A two year longitudinal study of mood disorders following stroke. Prevalence and duration at six months follow-up.
We are conducting a two year longitudinal study of mood disorders in a group of 103 stroke patients. During the first six month period following the stroke the prevalence of symptoms of major depression increased from 23 per cent to 34 per cent while the frequency of symptoms of dysthymic depression increased from 20 per cent to 26 per cent. In addition, 10 of 13 patients who had major depressive symptoms during the initial evaluation continued to have these symptoms at six months follow-up and 4 of 9 patients who had minor depression in hospital developed major depression, while 5 of 9 continued to have minor depressive symptoms. Thus, the duration of depression following stroke is more than six months and the prevalence of major depressive symptoms increases steadily for the first half year after a cerebral hemorrhage or ischemic lesion.


Hyperglycaemia and apoptosis of microglial cells in human septic shock.
The effect of hyperglycaemia on the brain cells of septic shock patients is unknown. The objective of this study was to evaluate the relationship between hyperglycaemia and apoptosis in the brains of septic shock patients. In a prospective study of 17 patients who died from septic shock, hippocampal tissue was assessed for neuronal ischaemia, neuronal and microglial apoptosis, neuronal Glucose Transporter (GLUT) 4, endothelial inducible Nitric Oxide Synthase (iNOS), microglial GLUT5 expression, microglial and astrocyte activation. Blood glucose (BG) was recorded five times a day from ICU admission to death. Hyperglycaemia was defined as a BG 200 mg/dL g/l and the area under the BG curve (AUBGC) > 2 g/l was assessed. Median BG over ICU stay was 2.2 g/l. Neuronal apoptosis was correlated with endothelial iNOS expression (rho = 0.68, P = 0.04), while microglial apoptosis was associated with AUBGC > 2 g/l (rho = 0.70; P = 0.002). Neuronal and microglial apoptosis correlated with each other (rho = 0.69, P = 0.006), but neither correlated with the duration of septic shock, nor with GLUT4 and 5 expression. Neuronal apoptosis and ischaemia tended to correlate with duration of hypotension. In patients with septic shock, neuronal apoptosis is rather associated with iNOS expression and microglial apoptosis with hyperglycaemia, possibly because GLUT5 is not downregulated. These data provide a mechanistic basis for understanding the neuroprotective effects of glycemic control.


Electronic Properties of Chlorine, Methyl, and Chloromethyl as Substituents to the Ethylene Group--Viewed from the Core of Carbon.
"Substituent effects" is an important and useful concept in organic chemistry. Although there are many approaches to parametrizing the electronic and steric effects of substituents, the physical basis for the parameters is often unclear. The purpose of the present work is to explore the properties of chemical shifts in carbon 1s energies as a well-defined basis for characterizing substituents to an ethylene C═C moiety. To this end, high-resolution carbon 1s photoelectron spectra of six chloro-substituted ethenes and seven chloro-substituted propenes have been measured in the gas phase. Site-specific adiabatic ionization energies have been determined from the spectra using theoretical ab initio calculations to predict the vibrational structures. For two molecules, 3-chloropropene and 2,3-dichloropropene, the spectral analyses give quantitative results for the conformer populations. The observed shifts have been analyzed in terms of initial-state (potential) and relaxation effects, and charge relaxation has also been analyzed by means of natural resonance theory. On the basis of core-level spectroscopy and models, chlorine, methyl, and chloromethyl have been characterized in terms of their effect on the carbon to which they are attached (α site) as well as the neighboring sp(2) carbon (β site). The derived spectroscopic substituent parameters are characterized by both inductive (electronegativity) effects and the ability of each substituent to engage in electron delocalization via the π system. Moreover, the adopted approach is extended to include substituent-substituent interaction parameters.


Increased Grik4 Gene Dosage Causes Imbalanced Circuit Output and Human Disease-Related Behaviors.
Altered glutamatergic neurotransmission is thought to contribute to mental disorders and neurodegenerative diseases. Copy-number variation in genes associated with glutamatergic synapses represents a source of genetic variability, possibly underlying neurological and mental disease susceptibility. The GRIK4 gene encodes a high-affinity kainate receptor subunit of essentially unknown function, although de novo duplication of the 11q23.3-q24.1 locus to which it maps has been detected in autism and other disorders. To determine how changes in the dose of Grik4 affect synaptic activity, we studied mice overexpressing this gene in the forebrain. A mild gain in Grik4 enhances synaptic transmission, causing a persistent imbalance in inhibitory and excitatory activity and disturbing the circuits responsible for the main amygdala outputs. These changes in glutamatergic activity reverse when Grik4 levels are normalized; thus, they may account for the behavioral abnormalities in disorders like autism or schizophrenia.


Nut consumption and type 2 diabetes risk: a systematic review and meta-analysis of observational studies.
Previous meta-analyses, with some methodological controversies, have assessed the relation between nut consumption and type 2 diabetes (T2D) risk and pointed to contradictory results, making desirable the performance of an updated meta-analysis. We aimed to systematically review and meta-analyze all the published studies investigating the relations of total nuts and different types of nuts-i.e., walnuts, peanuts, peanut butter, and total tree nuts-with the prevalence and incidence of T2D. A systematic search was conducted in the PubMed and Cochrane databases through 12 August, 2020. The inverse variance method with fixed-effect models was used to pool data across studies, expressed as risk ratios (RRs) or ORs and 95% CIs for prospective cohort and cross-sectional studies, respectively. The Cochran Q test and I2 statistics were used to test and quantify heterogeneity, respectively. Dose-response meta-analysis was also conducted. Eight studies (5 prospective and 3 cross-sectional) were included in the quantitative synthesis. Meta-analyses of cross-sectional studies and prospective cohort studies, comparing the highest with the lowest categories, revealed a nonsignificant association between total nut consumption and T2D. Meta-analyses of prospective cohort studies showed an inverse association between peanut butter consumption and T2D incidence (RR: 0.87; 95% CI: 0.77, 0.98; I2 = 50.6%; Pheterogeneity = 0.16), whereas no association was observed between peanuts or tree nuts and T2D. There was no evidence of a linear dose-response or nonlinear dose-response gradient for total nut and peanut consumption in prospective cohort studies. The certainty of the evidence using NutriGrade was very low for all the exposures. Current results do not demonstrate an association of total nut, peanut, or tree nut consumption with T2D. Peanut butter consumption may be inversely associated with this disease.This review protocol was registered at www.crd.york.ac.uk/prospero/ as CRD42020149756.


Osteitis of the axial border of the proximal sesamoid bones in horses: eight cases (1993-1999).
To determine clinical, radiographic, and scintigraphic abnormalities in and outcome of horses with septic or nonseptic osteitis of the axial border of the proximal sesamoid bones. Retrospective study. 8 horses. Data collected from medical records included signalment; history; horse use; severity and duration of lameness; results of perineural anesthesia, radiography, ultrasonography, and scintigraphy; and outcome following surgery. Five horses did not have any evidence of sepsis; the other 3 had sepsis of the metacarpophalangeal or metatarsophalangeal joint or the digital synovial sheath. All horses had a history of chronic unilateral lameness. Three of 5 horses improved after diagnostic anesthesia of the metacarpophalangeal or metatarsophalangeal joint; the other 2 improved only after diagnostic anesthesia of the digital synovial sheath. Nuclear scintigraphy was beneficial in localizing the source of the lameness to the proximal sesamoid bones in 4 horses. Arthroscopy of the palmar or plantar pouch of the joint or of the digital synovial sheath revealed intersesamoidean ligament damage and osteomalacia of the axial border of the proximal sesamoid bones in all horses. All 5 horses without sepsis and 1 horse with sepsis returned to their previous uses. Results suggest that osteitis of the axial border of the proximal sesamoid bones is a distinct entity in horses that typically is associated with inflammation of the associated metacarpointersesamoidean or metatarsointersesamoidean ligament and may be a result of sepsis or nonseptic inflammation. Arthroscopic debridement may allow horses without evidence of sepsis to return to their previous level of performance.


