understand the experiences of psychotic patients.


Food protein-derived anxiolytic peptides: their potential role in anxiety management.
About one in three people are affected by anxiety disorders during their lifetime. Anxiety episodes can be brief due to a stressful event, but anxiety disorders can last at least 6 months. A wide variety of therapeutic drugs are available for the treatment of anxiety disorders, but due to the associated side effects of these anxiolytics, it is interesting to find alternatives. Some food protein hydrolysates or active peptide fragments present in such hydrolysates provide a natural and promising mean for preventing certain forms of anxiety. To date, only a small number of hydrolysates or peptides from food proteins with anxiolytic-like activity have been characterized. Most of these hydrolysates or peptides have displayed potent anxiolytic profiles in animal or clinical studies. The results suggest that these molecules may exert their effects at different levels. This paper reviews the data of the structure/activity relationship, physiological effects displayed in in vitro and in vivo assays, bioavailability, and safety profiles of anxiolytic peptides.


Characterization of pleural effusion after left atrial appendage exclusion using the Lariat procedure.
The Lariat procedure is increasingly used for the exclusion of the left atrial appendage (LAA) in atrial fibrillation (AF) patients. There are anecdotal reports of pleural effusions after the Lariat procedure. However, the incidence, demographics, and pathophysiology of these effusions are largely unknown. Characterization of pleural effusions in patients who underwent LAA exclusion using the Lariat procedure. We report the incidence, demographics, and clinical and laboratory characteristics of patients from a multicenter prospective registry who underwent the Lariat procedure and subsequently developed pleural effusions. A total of 10 out of 310 (3.2%) patients developed significant pleural effusions after the Lariat procedure. The mean age of these patients was 67 ± 9, ranging from 52 to 78 years and included 5 (50%) males. Nine patients had persistent AF with median CHADS2 score of 2.7 ± 1.2. The LAA was successfully ligated in all these patients. Post-Lariat procedure, 6 patients developed bilateral and 4 patients developed left-sided pleural effusions. Pleural tap revealed transudative in 2 and exudative in 6 patients. The remaining 2 patients responded to active diuresis and behaved clinically like transudative effusions. There is a statistically significant difference between the onset of pleural effusion after the Lariat procedure between tPLE versus ePLE groups (14 ± 1.2 vs. 6 ± 6, P = 0.05). Incidence of clinically significant pleural effusion is uncommon after the Lariat procedure and can be either exudative or transudative in nature depending on the underlying mechanisms. More prospective studies are needed to study the pathophysiologic basis of development of pleural effusions after the Lariat procedure.


Development of CLL in individuals with mild lymphocytosis, without bone marrow infiltration, but with evidence of a monoclonally expanded population in peripheral blood.
Mild lymphocytosis (< 10 x 10(9)/L) is a common finding in routine blood tests. When it persists, it raises the question of whether this disorder is an early manifestation of chronic lymphocytic leukemia (CLL). If it is accompanied by bone marrow infiltration, it can be safely considered as a sign of CLL. The aim of this study was to analyze retrospectively the usefulness of immunophenotyping and immunogenotyping for early detection of lymphocyte clonality in ambiguous cases of lymphocytosis without bone marrow infiltration. Twenty-six healthy individuals, 47 to 77 years old, with an absolute lymphocyte count (ALC) at the "onset" of the disorder between 4 x 10(9)/L and 9 x 10(9)/L, without marrow infiltration, were studied and followed for a period of 31 to 51 months. CD19, CD20, CD5, CD2, CD4, CD8 surface markers and amplification of the Ig heavy chain CDR-3 locus were used for immunophenotypic and genotypic analysis, respectively. Our studies indicate that immunophenotyping alone is sufficient and superior to CDR-3 locus amplification for the early detection of lymphocyte clonality in peripheral blood. Furthermore, the high frequency of CLL development in individuals with established monoclonality strongly suggests that patients with mild borderline lymphocytosis, even without bone marrow infiltration, have to be followed for progression to CLL and its possible complications.


Modeling Linkage Disequilibrium Increases Accuracy of Polygenic Risk Scores.
Polygenic risk scores have shown great promise in predicting complex disease risk and will become more accurate as training sample sizes increase. The standard approach for calculating risk scores involves linkage disequilibrium (LD)-based marker pruning and applying a p value threshold to association statistics, but this discards information and can reduce predictive accuracy. We introduce LDpred, a method that infers the posterior mean effect size of each marker by using a prior on effect sizes and LD information from an external reference panel. Theory and simulations show that LDpred outperforms the approach of pruning followed by thresholding, particularly at large sample sizes. Accordingly, predicted R(2) increased from 20.1% to 25.3% in a large schizophrenia dataset and from 9.8% to 12.0% in a large multiple sclerosis dataset. A similar relative improvement in accuracy was observed for three additional large disease datasets and for non-European schizophrenia samples. The advantage of LDpred over existing methods will grow as sample sizes increase.


