The human B cell-associated antigen CD24 is a single chain sialoglycoprotein.
The CD24 human B cell-associated antigen was originally characterized in this laboratory with the use of monoclonal antibody BA-1 combined with a standard radioimmuneprecipitation technique, and was reported to be a three chain glycoprotein complex of 45, 55, and 65 kilodaltons. We have compared our standard radioimmuneprecipitation technique (BA-1 ascites followed by rabbit anti-mouse IgM-coated protein A-Sepharose) to BA-1 conjugated directly to CNBr-Sepharose (BA-1-Sepharose) and report striking differences in the electrophoretic profile of CD24 immuneprecipitates. The CD24 immuneprecipitate obtained with BA-1-Sepharose showed a single broad band with a relative mobility of 42 kilodaltons. A series of experiments performed with the two immuneprecipitation techniques, reducing or nonreducing electrophoretic conditions, and addition of preformed mock BA-1 immuneprecipitate to BA-1-Sepharose immuneprecipitates convincingly demonstrated that the previously described 55 and 65 kilodalton components were artifacts caused by co-migration of CD24 with IgG and IgM heavy chains, respectively. Because of the consistent association and co-migration of CD24 with IgG heavy chains, we investigated the possibility that CD24 might be related to the 45 kilodalton Fc-gamma receptor expressed on B cells and eosinophils. We found, however, no evidence for such a relationship by cross adsorption analysis with BA-1-Sepharose and IgG-Sepharose.


Development of a thyroperoxidase inhibition assay for high-throughput screening.
High-throughput screening (HTPS) assays to detect inhibitors of thyroperoxidase (TPO), the enzymatic catalyst for thyroid hormone (TH) synthesis, are not currently available. Herein, we describe the development of a HTPS TPO inhibition assay. Rat thyroid microsomes and a fluorescent peroxidase substrate, Amplex UltraRed (AUR), were employed in an end-point assay for comparison to the existing kinetic guaiacol (GUA) oxidation assay. Following optimization of assay metrics, including Z', dynamic range, and activity, using methimazole (MMI), the assay was tested with a 21-chemical training set. The potency of MMI-induced TPO inhibition was greater with AUR compared to GUA. The dynamic range and Z' score with MMI were as follows: 127-fold and 0.62 for the GUA assay, 18-fold and 0.86 for the 96-well AUR assay, and 11.5-fold and 0.93 for the 384-well AUR assay. The 384-well AUR assay drastically reduced animal use, requiring one-tenth of the rat thyroid microsomal protein needed for the GUA 96-well format assay. Fourteen chemicals inhibited TPO, with a relative potency ranking of MMI > ethylene thiourea > 6-propylthiouracil > 2,2',4,4'-tetrahydroxy-benzophenone > 2-mercaptobenzothiazole > 3-amino-1,2,4-triazole > genistein > 4-propoxyphenol > sulfamethazine > daidzein > 4-nonylphenol > triclosan > iopanoic acid > resorcinol. These data demonstrate the capacity of this assay to detect diverse TPO inhibitors. Seven chemicals acted as negatives: 2-hydroxy-4-methoxybenzophenone, dibutylphthalate, diethylhexylphthalate, diethylphthalate, 3,5-dimethylpyrazole-1-methanol, methyl 2-methyl-benzoate, and sodium perchlorate. This assay could be used to screen large numbers of chemicals as an integral component of a tiered TH-disruptor screening approach.


Quantitative process evaluation of a community-based HIV/AIDS behavioural intervention in rural Uganda.
This paper describes the implementation of a large community-based HIV/AIDS behavioural intervention in rural Uganda and presents 4 years' worth of quantitative process data. The intervention involved 560 field-based workers (57% male, 76% subsistence farmers, mean age 35 years), supervised by six central staff. Intervention channels included drama and video shows, Community Educators (CEs), as well as leaflet and condom distribution. Activities focused on one or more of 16 key topics. In total, 392 000 attendances (51% female) were recorded--a mean of over 6 for each of the 64 000 target adults--at 81 000 activities, with CEs attracting 71% of the total attendance; 164 000 leaflets and 242 000 condoms were also distributed. The annual cost of the intervention per target individual was approximately US$1.76. Our voluntary workforce experienced an annual attrition rate of 11%, with 'stable' workers more likely to be older, married or opinion leaders in their community than those who dropped out. We calculate that even a significant increase in the proportion of female field workers would have made little difference either to the sex ratio of attendees or to overall attendance. In spite of some initial resistance to the intervention, particularly in relation to condoms, we have demonstrated that people in rural Africa can accept and actively participate in the dissemination of HIV/AIDS prevention messages throughout their own communities.


Diffraction-unlimited all-optical imaging and writing with a photochromic GFP.
Lens-based optical microscopy failed to discern fluorescent features closer than 200 nm for decades, but the recent breaking of the diffraction resolution barrier by sequentially switching the fluorescence capability of adjacent features on and off is making nanoscale imaging routine. Reported fluorescence nanoscopy variants switch these features either with intense beams at defined positions or randomly, molecule by molecule. Here we demonstrate an optical nanoscopy that records raw data images from living cells and tissues with low levels of light. This advance has been facilitated by the generation of reversibly switchable enhanced green fluorescent protein (rsEGFP), a fluorescent protein that can be reversibly photoswitched more than a thousand times. Distributions of functional rsEGFP-fusion proteins in living bacteria and mammalian cells are imaged at <40-nanometre resolution. Dendritic spines in living brain slices are super-resolved with about a million times lower light intensities than before. The reversible switching also enables all-optical writing of features with subdiffraction size and spacings, which can be used for data storage.


Career development: graduate nurse views.
To explore recent Singapore nursing graduates' experience of and views about their career development and progress. The recruitment and retention of an adequate number of registered nurses is a continuing workforce issue in Singapore and other major cities. Survey of recent nursing graduates. Recent nursing graduates from the Bachelor programme (n = 147) were sent an individual survey; a response rate of 54% was achieved. Findings show that nurses rated their self-concept in a positive manner and were most satisfied (moderately to very) with helping patients and providing effective care, and the level of patient involvement. They were least satisfied (moderately to only a little) with prestige among the general medical community and the general public, hours of work, lifestyle factors and research opportunities. The following four factors were identified as significant impediments to career development; lack of support in the work place; perceived insufficient clinical career development opportunities; excessive work hours; and limited access to merit-based places in further education. Suggestions made to overcome perceived career development barriers are as follows: broad multifactorial healthcare system changes; decreased and more flexible working hours; and fairer access to further clinical and higher education. Results highlight the value clinical nurses place on having access to career development opportunities, merit-based further education and work place supports. These factors also have the potential to influence patient care and impact on the retention of nurses in their present job and satisfaction with their nursing career.


Bridging experiments, models and simulations: an integrative approach to validation in computational cardiac electrophysiology.
Computational models in physiology often integrate functional and structural information from a large range of spatiotemporal scales from the ionic to the whole organ level. Their sophistication raises both expectations and skepticism concerning how computational methods can improve our understanding of living organisms and also how they can reduce, replace, and refine animal experiments. A fundamental requirement to fulfill these expectations and achieve the full potential of computational physiology is a clear understanding of what models represent and how they can be validated. The present study aims at informing strategies for validation by elucidating the complex interrelations among experiments, models, and simulations in cardiac electrophysiology. We describe the processes, data, and knowledge involved in the construction of whole ventricular multiscale models of cardiac electrophysiology. Our analysis reveals that models, simulations, and experiments are intertwined, in an assemblage that is a system itself, namely the model-simulation-experiment (MSE) system. We argue that validation is part of the whole MSE system and is contingent upon 1) understanding and coping with sources of biovariability; 2) testing and developing robust techniques and tools as a prerequisite to conducting physiological investigations; 3) defining and adopting standards to facilitate the interoperability of experiments, models, and simulations; 4) and understanding physiological validation as an iterative process that contributes to defining the specific aspects of cardiac electrophysiology the MSE system targets, rather than being only an external test, and that this is driven by advances in experimental and computational methods and the combination of both.


Pretreatment neuropsychological deficits in children with brain tumors.
Treatment of childhood brain cancer has been associated with long-term cognitive morbidity in children. In the present study, the cognitive status of children with brain tumors was examined prior to any treatment to single out the role of tumor and tumor-related factors in cognitive deficits. Eighty-three children with newly diagnosed brain tumors (mean age, 8.6 years; range, 7 months to 16.6 years; median, 9.4 years) completed an extensive battery of age-related tests to assess cognitive function before any therapeutic intervention. Magnetic resonance imaging (MRI) was used to determine tumor site and volume and tumor-related factors. Performance under test was compared with symptom duration, neurological status, epilepsy, and MRI. Cognitive difficulties are detected at diagnosis in as many as 50% of patients for some cognitive domains; 6% of patients present with true-diagnosed mental retardation. The location of the tumor is the principal determinant of cognitive deficits, with major impairment in children with cortical tumors. Symptom duration and the presence of epilepsy are significantly associated with neuropsychological disabilities, while neuroradiological tumor-related variables do not correlate clearly with neurocognitive performance. The knowledge of the pre-existing cognitive deficits is critical to evaluate the results of treatment, providing a baseline for assessing the true impact of therapy in determining cognitive decline. In addition, the study suggests that some clinical variables require careful monitoring, because they could be specifically implicated in the neuropsychological outcome; the efforts to reduce the impact of these factors could ameliorate long-term prognosis.


The sea lamprey Petromyzon marinus: a model for evolutionary and developmental biology.
Sea lampreys (Petromyzon marinus) are cyclostomes, the most basal extant group of vertebrates, and are thought to have existed largely unchanged for more than 500 million years. They are aquatic, eel-shaped animals that spend a major part of their life as filter-feeding larvae called ammocoetes, inhabiting many freshwater bodies in the northern hemisphere. After metamorphosis, sea lampreys migrate to the ocean (or to the Great Lakes), where they feed on the blood and bodily fluids of salmonid fish and ultimately return to freshwater streams and rivers to spawn and die. The unique evolutionary position of lampreys and the relative ease of obtaining mature adults and embryos make this animal an ideal model for investigations into early vertebrate evolution. Studies of features shared between lampreys and jawed vertebrates, but distinct from those in nonvertebrate chordates, have provided information on the origin and evolution of hallmark vertebrate characteristics such as the neural crest, ectodermal placodes, and jaw. In addition, studies of features that are unique to lampreys (e.g., the variable lymphocyte receptor-mediated immune system) provide insights into mechanisms of parallel evolution (e.g., the adaptive immune system). With the establishment of techniques for the extended maintenance and spawning of lampreys in the laboratory, the sequencing of the lamprey genome, and the adaptation and optimization of many established molecular biology and histochemistry techniques for use in this species, P. marinus is poised to become an evolutionary developmental model of choice.