Liquid-crystalline calcium carbonate: biomimetic synthesis and alignment of nanorod calcite.
Liquid-crystalline CaCO3 has been prepared for the first time. The nanorods of CaCO3 calcite are obtained by bio-inspired crystallization through aqueous colloidal precursors of amorphous CaCO3 stabilized by poly(acrylic acid). The synthesized calcite nanocrystals have well-tuned morphologies that are preferable for formation of liquid-crystalline phases in concentrated aqueous colloidal solution. The one-dimensional alignment of calcite crystals is achieved by mechanical shearing of the aqueous colloidal solution showing liquid-crystalline phases. These CaCO3-based liquid crystals formed by a self-organization process in mild conditions may have great potential for use as environmentally friendly materials.


Inverse mapping the neuronal substrates of face categorizations.
Face perception is a complex process involving a network of brain structures, dynamically processing information to enable judgments about a face to be made (e.g., familiarity, identity, and expression). Here we introduce an analysis methodology that makes it possible to directly study this information processing in the brain from spatially and temporally resolved magnetoencephalographic signals. We apply our methodology to the study of 2 face categorization tasks, gender and expressiveness, and track the processing of 3 key visual features that underlie behavioral performance, over time and throughout the cortex. We find information processing correlates beginning from 90 ms following stimulus onset, where features are processed in isolation in occipital extrastriate regions. Over time, processing of successively more features and feature combinations takes place in occipitotemporal regions, with maximal information processing of visual information coinciding with the well-established face-selective M170 component at 170 ms. Later still, around 250-400 ms, cortical activity responds significantly more to task-specific features and their complex combinations. These results indicate a complex process of visual information processing during face perception with face parts processed in isolation at very early stages, and task-specific processing of combinations of features taking place within 300 ms. Crucially, our approach specifically establishes which information in the visual stimulus the brain signal is responding to and how this varies with time, cortical location, and task demands to establish a more precise tracking of information processing mechanisms in the cortex during face perception.


Comparative genomic analysis of the zebra finch degradome provides new insights into evolution of proteases in birds and mammals.
The degradome -the complete repertoire of proteases in an organism- is involved in multiple key biological and pathological processes. Previous studies in several organisms have yielded sets of curated protease sequences which may be used to characterize the degradome in a novel genome by similarity. Differences between degradomes can then be related to physiological traits of the species under study. Therefore, the sequencing of the zebra finch genome allows the comparison between the degradomes of mammals and birds and may help to understand the biological peculiarities of the zebra finch. A set of curated protease sequences from humans and chicken was used to predict the sequences of 460 protease and protease-like genes in the zebra finch genome. This analysis revealed important differences in the evolution of mammalian and bird degradomes, including genomic expansions and deletions of caspases, cytotoxic proteases, kallikreins, matrix metalloproteases, and trypsin-like proteases. Furthermore, we found several zebra finch-specific features, such as duplications in CASP3 and BACE, and a large genomic expansion of acrosin. We have compared the degradomes of zebra finch, chicken and several mammalian species, with the finding of multiple differences which illustrate the evolution of the protease complement of these organisms. Detailed analysis of these changes in zebra finch proteases has shown that they are mainly related to immunological, developmental, reproductive and neural functions.


A STUDY OF EPIDERMAL FIBRILS.
In certain animals, in addition to the protoplasmic fibrils of the stratum filamentosum that are present normally in human epidermis, there are well developed fibrils in the cells of the stratum germinativum. These fibrils are present in the human epidermis only under conditions of increased cell activity. These fibrils, that are prominent in the stratum germinativum of human epidermis under conditions of increased cell activity, seem to increase in direct ratio to rapidity of cell production. The differentiation of these fibrils of the stratum germinativum into the finer fibrils which connect the cells of the stratum filamentosum can be observed under proper conditions of growth and therefore there should be no separation of the fibrils into different groups with various names. This process of fibre production by the cells of the human epidermis is analogous to increased fibre production by various other cells under similar conditions, and may be of importance in identifying or classifying new growths of epithelial origin.


Prevalence of low back pain in Latin America: a systematic literature review.
Chronic low back pain is considered as a high-impact condition that affects the working population of Latin America, with long reaching social and economic repercussions. Its true frequency is unknown due to the absence of well-designed clinical trials that use standardized definitions and criteria. To evaluate the prevalence of chronic non-specific low back pain among the Latin American population. A systematic review of chronic non-specific low back pain in Latin America. Meeting of Change Pain Latin America, Mexico. Data sources included relevant literature identified through searches of published studies between August 30, 2002, and August 30, 2012, in 7 electronic databases: Cochrane BVS, Pubmed, Medline, Lilacs, Scielo, Hinari, and MedCarib. Publications dealing with low back pain of a post-traumatic, infectious, or malignant origin were excluded. Two reviewers selected in an independent manner all eligible studies using the MOOSE checklist and extracted data on both prevalence and risk factors associated with low back pain. A narrative synthesis of the results was drafted, which was later validated by a panel of clinical experts on pain. Twenty-eight studies were included in the review, comprising a total of 20,559 subjects from 7 countries in the region. Four of these studies, with significant methodological differences between them, measured the frequency of chronic low back pain with results that varied from 4.2% to 10.1%. Four studies are part of the Community Oriented Program for Control of Rheumatic Diseases (COPCORD) program reports, and were pooled and analyzed separately because of their particular design. Their prevalence estimations varied between 1.8% and 11.3%. The remaining 20 studies evaluated a total population of 6,992 subjects, and found a prevalence of low back pain of 31.3%. Based on an epidemiological model constructed on both times to resolution and low back pain recurrence rates, the prevalence of chronic low back pain in Latin America was estimated to be around 10.5%. Some risk factors reported by the authors are long working hours with the worker in the sitting position, obesity and overweight, pregnancy, smoking, advanced age, lifting and carrying heavy loads, domestic work, sedentary lifestyles, and duration of current employment. A subgroup analysis of the population under study yielded an estimated prevalence of low back pain of 16.7% for the population exposed to a lower number of risk factors and 65% for the higher risk subgroup. In this review, we made an exhaustive search of studies evaluating the epidemiology of chronic low back pain in the Latin America region. The large topographic and chronologic variability in definitions of low back pain, interviewer bias, and subject selection bias. Despite the sparse information and the methodological heterogeneity of the studies, pooled results allowed for an indirect estimation of the prevalence of low back pain in the region that was pretty consistent with the published results obtained from other settings. New studies need to be carried out to supplement and overcome the methodological weaknesses of those previously conducted.


Venezuelan equine encephalitis and Oropouche virus infections among Peruvian army troops in the Amazon region of Peru.
An outbreak of a febrile illness characterized by headache, ocular pain, myalgia, and arthralgia occurred during June 1994 among Peruvian army troops in Northern Peru. On June 14-16, 1994, clinical data and blood samples were obtained from eight soldiers with a febrile illness, and from 26 others who had a history of febrile illness during the past three months. A follow-up blood sample was obtained 107 days later from four of the febrile and seven of the afebrile soldiers. Serum samples were tested for dengue (DEN), Oropouche (ORO), and Venezuelan equine encephalitis (VEE) IgM and IgG antibodies by an enzyme-linked immunosorbent assay (ELISA). Virus isolation was performed by inoculation of newborn mice and Vero cell cultures. Viral isolates were identified by immunofluorescence, ELISA, and nucleotide sequencing. A VEE virus infection was confirmed in three of the eight febrile soldiers, two by virus isolation, and one by serology. Antigenic analysis indicated that one of the virus isolates was similar to VEE subtype I, variety ID, viruses previously isolated in Colombia and Venezuela. Nucleotide sequence data showed that both viral isolates were identical to one another and closely related to VEE ID viruses previously isolated in Peru, Colombia, and Venezuela. Serologic results showed that two of 26 afebrile soldiers had IgM antibody to VEE and four had IgG antibody to VEE; two febrile soldiers had IgG antibody in their first serum samples. Oropouche-specific IgM antibody was detected in one of the eight febrile and five of the afebrile soldiers, and 18 of the 34 soldiers had low titers of ORO IgG antibody titers, which did not meet the diagnostic criteria for confirmed cases. All soldiers were negative for DEN IgM antibody, and 10 had flavivirus IgG antibody that reacted with DEN antigens. These data indicated that VEE ID virus was one of the causes of illness among Peruvians soldiers and that this was the first association of this VEE subtype with human disease in Peru.