Additional impact of electrocardiographic over echocardiographic diagnosis of left ventricular hypertrophy for predicting the risk of ischemic stroke.
Patients with left ventricular hypertrophy (LVH) have an increased risk of ischemic stroke. Although echocardiography is commonly used for the diagnosis of LVH, there is little information about the potential role of electrocardiography in providing additional prognostic information. The purpose of this study is to determine if electrocardiographically derived criteria for LVH provide additional prognostic value over echocardiography for predicting ischemic stroke in a multiethnic population. A population-based, case-control study was conducted in 177 patients who had had a first ischemic stroke and in 246 control patients matched for age, gender, and race or ethnicity. Left ventricular mass was measured by using 2-dimensional transthoracic echocardiography. Logistic regression analysis was performed to assess the risk of stroke associated with the presence of LVH diagnosed by electrocardiography (defined by 4 established criteria) after adjustment for the presence of other stroke risk factors and for echocardiographically determined LVH. After adjustment for the presence of other established stroke risk factors, ECG-LVH was associated with ischemic stroke, using Sokolow-Lyon (odds ratio [OR] 2.12, 95% CI 1.05-4.30), Cornell voltage (OR 2.06, 95% CI, 1.26-3.35), and Cornell product criteria (OR 2.12, 95% CI, 1.13-3.97). Cornell voltage criterion (men, >2.8 mV; women, >2.0 mV) was associated with ischemic stroke even after adjustment for echocardiographically determined LVH (OR 1.73, 95% CI, 1.04-2.88). The combination of echo-LVH and a positive Cornell voltage criterion was associated with a 3.5-fold increase in stroke risk. Our study indicates that the presence of ECG-LVH is associated with an increased risk of ischemic stroke after adjustment for other stroke risk factors. For Cornell voltage criteria, this relationship persisted even after adjustment for echocardiographic LVH. Electrocardiographic results can provide independent information for left ventricular myocardial changes and should be considered together with echocardiographic results to fully assess the risk of ischemic stroke.


Induction of alopecia in mice exposed to cigarette smoke.
Besides being responsible for a high proportion of those chronic degenerative diseases that are the leading causes of death in the population, tobacco smoking has been associated with skin diseases. Smoke genotoxicants are metabolized in hair follicle cells, where they form DNA adducts and cause DNA damage. The suspicion was raised that, in humans, a link may exist between smoking and both premature grey hair and hair loss. In order to check this hypothesis, we carried out a study in C57BL/6 mice exposed whole-body to a mixture of sidestream and mainstream cigarette smoke. After 3 months exposure, most mice developed areas of alopecia and grey hair, while no such lesions occurred either in sham-exposed mice or in smoke-exposed mice receiving the chemopreventive agent N-acetylcysteine with drinking water. Cell apoptosis occurred massively in the hair bulbs at the edge of alopecia areas. Smoke-exposed mice had extensive atrophy of the epidermis, reduced thickness of the subcutaneous tissue, and scarcity of hair follicles. On the whole, exposure to smoke genotoxic components appears to alter the hair cycle with a dystrophic anagen pattern. Although this mechanism is different from that of genotoxic cytostatic drugs, N-acetylcysteine appears to exert protective effects in both conditions.


Branching patterns and symmetry of the course of the facial artery in Koreans.
The topography and the course of the facial artery were investigated in 47 Korean cadavers. The final branch of the facial artery was the lateral nasal branch in 44.0% whereas it was the angular branch in 36.3% of the cases. In 54.5% of the cases, the facial artery ended symmetrically. According to previous studies, variations in the distribution pattern of the facial artery have been regarded as racial difference. However, in this study we showed that the diverse pattern of the facial artery distribution demonstrates individual variation rather than racial difference. The superior and inferior labial arteries on the right side were more dominant than those on the left. The average distance between the branching points for the inferior alar branch and for the lateral nasal branch was 15.9 mm, and it was 25.2 mm between the points for the superior labial branch and for the inferior alar branch. The branching point of the inferior labial branch was 30.9 mm apart on average from that of the superior labial branch. The courses of the facial arteries showed no significant differences based on either laterality or gender.


Tobacco control in the Russian Federation--a policy analysis.
The Russian Federation (Russia) has one of the highest smoking rates in the world. The purpose of this study is to analyze past and current trends of the tobacco epidemic in the Russian Federation, review current tobacco control policy responses, and identify areas of opportunity for policy priorities. We used a policy triangle as analytical framework to examine content, context, and processes of Russian tobacco control policy. The analysis was based on secondary data on supply and demand sides of the Russian tobacco epidemic, tobacco-related economic and health effects during Russia's economic transition, and compliance of Russian tobacco policy with international standards and regulations. Tobacco-promoting strategies have specifically targeted women and youth. Russia's approval of a "National Tobacco Control Concept" and draft for a comprehensive tobacco control bill increasingly align national legislature with the WHO Framework Convention on Tobacco Control (FCTC). However, several structural and cultural factors represent substantial barriers to the policy process. The influence of transnational tobacco companies on policy processes in Russia has so far impeded a full implementation of the FCTC mandates. Several strategies have been identified as having the potential to reduce the prevalence of tobacco use in Russia and decrease tobacco-related national health and economic burden: adjusting national tobacco policy by raising tobacco tax from the current lowest level in Europe to at least 70%; consequent enforcement of a complete smoking ban in public places; marketing restrictions; and smoking cessation interventions integrated into primary care. Russia's tobacco control efforts need to target women and youths specifically to efficiently counter industry efforts.