Children's well-being 11 years after the Chornobyl catastrophe.
The psychological effects of technological disasters have rarely been studied in children. This study assessed the aftermath of the 1986 Chornobyl disaster in children evacuated to Kyiv from the contaminated zone surrounding the nuclear power facility. In 1997, we evaluated three hundred 10- to 12-year-old children in Kyiv who were in utero or infants at the time of the disaster and who had resided near Chornobyl (evacuees) and 300 sex-matched homeroom classmates who had never lived in a radiation-contaminated area. Response rates were 92% (evacuees) and 85% (classmates). Data were obtained from children, mothers, and teachers using standard measures of well-being and risk factors for childhood psychopathology. The children also received physical examinations and basic blood tests. The evacuees and classmates perceived their mental health similarly except for Chornobyl-related anxiety symptoms and perceived scholastic competence. No differences were found on the Iowa Conners' Teacher Rating Scale. Although the physical examination and blood test results were normal, the evacuee mothers rated their children's well-being as significantly worse, especially with respect to somatic symptoms on the Children's Somatization Inventory and Child Behavior Checklist. The most important risk factors for these ratings were maternal somatization and Chornobyl-related stress. Given the multiple stressful experiences to which evacuee families were exposed, the small differences in the children's self-reports suggest that there are protective factors in the lives of these children. The trauma experienced by the mothers was reflected in their perceptions of their children's well-being, particularly somatic symptoms, but was not transmitted to the children themselves.


Babies at double hazard: early development of infants at biologic and social risk.
From the population of a neonatal intensive care unit, 114 infants and their families were followed from birth to age 3 1/2 years. Infants showing massive brain damage at birth and/or severe mental retardation at 7 months of age were excluded from this analysis. The remainder were predominantly poor and nonwhite. The group showed normal cognitive development through age 15 months. By 28 months of age and thereafter, a severe decline in cognitive status proved to be associated with social class. In addition, serious behavioral maladjustment led to improverished cognitive development. The incidence of maladjustment was unrelated to social class. The impact of maladjustment on test scores was significant in all classes, but greater for the higher rather than the lower socioeconomic social groups. Neither neurologic pathology (excepting severe brain damage) nor gestational age (small for gestation age [SGA] vs appropriate for gestational age [AGA]) had a significant effect on IQ scores at 3 1/2 years of age. It is suggested that environmental deficits and stresses impair early cognitive and psychosocial development for both full-term and premature infants, but that the latter group is more vulnerable to environmental insufficiencies than are full-term babies.


Prevention of acute kidney injury by tauroursodeoxycholic acid in rat and cell culture models.
Acute kidney injury (AKI) has grave short- and long-term consequences. Often the onset of AKI is predictable, such as following surgery that compromises blood flow to the kidney. Even in such situations, present therapies cannot prevent AKI. As apoptosis is a major form of cell death following AKI, we determined the efficacy and mechanisms of action of tauroursodeoxycholic acid (TUDCA), a molecule with potent anti-apoptotic and pro-survival properties, in prevention of AKI in rat and cell culture models. TUDCA is particularly attractive from a translational standpoint, as it has a proven safety record in animals and humans. We chose an ischemia-reperfusion model in rats to simulate AKI in native kidneys, and a human kidney cell culture model to simulate AKI associated with cryopreservation in transplanted kidneys. TUDCA significantly ameliorated AKI in the test models due to inhibition of the mitochondrial pathway of apoptosis and upregulation of survival pathways. This study sets the stage for testing TUDCA in future clinical trials for prevention of AKI, an area that needs urgent attention due to lack of effective therapies.


Microsatellites are molecular clocks that support accurate inferences about history.
Microsatellite length mutations are often modeled using the generalized stepwise mutation process, which is a type of random walk. If this model is sufficiently accurate, one can estimate the coalescence time between alleles of a locus after a mathematical transformation of the allele lengths. When large-scale microsatellite genotyping first became possible, there was substantial interest in using this approach to make inferences about time and demography, but that interest has waned because it has not been possible to empirically validate the clock by comparing it with data in which the mutation process is well understood. We analyzed data from 783 microsatellite loci in human populations and 292 loci in chimpanzee populations, and compared them with up to one gigabase of aligned sequence data, where the molecular clock based upon nucleotide substitutions is believed to be reliable. We empirically demonstrate a remarkable linearity (r(2) > 0.95) between the microsatellite average square distance statistic and sequence divergence. We demonstrate that microsatellites are accurate molecular clocks for coalescent times of at least 2 million years (My). We apply this insight to confirm that the African populations San, Biaka Pygmy, and Mbuti Pygmy have the deepest coalescent times among populations in the Human Genome Diversity Project. Furthermore, we show that microsatellites support unbiased estimates of population differentiation (F(ST)) that are less subject to ascertainment bias than single nucleotide polymorphism (SNP) F(ST). These results raise the prospect of using microsatellite data sets to determine parameters of population history. When genotyped along with SNPs, microsatellite data can also be used to correct for SNP ascertainment bias.


A randomized controlled clinical trial evaluating quality of life when using a simple acupressure protocol in women with primary dysmenorrhea.
To evaluate a simple acupressure protocol in LIV3 and LI4 acupoints in women with primary dysmenorrhea. This paper reports a randomized, single blinded clinical trial. 90 young women with dysmenorrhea were recruited to three groups to receive 20min acupressure every day in either LIV3 or LI4, or placebo points. Acupressure was timed five days before menstruation for three successive menstrual cycles. On menstruation, each participant completed the Wong Baker faces pain scale, and the quality of life short form -12 (QOL SF-12). Intensity and duration of pain between the three groups in the second and third cycles during the intervention (p<0.05) differed significantly. Significant differences were seen in all domains of QOL except for mental health (p=0.4), general health (p=0.7) and mental subscale component (p=0.12) in the second cycle, and mental health (p=0.9), and mental subscale component (p=0.14) in the third cycle. Performing the simple acupressure protocol is an effective method to decrease the intensity and duration of dysmenorrhea, and improve the QOL. Registration ID in IRCT: IRCT2016052428038N1.


Magnetoglow: a new geophysical resource.
Evidence has been found that the earth is immersed in a large volume of glowing helium ions. This ionic glow, at 304 angstroms, is similar to the geocoronal hydrogen glow in that it extends to very high altitudes, but it is unique in that it is largely confined to the closed field line portion of the magnetosphere. Because of its magnetic containment, this ionic radiation is called the "magnetoglow." Observations of the magnetoglow from inside and outside the magnetospheric cavity promise to provide a valuable means of studying the structural dynamics of the magnetosphere.


The cumulative risk of jail incarceration.
Research on incarceration has focused on prisons, but jail detention is far more common than imprisonment. Jails are local institutions that detain people before trial or incarcerate them for short sentences for low-level offenses. Research from the 1970s and 1980s viewed jails as "managing the rabble," a small and deeply disadvantaged segment of urban populations that struggled with problems of addiction, mental illness, and homelessness. The 1990s and 2000s marked a period of mass criminalization in which new styles of policing and court processing produced large numbers of criminal cases for minor crimes, concentrated in low-income communities of color. In a period of widespread criminal justice contact for minor offenses, how common is jail incarceration for minority men, particularly in poor neighborhoods? We estimate cumulative risks of jail incarceration with an administrative data file that records all jail admissions and discharges in New York City from 2008 to 2017. Although New York has a low jail incarceration rate, we find that 26.8% of Black men and 16.2% of Latino men, in contrast to only 3% of White men, in New York have been jailed by age 38 y. We also find evidence of high rates of repeated incarceration among Black men and high incarceration risks in high-poverty neighborhoods. Despite the jail's great reach in New York, we also find that the incarcerated population declined in the study period, producing a large reduction in the prevalence of jail incarceration for Black and Latino men.


Diminished pulmonary function in long-term workers exposed to cotton dust determined in a cross-sectional study in small Pakistani enterprises.
While large cotton industry plants producing for the foreign market have undergone improvements of occupational hygiene lately, working conditions in power looms producing mainly for the local market are still poor. The respiratory health in workers of power looms has not been studied so far. In a cross-sectional study, 51 male cotton workers from power looms, 58 from large cotton factories and 52 controls were investigated by spirometry and a symptoms questionnaire. Spirometric measurements and respiratory symptoms were related to exposure duration in large and small factories by multivariate linear regression and logistic regression, respectively, with smoking, age and anthropometric parameters considered as confounders. Work duration in power looms was associated with declines in forced expiratory volume in 1 s, peak flow, maximum midexpiratory flow and forced expiratory flow 75 and an increased risk of obstruction (OR 1.09/year; 95% CI: 1.002 to 1.18) by applying the Global Lung Initiative equations. Except cough that was only associated with smoking, all respiratory symptoms (chest tightness, shortness of breath, fever) increased as a function of duration of exposure in power looms but not in large factories. Despite limitations inherent in the cross-sectional approach, there is evidence for workers in small weaving factories to face an increased risk of pulmonary function loss and respiratory symptoms. Industrial hygiene measures in these small enterprises should be encouraged. Failure to detect adverse respiratory outcomes from occupation in large factories could be due to a 'survivor' (healthy worker) effect.