Mixed methods study of clinicians' perspectives on barriers to implementation of treat to target in psoriatic arthritis.
In treat to target (T2T), the patient is treated to reach and maintain specified and sequentially measured goals, such as remission or low disease activity. T2T in psoriatic arthritis (PsA) has demonstrated improved clinical and patient-reported outcomes and is recommended in European guidelines. However, most clinicians do not use T2T in PsA. This study examined the barriers and enablers to implementation in practice. Sequential mixed methods comprising a qualitative design (interviews and focus group) to inform a quantitative design (survey). Qualitative data were analysed thematically, and quantitative statistics were analysed descriptively. Nineteen rheumatology clinicians participated in telephone interviews or a face-to-face focus group. An overarching theme 'Complexity' (including 'PsA vs Rheumatoid Arthritis', 'Measurement' and 'Resources') and an underpinning theme 'Changes to current practice' (including 'Reluctance due to organisational factors' and 'Individual determination to make changes') were identified. 153 rheumatology clinicians responded to an online survey. Barriers included limited clinical appointment time to collect outcome data (54.5%) and lack of training in assessing skin disease (35%). Enablers included provision of a protocol (86.4%), a local implementation lead (80.9%), support in clinic to measure outcomes (83.3%) and training in T2T (69.8%). The importance of regular audit with feedback, specialist PsA clinics and a web-based electronic database linked to hospital/national information technology (IT) systems were also identified as enablers. Implementation of T2T in PsA requires an integrated approach to address the support, training and resource needs of individual clinicians, rheumatology teams, local IT systems and service providers to maximise success.


Self-reported and performance based perfectionism in mothers of individuals with Anorexia Nervosa: a pilot study.
Perfectionism is implicated in the development and maintenance of Anorexia Nervosa and there is some evidence that perfectionism may be elevated in family members. However to date there are no studies investigating behavioural aspects of perfectionism in unaffected mothers. Forty-one participants took part in this pilot study: 21 unaffected mothers of individuals with Anorexia Nervosa and 20 healthy control mothers. Participants completed two performance based tasks assessing perfectionism-a text replication task and a bead sorting task-along with self-report measures of perfectionism. No group differences were found between unaffected AN mothers and HC mothers on performance measures of perfectionism. The findings are discussed in relation to existing studies and clinical implications explored.


Electronic medical records and efficiency and productivity during office visits.
To estimate the relationship between electronic medical record (EMR) use and efficiency of utilization and provider productivity during visits to US office-based physicians. Cross-sectional analysis of the 2006-2007 National Ambulatory Medical Care Survey. The sample included 62,710 patient visits to 2625 physicians. EMR systems included demographics, clinical notes, prescription orders, and laboratory and imaging results. Efficiency was measured as utilization of examinations, laboratory tests, radiology procedures, health education, nonmedication treatments, and medications. Productivity was measured as total services provided per 20-minute period. Survey-weighted regressions estimated association of EMR use with services provided, visit intensity/duration, and productivity. Marginal effects were estimated by averaging across all visits and by major reason for visit. EMR use was associated with higher probability of any examination (7.7%, 95% confidence interval [CI] = 2.4%, 13.1%); any laboratory test (5.7%, 95% CI = 2.6%, 8.8%); any health education (4.9%, 95% CI = 0.2%, 9.6%); and fewer laboratory tests (-7.1%, 95% CI = -14.2%, -0.1%). During pre/post surgery visits, EMR use was associated with 7.3% (95% CI= -12.9%, -1.8%) fewer radiology procedures. EMR use was not associated with utilization of nonmedication treatments and medications, or visit duration. During routine visits for a chronic problem, EMR use was associated with 11.2% (95% CI = 5.7%, 16.8%) more diagnostic/screening services provided per 20-minute period. EMR use had a mixed association with efficiency and productivity during office visits. EMRs may improve provider productivity, especially during visits for a new problem and routine chronic care.


Characterization of new-generation aminoglycoside promoting premature termination codon readthrough in cancer cells.
Nonsense mutations, generating premature termination codons (PTCs), account for 10% to 30% of the mutations in tumor suppressor genes. Nonsense translational suppression, induced by small molecules including gentamicin and G418, has been suggested as a potential therapy to counteract the deleterious effects of nonsense mutations in several genetic diseases and cancers. We describe here that NB124, a synthetic aminoglycoside derivative recently developed especially for PTC suppression, strongly induces apoptosis in human tumor cells by promoting high level of PTC readthrough. Using a reporter system, we showed that NB124 suppressed several of the PTCs encountered in tumor suppressor genes, such as the p53 and APC genes. We also showed that NB124 counteracted p53 mRNA degradation by nonsense-mediated decay (NMD). Both PTC suppression and mRNA stabilization contributed to the production of a full-length p53 protein capable of activating p53-dependent genes, thereby specifically promoting high levels of apoptosis. This new-generation aminoglycoside thus outperforms the only clinically available readthrough inducer (gentamicin). These results have important implications for the development of personalised treatments of PTC-dependent diseases and for the development of new drugs modifying translation fidelity.


Screening and hit evaluation of a chemical library against blood-stage Plasmodium falciparum.
In view of the need to continuously feed the pipeline with new anti-malarial agents adapted to differentiated and more stringent target product profiles (e.g., new modes of action, transmission-blocking activity or long-duration chemo-protection), a chemical library consisting of more than 250,000 compounds has been evaluated in a blood-stage Plasmodium falciparum growth inhibition assay and further assessed for chemical diversity and novelty. The selection cascade used for the triaging of hits from the chemical library started with a robust three-step in vitro assay followed by an in silico analysis of the resulting confirmed hits. Upon reaching the predefined requirements for selectivity and potency, the set of hits was subjected to computational analysis to assess chemical properties and diversity. Furthermore, known marketed anti-malarial drugs were co-clustered acting as 'signposts' in the chemical space defined by the hits. Then, in cerebro evaluation of the chemical structures was performed to identify scaffolds that currently are or have been the focus of anti-malarial medicinal chemistry programmes. Next, prioritization according to relaxed physicochemical parameters took place, along with the search for structural analogues. Ultimately, synthesis of novel chemotypes with desired properties was performed and the resulting compounds were subsequently retested in a P. falciparum growth inhibition assay. This screening campaign led to a 1.25% primary hit rate, which decreased to 0.77% upon confirmatory repeat screening. With the predefined potency (EC₅₀ < 1 μM) and selectivity (SI > 10) criteria, 178 compounds progressed to the next steps where chemical diversity, physicochemical properties and novelty assessment were taken into account. This resulted in the selection of 15 distinct chemical series. A selection cascade was applied to prioritize hits resulting from the screening of a medium-sized chemical library against blood-stage P. falciparum. Emphasis was placed on chemical novelty whereby computational clustering, data mining of known anti-malarial chemotypes and the application of relaxed physicochemical filters, were key to the process. This led to the selection of 15 chemical series from which ten confirmed their activity when newly synthesized sample were tested.


Ovarian cancer in the world: epidemiology and risk factors.
Aim: Ovarian cancer is one of the most common gynecologic cancers that has the highest mortality rate. Considering the fact that knowledge on the incidence, mortality of ovarian cancer, as well as its risk factors is necessary for planning and preventing complications, this study was conducted with the aim of examining the epidemiology and risk factors of ovarian cancer in the world. Materials and methods: In order to access the articles, Medline, Web of Science Core Collection, and Scopus databases were searched from their start to the year 2018. Full-text, English observational studies that referred to various aspects of ovarian cancer were included in the study. Results: In total, 125 articles that had been published during the years 1925-2018 were entered into the study. Ovarian cancer is the seventh most common cancer among women. Increased risk factors of cancer have led to an upward trend in the incidence of cancer around the world. In 2018, 4.4% of entire cancer-related mortality among women was attributed to ovarian cancer. Although the incidence of cancer is higher among high Human Development Index (HDI) countries, the trend of mortality rate tends to be reversing. Various factors affect the occurrence of ovarian cancer, from which genetic factor are among the most important ones. Pregnancy, lactation, and oral contraceptive pills play a role in reducing the risk of this disease. Conclusion: This study provides significant evidence about ovarian cancer. Considering the heavy burden of ovarian cancer on women's health, preventive measures as well as health education and early detection in high risk groups of women are highly recommended. Although some risk factors cannot be changed, a focus on preventable risk factors may reduce the risk of ovarian cancer. More studies are needed to explore the role of unclear risk factors in ovarian cancer occurrence.