Pontine regulation of REM sleep components in cats: integrity of the pedunculopontine tegmentum (PPT) is important for phasic events but unnecessary for atonia during REM sleep.
Transection, lesion and unit recording studies have localized rapid eye movement (REM) sleep mechanisms to the pons. Recent work has emphasized the role of pontine cholinergic cells, especially those of the pedunculopontine tegmentum (PPT). The present study differentiated REM sleep deficits associated with lesions of the PPT from other pontine regions implicated in REM sleep generation, including those with predominantly cholinergic vs non-cholinergic cells. Twelve hour polygraphic recordings were obtained in 18 cats before and 1-2 weeks after bilateral electrolytic or radio frequency lesions of either: (1) PPT, which contains the dorsolateral pontine cholinergic cell column; (2) laterodorsal tegmental nucleus (LDT), which contains the dorsomedial pontine cholinergic cell column; (3) locus ceruleus (LC), which contains mostly noradrenergic cells; or (4) subceruleus (LC alpha, peri-LC alpha and the lateral tegmental field), which also contains predominantly noncholinergic cells. There were three main findings: (i) Only lesions of PPT and subceruleus significantly affected REM sleep time. These lesions produced comparable reductions in REM sleep time but influenced REM sleep components quite differently: (ii) PPT lesions, estimated to damage 90 +/- 4% of cholinergic cells, reduced the number of REM sleep entrances and phasic events, including ponto-geniculooccipital (PGO) spikes and rapid eye movements (REMs), but did not prevent complete atonia during REM sleep: (iii) Subceruleus lesions eliminated atonia during REM sleep. Mobility appeared to arouse the cat prematurely from REM sleep and may explain the brief duration of REM sleep epochs seen exclusively in this group. Despite the reduced amount of REM sleep, the total number of PGO spikes and REM sleep entrances increased over baseline values. Collectively, the results distinguish pontine loci regulating phasic events vs atonia. PPT lesions reduced phasic events, whereas subceruleus lesions created REM sleep without atonia. Severe REM sleep deficits after large pontine lesions, including PPT and subceruleus, might be explained by simultaneous production of both REM sleep syndromes. However, extensive loss of ACh neurons in the PPT does not disrupt REM sleep atonia.


p53MVA therapy in patients with refractory gastrointestinal malignancies elevates p53-specific CD8+ T-cell responses.
To conduct a phase I trial of a modified vaccinia Ankara (MVA) vaccine delivering wild-type human p53 (p53MVA) in patients with refractory gastrointestinal cancers. Three patients were vaccinated with 1.0×10(8) plaque-forming unit (pfu) p53MVA followed by nine patients at 5.6×10(8) pfu. Toxicity was classified using the NCI Common Toxicity Criteria and clinical responses were assessed by CT scan. Peripheral blood samples were collected pre- and post-immunization for immunophenotyping, monitoring of p53MVA-induced immune response, and examination of PD1 checkpoint inhibition in vitro. p53MVA immunization was well tolerated at both doses, with no adverse events above grade 2. CD4+ and CD8+ T cells showing enhanced recognition of a p53 overlapping peptide library were detectable after the first immunization, particularly in the CD8+ T-cell compartment (P=0.03). However, in most patients, this did not expand further with the second and third immunization. The frequency of PD1+ T cells detectable in patients' peripheral blood mononuclear cells (PBMC) was significantly higher than in healthy controls. Furthermore, the frequency of PD1+ CD8+ T cells showed an inverse correlation with the peak CD8+ p53 response (P=0.02) and antibody blockade of PD1 in vitro increased the p53 immune responses detected after the second or third immunizations. Induction of strong T-cell and antibody responses to the MVA backbone were also apparent. p53MVA was well tolerated and induced robust CD8+ T-cell responses. Combination of p53MVA with immune checkpoint inhibition could help sustain immune responses and lead to enhanced clinical benefit.


Inflammation, oxidative stress, and obesity.
Obesity is a chronic disease of multifactorial origin and can be defined as an increase in the accumulation of body fat. Adipose tissue is not only a triglyceride storage organ, but studies have shown the role of white adipose tissue as a producer of certain bioactive substances called adipokines. Among adipokines, we find some inflammatory functions, such as Interleukin-6 (IL-6); other adipokines entail the functions of regulating food intake, therefore exerting a direct effect on weight control. This is the case of leptin, which acts on the limbic system by stimulating dopamine uptake, creating a feeling of fullness. However, these adipokines induce the production of reactive oxygen species (ROS), generating a process known as oxidative stress (OS). Because adipose tissue is the organ that secretes adipokines and these in turn generate ROS, adipose tissue is considered an independent factor for the generation of systemic OS. There are several mechanisms by which obesity produces OS. The first of these is the mitochondrial and peroxisomal oxidation of fatty acids, which can produce ROS in oxidation reactions, while another mechanism is over-consumption of oxygen, which generates free radicals in the mitochondrial respiratory chain that is found coupled with oxidative phosphorylation in mitochondria. Lipid-rich diets are also capable of generating ROS because they can alter oxygen metabolism. Upon the increase of adipose tissue, the activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), was found to be significantly diminished. Finally, high ROS production and the decrease in antioxidant capacity leads to various abnormalities, among which we find endothelial dysfunction, which is characterized by a reduction in the bioavailability of vasodilators, particularly nitric oxide (NO), and an increase in endothelium-derived contractile factors, favoring atherosclerotic disease.


Concomitancy of mutation in FRDA gene and FMR1 premutation in 58 year-old woman.
DNA testing broadens diagnostic tools available for hereditary ataxias. However, together with current knowledge of genes and their mutations crop up new phenotype figures of diseases already well known. Diagnostic problems in practice can consist in part due to the very similar symptoms of hereditary ataxias and acquaintance in or availability of new techniques such as DNA testing and result in misdiagnosis. We present a case study of a 57 year-old woman with both expansion of the triplet repetitive sequence of FRDA gene and a premutation in FMR1 gene. At present we diagnose her with Very Late Onset Friedreich s ataxia, but we advise of possible combinations or aggravations of her symptoms due to manifestation of Fragile X premutation tremor/ataxia syndrome. In nontypical phenotypes of DNA verifying hereditary ataxias we recommend searching of comorbidity, specifically from a range of hereditary ataxias with very similar spectra of symptoms.