An Overview of Different Interdental Cleaning Aids and Their Effectiveness.
Optimisation of plaque control is essential for the success of non-surgical and surgical periodontal therapy. This cannot be achieved with brushing alone; hence, there is a need for adjunctive interdental cleaning aids. The aim of this paper is to provide an overview of different interdental cleaning aids and review the literature for consensus on their effectiveness. A literature search of articles in English, up to December 2018, was conducted in Pubmed. High-quality flossing is difficult to achieve, and ineffective routine use of floss may not confer significant benefits over brushing alone. Interdental brushes are more effective than brushing as a monotherapy. They are at least as good if not superior to floss in reducing plaque and gingivitis. Although they are effective for patients regardless of their periodontal status (healthy or active), they are especially indicated in periodontal patients where widened embrasures are common. Added benefits include ease of use, patient acceptance, and recontouring of interdental tissues. Rubberpiks do not demonstrate inferiority to conventional interdental brushes. Wooden interdental aids appear to offer no significant advantage over brushing with respect to plaque removal; they may, however, reduce gingival bleeding. Oral irrigators are a promising tool for reducing gingival inflammation, despite minimal changes to plaque levels. For cleaning around dental implants, oral irrigators and interdental brushes are preferred over floss.


The regulation of hepatic fatty acid synthesis and partitioning: the effect of nutritional state.
Nonalcoholic fatty liver disease (NAFLD) is an increasing global public health burden. NAFLD is strongly associated with type 2 diabetes mellitus, obesity and cardiovascular disease and begins with intrahepatic triacylglycerol accumulation. Under healthy conditions, the liver regulates lipid metabolism to meet systemic energy needs in the fed and fasted states. The processes of fatty acid uptake, fatty acid synthesis and the intracellular partitioning of fatty acids into storage, oxidation and secretion pathways are tightly regulated. When one or more of these processes becomes dysregulated, excess lipid accumulation can occur. Although genetic and environmental factors have been implicated in the development of NAFLD, it remains unclear why an imbalance in these pathways begins. The regulation of fatty acid partitioning occurs at several points, including during triacylglycerol synthesis, lipid droplet formation and lipolysis. These processes are influenced by enzyme function, intake of dietary fats and sugars and whole-body metabolism, and are further affected by the presence of obesity or insulin resistance. Insight into how the liver controls fatty acid metabolism in health and how these processes might be affected in disease would offer the potential for new therapeutic treatments for NAFLD to be developed.


Inhibition of IL-6 and IL-10 signaling and Stat activation by inflammatory and stress pathways.
The development and resolution of an inflammatory process are regulated by a complex interplay among cytokines that have pro- and anti-inflammatory effects. Effective and sustained action of a proinflammatory cytokine depends on synergy with other inflammatory cytokines and antagonism of opposing cytokines that are often highly expressed at inflammatory sites. We analyzed the effects of the inflammatory and stress agents, IL-1, TNF-alpha, LPS, sorbitol, and H(2)O(2), on signaling by IL-6 and IL-10, pleiotropic cytokines that activate the Jak-Stat signaling pathway and have both pro- and anti-inflammatory actions. IL-1, TNF-alpha, and LPS blocked the activation of Stat DNA binding and tyrosine phosphorylation by IL-6 and IL-10, but not by IFN-gamma, in primary macrophages. Inhibition of Stat activation correlated with inhibition of expression of IL-6-inducible genes. The inhibition was rapid and independent of de novo gene induction and occurred when the expression of suppressor of cytokine synthesis-3 was blocked. Inhibition of IL-6 signaling was mediated by the p38 subfamily of stress-activated protein kinases. Jak1 was inhibited at the level of tyrosine phosphorylation, indicating that inhibition occurred at least in part upstream of Stats in the Jak-Stat pathway. Experiments using Stat3 mutated at serine 727 and using truncated IL-6Rs suggested that the target of inhibition is contained within the membrane-proximal region of the cytoplasmic domain of the gp130 subunit of the IL-6 receptor and is different from the SH2 domain-containing protein-tyrosine phosphatase/suppressor of cytokine synthesis-3 docking site. These results identify a new level at which IL-1 and TNF-alpha modulate signaling by pleiotropic cytokines such as IL-6 and IL-10 and provide a molecular basis for the previously described antagonism of certain IL-6 actions by IL-1.


The effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta-lactamase.
For a number of years a major limitation in genetic analysis of protein function has been the inability to introduce multiple substitutions at distant sites that would enable the selection of clusters of mutations required for improved or novel biological functions. In order to achieve this, we have recently developed a novel mutagenesis procedure in which the triphosphate derivatives of a pyrimidine (6-(2-deoxy-beta-d-ribofuranosyl)-3, 4-dihydro-8H-pyrimido-[4,5-c][1,2]oxazin-7-one; dP) and a purine (8-oxo-2'-deoxyguanosine; 8-oxodG) nucleoside analogue are employed in DNA synthesis reactions in vitro. The procedure allows control of the mutational load and can yield frequencies of amino acid residue substitutions at least one order of magnitude greater than those previously achieved. Here we report the results of an experiment in which we have hypermutated the bacterial enzyme TEM-1 beta-lactamase and selected small pools (<1.5x10(5)) of clones for enzymatic activity against the beta-lactam antibiotic cefotaxime. The experiment resulted in the isolation of a number of TEM-1 mutants with greatly improved activity against cefotaxime. Among these, clone 3D.5 (E104K:M182T:G238S) exhibited a minimum inhibitory concentration for cefotaxime 20,000-fold higher than wild-type TEM-1 and a catalytic efficiency (kcat/Km) 2383 times higher than the wild-type enzyme. Thus, small pools of hypermutated sequences enabled the selection of one of the most active extended beta-lactamases described so far. These results argue against the accepted view that multiple rounds of low-rate mutagenesis and stepwise selection are essential for in vitro protein evolution and extend the scope of directed molecular evolution to proteins for which no genetic selection is available.


Surface water mineralization of isopyrazam according to OECD 309: observations on implementation of the new data requirement within agrochemical regulation.
A surface water mineralization study (according to the Organisation for Economic Co-operation and Development [OECD] guideline OECD 309) is a new requirement in European Union agrochemical regulations; therefore, industry has little experience with this test. The guideline allows for a number of options within the test design, notably the options to conduct the study under diffuse light and to include an inoculum of suspended sediment. The present study was designed to investigate the potential impact of these options on the degradation rate of a representative compound. The fungicide, isopyrazam, was chosen as it was previously shown to be susceptible to metabolism by phototrophic organisms under a fluorescent light-dark cycle. The impact of diffuse light was investigated at light intensities representative of those at depth in large, open water bodies (<7% of the incident intensity), and it was demonstrated that metabolism of isopyrazam by phototrophic microorganisms was rapid (median degradation time for 50% of the test compound [DT50] < 50 d), whereas degradation in continuous darkness was negligible. Furthermore, investigation at 2 different light intensities resulted in similar degradation rates, indicating that this transformation mechanism was not proportional to light intensity, provided that there was sufficient light for photosynthesis to occur. Inclusion of suspended sediment did not have a significant impact on the degradation rate of isopyrazam, except at extremely high sediment concentrations, which were not considered representative of conditions in large, open water bodies.


Simplifying mixture models using the unscented transform.
Mixture of Gaussians (MoG) model is a useful tool in statistical learning. In many learning processes that are based on mixture models, computational requirements are very demanding due to the large number of components involved in the model. We propose a novel algorithm for learning a simplified representation of a Gaussian mixture, that is based on the Unscented Transform which was introduced for filtering nonlinear dynamical systems. The superiority of the proposed method is validated on both simulation experiments and categorization of a real image database. The proposed categorization methodology is based on modeling each image using a Gaussian mixture model. A category model is obtained by learning a simplified mixture model from all the images in the category.


Autoimmune diseases, gastrointestinal disorders and the microbiome in schizophrenia: more than a gut feeling.
Autoimmunity, gastrointestinal (GI) disorders and schizophrenia have been associated with one another for a long time. This paper reviews these connections and provides a context by which multiple risk factors for schizophrenia may be related. Epidemiological studies strongly link schizophrenia with autoimmune disorders including enteropathic celiac disease. Exposure to wheat gluten and bovine milk casein also contribute to non-celiac food sensitivities in susceptible individuals. Co-morbid GI inflammation accompanies humoral immunity to food antigens, occurs early during the course of schizophrenia and appears to be independent from antipsychotic-generated motility effects. This inflammation impacts endothelial barrier permeability and can precipitate translocation of gut bacteria into systemic circulation. Infection by the neurotropic gut pathogen, Toxoplasma gondii, will elicit an inflammatory GI environment. Such processes trigger innate immunity, including activation of complement C1q, which also functions at synapses in the brain. The emerging field of microbiome research lies at the center of these interactions with evidence that the abundance and diversity of resident gut microbiota contribute to digestion, inflammation, gut permeability and behavior. Dietary modifications of core bacterial compositions may explain inefficient gluten digestion and how immigrant status in certain situations is a risk factor for schizophrenia. Gut microbiome research in schizophrenia is in its infancy, but data in related fields suggest disease-associated altered phylogenetic compositions. In summary, this review surveys associative and experimental data linking autoimmunity, GI activity and schizophrenia, and proposes that understanding of disrupted biological pathways outside of the brain can lend valuable information regarding pathogeneses of complex, polygenic brain disorders.


A single amino acid change in histone H4 enhances UV survival and DNA repair in yeast.
Single amino acid changes at specific DNA contacts of histones H3 and H4 generate SWI/SNF-independent (Sin) mutants in yeast. We have analyzed the effect of the Sin mutation at R45 of histone H4 on cell survival following UV irradiation, nucleotide excision repair (NER) and chromatin structure. We find that this mutation renders yeast cells more resistant to UV damage and enhances NER at specific chromatin loci. In the transcriptionally silent HML, repressed GAL10 and the constitutively active RPB2 loci, H4 R45 mutants exhibit enhanced repair of UV-induced cyclobutane pyrimidine dimers (CPDs) compared to wild-type (wt). However, the H4 R45 mutation does not increase the transcription of NER genes, disrupt transcriptional silencing of the HML locus or alter repression in the GAL10 locus. We have further shown that the H4 R45C mutation increases the accessibility of nucleosome DNA in chromatin to exogenous nucleases and may expedite nucleosome rearrangements during NER. Taken together, our results indicate that the increased repair observed in Sin mutants is a direct effect of the altered chromatin landscape caused by the mutation, suggesting that such subtle changes in the conserved histone residues can influence the accessibility of DNA repair factors in chromatin.