An individual difference analysis of false recognition.
Two studies with moderately large samples of participants were conducted to examine correlates of false recognition. In Experiment 1 false recognition of words was found to be a robust and reliable phenomenon at the level of individuals, and the tendency to classify critical lures as old was more closely related to the correct classification of old items as old than to the incorrect classification of unrelated new items as old. False recognition was not significantly related to any of the cognitive abilities that were assessed, including episodic memory, or to other factors such as personality and chronic mood. In Experiment 2 these findings were extended to include dot pattern and face stimuli. Although measures of veridical memory were significantly correlated across the different types of stimulus material, false recognition rates only had modest and generally not significant correlations, which suggests that the tendency to produce false recognitions may be a task-specific characteristic of individuals.


Improved Oral Pharmacokinetics of Pentoxifylline with Palm Oil and Capmul® MCM Containing Self-Nano-Emulsifying Drug Delivery System.
Pentoxifylline (PTX), an anti-hemorrhage drug used in the treatment of intermittent claudication, is extensively metabolized by the liver resulting in a reduction of the therapeutic levels within a short duration of time. Self-nano-emulsifying drug delivery system (SNEDDS) is well reported to enhance the bio-absorption of drugs by forming nano-sized globules upon contact with the biological fluids after oral administration. The present study aimed to formulate, characterize, and improve the oral bioavailability of PTX using SNEDDS. The formulated SNEDDS consisted of palm oil, Capmul® MCM, and Tween® 80 as oil, surfactant, and co-surfactant, respectively. The mixture design module under the umbrella of the design of experiments was used for the optimization of SNEDDS. The dynamic light-scattering technique was used to confirm the formation of nanoemulsion based on the globule size, in addition to the turbidity measurements. In vivo bioavailability studies were carried out on male Wistar rats. The pharmacokinetic parameters upon oral administration were calculated using the GastroPlus software. The optimized SNEDDS had a mean globule size of 165 nm with minimal turbidity in an aqueous medium. Bioavailability of PTX increased 1.5-folds (AUC = 1013.30 ng h/mL) as SNEDDS than the pure drug with an AUC of 673.10 ng h/mL. In conclusion, SNEDDS was seen to enhance the bioavailability of PTX and can be explored to effectively control the incidents of intermittent claudication.


Differential accumulation and function of proinflammatory 6-sulfo LacNAc dendritic cells in lymph node and colon of Crohn's versus ulcerative colitis patients.
Human Slan DCs have been studied in patients with psoriasis, rheumatoid arthritis, cancer, and autoimmune diseases. In this study, we investigated the frequency, phenotype, and function of Slan DCs in blood, colon, as well as mLNs of patients with IBD. We first show that the frequency of circulating CD14(dull)Slan DCs was reduced in CD patients refractory to immunosuppressive drugs or TNF-α blockers relative to untreated CD, UC, and healthy subjects. In blood of CD patients, Slan DCs expressed CD172a, as detected by CD47 fusion protein binding, when compared with its lack of expression in control subjects. Next, we demonstrate that CD172a(+)Slan DCs that produced IL-1β and TNF-α accumulated in mLNs and colons of CD patients. The CD172a(+)Slan DCs up-regulated their expression of CD14 in CD tissues and the proinflammatory cytokines were produced in CD14(bright)CD172a(+)Slan DCs. By contrast, no difference was noted in the frequency of Slan DCs between inflamed, noninflamed colonic mucosa of UC patients and control, non-IBD donors. Finally, the percentage of cytokine-producing Slan DCs also augmented in response to TLR2 and NOD2 in in vitro stimulation in PBMCs of CD, but not UC, patients. In conclusion, we propose that proinflammatory CD14(bright)CD172a(+)Slan DCs are a distinguishing feature between CD and UC, as these cells accumulate uniquely in mLNs and colonic mucosa of CD patients. Thus, Slan DCs may contribute to CD immunopathogenesis.


There are three major Neisseria gonorrhoeae β-lactamase plasmid variants which are associated with specific lineages and carry distinct TEM alleles.
Neisseria gonorrhoeae is a significant threat to global health with an estimated incidence of over 80 million cases each year and high levels of antimicrobial resistance. The gonococcal β-lactamase plasmid, pbla, carries the TEM β-lactamase, which requires only one or two amino acid changes to become an extended-spectrum β-lactamase (ESBL); this would render last resort treatments for gonorrhoea ineffective. Although pbla is not mobile, it can be transferred by the conjugative plasmid, pConj, found in N. gonorrhoeae. Seven variants of pbla have been described previously, but little is known about their frequency or distribution in the gonococcal population. We characterised sequences of pbla variants and devised a typing scheme, Ng_pblaST that allows their identification from whole genome short-read sequences. We implemented Ng_pblaST to assess the distribution of pbla variants in 15 532 gonococcal isolates. This demonstrated that only three pbla variants commonly circulate in gonococci, which together account for >99 % of sequences. The pbla variants carry different TEM alleles and are prevalent in distinct gonococcal lineages. Analysis of 2758 pbla-containing isolates revealed the co-occurrence of pbla with certain pConj types, indicating co-operativity between pbla and pConj variants in the spread of plasmid-mediated AMR in N. gonorrhoeae. Understanding the variation and distribution of pbla is essential for monitoring and predicting the spread of plasmid-mediated β-lactam resistance in N. gonorrhoeae.


Regional Mortality Disparities in Germany: Long-Term Dynamics and Possible Determinants.
While regional mortality inequalities in Germany tend to be relatively stable in the short run, over the course of the past century marked changes have occurred in the country's regional mortality patterns. These changes include not only the re-emergence of stark differences between eastern and western Germany after 1970, which have almost disappeared again in the decades after the reunification of Germany in 1990; but also substantial changes in the disparities between northern and southern Germany. At the beginning of the twentieth century, the northern regions in Germany had the highest life expectancy levels, while the southern regions had the lowest. Today, this mortality pattern is reversed. In this paper, we study these long-term trends in spatial mortality disparities in Germany since 1910, and link them with theoretical considerations and existing research on the possible determinants of these patterns. Our findings support the view that the factors which contributed to shape spatial mortality variation have changed substantially over time, and suggest that the link between regional socioeconomic conditions and recorded mortality levels strengthened over the last 100 years.


Genetic variants associated with longer telomere length are associated with increased lung cancer risk among never-smoking women in Asia: a report from the female lung cancer consortium in Asia.
Recent evidence from several relatively small nested case-control studies in prospective cohorts shows an association between longer telomere length measured phenotypically in peripheral white blood cell (WBC) DNA and increased lung cancer risk. We sought to further explore this relationship by examining a panel of seven telomere-length associated genetic variants in a large study of 5,457 never-smoking female Asian lung cancer cases and 4,493 never-smoking female Asian controls using data from a previously reported genome-wide association study. Using a group of 1,536 individuals with phenotypically measured telomere length in WBCs in the prospective Shanghai Women's Health study, we demonstrated the utility of a genetic risk score (GRS) of seven telomere-length associated variants to predict telomere length in an Asian population. We then found that GRSs used as instrumental variables to predict longer telomere length were associated with increased lung cancer risk (OR = 1.51 (95% CI = 1.34-1.69) for upper vs. lower quartile of the weighted GRS, p value = 4.54 × 10(-14) ) even after removing rs2736100 (p value = 4.81 × 10(-3) ), a SNP in the TERT locus robustly associated with lung cancer risk in prior association studies. Stratified analyses suggested the effect of the telomere-associated GRS is strongest among younger individuals. We found no difference in GRS effect between adenocarcinoma and squamous cell subtypes. Our results indicate that a genetic background that favors longer telomere length may increase lung cancer risk, which is consistent with earlier prospective studies relating longer telomere length with increased lung cancer risk.