Selective elimination of anti-DNA antibody-producing cells by antiidiotypic antibody conjugated with neocarzinostatin.
A new strategy was shown for the manipulation of autoantibody production in humans. Antiidiotypic antibody to human anti-DNA autoantibody was conjugated with neocarzinostatin (NCS), a cytotoxic agent, by using N-succimidyl 3-(2-pyridyldithio) propionate as a coupling agent. Human B cell clones, which produce anti-DNA autoantibodies, were killed by in vitro treatment with antiidiotype (Id)-NCS conjugates, while clones expressing an Id with irrelevant specificity were unaffected. These results indicate that treatment with anti-Id-NCS conjugates can act as a potent and specific means of generating immunosuppression of autoantibody production. This approach will have a significant advantage in aborting clones that are not effectively suppressed for the autoantibodies by anti-Id antibodies alone, and will result in a potential therapeutic treatment for systemic lupus erythematosus.


Tranexamic Acid in Shoulder Arthroplasty: A Systematic Review and Meta-Analysis.
The role of tranexamic acid (TXA) in reducing blood loss following primary shoulder arthroplasty has been demonstrated in small retrospective and controlled clinical trials. This study comprehensively evaluates current literature on the efficacy of TXA to reduce perioperative blood loss and transfusion requirements following shoulder arthroplasty. PubMed, MEDLINE, CENTRAL, and Embase were searched from the database inception date through October 27, 2016, for all articles evaluating TXA in shoulder arthroplasty. Two reviewers independently screened articles for eligibility and extracted data for analysis. A methodological quality assessment was completed for all included studies, including assessment of the risk of bias and strength of evidence. The primary outcome was change in hemoglobin and the secondary outcomes were drain output, transfusion requirements, and complications. Pooled outcomes assessing changes in hemoglobin, drain output, and transfusion requirements were determined. Five articles (n = 629 patients), including 3 Level-I and 2 Level-III studies, were included. Pooled analysis demonstrated a significant reduction in hemoglobin change (mean difference [MD], -0.64 g/dL; 95% confidence interval [CI], -0.84 to -0.44 g/dL; p < 0.00001) and drain output (MD, -116.80 mL; 95% CI, -139.20 to -94.40 mL; p < 0.00001) with TXA compared with controls. TXA was associated with a point estimate of the treatment effect suggesting lower transfusion requirements (55% lower risk); however, the wide CI rendered this effect statistically nonsignificant (risk ratio, 0.45; 95% CI, 0.18 to 1.09; p = 0.08). Findings were robust with sensitivity analysis of pooled outcomes from only Level-I studies. Moderate-strength evidence supports use of TXA for decreasing blood loss in primary shoulder arthroplasty. Further research is necessary to evaluate the efficacy of TXA in revision shoulder arthroplasty and to identify the optimal dosing and route of administration of TXA in shoulder arthroplasty. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


The Impact of CKD on Perioperative Risk and Mortality after Bariatric Surgery.
Twenty percent of patients with CKD in the United States have a body mass index (BMI) ≥35 kg/m2. Bariatric surgery reduces progression of CKD to ESKD, but the risk of perioperative complications remains a concern. The 24-month data spanning 2017-2018 were obtained from the Metabolic and Bariatric Surgery Quality Improvement Program (MBSAQIP) database and analyzed. Surgical complications were assessed on the basis of the length of hospital stay, mortality, reoperation, readmission, surgical site infection (SSI), and worsening of kidney function during the first 30 days after surgery. The 277,948 patients who had primary bariatric procedures were 44±11.9 (mean ± SD) years old, 79.6% were women, and 71.2% were White. Mean BMI was 45.7±7.6 kg/m2. Compared with patients with an eGFR≥90 ml/min per BSA, those with stage 5 CKD/ESKD were 1.91 times more likely to be readmitted within 30 days of a bariatric procedure (95% CI, 1.37 to 2.67; P<0.001). Similarly, length of hospital stay beyond 2 days was 2.05-fold (95% CI, 1.64 to 2.56; P<0.001) higher and risk of deep incisional SSI was 6.92-fold (95% CI, 1.62 to 29.52; P=0.009) higher for those with stage 5 CKD/ESKD. Risk of early postoperative mortality increased with declining preoperative eGFR, such that patients with stage 3b CKD were 3.27 (95% CI, 1.82 to 5.89; P<0.001) times more likely to die compared with those with normal kidney function. However, absolute mortality rates remained relatively low at 0.53% in those with stage 3b CKD. Furthermore, absolute mortality rates were <0.5% in those with stages 4 and 5 CKD, and these advanced CKD stages were not independently associated with an increased risk of early postoperative mortality. Increased severity of kidney disease was associated with increased complications after bariatric surgery. However, even for the population with advanced CKD, the absolute rates of postoperative complications were low. The mounting evidence for bariatric surgery as a renoprotective intervention in people with and without established kidney disease suggests that bariatric surgery should be considered a safe and effective option for patients with CKD.


Direct observation of Anderson localization of matter waves in a controlled disorder.
In 1958, Anderson predicted the localization of electronic wavefunctions in disordered crystals and the resulting absence of diffusion. It is now recognized that Anderson localization is ubiquitous in wave physics because it originates from the interference between multiple scattering paths. Experimentally, localization has been reported for light waves, microwaves, sound waves and electron gases. However, there has been no direct observation of exponential spatial localization of matter waves of any type. Here we observe exponential localization of a Bose-Einstein condensate released into a one-dimensional waveguide in the presence of a controlled disorder created by laser speckle. We operate in a regime of pure Anderson localization, that is, with weak disorder-such that localization results from many quantum reflections of low amplitude-and an atomic density low enough to render interactions negligible. We directly image the atomic density profiles as a function of time, and find that weak disorder can stop the expansion and lead to the formation of a stationary, exponentially localized wavefunction-a direct signature of Anderson localization. We extract the localization length by fitting the exponential wings of the profiles, and compare it to theoretical calculations. The power spectrum of the one-dimensional speckle potentials has a high spatial frequency cutoff, causing exponential localization to occur only when the de Broglie wavelengths of the atoms in the expanding condensate are greater than an effective mobility edge corresponding to that cutoff. In the opposite case, we find that the density profiles decay algebraically, as predicted in ref. 13. The method presented here can be extended to localization of atomic quantum gases in higher dimensions, and with controlled interactions.