Pathophysiological Mechanisms of Sino-Atrial Dysfunction and Ventricular Conduction Disease Associated with SCN5A Deficiency: Insights from Mouse Models.
Genetically modified mice provide a number of models for studying cardiac channelopathies related to cardiac Na(+) channel (SCN5A) abnormalities. We review key pathophysiological features in these murine models that may underlie clinical features observed in sinus node dysfunction and progressive cardiac conduction disease, thereby providing insights into their pathophysiological mechanisms. We describe loss of Na(+) channel function and fibrotic changes associated with both loss and gain-of-function Na(+) channel mutations. Recent reports further relate the progressive fibrotic changes to upregulation of TGF-β1 production and the transcription factors, Atf3, a stress-inducible gene, and Egr1, to the presence of heterozygous Scn5a gene deletion. Both changes are thus directly implicated in the clinically observed disruptions in sino-atrial node pacemaker function, and sino-atrial and ventricular conduction, and their progression with age. Murine systems with genetic modifications in Scn5a thus prove a useful tool to address questions concerning roles of genetic and environmental modifiers on human SCN5A disease phenotypes.


Temperature Effects on Race Determination in Heterodera glycines.
Currently there are 16 possible races for Heterodera glycines, and these are differentiated based on ability of a nematode population to develop on a set of four differential soybean genotypes. Because results are based on numbers of nematode females that develop to a specific stage rather than on the reproductive capability of these females, race determinations based on female indices may not represent results obtained after several reproductive cycles of H. glycines. Counting numbers of eggs and juveniles, and then developing corresponding indices, would allow reproduction to be considered in making race determinations. Our objectives were to compare the present race identification scheme for H. glycines based on female indices with those using egg and juvenile indices and to examine the effect of temperature on race designations using female, egg, and juvenile indices. Race designations for H. glycines populations from two locations in Illinois were determined at 20, 27, and 30 degrees C in a water bath. The numbers of females, eggs, and juveniles (at 19 days) were recorded, and an index based on each life stage was calculated. Race determinations based on female, egg, or juvenile indices were inconsistent when conducted at 20 degrees C, which demonstrates that this temperature is not suitable for identifying races of H. glycines. However race designations at 27 and 30 degrees C were consistent for all three indices. This indicates that counting females, eggs, or juveniles should be equally reliable when race determinations are conducted at these two temperatures, and choice of method would depend on investigator preference or research objective.


Ubiquitin ligases: guardians of mammalian development.
Mammalian development demands precision. Millions of molecules must be properly located in temporal order, and their function regulated, to orchestrate important steps in cell cycle progression, apoptosis, migration and differentiation, to shape developing embryos. Ubiquitin and its associated enzymes act as cellular guardians to ensure precise spatio-temporal control of key molecules during each of these important cellular processes. Loss of precision results in numerous examples of embryological disorders or even cancer. This Review discusses the crucial roles of E3 ubiquitin ligases during key steps of early mammalian development and their roles in human disease, and considers how new methods to manipulate and exploit the ubiquitin regulatory machinery - for example, the development of molecular glues and PROTACs - might facilitate clinical therapy.


Isolation of chikungunya virus contaminating an Aedes albopictus cell line.
An Aedes albopictus cell line was found contaminated with structures morphologically compatible with an alphavirus. Rapid isolation of a cytopathic virus was effected by combining sonication, concentration with Aquacide II-r, rate zonal sedimentation and subsequent plating of fractions on Vero cells under agar overlay. The virus caused neithedeath nor disease on inoculation into infant and adult mice. It produced a c.p.e. in Vero and BHK 21 cells, and multiplied in Singh's Aedes aegypti cells. The virus was identified serologically as chikungunya by complement-fixation and plaque reduction netralization test. Virus was not detected in a single attempt by these methods in the American Type Culture Collection A. albopicturs line. The presence of chikungunya virus in A. albopictus cells is not easily recognized and may complicate interpretation of experimental results.


Neuroprotective effect of caffeic acid phenethyl ester in 3-nitropropionic acid-induced striatal neurotoxicity.
Caffeic acid phenethyl ester (CAPE), derived from honeybee hives, is a bioactive compound with strong antioxidant activity. This study was designed to test the neuroprotective effect of CAPE in 3-nitropropionic acid (3NP)-induced striatal neurotoxicity, a chemical model of Huntington's disease (HD). Initially, to test CAPE's antioxidant activity, a 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) antioxidant assay was employed, and CAPE showed a strong direct radical-scavenging eff ect. In addition, CAPE provided protection from 3NP-induced neuronal cell death in cultured striatal neurons. Based on these observations, the in vivo therapeutic potential of CAPE in 3NP-induced HD was tested. For this purpose, male C57BL/6 mice were repeatedly given 3NP to induce HD-like pathogenesis, and 30 mg/kg of CAPE or vehicle (5% dimethyl sulfoxide and 95% peanut oil) was administered daily. CAPE did not cause changes in body weight, but it reduced mortality by 29%. In addition, compared to the vehicle-treated group, robustly reduced striatal damage was observed in the CAPE-treated animals, and the 3NP-induced behavioral defi cits on the rotarod test were signifi cantly rescued after the CAPE treatment. Furthermore, immunohistochemical data showed that immunoreactivity to glial fibrillary acidic protein (GFAP) and CD45, markers for astrocyte and microglia activation, respectively, were strikingly reduced. Combined, these data unequivocally indicate that CAPE has a strong antioxidant eff ect and can be used as a potential therapeutic agent against HD.


Alpha-pyridylation of chiral amines via urea coupling, lithiation and rearrangement.
2-, 3- and 4-Bromopyridine, along with other brominated electron-deficient arenes, undergo palladium-catalyzed coupling with ureas of structure RNMeCONHMe. Where R is a chiral, alpha-substituted benzyl group, treatment with LDA leads to a benzylic organolithium which undergoes rearrangement with stereospecific and regiospecific transfer of the pyridyl group, generating a quaternary stereogenic center with high enantioselectivity. Alcoholysis of the urea under neutral conditions reveals the pyridyl amine.


Diagnostic value of peritumoral minimum apparent diffusion coefficient for differentiation of glioblastoma multiforme from solitary metastatic lesions.
In glioblastoma multiforme, the peritumoral region may be infiltrated with malignant cells in addition to vasogenic edema, whereas in a metastatic deposit, the peritumoral areas comprise predominantly vasogenic edema. The purpose of this study was to determine whether the minimum apparent diffusion coefficient (ADC) can be used to differentiate glioblastoma from solitary metastasis on the basis of cellularity levels in the enhancing tumor and in the peritumoral region. Seventy-three patients underwent conventional MRI and diffusion-weighted imaging (DWI) before undergoing treatment. The minimum ADC was measured in the enhancing tumor, peritumoral region, and contralateral normal white matter. To determine whether there was a statistical difference between metastasis and glioblastoma, we analyzed patient age and sex, minimum ADC value, and ADC ratio of the two groups. A receiver operating characteristic (ROC) curve analysis was used to determine the cutoff value of the minimum ADC that had the best combination of sensitivity and specificity for distinguishing between glioblastoma and metastasis. The mean minimum ADC values and mean ADC ratios in the peritumoral regions of glioblastomas were significantly higher than those in metastases. However, the mean minimum ADC values and mean ADC ratios in enhancing tumors showed no statistically significant difference between the two groups. According to ROC curve analysis, a cutoff value of 1.302 × 10(-3) mm(2)/s for the minimum peritumoral ADC value generated the best combination of sensitivity (82.9%) and specificity (78.9%) for distinguishing between glioblastoma and metastasis. Although the characteristics of solitary metastasis and glioblastoma multiforme may be similar on conventional MRI, DWI can offer diagnostic information to distinguish between the tumors.


Elevated risk for autoimmune disorders in iraq and afghanistan veterans with posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD) is associated with endocrine and immune abnormalities that could increase risk for autoimmune disorders. However, little is known about the risk for autoimmune disorders among individuals with PTSD. We conducted a retrospective cohort study of 666,269 Iraq and Afghanistan veterans under age 55 who were enrolled in the Department of Veterans Affairs health care system between October 7, 2001, and March 31, 2011. Generalized linear models were used to examine if PTSD, other psychiatric disorders, and military sexual trauma exposure increased risk for autoimmune disorders, including thyroiditis, inflammatory bowel disease, rheumatoid arthritis, multiple sclerosis, and lupus erythematosus, adjusting for age, gender, race, and primary care visits. PTSD was diagnosed in 203,766 veterans (30.6%), and psychiatric disorders other than PTSD were diagnosed in an additional 129,704 veterans (19.5%). Veterans diagnosed with PTSD had significantly higher adjusted relative risk (ARR) for diagnosis with any of the autoimmune disorders alone or in combination compared with veterans with no psychiatric diagnoses (ARR = 2.00; 95% confidence interval, 1.91-2.09) and compared with veterans diagnosed with psychiatric disorders other than PTSD (ARR = 1.51; 95% confidence interval, 1.43-1.59; p < .001). The magnitude of the PTSD-related increase in risk for autoimmune disorders was similar in women and men, and military sexual trauma exposure was independently associated with increased risk in both women and men. Trauma exposure and PTSD may increase risk for autoimmune disorders. Altered immune function, lifestyle factors, or shared etiology may underlie this association.


An analysis of the accuracy of magnetic resonance flip angle measurement methods.
Several methods of flip angle mapping for magnetic resonance imaging have been proposed. We evaluated the accuracy of five methods of flip angle measurement in the presence of measurement noise. Our analysis was performed in a closed form by propagation of probability density functions (PDFs). The flip angle mapping methods compared were (1) the phase-sensitive method, (2) the dual-angle method using gradient recalled echoes (GRE), (3) an extended version of the GRE dual-angle method incorporating phase information, (4) the AFI method and (5) an extended version of the AFI method incorporating phase information. Our analysis took into account differences in required imaging time for these methods in the comparison of noise efficiency. PDFs of the flip angle estimate for each method for each value of true flip angle were calculated. These PDFs completely characterize the performance of each method. Mean bias and standard deviation were computed from these PDFs to more simply quantify the relative accuracy of each method over its range of measurable flip angles. We demonstrate that the phase-sensitive method provides the lowest mean bias and standard deviation of flip angle estimate of the five methods evaluated over a wide range of flip angles.