Hydrodynamic stimulation of dinoflagellate bioluminescence: a computational and experimental study.
Dinoflagellate bioluminescence provides a near-instantaneous reporter of cell response to flow. Although both fluid shear stress and acceleration are thought to be stimulatory, previous studies have used flow fields dominated by shear. In the present study, computational and experimental approaches were used to assess the relative contributions to bioluminescence stimulation of shear stress and acceleration in a laminar converging nozzle. This flow field is characterized by separate regions of pronounced acceleration away from the walls, and shear along the wall. Bioluminescence of the dinoflagellates Lingulodinium polyedrum and Ceratocorys horrida, chosen because of their previously characterized different flow sensitivities, was imaged with a low-light video system. Numerical simulations were used to calculate the position of stimulated cells and the levels of acceleration and shear stress at these positions. Cells were stimulated at the nozzle throat within the wall boundary layer where, for that downstream position, shear stress was relatively high and acceleration relatively low. Cells of C. horrida were always stimulated significantly higher in the flow field than cells of L. polyedrum and at lower flow rates, consistent with their greater flow sensitivity. For both species, shear stress levels at the position of stimulated cells were similar to but slightly greater than previously determined response thresholds using independent flow fields. L. polyedrum did not respond in conditions where acceleration was as high as 20 g. These results indicate that shear stress, rather than acceleration, was the stimulatory component of flow. Thus, even in conditions of high acceleration, dinoflagellate bioluminescence is an effective marker of shear stress.


The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release.
In mammals, mitochondria have been shown to play a key intermediary role in apoptosis, a morphologically distinct form of programmed cell death (PCD), for example, through the release of cytochrome c, which activates a proteolytic enzyme cascade, resulting in specific nuclear DNA degradation and cell death. In plants, PCD is a feature of normal development, including the penultimate stage of anther development, leading to dehiscence and pollen release. However, there is little evidence that plant mitochondria are involved in PCD. In a wide range of plant species, anther and/or pollen development is disrupted in a class of mutants termed CMS (for cytoplasmic male sterility), which is associated with mutations in the mitochondrial genome. On the basis of the manifestation of a number of morphological and biochemical markers of apoptosis, we have shown that the PET1-CMS cytoplasm in sunflower causes premature PCD of the tapetal cells, which then extends to other anther tissues. These features included cell condensation, oligonucleosomal cleavage of nuclear DNA, separation of chromatin into delineated masses, and initial persistence of mitochondria. In addition, immunocytochemical analysis revealed that cytochrome c was released partially from the mitochondria into the cytosol of tapetal cells before the gross morphological changes associated with PCD. The decrease in cytochrome c content in mitochondria isolated from male sterile florets preceded a decrease in the integrity of the outer mitochondrial membrane and respiratory control ratio. Our data suggest that plant mitochondria, like mammalian mitochondria, play a key role in the induction of PCD. The tissue-specific nature of the CMS phenotype is discussed with regard to cellular respiratory demand and PCD during normal anther development.


Availability of breastfeeding peer support in the United Kingdom: A cross-sectional study.
Peer support is recommended by the World Health Organization for the initiation and continuation of breastfeeding, and this recommendation is included in United Kingdom (U.K.) guidance. There is a lack of information about how, when, and where breastfeeding peer support was provided in the U.K. We aimed to generate an overview of how peer support is delivered in the U.K. and to gain an understanding of challenges for implementation. We surveyed all U.K. infant feeding coordinators (n = 696) who were part of U.K.-based National Infant Feeding Networks, covering 177 National Health Service (NHS) organisations. We received 136 responses (individual response rate 19.5%), covering 102 U.K. NHS organisations (organisational response rate 58%). We also searched NHS organisation websites to obtain data on the presence of breastfeeding peer support. Breastfeeding peer support was available in 56% of areas. However, coverage within areas was variable. The provision of training and ongoing supervision, and peer-supporter roles, varied significantly between services. Around one third of respondents felt that breastfeeding peer-support services were not well integrated with NHS health services. Financial issues were commonly reported to have a negative impact on service provision. One quarter of respondents stated that breastfeeding peer support was not accessed by mothers from poorer social backgrounds. Overall, there was marked variation in the provision of peer-support services for breastfeeding in the U.K. A more robust evidence base is urgently needed to inform guidance on the structure and provision of breastfeeding peer-support services.


Carbonic anhydrase II is a tumor vessel endothelium-associated antigen targeted by dendritic cell therapy.
Tumor-associated antigens are promising candidates as target molecules for immunotherapy and a wide variety of tumor-associated antigens have been discovered through the presence of serum antibodies in cancer patients. We previously conducted dendritic cell therapy on 10 malignant melanoma patients and shrinkage or disappearance of metastatic tumors with massive necrosis occurred in two patients. In this study, we found a 29-kDa protein against which antibody was elicited by dendritic cell therapy in one of the two patients. Matrix-assisted laser desorption ionization-time of flight/mass spectrometry analysis of the protein isolated by two-dimensional electrophoresis combined with Western blots revealed that the 29-kDa protein was carbonic anhydrase II (CA-II). Immunohistochemistry of the tumors and normal tissues showed that CA-II was expressed in the tumor vessel but not in normal vessel endothelium. CA-II expression in tumor endothelium was observed as well in other cancers including esophageal, renal, and lung cancers. In an in vitro angiogenesis model, CA-II expression of normal human vein endothelial cells was significantly up-regulated when cells were cultured in the acidic and hypoxic conditions indicative of a tumor environment. These findings suggest that CA-II is a tumor vessel endothelium-associated antigen in melanoma and other cancers, and elicitation of serum anti-CA-II antibody by dendritic cell therapy may be associated with good clinical outcome including tumor reduction.


A single dose of the DENV-1 candidate vaccine rDEN1Δ30 is strongly immunogenic and induces resistance to a second dose in a randomized trial.
Dengue is an emerging infectious disease that has become the most important arboviral infection worldwide. There are four serotypes of dengue virus, DENV-1, DENV-2, DENV-3, and DENV-4, each capable of causing the full spectrum of disease. rDEN1Δ30 is a live attenuated investigational vaccine for the prevention of DENV-1 illness and is also a component of an investigational tetravalent DENV vaccine currently in Phase I evaluation. A single subcutaneous dose of rDEN1Δ30 was previously shown to be safe and immunogenic in healthy adults. In the current randomized placebo-controlled trial, 60 healthy flavivirus-naive adults were randomized to receive 2 doses of rDEN1Δ30 (N = 50) or placebo (N = 10), either on study days 0 and 120 (cohort 1) or 0 and 180 (cohort 2). We sought to evaluate the safety and immunogenicity of this candidate vaccine in 50 additional vaccinees and to test whether the humoral immune response could be boosted by a second dose administered 4 or 6 months after the first dose. The first dose of vaccine was well tolerated, infected 47/50 vaccinees and induced seroconversion in 46/50 vaccinees. Irrespective of dosing interval, the second dose of vaccine was also well tolerated but did not induce any detectable viremia or ≥4-fold rise in serum neutralizing antibody titer.Only five subjects had an anamnestic antibody response detectable by ELISA following a second dose of vaccine, demonstrating that the vaccine induced sterilizing humoral immunity in most vaccinees for at least six months following primary vaccination.The promising safety and immunogenicity profile of this vaccine confirms its suitability for inclusion in a tetravalent dengue vaccine.


The ClinicalTrials.gov results database--update and key issues.
The ClinicalTrials.gov trial registry was expanded in 2008 to include a database for reporting summary results. We summarize the structure and contents of the results database, provide an update of relevant policies, and show how the data can be used to gain insight into the state of clinical research. We analyzed ClinicalTrials.gov data that were publicly available between September 2009 and September 2010. As of September 27, 2010, ClinicalTrials.gov received approximately 330 new and 2000 revised registrations each week, along with 30 new and 80 revised results submissions. We characterized the 79,413 registry and 2178 results of trial records available as of September 2010. From a sample cohort of results records, 78 of 150 (52%) had associated publications within 2 years after posting. Of results records available publicly, 20% reported more than two primary outcome measures and 5% reported more than five. Of a sample of 100 registry record outcome measures, 61% lacked specificity in describing the metric used in the planned analysis. In a sample of 700 results records, the mean number of different analysis populations per study group was 2.5 (median, 1; range, 1 to 25). Of these trials, 24% reported results for 90% or less of their participants. ClinicalTrials.gov provides access to study results not otherwise available to the public. Although the database allows examination of various aspects of ongoing and completed clinical trials, its ultimate usefulness depends on the research community to submit accurate, informative data.