State-dependent sentinels: an experimental study in the Arabian babbler.
Natural patterns of cooperative sentinel behaviour in Arabian babblers, Turdoides squamiceps, have proven consistent with state-dependent models of individually selfish anti-predator strategies. Here we demonstrate experimentally that sentinel effort within groups is determined simply by individual state. The two highest-ranking males in eight groups were separately fed a supplement of mealworms, each for one day at a time. Control days before and after each treatment confirmed that no carry-over effects occurred, and that most normal sentinel activity was carried out by alpha males, then beta males and then by the rest of the group. When supplemented, both alpha and beta males exhibited similar marked increases in sentinel activity, relative to control days. Unsupplemented males and the rest of the group incompletely compensated for these increases with reductions in sentinel effort. Differences in individual body mass within groups reflected natural and experimental variation in sentinel effort. Alpha and beta males weighed more than other group members, and gained mass only when supplementally fed. There was no evidence either for competitive sentinel behaviour, nor for any increased interference between males during the supplementation treatments. These results therefore provide strong evidence in support of the state-dependent approach to cooperative sentinel behaviour.


Left sinus of Valsalva-Electroanatomic basis and outcomes with ablation for outflow tract arrhythmias.
The ablation of outflow tract premature ventricular contractions (PVCs) is generally safe and effective. In some patients, successful ablation sites may not correlate with the earliest activation. We sought to evaluate mechanistic and anatomic relevance of the region below the left sinus of Valsalva in variable morphology outflow tract ventricular arrhythmias. PVC cases where ablation was in the region inferior to the left sinus of Valsalva were identified. Procedural and demographic information and long-term outcomes were obtained. Cadaver dissections to evaluate regional anatomy were done as well. A total of 51 cases were included (age 53 ± 10; 37 [73%] males). Ablation was done for high PVC burden (>20%; mean 27% ± 8%) and presence of symptoms (73%) or ejection fraction less than 50% (78%). QRS morphology included either R wave (8; 16%), Rs (9; 18%), or rS (67%) in lead I, no precordial transition (40; 78%), V2 transition, (7; 14%), or V3 transition (4; 8%). In 31 (61%), the site just below the left coronary cusp was the earliest site, while the remainder had another site earlier. Ablation was acutely successful in 50 of 51 (98%). After 3 months, success was noted in 48 of 51 (94%). In two patients, repeat ablation in the same region resulted in durable suppression. The cases presented emphasize the importance of a region centered below the left sinus of Valsalva, where multivariable morphology QRS may be successfully ablated. Consideration of mapping and ablation even when signals are late in this region may be warranted in previously failed ablation attempts or first-line evaluation.


A randomized clinical trial of high-dose epinephrine and norepinephrine vs standard-dose epinephrine in prehospital cardiac arrest.
To determine the relative efficacy of high- vs standard-dose catecholamines in initial treatment of prehospital cardiac arrest. Randomized, prospective, double-blind clinical trial. Prehospital emergency medical system of a major US city. All adults in nontraumatic cardiac arrest, treated by paramedics, who would receive epinephrine according to American Heart Association advanced cardiac life support guidelines. High-dose epinephrine (HDE, 15 mg), high-dose norepinephrine bitartrate (NE, 11 mg), or standard-dose epinephrine (SDE, 1 mg) was blindly substituted for advanced cardiac life support doses of epinephrine. Restoration of spontaneous circulation in the field, admission to hospital, hospital discharge, and Cerebral Performance Category score. Of 2694 patients with cardiac arrests during the study period, resuscitation was attempted on 1062 patients. Of this total, 816 patients met study criteria and were enrolled. In the entire cardiac arrest population, 63% of the survivors were among the 11% of patients who were defibrillated by first responders. The three drug treatment groups were similar for all independent variables. Thirteen percent of patients receiving HDE regained a pulse in the field vs 8% of those receiving SDE (P = .01), and 18% of HDE patients were admitted to the hospital vs 10% of SDE patients who were admitted to the hospital (P = .02). Similar trends for NE were not significant. There were 18 survivors; 1.7% of HDE patients and 2.6% of NE patients were discharged from the hospital compared with 1.2% of SDE patients, but this was not significant (P = .37; beta = .38). There was a nonsignificant trend for Cerebral Performance Category scores to be worse for HDE (3.2) and NE patients (3.7) than for SDE patients (2.3) (P = .10; beta = .31). No significant complications were identified. High-dose epinephrine did not produce longer hospital or critical care unit stays. High-dose epinephrine significantly improves the rate of return of spontaneous circulation and hospital admission in patients who are in prehospital cardiac arrest without increasing complications. However, the increase in hospital discharge rate is not statistically significant, and no significant trend could be determined for neurological outcome. No benefit of NE compared with HDE was identified. Further study is needed to determine the optimal role of epinephrine in prehospital cardiac arrest.