Sleep to Lower Elevated Blood Pressure: A Randomized Controlled Trial (SLEPT).
Impaired sleep quality is common and associated with an increased risk of cardiovascular disease (CVD), thought to be mediated through adverse effects on established vascular risk factors, particularly hypertension. We determined if a web-delivered sleep intervention (sleep-hygiene education, stimulus control, and cognitive behavioral therapy) reduces blood pressure compared to vascular risk factor education (standard care) alone. Phase II randomized, blinded, controlled trial of 134 participants without CVD with mild sleep impairment and blood pressure 130-160/<110 mm Hg. The primary outcome was the difference in the mean change in 24-hour ambulatory systolic blood pressure (SBP) over 8 weeks between intervention and control groups. Secondary outcomes included measures of sleep quality and psychosocial health, namely Insomnia Severity Index (ISI), Pittsburgh Sleep Quality Index (PSQI), Beck Depression Inventory (BDI), and Beck Anxiety Inventory (BAI). Participants in the sleep intervention group showed significantly greater improvements in sleep quality, including ISI [difference in mean improvement 2.8; 95% confidence interval (CI), 1.3-4.4], PSQI (1.1; 95% CI, 0.1-2.2), sleep condition indicator (0.8; 95% CI, 0.2-1.4), and psychosocial health, including BDI (2.0; 95% CI, 0.3-3.7) and BAI (1.4; 95% CI, 0.02-2.8). The mean improvement in 24-hour ambulatory SBP did not differ between the sleep intervention (0.9 mm Hg) and control (0.8 mm Hg) arms, (difference in mean improvement 0.1; 95% CI, -3.4 to 3.2). A simple, low-cost, web-delivered sleep intervention is feasible and significantly improves sleep quality and measures of psychosocial health in individuals with mild sleep impairment but does not result in short-term improvements in blood pressure.


Pathogenic properties of Alzheimer's β-amyloid identified from structure-property patient-phenotype correlations.
β-Amyloid (Aβ) plays a central role in Alzheimer's disease (AD), but the specific molecular mechanism and associated structures remain unknown. We compiled patient data for carriers of genetic variants of Aβ that cause AD and correlated these data against chemical properties for 56 mutant conformations derived from four published experimental conformations of Aβ of variable structure and chemical environment. Disease onset of variants is significantly (p ∼ 0.006) correlated to hydrophobic surfaces of disordered conformations (2LFM), whereas structured conformations yielded no correlations. Correlation also applied (p < 0.03) to in vitro steady-state Aβ levels. We conclude that disordered monomers are likely to be pathogenically important in contrast to structured conformations and that hydrophobic surface correlates with pathogenesis. This first established correlation between clinical and chemical data suggests that specific exposed, disordered monomers are viable targets for AD therapy.


Regional specificity of synaptic plasticity deficits in a knock-in mouse model of DYT1 dystonia.
DYT1 dystonia is a movement disorder caused by a deletion in the C-terminal of the protein torsinA. It is unclear how torsinA mutation might disrupt cellular processes encoding motor activity, and whether this impairment occurs in specific brain regions. Here, we report a selective impairment of corticostriatal synaptic plasticity in knock-in mice heterozygous for Δ-torsinA (Tor1a(+/Δgag) mice) as compared to controls (Tor1a(+/+) mice). In striatal spiny neurons from Tor1a(+/Δgag) mice, high-frequency stimulation failed to induce long-term depression (LTD), whereas long-term potentiation (LTP) exhibited increased amplitude. Of interest, blockade of D2 dopamine receptors (D2Rs) increased LTP in Tor1a(+/+) mice to a level comparable to that measured in Tor1a(+/Δgag) mice and normalized the levels of potentiation across mouse groups. A low-frequency stimulation (LFS) protocol was unable to depotentiate corticostriatal synapses in Tor1a(+/Δgag) mice. Muscarinic M1 acetylcholine receptor (mAChR) blockade rescued plasticity deficits. Additionally, we found an abnormal responsiveness of cholinergic interneurons to D2R activation, consisting in an excitatory response rather than the expected inhibition, further confirming an imbalance between dopaminergic and cholinergic signaling in the striatum. Conversely, synaptic activity and plasticity in the CA1 hippocampal region were unaltered in Tor1a(+/Δgag) mice. Importantly, the M1 mAChR-dependent enhancement of hippocampal LTP was unaffected in both genotypes. Similarly, both basic properties of dopaminergic nigral neurons and their responses to D2R activation were normal. These results provide evidence for a regional specificity of the electrophysiological abnormalities observed and demonstrate the reproducibility of such alterations in distinct models of DYT1 dystonia.


Health system responsiveness: a systematic evidence mapping review of the global literature.
The World Health Organisation framed responsiveness, fair financing and equity as intrinsic goals of health systems. However, of the three, responsiveness received significantly less attention. Responsiveness is essential to strengthen systems' functioning; provide equitable and accountable services; and to protect the rights of citizens. There is an urgency to make systems more responsive, but our understanding of responsiveness is limited. We therefore sought to map existing evidence on health system responsiveness. A mixed method systemized evidence mapping review was conducted. We searched PubMed, EbscoHost, and Google Scholar. Published and grey literature; conceptual and empirical publications; published between 2000 and 2020 and English language texts were included. We screened titles and abstracts of 1119 publications and 870 full texts. Six hundred twenty-one publications were included in the review. Evidence mapping shows substantially more publications between 2011 and 2020 (n = 462/621) than earlier periods. Most of the publications were from Europe (n = 139), with more publications relating to High Income Countries (n = 241) than Low-to-Middle Income Countries (n = 217). Most were empirical studies (n = 424/621) utilized quantitative methodologies (n = 232), while qualitative (n = 127) and mixed methods (n = 63) were more rare. Thematic analysis revealed eight primary conceptualizations of 'health system responsiveness', which can be fitted into three dominant categorizations: 1) unidirectional user-service interface; 2) responsiveness as feedback loops between users and the health system; and 3) responsiveness as accountability between public and the system. This evidence map shows a substantial body of available literature on health system responsiveness, but also reveals evidential gaps requiring further development, including: a clear definition and body of theory of responsiveness; the implementation and effectiveness of feedback loops; the systems responses to this feedback; context-specific mechanism-implementation experiences, particularly, of LMIC and fragile-and conflict affected states; and responsiveness as it relates to health equity, minority and vulnerable populations. Theoretical development is required, we suggest separating ideas of services and systems responsiveness, applying a stronger systems lens in future work. Further agenda-setting and resourcing of bridging work on health system responsiveness is suggested.


Intraventricular Thrombus Formation and Embolism in Takotsubo Syndrome: Insights From the International Takotsubo Registry.
Takotsubo syndrome (TTS) is characterized by acute left ventricular dysfunction, which can contribute to intraventricular thrombus and embolism. Still, prevalence and clinical impact of thrombus formation and embolic events on outcome of TTS patients remain unclear. This study aimed to investigate clinical features and outcomes of patients with and without intraventricular thrombus or embolism. Additionally, factors associated with thrombus formation or embolism, as well as predictors for mortality, were identified. Approach and Results: TTS patients enrolled in the International Takotsubo Registry at 28 centers in Australia, Europe, and the United States were dichotomized according to the occurrence/absence of intraventricular thrombus or embolism. Patients with intraventricular thrombus or embolism were defined as the ThrombEmb group. Of 1676 TTS patients, 56 (3.3%) patients developed intraventricular thrombus and/or embolism following TTS diagnosis (median time interval, 2.0 days [range, 0-38 days]). Patients in the ThrombEmb group had a different clinical profile including lower left ventricular ejection fraction, higher prevalence of the apical type, elevated levels of troponin and inflammatory markers, and higher prevalence of vascular disease. In a Firth bias-reduced penalized-likelihood logistic regression model apical type, left ventricular ejection fraction ≤30%, previous vascular disease, and a white blood cell count on admission >10×103 cells/μL emerged as independent predictors for thrombus formation or embolism. Intraventricular thrombus or embolism occur in 3.3% of patients in the acute phase of TTS. A simple risk score including clinical parameters associated with intraventricular thrombus formation or embolism identifies patients at increased risk. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01947621.


Abundance of cladocerans in the littoral regions of two environments of the Upper Paraná River floodplain, Mato Grosso do Sul, Brazil.
The abundance of cladocerans recorded in plankton samples from the littoral regions of two environments (Baía River and Guaraná lake), the Upper Paraná River floodplain and some limnological variables important in controlling their abundance were investigated at three sites in the lake and two in the river, from March/93 to January/94. Abundance was highest in the lake, chiefly during high water. Bosminidae dominated in both environments and hydrological periods (high and low water). Chydoridae were more important in the lake, and Sididae in the river. A higher number of Pearson's correlations significative (p < 0.03) between the densities of the ten most important species and the limnological variables was observed in the river. This suggests that limnological variables are more important in controlling the abundance of cladocerans in the river. On the other hand, in the lake their abundance was probably associated with the physical structure of the environment, the abundance of aquatic macrophytes, and biotic relationships such as competition and predation.


Mutational clusters generated by non-processive polymerases: A case study using DNA polymerase betain vitro.
Available DNA mutational spectra reveal that the number of mutants with multiple mutations ("multiples") is usually greater than expected from a random distribution of mutations among mutants. These overloads imply the occurrence of non-random clusters of mutations, probably generated during episodes of low-fidelity DNA synthesis. Excess multiples have been reported not only for viruses, bacteria, and eukaryotic cells but also for the DNA polymerases of phages T4 and RB69 in vitro. In the simplest case of a purified polymerase, non-random clusters may be generated by a subfraction of phenotypic variants able to introduce more errors per cycle of DNA synthesis than the normal enzyme. According to this hypothesis, excess multiples are not expected with non-processive polymerases even if they harbor rare mutator variants. DNA polymerase beta (Pol beta) is a mammalian DNA-repair polymerase with very low processivity. Although several Pol beta mutational spectra have been described, there is conflicting evidence on whether or not excess multiples occur, with spectra based on the HSV-tk system tending to show excess multiples. Excess multiples generated by Pol beta or any of its mutants might imply that the excesses of multiples observed in numerous other systems, especially those with processive polymerases, could be artifactual. Here, the distributions of mutations generated by native and recombinant rat Pol beta and by the Pol beta(Y265C) mutator were analyzed in the M13mp2 lacZalpha system. Our results present no evidence for a significant excess of multiples over the expected numbers with any of the Pol beta enzymes tested in this system. The reported excess of Pol beta-generated multiples in the HSV-tk system may reflect a reduced efficiency of detection of base substitutions that cause weak phenotypes, which in turn may artifactually increase the frequency of multiples.