Risk of stricture after endoscopic submucosal dissection for large rectal neoplasms.
Improvements in the endoscopic submucosal dissection (ESD) technique have made circumferential ESD in the rectum possible. However, little is known about the clinical course after extensive ESD in the rectum. The aim of this study was to determine the stricture risk in the rectum after total or subtotal circumferential ESD. A total of 69 patients with 69 rectal tumors that required ≥ 75 % circumferential resection were identified at Kobe University Hospital and an affiliated hospital between April 2005 and May 2014. Among the patients, 61 were available for evaluation of stricture development, either by follow-up colonoscopy or by surgical specimens. The rate and possible risk factors of post-ESD strictures were investigated. Post-ESD rectal strictures developed in 12 patients (19.7 %). Patients who underwent total circumferential ESD developed a stricture (5/7, 71.4 %) more frequently than those with subtotal (≥ 90 %) ESD (7/16, 43.8 %). Patients undergoing an ESD procedure that involved < 90 % of the circumference did not develop strictures. The strictures were membranous or < 10 mm long in all cases. Of the patients with stricture, 11 received endoscopic balloon dilation and one received bougie with short-caliber-tip transparent hood; all strictures improved following dilation therapy. Statistical analysis revealed that ≥ 90 % circumferential resection was an independent risk factor for stricture, whereas morphology and size were not. Patients who underwent total or subtotal circumferential ESD of a rectal tumor had a high risk of stricture formation. Dilation helped to alleviate the stenosis.Study registered at University Hospital Medical Information Network (UMIN 000016559).


Paradoxical effects of thought suppression: a meta-analysis of controlled studies.
Research has shown that attempts to suppress a thought can cause an increase in the frequency of the thought. These paradoxical effects of thought suppression play a key role in cognitive-behavioral models of several emotional disorders. Laboratory studies of this phenomenon, however, have yielded mixed results; and narrative summaries of the literature have not been able to draw firm conclusions about the effects of thought suppression. We used meta-analysis to quantitatively examine the magnitude of thought suppression effects across controlled studies. Moreover, we explored whether the variability in effect sizes could be explained by methodological differences within and between studies. Results indicated a small to moderate rebound effect of thought suppression that varied in magnitude depending on the nature of the target thought and the method by which thought frequency was measured. Participants with clinical diagnoses did not show larger rebound effects than nonclinical or analogue participants, however, only a few studies included clinical samples. Findings are discussed in terms of implications for the ironic process theory of thought suppression, and avenues for future research on this phenomenon.


Physical and mental health-related correlates of physical function in community dwelling older adults: a cross sectional study.
Physical function is the ability to perform both basic and instrumental activities of daily living, and the ability of older adults to reside in the community depends to a large extent on their level of physical function. Multiple physical and health-related variables may differentially affect physical function, but they have not been well characterized. The purpose of this investigation was to identify and examine physical and mental health-related correlates of physical function in a sample of community-dwelling older adults. Nine hundred and four community dwelling older men (n = 263) and women (n = 641) with a mean (95% Confidence Interval) age of 76.6 (76.1, 77.1) years underwent tests of physical function (Timed Up and Go; TUG), Body Mass Index (BMI) was calculated from measured height and weight, and data were collected on self-reported health quality of life (SF-36), falls during the past 6 months, number of medications per day, depression (Geriatric Depression Scale; GDS), social support, and sociodemographic variables. Subjects completed the TUG in 8.7 (8.2, 9.2) seconds and expended 6,976 (6,669, 7,284) Kcal.wk-1 in physical activity. The older persons had a mean BMI of 27. 6 (27.2, 28.0), 62% took 3 or more medications per day, and 14.4% had fallen one or more times over the last 6 months. Mean scores on the Mental Component Summary (MCS) was 50.6 (50.2, 51,0) and the Physical Component Summary (PCS) was 41.3 (40.8, 41.8).Multiple sequential regression analysis showed that, after adjustment for TUG floor surface correlates of physical function included age, sex, education, physical activity (weekly energy expenditure), general health, bodily pain, number of medications taken per day, depression and Body Mass Index. Further, there is a dose response relationship such that greater degree of physical function impairment is associated with poorer scores on physical health-related variables. Physical function in community-dwelling older adults is associated with several physical and mental health-related factors. Further study examining the nature of the relationships between these variables is needed.


TOMM40 poly-T variants and cerebrospinal fluid amyloid beta levels in the elderly.
A variable poly-T polymorphism in the TOMM40 gene, which is in linkage disequilibrium with APOE, was recently implicated with increased risk and earlier onset age for late-onset Alzheimer's disease in APOE ε3 carriers. To elucidate potential neurobiological mechanisms underlying this association, we compared the effect of TOMM40 poly-T variants to the effect of APOE, an established LOAD-risk modulator, on cerebrospinal fluid (CSF) amyloid beta (Aβ) and tau levels, in cognitively intact elderly subjects. APOE ε4 carriers showed significant reductions in Aβ 1-42 levels compared to non-ε4 carriers, but no differences were detected across TOMM40 variants. Neither Aβ 1-40 nor tau levels were affected by APOE or TOMM40.


Decoding and targeting the molecular basis of MACC1-driven metastatic spread: Lessons from big data mining and clinical-experimental approaches.
Metastasis remains the key issue impacting cancer patient survival and failure or success of cancer therapies. Metastatic spread is a complex process including dissemination of single cells or collective cell migration, penetration of the blood or lymphatic vessels and seeding at a distant organ site. Hundreds of genes involved in metastasis have been identified in studies across numerous cancer types. Here, we analyzed how the metastasis-associated gene MACC1 cooperates with other genes in metastatic spread and how these coactions could be exploited by combination therapies: We performed (i) a MACC1 correlation analysis across 33 cancer types in the mRNA expression data of TCGA and (ii) a comprehensive literature search on reported MACC1 combinations and regulation mechanisms. The key genes MET, HGF and MMP7 reported together with MACC1 showed significant positive correlations with MACC1 in more than half of the cancer types included in the big data analysis. However, ten other genes also reported together with MACC1 in the literature showed significant positive correlations with MACC1 in only a minority of 5 to 15 cancer types. To uncover transcriptional regulation mechanisms that are activated simultaneously with MACC1, we isolated pan-cancer consensus lists of 1306 positively and 590 negatively MACC1-correlating genes from the TCGA data and analyzed each of these lists for sharing transcription factor binding motifs in the promotor region. In these lists, binding sites for the transcription factors TELF1, ETS2, ETV4, TEAD1, FOXO4, NFE2L1, ELK1, SP1 and NFE2L2 were significantly enriched, but none of them except SP1 was reported in combination with MACC1 in the literature. Thus, while some of the results of the big data analysis were in line with the reported experimental results, hypotheses on new genes involved in MACC1-driven metastasis formation could be generated and warrant experimental validation. Furthermore, the results of the big data analysis can help to prioritize cancer types for experimental studies and testing of combination therapies.


A migrant study of pubertal timing and tempo in British-Bangladeshi girls at varying risk for breast cancer.
Earlier menarche is related to subsequent breast cancer risk, yet international differences in the age and tempo of other pubertal milestones and their relationships with body mass index (BMI) are not firmly established in populations at differing risk for breast cancer. We compared age and tempo of adrenarche, thelarche, pubarche, and menarche in a migrant study of Bangladeshi girls to the United Kingdom (UK) and assessed whether differences by migration were explained by differences in BMI. Included were groups of Bangladeshi (n =168), British-Bangladeshi (n =174) and white British (n =54) girls, aged 5 to 16 years. Interviewer-administered questionnaires obtained pubertal staging; height and weight were measured. Salivary dehydroepiandrosterone-sulfate concentrations >400 pg/ml defined adrenarche. Median ages of pubertal milestones and hazard ratios (HR) with 95% confidence intervals (CI) were estimated from Weibull survival models. In all three groups, adrenarche occurred earliest, followed by thelarche, pubarche, and finally menarche. Neither median age at adrenarche (Bangladeshi = 7.2, British-Bangladeshi = 7.4, white British = 7.1; P-trend = 0.70) nor at menarche (Bangladeshi = 12.5, British-Bangladeshi = 12.1, white British = 12.6; P-trend = 0.70) differed across groups. In contrast, median age at thelarche (Bangladeshi = 10.7, British-Bangladeshi = 9.6, white British = 8.7; P-trend <0.01) occurred earlier among girls living in the UK. Compared with Bangladeshi girls, HRs (95% CI) for earlier thelarche were 1.6 (1.1 to 2.4) for British-Bangladeshi girls and 2.6 (1.5 to 4.4) for white British girls (P-trend <0.01), but were attenuated after adjustment for BMI (British-Bangladeshi = 1.1 (0.7 to 1.8), white British = 1.7(1.0 to 3.1); P-trend =0.20). Thelarche occurred earlier, but puberty progressed slower with increasing exposure to the UK environment; differences were partially explained by greater BMI. The growth environment might account for much of the ethnic differences in pubertal development observed across and within countries.