Novel anticancer therapeutics targeting telomerase.
Telomeres are protective caps at the ends of human chromosomes. Telomeres shorten with each successive cell division in normal human cells whereas, in tumors, they are continuously elongated by human telomerase reverse transcriptase (hTERT). Telomerase is overexpressed in 80-95% of cancers and is present in very low levels or is almost undetectable in normal cells. Because telomerase plays a pivotal role in cancer cell growth it may serve as an ideal target for anticancer therapeutics. Inhibition of telomerase may lead to a decrease of telomere length resulting in cell senescence and apoptosis in telomerase positive tumors. Several strategies of telomerase inhibition are reviewed, including small molecule inhibitors, antisense oligonucleotides, immunotherapies and gene therapies, targeting the hTERT or the ribonucleoprotein subunit hTER. G-quadruplex stabilizers, tankyrase and HSP90 inhibitors targeting telomere and telomerase assembly, and T-oligo approach are also covered. Based on this review, the most promising current telomerase targeting therapeutics are the antisense oligonucleotide inhibitor GRN163L and immunotherapies that use dendritic cells (GRVAC1), hTERT peptide (GV1001) or cryptic peptides (Vx-001). Most of these agents have entered phase I and II clinical trials in patients with various tumors, and have shown good response rates as evidenced by a reduction in tumor cell growth, increased overall disease survival, disease stabilization in advanced staged tumors and complete/partial responses. Most therapeutics have shown to be more effective when used in combination with standard therapies, resulting in concomitant telomere shortening and tumor mass shrinkage, as well as preventing tumor relapse and resistance to single agent therapy.


School-based physical education programs: evidence-based physical activity interventions for youth in Latin America.
This article focuses on results of the systematic review from the Guide for Useful Interventions for Activity in Latin America project related to school-based physical education (PE) programs in Latin America. The aims of the article are to describe five school-based PE programs from Latin America, discuss implications for effective school-based PE recommendations, propose approaches for implementing these interventions, and identify gaps in the research literature related to physical activity promotion in Latin American youth. Following the US Community Guide systematic review process, five school-based PE intervention studies with sufficient quality of design, execution and detail of intervention and outcomes were selected for full abstraction. One study was conducted in Brazil, two studies were conducted in Chile and two studies were conducted on the US/Mexico border. While studies presented assorted outcomes, methods and duration of interventions, there were consistent positive increases in physical activity levels for all outcomes measured during PE classes, endurance and active transportation to school in all three randomized studies. Except for one cohort from one study, the non-randomized studies showed positive intervention effects for moderate and vigorous physical activity levels during PE classes. The core elements of these five interventions included capacity building and staff training (PE specialists and/or classroom teachers); changes in the PE curricula; provision of equipment and materials; and adjustment of the interventions to specific target populations. In order to translate the strong evidence for school-based PE into practice, systematic attention to policy and implementation issues is required.


Stress Physiology in Infancy and Early Childhood: Cortisol Flexibility, Attunement and Coordination.
Research on stress physiology in infancy has assumed increasing importance due to its lifelong implications. In this review, we focus on measurement of hypothalamic-pituitary-adrenal (HPA) function, in particular, and on complementary autonomic processes. We suggest that the measure of HPA function has been overly exclusive, focusing on individual reactivity to single, pragmatically selected laboratory challenges. We advocate use of multiple, strategically chosen challenges and within-subject designs. By administering one challenge that typically does not provoke reactivity and another that does, it is possible to represent allostatic load in terms of "flexibility," the capacity to titrate response to challenge. We also recommend assessing infant reactivity in the context of the primary caregiver's physiological function. Infant-mother "attunement" is central to developmental psychology, permeating diverse developmental domains with varied consequences. A review of adrenocortical attunement suggests that attunement is a reliable process, manifest across varied populations. However, attunement appears stronger in the context of more highly stressful circumstances, such that administration of multiple, selected challenges may help evaluate the degree to which individuals titrate attunement to challenge and determine the correlates of this differential attunement. Finally, we advocate studying the "coordination" of HPA function with other aspects of stress physiology and variation in the degree of this coordination. The use of multiple stressors is important here because each stress system is differentially sensitive to different types of challenge. Therefore, use of single stressors in between-subject designs impedes full recognition of the role played by each system. Overall, we recommend measure of flexibility, attunement, and coordination in the context of multiple challenges to capture allostasis in environmental and physiological context. The simultaneous use of such inclusive and integrative metrics may yield more reliable findings than has hitherto been the case. The interrelation of these metrics can be understood in the context of the adaptive calibration model..


Renal expression of Na+-phosphate cotransporter mRNA and protein: effect of the Gy mutation and low phosphate diet.
The X-linked Gy mutation is closely linked, but not allelic, to Hyp and is characterized by rickets, hypophosphatemia, decreased renal tubular maximum for phosphate (Pi) reabsorption (TmP) and a specific reduction in renal brush-border membrane (BBM) Na+-Pi cotransport. Gy mice, like their normal littermates, respond to a low-Pi diet with an increase in BBM Na+-Pi cotransport, but fail to show an adaptive increase in Tmp. Using an antibody raised against the NH2 terminal peptide of the rat renal-specific Na+-Pi cotransporter (NaPi-2) and a NaPi-2 cDNA probe, we examined the effect of the Gy mutation and low-Pi diet (0.03% Pi) on NaPi-2 protein and mRNA abundance. The reduction in BBM Na+-Pi cotransport in Gy mice (51 +/- 5% of normal, P < 0.05) was associated with a decrease in NaPi-2 protein (46 +/- 12% of normal, P < 0.05) and mRNA abundance (76 +/- 5%, P < 0.05). The low-Pi diet elicited a two- to three-fold increase in Na+-Pi cotransport in both normal and Gy mice that was accompanied by a large increase in NaPi-2 protein (10.2-fold in normal and 16.9-fold in Gy mice) and a modest increase in NaPi-2 mRNA (1.3-fold in both mouse strains, P < 0.05). The present data demonstrate that (1) the renal defect in BBM Pi transport in Gy mice can be ascribed to a deficit in NaPi-2 protein and mRNA abundance, (2) both normal and Gy mice respond to low Pi with an adaptive increase in NaPi-2 protein that exceeds the increase in Na+-Pi cotransport activity and NaPi-2 mRNA, (3) the adaptive increase in NaPi-2 protein and mRNA are not sufficient for the overall increase in TmP following Pi restriction.