Systematic review: unmet supportive care needs in people diagnosed with chronic liver disease.
People with chronic liver disease, particularly those with decompensated cirrhosis, experience several potentially debilitating complications that can have a significant impact on activities of daily living and quality of life. These impairments combined with the associated complex treatment mean that they are faced with specific and high levels of supportive care needs. We aimed to review reported perspectives, experiences and concerns of people with chronic liver disease worldwide. This information is necessary to guide development of policies around supportive needs screening tools and to enable prioritisation of support services for these patients. Systematic searches of PubMed, MEDLINE, CINAHL and PsycINFO from the earliest records until 19 September 2014. Data were extracted using standardised forms. A qualitative, descriptive approach was utilised to analyse and synthesise data. The initial search yielded 2598 reports: 26 studies reporting supportive care needs among patients with chronic liver disease were included, but few of them were patient-reported needs, none used a validated liver disease-specific supportive care need assessment instrument, and only three included patients with cirrhosis. Five key domains of supportive care needs were identified: informational or educational (eg, educational material, educational sessions), practical (eg, daily living), physical (eg, controlling pruritus and fatigue), patient care and support (eg, support groups), and psychological (eg, anxiety, sadness). While several key domains of supportive care needs were identified, most studies included hepatitis patients. There is a paucity of literature describing the supportive care needs of the chronic liver disease population likely to have the most needs--namely those with cirrhosis. Assessing the supportive care needs of people with chronic liver disease have potential utility in clinical practice for facilitating timely referrals to support services.


Identification of subtypes of problem drinkers based on neuropsychological performance.
The key issue addressed in this paper is: Can specific subtypes of drinkers be identified on the basis of their neuropsychological performance? A multivariate model of neuropsychological deficits related to alcohol abuse was proposed and cluster analysis was used to see if subtypes could be identified which matched those indicated in the multivariate model. A neuropsychological cognitive assessment battery was given to a wide variety of drinkers (N = 88). Factor analysis yielded scores on four factors which formed the basis for the cluster analysis. Seven stable clusters were identified based on cognitive performance alone. Additionally, clusters were significantly differentiated by age, IQ, education, number of units of alcohol consumed on a heavy drinking day, nutritional status, stress and the Eysenck Personality Questionnaire Lie score. The seven clusters were eventually profiled as healthy males, healthy females, males with stress-related deficits, females with stress-related deficits, mildly impaired males, deficits related to liver dysfunction and mild alcoholic Korsakoff syndrome. The clusters successfully mapped onto the proposed model reinforcing the need for a multivariate approach to the study of neuropsychological deficits in problem drinkers.


Constituents of human neutrophils that mediate enhanced adherence to surfaces: purification and identification as acidic proteins of the specific granules.
The endogenous constituents of human neutrophils that enhance the adherence of the neutrophils to surfaces have been isolated from sonicates of purified neutrophils. The predominant adherence-enhancing activity in the neutrophil sonicates cofiltered on Sephadex G-75 with a major peak of chemotactic inhibitory activity and exhibited approximately 30,000 mol wt. Sequential isoelectric focusing and electrophoresis in glycerol gradients of the 30,000-mol wt activities resolved two distinct acidic protein with isoelectric points of 3.6-3.8 and 3.3-3.4 that were designated the neutrophil adherence factor (NAF) I and II, respectively. Glutamic acid and aspartic acid together accounted for a total of 18 and 19% of the amino acids in purified preparations of NAF I and NAF II, respectively, whereas the basic amino acids lysine, arginine, and histidine represented <2 and 3% of the total residues. The preincubation of portions of 2 x 10(6) neutrophils with as little as 6 pmol of NAF I or 9 pmol of NAF II enhanced adherence to plastic petri dishes and inhibited chemotactic migration to a maximal extent, with comparable dose-response relationships for the two effects. Neither of the NAF was cytotoxic, exhibited substantial neutrophil chemotactic or chemokinetic activity, or influenced the phagocytosis of sheep erythrocytes sensitized with immunoglobulin (Ig)G. Analyses of subcellular fractions of neutrophils indicated that the NAF are contained predominantly in the specific granules. These distinctive acidic proteins of the specific granules of human neutrophils represent a new class of endogenous constituents that may regulate the involvement of neutrophils in inflammation.


Geographic and species association of hepatitis B virus genotypes in non-human primates.
Infection with hepatitis B virus (HBV) has been detected in human populations throughout the world, as well as in a number of ape species (Pan troglodytes, Gorilla gorilla, gibbons [Nomascus and Hylobates species] and Pongo pygmaeus). To investigate the distribution of naturally occurring HBV infection in these species and other African Old World monkey species (Cercopithecidae), we screened 137 plasma samples from mainly wild caught animals by polymerase chain reaction (PCR) using several of highly conserved primers from the HB surface (HBs) gene, and for HBs antigen (HBsAg) by ELISA. None of the 93 Cercopithecidae screened (6 species) showed PCR or serology evidence for HBV infection; in contrast 2 from 8 chimpanzees and 5 from 22 gibbons were PCR-positive with each set of primers. Complete genome sequences from each of the positive apes were obtained and compared with all previously published complete and surface gene sequences. This extended phylogenetic analysis indicated that HBV variants from orangutans were interspersed by with HBV variants from southerly distributed gibbon species (H. agilis and H. moloch) occupying overlapping or adjacent habitat ranges with orangutans; in contrast, HBV variants from gibbon species in mainland Asia were phylogenetically distinct. A geographical rather than (sub)species association of HBV would account for the distribution of HBV variants in different subspecies of chimpanzees in Africa, and explain the inlier position of the previously described lowland gorilla sequence in the chimpanzee clade. These new findings have a number of implication for understanding the origins and epidemiology of HBV infection in non-human primates.


A new achievement in green degradation of aqueous organic pollutants under visible-light irradiation.
The global attention has been focused on degradation of the environmental organic pollutants through green methods such as advanced oxidation processes (AOPs) under sunlight. However, AOPs have not yet been efficient in function of the photocatalyst that has been used. In this work, firstly, CaCu3Ti4O12 nanocomposite was simultaneously synthesized and decorated in different amounts of graphene oxide to enhance photodegradation of the organics. The result of the photocatalyst characterization showed that the sample with 8% graphene presented optimum photo-electrical properties such as low band gap energy and a great surface area. Secondly, the photocatalyst was applied for photodegradation of an organic model in a batch photoreactor. Thirdly, to scale up the process and optimize the efficiency, the photodegradation was modeled by multivariate semi-empirical methods. As the optimized condition showed, 45 mg/L of the methyl-orange has been removed at pH 5.8 by 0.96 g/L of the photocatalyst during 288 min of the light irradiation. Moreover, the photodegradation has been scaled up for industrial applications by determining the importance of the input effective variables according to the following organics order > photocatalyst > pH > irradiation time.


One-step implementation of a multiqubit phase gate with one control qubit and multiple target qubits in coupled cavities.
We propose a one-step scheme to implement a multiqubit controlled phase gate with one qubit simultaneously controlling multiple qubits with three-level atoms at distant nodes in coupled cavity arrays. Selective qubit-qubit couplings are achieved by adiabatically eliminating the atomic excited states and photonic states, and the required phase shifts between the control qubit and any target qubit can be realized through suitable choices of the parameters of the external fields. Moreover, the effective model is robust against decoherence because neither the atoms nor the field modes are excited during the gate operation, leading to a useful step toward scalable quantum computing networks.


Delirium in elderly patients and the risk of postdischarge mortality, institutionalization, and dementia: a meta-analysis.
Delirium is a common and serious complication in elderly patients. Evidence suggests that delirium is associated with long-term poor outcome but delirium often occurs in individuals with more severe underlying disease. To assess the association between delirium in elderly patients and long-term poor outcome, defined as mortality, institutionalization, or dementia, while controlling for important confounders. A systematic search of studies published between January 1981 and April 2010 was conducted using the databases of MEDLINE, EMBASE, PsycINFO, and CINAHL. Observational studies of elderly patients with delirium as a study variable and data on mortality, institutionalization, or dementia after a minimum follow-up of 3 months, and published in the English or Dutch language. Titles, abstracts, and articles were reviewed independently by 2 of the authors. Of 2939 references in the original search, 51 relevant articles were identified. Information on study design, characteristics of the study population, and outcome were extracted. Quality of studies was assessed based on elements of the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist for cohort studies. The primary analyses included only high-quality studies with statistical control for age, sex, comorbid illness or illness severity, and baseline dementia. Pooled-effect estimates were calculated with random-effects models. The primary analysis with adjusted hazard ratios (HRs) showed that delirium is associated with an increased risk of death compared with controls after an average follow-up of 22.7 months (7 studies; 271/714 patients [38.0%] with delirium, 616/2243 controls [27.5%]; HR, 1.95 [95% confidence interval {CI}, 1.51-2.52]; I(2), 44.0%). Moreover, patients who had experienced delirium were also at increased risk of institutionalization (7 studies; average follow-up, 14.6 months; 176/527 patients [33.4%] with delirium and 219/2052 controls [10.7%]; odds ratio [OR], 2.41 [95% CI, 1.77-3.29]; I(2), 0%) and dementia (2 studies; average follow-up, 4.1 years; 35/56 patients [62.5%] with delirium and 15/185 controls [8.1%]; OR, 12.52 [95% CI, 1.86-84.21]; I(2), 52.4%). The sensitivity, trim-and-fill, and secondary analyses with unadjusted high-quality risk estimates stratified according to the study characteristics confirmed the robustness of these results. This meta-analysis provides evidence that delirium in elderly patients is associated with poor outcome independent of important confounders, such as age, sex, comorbid illness or illness severity, and baseline dementia.