FtsZ filament dynamics at steady state: subunit exchange with and without nucleotide hydrolysis.
We have measured three aspects of FtsZ filament dynamics at steady state: rates of GTP hydrolysis, subunit exchange between protofilaments, and disassembly induced by dilution or excess GDP. All three reactions were slowed with an increase in the potassium concentration from 100 to 500 mM, via replacement of potassium with rubidium, or with an increase in the magnesium concentration from 5 to 20 mM. Electron microscopy showed that the polymers assembled under the conditions of fastest assembly were predominantly short, one-stranded protofilaments, whereas under conditions of slower dynamics, the protofilaments tended to associate into long, thin bundles. We suggest that exchange of subunits between protofilaments at steady state involves two separate mechanisms: (1) fragmentation or dissociation of subunits from protofilament ends following GTP hydrolysis and (2) reversible association and dissociation of subunits from protofilament ends independent of hydrolysis. Exchange of nucleotides on these recycling subunits could give the appearance of exchange directly into the polymer. Several of our observations suggest that exchange of nucleotide can take place on these recycling subunits, but not directly into the FtsZ polymer. Annealing of protofilaments was demonstrated for the L68W mutant in EDTA buffer but not in Mg buffer, where rapid cycling of subunits may obscure the effect of annealing. We also reinvestigated the nucleotide composition of FtsZ polymers at steady state. We found that the GDP:GTP ratio was 50:50 for concentrations of GTP >100 microM, significantly higher than the 20:80 ratio previously reported at 20 microM GTP.


Evaluation of two mitochondrial DNA biomarkers for prostate cancer detection.
A 3.4kb deletion (3.4kbΔ ) in mitochondrial DNA (mtDNA) found in histologically normal prostate biopsy specimens has been reported to be a biomarker for the increased probability of prostate cancer. Increased mtDNA copy number is also reported as associated with cancer. Independent evaluation of these two potential prostate cancer biomarkers using formalin-fixed paraffin-embedded (FFPE) prostate tissue and matched urine and serum from a high risk cohort of men with and without prostate cancer. Biomarker levels were detected via qPCR. Both 3.4kbΔ and mtDNA levels were significantly higher in cancer patient FFPE cores (p= 0.045 and p= 0.070 respectively at > 90% confidence). Urine from cancer patients contained significantly higher levels of mtDNA (p= 0.006, 64.3% sensitivity, 86.7% specificity). Combining the 3.4kbΔ and mtDNA gave better performance of detecting prostate cancer than either biomarker alone (FFPE 73.7% sensitivity, 65% specificity; urine 64.3% sensitivity, 100% specificity). In serum, there was no difference for any of the biomarkers. This is the first report on detecting the 3.4kbΔ in urine and evaluating mtDNA levels as a prostate cancer biomarker. A confirmation study with increased sample size and possibly with additional biomarkers would need to be conducted to corroborate and extend these observations.


The Deletion of GluK2 Alters Cholinergic Control of Neuronal Excitability.
Kainate receptors (KARs) are key regulators of synaptic circuits by acting at pre- and postsynaptic sites through either ionotropic or metabotropic actions. KARs can be activated by kainate, a potent neurotoxin, which induces acute convulsions. Here, we report that the acute convulsive effect of kainate mostly depends on GluK2/GluK5 containing KARs. By contrast, the acute convulsive activity of pilocarpine and pentylenetetrazol is not alleviated in the absence of KARs. Unexpectedly, the genetic inactivation of GluK2 rather confers increased susceptibility to acute pilocarpine-induced seizures. The mechanism involves an enhanced excitability of GluK2-/- CA3 pyramidal cells compared with controls upon pilocarpine application. Finally, we uncover that the absence of GluK2 increases pilocarpine modulation of Kv7/M currents. Taken together, our findings reveal that GluK2-containing KARs can control the excitability of hippocampal circuits through interaction with the neuromodulatory cholinergic system.


Sudden infant death syndrome caused by cardiac arrhythmias: only a matter of genes encoding ion channels?
Sudden infant death syndrome is the unexpected demise of a child younger than 1 year of age which remains unexplained after a complete autopsy investigation. Usually, it occurs during sleep, in males, and during the first 12 weeks of life. The pathophysiological mechanism underlying the death is unknown, and the lethal episode is considered multifactorial. However, in cases without a conclusive post-mortem diagnosis, suspicious of cardiac arrhythmias may also be considered as a cause of death, especially in families suffering from any cardiac disease associated with sudden cardiac death. Here, we review current understanding of sudden infant death, focusing on genetic causes leading to lethal cardiac arrhythmias, considering both genes encoding ion channels as well as structural proteins due to recent association of channelopathies and desmosomal genes. We support a comprehensive analysis of all genes associated with sudden cardiac death in families suffering of infant death. It allows the identification of the most plausible cause of death but also of family members at risk, providing cardiologists with essential data to adopt therapeutic preventive measures in families affected with this lethal entity.


Inactivation of SAM-methyltransferase is the mechanism of attenuation of a historic louse borne typhus vaccine strain.
Louse borne typhus (also called epidemic typhus) was one of man's major scourges, and epidemics of the disease can be reignited when social, economic, or political systems are disrupted. The fear of a bioterrorist attack using the etiologic agent of typhus, Rickettsia prowazekii, was a reality. An attenuated typhus vaccine, R. prowazekii Madrid E strain, was observed to revert to virulence as demonstrated by isolation of the virulent revertant Evir strain from animals which were inoculated with Madrid E strain. The mechanism of the mutation in R. prowazekii that affects the virulence of the vaccine was not known. We sequenced the genome of the virulent revertant Evir strain and compared its genome sequence with the genome sequences of its parental strain, Madrid E. We found that only a single nucleotide in the entire genome was different between the vaccine strain Madrid E and its virulent revertant strain Evir. The mutation is a single nucleotide insertion in the methyltransferase gene (also known as PR028) in the vaccine strain that inactivated the gene. We also confirmed that the vaccine strain E did not cause fever in guinea pigs and the virulent revertant strain Evir caused fever in guinea pigs. We concluded that a single nucleotide insertion in the methyltransferase gene of R. prowazekii attenuated the R. prowazekii vaccine strain E. This suggested that an irreversible insertion or deletion mutation in the methyl transferase gene of R. prowazekii is required for Madrid E to be considered a safe vaccine.


Superhalogens as building blocks of two-dimensional organic-inorganic hybrid perovskites for optoelectronics applications.
Organic-inorganic hybrid perovskites, well known for their potential as the next generation solar cells, have found another niche application in optoelectronics. This was demonstrated in a recent experiment (L. Dou, et al., Science, 2015, 349, 1518) on atomically thin (C4H9NH3)2PbBr4, where, due to quantum confinement, the bandgap and the exciton binding energy are enhanced over their corresponding values in the three-dimensional bulk phase. Using density functional theory we show that when halogen atoms (e.g. I) are sequentially replaced with superhalogen molecules (e.g. BH4) the bandgap and exciton binding energy increase monotonically with the superhalogen content with the exciton binding energy of (C4H9NH3)2Pb(BH4)4 approaching the value in monolayer black phosphorus. Lead-free admixtures (C4H9NH3)2MI4-x(BH4)x (M = Sn and Ge; x = 0-4) also show a similar trend. Thus, a combination of quantum confinement and compositional change can be used as an effective strategy to tailor the bandgap and the exciton binding energy of two-dimensional hybrid perovskites, making them promising candidates for optoelectronic applications.