STUDIES OF TWO KINDS OF VIRUS PARTICLES WHICH COMPRISE INFLUENZA A2 VIRUS STRAINS : I. CHARACTERIZATION OF STABLE HOMOGENEOUS SUBSTRAINS IN REACTIONS WITH SPECIFIC ANTIBODY, MUCOPROTEIN INHIBITORS, AND ERYTHROCYTES.
Two kinds of virus particles have been found in varying proportions in influenza A2 strains isolated during the 1957 pandemic. Pure populations of the different particles were obtained, and these substrains were genetically stable on serial passage in the chick embryo. The two virus particles differ markedly in several biological properties though they are antigenically similar. One kind of particle, designated "+," is relatively sensitive to specific antibody, is highly sensitive to inhibition by serum inhibitors and urinary mucoprotein, fails to elute or elutes very slowly from human erythrocytes, and is capable of agglutinating erythrocytes treated extensively with V. cholerae filtrate. The other particle, designated "-," is relatively insensitive to antibodies and urinary mucoprotein, completely insensitive to serum inhibitors, elutes rapidly from erythrocytes, and can agglutinate erythrocytes treated extensively with V. cholerae filtrate. Both "+" and "-" particles destroy virus receptors on urinary mucoprotein. The relative proportions of these two particles determine the characteristics of parent strains in reactions with specific antibody, mucoprotein inhibitors, and erythrocytes. The "+" and "-" particles with several easily identifiable markers are well suited for genetic studies.


Effects of vitamin E, vitamin C and polyphenols on the rate of blood pressure variation: results of two randomised controlled trials.
High blood pressure (BP) variability, which may be an important determinant of hypertensive end-organ damage, is emerging as an important predictor of cardiovascular health. Dietary antioxidants can influence BP, but their effects on variability are yet to be investigated. The aim of the present study was to assess the effects of vitamin E, vitamin C and polyphenols on the rate of daytime and night-time ambulatory BP variation. To assess these effects, two randomised, double-blind, placebo-controlled trials were performed. In the first trial (vitamin E), fifty-eight individuals with type 2 diabetes were given 500 mg/d of RRR-α-tocopherol, 500 mg/d of mixed tocopherols or placebo for 6 weeks. In the second trial (vitamin C-polyphenols), sixty-nine treated hypertensive individuals were given 500 mg/d of vitamin C, 1000 mg/d of grape-seed polyphenols, both vitamin C and polyphenols, or neither (placebo) for 6 weeks. At baseline and at the end of the 6-week intervention, 24 h ambulatory BP and rate of measurement-to-measurement BP variation were assessed. Compared with placebo, treatment with α-tocopherol, mixed tocopherols, vitamin C and polyphenols did not significantly alter the rate of daytime or night-time systolic BP, diastolic BP or pulse pressure variation (P>0·05). Treatment with the vitamin C and polyphenol combination resulted in higher BP variation: the rate of night-time systolic BP variation (P= 0·022) and pulse pressure variation (P= 0·0036) were higher and the rate of daytime systolic BP variation was higher (P= 0·056). Vitamin E, vitamin C or grape-seed polyphenols did not significantly alter the rate of BP variation. However, the increase in the rate of BP variation suggests that the combination of high doses of vitamin C and polyphenols could be detrimental to treated hypertensive individuals.


Identification and Mechanistic Understanding of Dihydroorotate Dehydrogenase Point Mutations in Plasmodium falciparum that Confer in Vitro Resistance to the Clinical Candidate DSM265.
Malaria is one of the most challenging human infectious diseases, and both prevention and control have been hindered by the development of Plasmodium falciparum resistance to existing therapies. Several new compounds with novel mechanisms are in clinical development for the treatment of malaria, including DSM265, an inhibitor of Plasmodium dihydroorotate dehydrogenase. To explore the mechanisms by which resistance might develop to DSM265 in the field, we selected for DSM265-resistant P. falciparum parasites in vitro. Any of five different amino acid changes led to reduced efficacy on the parasite and to decreased DSM265 binding to P. falciparum DHODH. The DSM265-resistant parasites retained full sensitivity to atovaquone. All but one of the observed mutations were in the DSM265 binding site, and the remaining C276F was in the adjacent flavin cofactor site. The C276F mutation was previously identified in a recrudescent parasite during a Phase IIa clinical study. We confirmed that this mutation (and the related C276Y) accounted for the full level of observed DSM265 resistance by regenerating the mutation using CRISPR/Cas9 genome editing. X-ray structure analysis of the C276F mutant enzyme showed that conformational changes of nearby residues were required to accommodate the larger F276 residue, which in turn led to a restriction in the size of the DSM265 binding pocket. These findings underscore the importance of developing DSM265 as part of a combination therapy with other agents for successful use against malaria.


Older persons' lived experiences of depression and self-management.
Mental ill-health, such as depression in the elderly, is a complex issue that is influenced by the life-world perspective of older persons. Their self-management ability should be strengthened based on an understanding of their situation, perspectives, and vulnerability. The aim of this study was to explore and increase understanding of old persons' lived experiences of depression and self-management using an interpretative explorative design. Understanding was developed by means of hermeneutic interpretation. One theme, Relationships and Togetherness, and four subthemes, A Sense of Carrying a Shoulder Bag, Walking on Eggshells, Holding the Reins, and Estrangement--a Loss of Togetherness, emerged. A collaborative approach can be important for empowering older persons through self-development and management. Although the findings of the present study cannot be considered conclusive or definitive, they nevertheless contribute new knowledge of older persons' lived experiences of depression in everyday life.