RAD51B in Familial Breast Cancer.
Common variation on 14q24.1, close to RAD51B, has been associated with breast cancer: rs999737 and rs2588809 with the risk of female breast cancer and rs1314913 with the risk of male breast cancer. The aim of this study was to investigate the role of RAD51B variants in breast cancer predisposition, particularly in the context of familial breast cancer in Finland. We sequenced the coding region of RAD51B in 168 Finnish breast cancer patients from the Helsinki region for identification of possible recurrent founder mutations. In addition, we studied the known rs999737, rs2588809, and rs1314913 SNPs and RAD51B haplotypes in 44,791 breast cancer cases and 43,583 controls from 40 studies participating in the Breast Cancer Association Consortium (BCAC) that were genotyped on a custom chip (iCOGS). We identified one putatively pathogenic missense mutation c.541C>T among the Finnish cancer patients and subsequently genotyped the mutation in additional breast cancer cases (n = 5259) and population controls (n = 3586) from Finland and Belarus. No significant association with breast cancer risk was seen in the meta-analysis of the Finnish datasets or in the large BCAC dataset. The association with previously identified risk variants rs999737, rs2588809, and rs1314913 was replicated among all breast cancer cases and also among familial cases in the BCAC dataset. The most significant association was observed for the haplotype carrying the risk-alleles of all the three SNPs both among all cases (odds ratio (OR): 1.15, 95% confidence interval (CI): 1.11-1.19, P = 8.88 x 10-16) and among familial cases (OR: 1.24, 95% CI: 1.16-1.32, P = 6.19 x 10-11), compared to the haplotype with the respective protective alleles. Our results suggest that loss-of-function mutations in RAD51B are rare, but common variation at the RAD51B region is significantly associated with familial breast cancer risk.


Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors.
We amplified various amounts of DNA derived from frozen SF210 and U251NCI human glioblastoma cells, carried out comparative genomic hybridization (CGH) using degenerate oligonucleotide primed-PCR (DOP-PCR) products as test probes, and compared results to analyses performed with probes prepared by standard nick translation. Next we extracted DNA from hematoxylin-eosin (HE)- and methyl green (MG)-stained, microdissected sections of formalin-fixed and paraffin-embedded U251NCI cells, amplified and labeled it by DOP-PCR, and subjected it to CGH. Finally, we used the same methods in multiple samples from a single human mixed glioma tissue. DOP-PCR products from 50 pg to 250 ng of DNA were equally effective in generating the same CGH profiles as the standard method. DOP-PCR products from microdissected pieces of MG-stained cells were effective probes for CGH, but HE-stained samples were not desirable. As the proportion of HE-stained sample increased, CGH profiles deteriorated. DOP-PCR products from microdissected pieces of MG-stained paraffin sections of glioma tissue produced CGH profiles compatible with their histological features. CGH performed with DOP-PCR products from microdissected paraffin blocks allows for the accurate investigation of the cytogenetic characteristics from invasive tumors and of cytogenetic heterogeneity within neoplastic tissue.


An injectable and physical levan-based hydrogel as a dermal filler for soft tissue augmentation.
The use of injectable materials as a biofiller for soft tissue augmentation has been increasing worldwide. Levan is a biocompatible and inexpensive polysaccharide with great potential in biomaterial applications, but it has not been extensively studied. In this study, we evaluated the potential of levan as a new material for dermal fillers and prepared an injectable and physical levan-based hydrogel by combining levan with Pluronic and carboxymethyl cellulose (CMC). A sol state was prepared by mixing the polymers in a specific ratio at 4 °C for 2 days and the hydrogel was formed by increasing the temperature to 37 °C. The elastic modulus of the levan hydrogel was higher than that of a hyaluronic acid (HA)-based hydrogel. The SEM images of the levan hydrogel showed an interconnected porous structure, similar to the HA hydrogel. Levan showed non-cytotoxicity, enhanced cell proliferation, and higher amount of collagen synthesis in human dermal fibroblast cells compared to HA. The injected levan hydrogel was biocompatible and stable over 2 weeks in vivo, longer than the Pluronic F127 hydrogel or HA hydrogel. Also, the levan hydrogel showed a higher amount of collagen production than the HA hydrogel in vivo. More importantly, the levan hydrogel showed enhanced anti-wrinkle efficacy compared to the HA hydrogel in a wrinkle model mouse. Thus, the levan hydrogel with injectability, biocompatibility, and an anti-wrinkle effect has high potential as an alternative to existing commercial dermal fillers.


The Impact of Low-Grade Germinal Matrix-Intraventricular Hemorrhage on Neurodevelopmental Outcome of Very Preterm Infants.
Very preterm infants often show germinal matrix-intraventricular hemorrhage (GMH-IVH) on cranial ultrasound (cUS). To determine the impact of low-grade GMH-IVH on early neurodevelopmental outcome in very preterm infants. A retrospective case-control study in very preterm infants with and without low-grade GMH-IVH on cUS. Additional magnetic resonance imaging (MRI) was available in all infants with a gestational age (GA) <28 weeks and high-risk infants >28 weeks. Infants were seen at 2 years' corrected age to assess neurodevelopment. In total, 136 infants (GA 24-32 weeks) with low-grade GMH-IVH on cUS were matched with 255 controls. Outcome data was available for 342 (87%) infants. Adverse outcome (i.e., cerebral palsy [CP], neurodevelopmental delay) was present in 11 (9%) cases and 20 (9%) controls. No statistically significant differences in outcome were found between cases and controls. Additional MRI was performed in 165/391 infants (42%) and showed additional lesions in 73 (44%) infants that could explain subsequent development of CP in 2 out of 5 infants and epilepsy in 1 of 2 infants. Very preterm infants with low-grade GMH-IVH on cUS have a similar early neurodevelopmental outcome compared with controls. Additional MRI showed mostly subtle abnormalities that were missed with cUS, but these could not explain subsequent development of CP and developmental delay in all infants.


Intracranial stents being modeled as a porous medium: flow simulation in stented cerebral aneurysms.
Intracranial aneurysms may be treated by flow diverters, alternatively to stents and coils combination. Numerical simulation allows the assessment of the complex nature of aneurismal flow. Endovascular devices present a rather dense and fine strut network, increasing the complexity of the meshing. We propose an alternative strategy, which is based on the modeling of the device as a porous medium. Two patient-specific aneurysm data sets were reconstructed using conventional clinical setups. The aneurysms selection was done so that intra-aneurismal flow was shear driven in one and inertia driven in the other. Stents and their porous medium analog were positioned at the aneurysm neck. Physiological flow and standard boundary conditions were applied. The comparison between both approaches was done by analyzing the velocity, vorticity, and shear rate magnitudes inside the aneurysm as well as the wall shear stress (WSS) at the aneurysm surface. Simulations without device were also computed. The average flow reduction reaches 76 and 41% for the shear and inertia driven flow models, respectively. When comparing the two approaches, results show a remarkable similarity in the flow patterns and magnitude. WSS, iso-velocity surfaces and velocity on a trans-sectional plane are in fairly good agreement. The root mean squared error on the investigated parameters reaches 20% for aneurysm velocity, 30.6% for aneurysm shear rate, and 47.4% for aneurysm vorticity. It reaches 20.6% for WSS computed on the aneurysm surface. The advantages of this approach reside in its facility to implement and in the gain in computational time. Results predicted by the porous medium approach compare well with the real stent geometry model and allow predicting the main effects of the device on intra-aneurismal flow, facilitating thus the analysis.


Relationship between perceived exertion and physiologic markers during arm exercise with able-bodied participants and participants with poliomyelitis.
To investigate the strength of the relationship between ratings of perceived exertion (RPE) and oxygen uptake (Vo(2)), heart rate, ventilation (Ve) and power output (PO) during an arm-crank ramped exercise test to volitional exhaustion in men and women who differed in physical status. Each participant completed an arm-crank ramp exercise test to volitional exhaustion. PO was increased by 15 W.min(-1) and 6 W.min(-1) for men and women able-bodied participants, respectively; for the poliomyelitis participants, 9 W.min(-1) and 6 W.min(-1) increments were used for men and women, respectively. Laboratory facilities at a university. Able-bodied participants (n=16; 9 men, 7 women) and participants with poliomyelitis (n=15, 8 men, 7 women) volunteered for the study. Strength of the relationship (R(2) values) between RPE and Vo(2), heart rate, Ve and PO. There were significantly higher values for maximum Vo(2) and maximum PO for able-bodied men compared with their counterparts with poliomyelitis (P<.05). However, when the data were controlled for age, there were no significant differences in these values (P>.05). Similar results were observed for the women who were able-bodied as well as for the women who had poliomyelitis (P>.05). The relationships between heart rate and RPE and Ve and RPE for able-bodied patients and patients with poliomyelitis were similar (R(2)>.87). The relationship between Vo(2) and RPE was stronger in the able-bodied participants compared wih the participants with poliomyelitis, regardless of sex (P<.05). However, when the data were controlled for age, there was no significant difference in the strength of this relationship between able-bodied participants and those with poliomyelitis, regardless of sex (P>.05). RPE is strongly related to physiologic markers of exercise intensity during arm exercise, irrespective of sex or participant's poliomyelitis status.


Regeneration of optic fibres through the chiasma in Xenopus laevis tadpoles.
The path through the chiasma followed by regenerating optic nerve fibres in Xenopus tadpoles was studied at light- and electron-microscopic levels, and with horseradish peroxidase as a fibre label. Over the period (5-10 days) in which regenerating fibres reach and cross the chiasma, they did not follow residual deep fibres through the chiasma, nor were they associated with the trail of degeneration in the chiasma which represented the remains of the deeper (older) parts of the original projection. The regenerating optic fibres were always seen in close association with the inner surfaces of the ependymoglial endfeet, or in the extracellular spaces that lie close to the endfeet in the most superficial part of the chiasma, where newly-growing fibres from the retinal margin are normally to be found.


Synchronization in time-varying networks.
We study the stability of the synchronized state in time-varying complex networks using the concept of basin stability, which is a nonlocal and nonlinear measure of stability that can be easily applied to high-dimensional systems [P. J. Menck, J. Heitzig, N. Marwan, and J. Kurths, Nature Phys. 9, 89 (2013)]. The time-varying character is included by stochastically rewiring each link with the average frequency f. We find that the time taken to reach synchronization is lowered and the stability range of the synchronized state increases considerably in dynamic networks. Further we uncover that small-world networks are much more sensitive to link changes than random ones, with the time-varying character of the network having a significant effect at much lower rewiring frequencies. At very high rewiring frequencies, random networks perform better than small-world networks and the synchronized state is stable over a much wider window of coupling strengths. Lastly we show that the stability range of the synchronized state may be quite different for small and large perturbations, and so the linear stability analysis and the basin stability criterion provide complementary indicators of stability.


Study of the developmental pattern of heme catabolism in liver and the effects of cobalt on cytochrome P-450 and the rate of heme oxidation during the neonatal period.
The comparative development patterns of heme oxidation andof cytochrome P-450 dependent drug oxidation in rat liver were examined. High levels of heme oxygenase activity were present in whole embryo preparations at day 13 of gestation. At birth this enzyme activity in liver was approximately equal to that of normal adult liver. In the immediate postnatal period the rate of hepatic heme oxidation increased sharply, reaching levels 3-5 times normal during the first week postpartum. Thereafter, this enzyme activity progressively decreased and returned to normal adult levels by the 28th postpartum day. The development of microsomal heme oxidation and of P-450 dependent drug oxidation exhibited reciprocal patterns, with the latter being at low levels of activity during the immediate postnatal period and reaching adult activity only 4 or more wk after birth. Cobalt injected into pregnant animals or in to nursing mothers did not induce heme oxygenase in the fetus or suckling neonate. However, when treated directly with the metal, 4-day old neonates exhibited a small induction response of this enzyme; and the inducibility of heme oxygenase increased gradually to fully adult levels by the end of the 4th postpartum week. Cobalt at all postnatal developmental stages was capable of diminishing hepatic contents of total microsomal heme and P-450; however this effect of the metal was small in the immediate period after birth and increased progressively with maturation. These findings demonstrate that the patterns of development of hepatic capacity for carrying out the oxidation of heme and the P-450 dependent oxidation of drugs are different and thus provide further evidence that these microsomal enzyme systems are distinct from each other and under separate regulatory mechanisms. The degree of induction response for hepatic heme oxygenase evoked by the trace metal, cobalt, was also shown to have developmental determinants as did the susceptibility of hepatic cytochrome P-450 to degradation by this metal. The very high levels of hepatic heme oxygenase activity which characterize neonates during the first week of life indicate that over-production of bilirubin contributes significantly to the mechanism of neonatal jaundice.


SU-E-T-439: First Experience of Three Dimensional Conformai Radiotherapy (3DCRT) Planning with Helical Tomotherapy.
A three-dimensional conformal radiotherapy (3DCRT) has been recently introduced to helical tomotherapy, allowing the user to plan and treat patients that do not require sophisticated IMRT planning and delivery. This study aims to test treatment planning on this modality and evaluate its performance by comparing to conventional LINAC-based 3DCRT planning. Four clinical cases (whole brain, extremity, lung, and partial breast irradiation) were retrospectively selected from a Pinnacle planning system (Philips Medical System, Fitchburg, WI) and planned on Tomotherapy (Accuray Inc., Sunnyvale, CA). Computed tomography (CT) images together with contours of target and critical structures were exported from Pinnacle to the Tomotherapy planning station. The same prescription and fractionation scheme was adopted. The pitch factor for all clinical cases was set to 0.287. A 2.5 cm jaw was employed except in the lung case the field size was set to 1.0 cm for better dose conformity. The dose grid size was chosen to be half of that of the planning CT images. On Pinnacle 100% prescription dose was delivered to the treatment isocenter while onTomotherapy it was stipulated that at least 95% of the target volume received the prescribed dose. Comparison between two planning strategies was performed, in terms of dose volume histograms (DVH), dosimetric and radiobiological parameters, for plan quality assessment. Comparison of DVHs reveals that up to 25% healthy tissue sparing in volume can be accomplished with Tomotherapy 3DCRT while the same target coverage is ensured. Dosimetric and radiobiological indices between Tomotherapy and Pinnacle planning agree to within 3.0%. Additional beam modifiers and non-coplanar beams associated with LINAC-based 3DCRT are not needed on Tomotherapy, making it more favorable. Tomotherapy 3DCRT has similar dosimetric performance when compared to conventional LINAC-based 3DCRT while it is substantially easier to use.


Drug- or alcohol-facilitated, incapacitated, and forcible rape in relationship to mental health among a national sample of women.
Rape is a well-established risk factor for mental health disorders such as posttraumatic stress disorder (PTSD) and depression. However, most studies have focused on forcible rape tactics and have not distinguished these from tactics that involve drug or alcohol intoxication. The authors' aim was to examine correlates of PTSD and depression in a community sample of women, with particular emphasis on evaluating the unique effects of lifetime exposure to three specific rape tactics. A nationally representative sample of 3,001 noninstitutionalized, civilian, English- or Spanish-speaking women (aged 18-86 years) participated in a structured telephone interview by use of Computer-Assisted Telephone Interviewing (CATI) technology. Multivariable models showed that history of drug or alcohol-facilitated rape tactics (OR = 1.87, p < .05) and history of forcible rape tactics (OR = 3.46, p < .001) were associated with PTSD. History of forcible rape was associated with depression (OR = 3.65, p < .001). Forcible rape tactics were associated with a number of factors that may have contributed to their stronger association with mental health outcomes, including force, injury, lower income, revictimization history, and labeling the event as rape. The results underscore the importance of using a behaviorally specific assessment of rape history, as rape tactic and multiple rape history differentially predicted psychopathology outcomes. The association between drug- or alcohol-facilitated rape tactics and PTSD suggests that these are important rape tactics to include in assessments and future studies.


The molecular basis of human retinal and vitreoretinal diseases.
During the last two to three decades, a large body of work has revealed the molecular basis of many human disorders, including retinal and vitreoretinal degenerations and dysfunctions. Although belonging to the group of orphan diseases, they affect probably more than two million people worldwide. Most excitingly, treatment of a particular form of congenital retinal degeneration is now possible. A major advantage for treatment is the unique structure and accessibility of the eye and its different components, including the vitreous and retina. Knowledge of the many different eye diseases affecting retinal structure and function (night and colour blindness, retinitis pigmentosa, cone and cone rod dystrophies, photoreceptor dysfunctions, as well as vitreoretinal traits) is critical for future therapeutic development. We have attempted to present a comprehensive picture of these disorders, including biological, clinical, genetic and molecular information. The structural organization of the review leads the reader through non-syndromic and syndromic forms of (i) rod dominated diseases, (ii) cone dominated diseases, (iii) generalized retinal degenerations and (iv) vitreoretinal disorders, caused by mutations in more than 165 genes. Clinical variability and genetic heterogeneity have an important impact on genetic testing and counselling of affected families. As phenotypes do not always correlate with the respective genotypes, it is of utmost importance that clinicians, geneticists, counsellors, diagnostic laboratories and basic researchers understand the relationships between phenotypic manifestations and specific genes, as well as mutations and pathophysiologic mechanisms. We discuss future perspectives.


Multi-material digital light processing bioprinting of hydrogel-based microfluidic chips.
Recent advancements in digital-light-processing (DLP)-based bioprinting and hydrogel engineering have enabled novel developments in organs-on-chips. In this work, we designed and developed a multi-material, DLP-based bioprinter for rapid, one-step prototyping of hydrogel-based microfluidic chips. A composite hydrogel bioink based on poly-ethylene-glycol-diacrylate (PEGDA) and gelatin methacryloyl (GelMA) was optimized through varying the bioprinting parameters such as light exposure time, bioink composition, and layer thickness. We showed a wide range of mechanical properties of the microfluidic chips for various ratios of PEGDA:GelMA. Microfluidic features of hydrogel-based chips were then tested using dynamic flow experiments. Human-derived tumor cells were encapsulated in 3D bioprinted structures to demonstrate their bioactivity and cell-friendly environment. Cell seeding experiments then validated the efficacy of the selected bioinks for vascularized micro-tissues. Our biofabrication approach offers a useful tool for the rapid integration of micro-tissue models into organs-on-chips and high-throughput drug screening platforms.


The relative importance of nitric oxide and nitric oxide-independent mechanisms in acetylcholine-evoked dilatation of the rat mesenteric bed.
1. The relative contribution of nitric oxide (NO) to acetylcholine-induced smooth muscle relaxation was investigated in the rat perfused mesenteric vasculature and in isolated segments of second, third and fourth order arterial branches. 2. The EC50 values and maximal relaxation to acetylcholine were not significantly different in the sequential arterial branches, being approximately 0.05 microM and 85%, respectively. 3. The NO synthase inhibitor L-NG-nitro-L-arginine methyl ester (L-NAME; 100 microM) reduced acetylcholine-evoked endothelium-dependent dilatation and relaxation in the perfused mesenteric bed and in isolated arterial segments. The maximum response to acetylcholine in both preparations was reduced by between 35% to 40% while the EC50 values were increased by 5-6 fold. L-NAME had no effect on basal smooth muscle tone in either case. 4. In contrast, endothelium-dependent dilatation of the perfused mesenteric bed evoked by A23187 (0.002-20 nmol), was unaffected by exposure to L-NAME. The EC50 values and maximal responses elicited by A23187 (20 nmol) before and after exposure to L-NAME were 0.96 +/- 0.5 nmol and 67.0 +/- 7.0% (n = 4), and 0.7 +/- 0.4 nmol and 70.0 +/- 5.0% (n = 4; P > 0.01), respectively. 5. Perfusion of the isolated mesenteric bed with raised K(+)-Krebs buffer (25 mM) had no effect on basal tone, but reduced the amplitude of both acetylcholine- and A23187-evoked dilatation. The maximum responses to acetylcholine (2 micromol) and A23187 (20 nmol) were reduced from 67.5 +/- 7.3% and 65.4+/-8.2% to 18.9 +/-11.0% (n=5; P<0.01) and 13.5 +/-12.0% (n=4; P<0.01), respectively.6. Exposure of the mesenteric bed to L-NAME in the presence of raised K+-Krebs further reduced the maximal response elicited by acetylcholine to only 8.9 +/- 2.8% (n =4; P< 0.01).7. These results indicate that acetylcholine-evoked vasodilatation of the rat mesenteric vasculature is mediated by both NO-dependent and -independent mechanisms. The relative contribution made by these mechanisms does not appear to differ in sequential branches of the mesenteric artery. In contrast,A23187-evoked vasodilatation appears to be mediated predominantly by a NO-independent mechanism which is sensitive to increases in the extracellular potassium concentration and may reflect the action of endothelium-derived hyperpolarizing factor (EDHF).