Microfluidic system with integrated electroosmotic pumps, concentration gradient generator and fish cell line (RTgill-W1)--towards water toxicity testing.
This study presents a microfluidic system that incorporates electroosmotic pumps, a concentration gradient generator and a fish cell line (rainbow trout gill) to perform toxicity testing on fish cells seeded in the system. The system consists of three mechanical components: (1) a toxicity testing chip containing a microfluidic gradient generator which creates a linear concentration distribution of toxicant in a cell test chamber, (2) an electroosmotic (EO) pump chip that controls the flow rate and operation of the toxicity chip, and (3) indirect reservoirs that connect the two chips allowing for the toxicant solution to be pumped separately from the electroosmotic pump solution. The flow rate and stability of the EO pumps was measured and tested by monitoring the gradient generator using fluorescence microscopy. Furthermore, a lethality test was performed with this system setup using a rainbow trout gill cell line (RTgill-W1) as the test cells and sodium dodecyl sulfate as a model toxicant. A gradient of sodium dodecyl sulfate, from 0 to 50 microg mL(-1), was applied for 1 hr to the attached cells, and the results were quantified using a Live/Dead cell assay. This work is a preliminary study on the application of EO pumps in a living cell assay, with the potential to use the pumps in portable water quality testing devices with RTgill-W1 cells as the biosensors.


Stable oxygen isotope and flux partitioning demonstrates understory of an oak savanna contributes up to half of ecosystem carbon and water exchange.
Semi-arid ecosystems contribute about 40% to global net primary production (GPP) even though water is a major factor limiting carbon uptake. Evapotranspiration (ET) accounts for up to 95% of the water loss and in addition, vegetation can also mitigate drought effects by altering soil water distribution. Hence, partitioning of carbon and water fluxes between the soil and vegetation components is crucial to gain mechanistic understanding of vegetation effects on carbon and water cycling. However, the possible impact of herbaceous vegetation in savanna type ecosystems is often overlooked. Therefore, we aimed at quantifying understory vegetation effects on the water balance and productivity of a Mediterranean oak savanna. ET and net ecosystem CO2 exchange (NEE) were partitioned based on flux and stable oxygen isotope measurements and also rain infiltration was estimated. The understory vegetation contributed importantly to total ecosystem ET and GPP with a maximum of 43 and 51%, respectively. It reached water-use efficiencies (WUE; ratio of carbon gain by water loss) similar to cork-oak trees. The understory vegetation inhibited soil evaporation (E) and, although E was large during wet periods, it did not diminish WUE during water-limited times. The understory strongly increased soil water infiltration, specifically following major rain events. At the same time, the understory itself was vulnerable to drought, which led to an earlier senescence of the understory growing under trees as compared to open areas, due to competition for water. Thus, beneficial understory effects are dominant and contribute to the resilience of this ecosystem. At the same time the vulnerability of the understory to drought suggests that future climate change scenarios for the Mediterranean basin threaten understory development. This in turn will very likely diminish beneficial understory effects like infiltration and ground water recharge and therefore ecosystem resilience to drought.


Many shades of gray-The context-dependent performance of organic agriculture.
Organic agriculture is often proposed as a more sustainable alternative to current conventional agriculture. We assess the current understanding of the costs and benefits of organic agriculture across multiple production, environmental, producer, and consumer dimensions. Organic agriculture shows many potential benefits (including higher biodiversity and improved soil and water quality per unit area, enhanced profitability, and higher nutritional value) as well as many potential costs including lower yields and higher consumer prices. However, numerous important dimensions have high uncertainty, particularly the environmental performance when controlling for lower organic yields, but also yield stability, soil erosion, water use, and labor conditions. We identify conditions that influence the relative performance of organic systems, highlighting areas for increased research and policy support.


Eyelid carcinoma: An experience from a tertiary cancer center.
Eyelid carcinoma is rare tumors of the head and neck. They are rarely lethal but can be associated with significant morbidity if not treated early and appropriately. There are limited data available from world over and in particular the Indian subcontinent regarding eyelid carcinoma and its prognostic factors influencing treatment outcomes. Setting and Design:Retrospective study of patients treated in a tertiary cancer center between 2005 and 2016. In this study, 51 patients with eyelid carcinoma treated at single tertiary cancer center were included. The demographic, clinical data, which includes the treatment received, histopathology report and follow-up, were recorded. All the relevant variables influencing disease-free survival (DFS) were analyzed. Sebaceous carcinoma was the most common eyelid carcinoma followed by squamous cell carcinoma and basal cell carcinoma in descending order in this series. Lower eyelid was involved most often. The incidence of nodal metastasis was low (14%). Multivariate analysis revealed that margin status influenced the DFS (P= 0.001) (hazard ratios = 15.9 [95% confidence interval: 1.8-135.2]). The 5 years' DFS was 70%. Eyelid tumors are less common cancer with good prognosis if treated appropriately. The morbidity associated with treatment can be reduced if treated early.


Forward error correction supported 150 Gbit/s error-free wavelength conversion based on cross phase modulation in silicon.
We build a forward error correction (FEC) module and implement it in an optical signal processing experiment. The experiment consists of two cascaded nonlinear optical signal processes, 160 Gbit/s all optical wavelength conversion based on the cross phase modulation (XPM) in a silicon nanowire and subsequent 160 Gbit/s-to-10 Gbit/s demultiplexing in a highly nonlinear fiber (HNLF). The XPM based all optical wavelength conversion in silicon is achieved by off-center filtering the red shifted sideband on the CW probe. We thoroughly demonstrate and verify that the FEC code operates correctly after the optical signal processing, yielding truly error-free 150 Gbit/s (excl. overhead) optically signal processed data after the two cascaded nonlinear processes.