Structural basis of chemokine sequestration by CrmD, a poxvirus-encoded tumor necrosis factor receptor.
Pathogens have evolved sophisticated mechanisms to evade detection and destruction by the host immune system. Large DNA viruses encode homologues of chemokines and their receptors, as well as chemokine-binding proteins (CKBPs) to modulate the chemokine network in host response. The SECRET domain (smallpox virus-encoded chemokine receptor) represents a new family of viral CKBPs that binds a subset of chemokines from different classes to inhibit their activities, either independently or fused with viral tumor necrosis factor receptors (vTNFRs). Here we present the crystal structures of the SECRET domain of vTNFR CrmD encoded by ectromelia virus and its complex with chemokine CX3CL1. The SECRET domain adopts a β-sandwich fold and utilizes its β-sheet I surface to interact with CX3CL1, representing a new chemokine-binding manner of viral CKBPs. Structure-based mutagenesis and biochemical analysis identified important basic residues in the 40s loop of CX3CL1 for the interaction. Mutation of corresponding acidic residues in the SECRET domain also affected the binding for other chemokines, indicating that the SECRET domain binds different chemokines in a similar manner. We further showed that heparin inhibited the binding of CX3CL1 by the SECRET domain and the SECRET domain inhibited RAW264.7 cell migration induced by CX3CL1. These results together shed light on the structural basis for the SECRET domain to inhibit chemokine activities by interfering with both chemokine-GAG and chemokine-receptor interactions.


Reengineering the glaucoma quality of life-15 questionnaire with rasch analysis.
To investigate, using Rasch analysis, whether the 15-item Glaucoma Quality of Life-15 (GQL-15) forms a valid scale and to optimize its psychometric properties. One hundred eighteen glaucoma patients (mean age, 65.7 years) completed the German-version of the GQL-15. Rasch analysis was performed to assess category function (how respondents differentiated between the response options), measurement precision (discriminative ability), unidimensionality (whether items measure a single construct), targeting (whether items are of appropriate difficulty for the sample), and differential item functioning (whether comparable subgroups respond differently to an individual item). Where any of these attributes were outside acceptable ranges, steps were taken to improve the instrument. The five-response categories of the GQL-15 were well differentiated by respondents, as demonstrated by ordered and well-spaced category thresholds. The GQL-15 had an excellent measurement precision but demonstrated poor targeting of item difficulty to person ability and multidimensionality, indicating that it was measuring more than one construct. Removal of six misfitting items created a nine-item unidimensional instrument with good measurement precision and no differential item functioning but poor targeting. A new name, the Glaucoma Activity Limitation (GAL-9) questionnaire, is proposed for the short version, which better reflects the construct under measurement. The GAL-9 has superior psychometric properties over the GQL-15. Its only limitation is poor targeting of item difficulty to person ability, which is an inevitable attribute of a vision-related activity limitation instrument for glaucoma patients, most of whom have only peripheral visual field defects and little difficulty with daily activities.


Screened coulomb hybrid DFT investigation of band gap and optical absorption predictions of CuVO3, CuNbO3 and Cu5Ta11O30 materials.
We present a joint theoretical and experimental investigation of the optoelectronic properties of CuVO3, CuNbO3 and Cu5Ta11O30 materials for potential photocatalytic and solar cell applications. In addition to the experimental results obtained by powder X-ray diffraction and UV-Vis spectroscopy of the materials synthesized under flowing N2 gas at atmospheric pressure via solid-state reactions, the electronic structure and the UV-Vis optical absorption coefficient of these compounds are predicted with high accuracy using advanced first-principles quantum methods based on DFT (including the perturbation theory approach DFPT) within the screened coulomb hybrid HSE06 exchange-correlation formalism. The calculated density of states are found to be in agreement with the UV-Vis diffuse reflectance spectra, predicting a small indirect band gap of 1.4 eV for CuVO3, a direct band gap of 2.6 eV for CuNbO3, and an indirect (direct) band gap of 2.1 (2.6) eV for Cu5Ta11O30. It is confirmed that the Cu(I)-based multi-metal oxides possess a strong contribution of filled Cu(I) states in the valence band and of empty d(0) metal states in the conduction band. Interestingly, CuVO3 with its predicted small indirect band gap of 1.4 eV shows the highest absorption coefficient in the visible range with a broad absorption edge extending to 886 nm. This novel result offers a great opportunity for this material to be an excellent candidate for solar cell applications.


Metabolic and inflammatory profiles of biomarkers in obesity, metabolic syndrome, and diabetes in a Mediterranean population. DARIOS Inflammatory study.
There is a paucity of data regarding the differences in the biomarker profiles of patients with obesity, metabolic syndrome, and diabetes mellitus as compared to a healthy, normal weight population. We aimed to study the biomarker profile of the metabolic risk continuum defined by the transition from normal weight to obesity, metabolic syndrome, and diabetes mellitus. We performed a pooled analysis of data from 7 cross-sectional Spanish population-based surveys. An extensive panel comprising 20 biomarkers related to carbohydrate metabolism, lipids, inflammation, coagulation, oxidation, hemodynamics, and myocardial damage was analyzed. We employed age- and sex-adjusted multinomial logistic regression models for the identification of those biomarkers associated with the metabolic risk continuum phenotypes: obesity, metabolic syndrome, and diabetes mellitus. A total of 2851 subjects were included for analyses. The mean age was 57.4 (8.8) years, 1269 were men (44.5%), and 464 participants were obese, 443 had metabolic syndrome, 473 had diabetes mellitus, and 1471 had a normal weight (healthy individuals). High-sensitivity C-reactive protein, apolipoprotein B100, leptin, and insulin were positively associated with at least one of the phenotypes of interest. Apolipoprotein A1 and adiponectin were negatively associated. There are differences between the population with normal weight and that having metabolic syndrome or diabetes with respect to certain biomarkers related to the metabolic, inflammatory, and lipid profiles. The results of this study support the relevance of these mechanisms in the metabolic risk continuum. When metabolic syndrome and diabetes mellitus are compared, these differences are less marked.


Hypoxia-inducible MiR-210 is an independent prognostic factor and contributes to metastasis in colorectal cancer.
MicroRNA-210 (miR-210), the master hypoxamir, plays pleiotropic roles in certain cancers; however, its role in the development of human colorectal cancer remains unclear. Herein, we report that miR-210 is frequently up-regulated in colorectal cancer tissues, with high miR-210 expression significantly correlating with large tumor size, lymph node metastasis, advanced clinical stage and poor prognosis. Functionally, miR-210 overexpression promotes the migration and invasion of colorectal cancer cells. Furthermore, miR-210 can be induced by hypoxia and mediates the hypoxia-induced metastasis of colorectal cancer cells. In addition, vacuole membrane protein 1 (VMP1) is identified as the direct and functional target of miR-210. Thus, miR-210 is a useful biomarker for hypoxic tumor cells and a prognostic factor that plays an essential role in colorectal cancer metastasis.


Risk assessment tools in criminal justice and forensic psychiatry: The need for better data.
Violence risk assessment tools are increasingly used within criminal justice and forensic psychiatry, however there is little relevant, reliable and unbiased data regarding their predictive accuracy. We argue that such data are needed to (i) prevent excessive reliance on risk assessment scores, (ii) allow matching of different risk assessment tools to different contexts of application, (iii) protect against problematic forms of discrimination and stigmatisation, and (iv) ensure that contentious demographic variables are not prematurely removed from risk assessment tools.


[Pain and opioids in preterm and newborns].
Despite the fact that neonates and infants are not capable of expressing their subjective pain sensations, it has become clear that they do perceive nociception, as pain correlates to hormonal, metabolic, immune, and cardiovascular changes. New findings support the notion that repetitive painful stimuli result in long term psycho-physiological effects with ensuing decreased attentiveness and orientation, poor regulation of behavioral state and motor processes, increase in irritability as well as an altered pattern of feeding and sleeping. These sequelae of repetitive painful experiences with an increase in sensitization of sensory afferent input supports the view of a sufficient analgesia during all kinds of painful procedures in the preterm and neonate. In order to sufficiently diminish nociceptive afferent input during surgery opioids are the drugs of choice aside from local anesthetics. However, the use of opioids in neonates and especially preterm infants must be considered in the light of certain pharmacokinetic and pharmacodynamic differences when compared to adults: 1. There is a longer elimination rate, which may result in post-operative overhang of respiratory depression, especially when opioids are given repetitively, resulting in an accumulation and an increased duration of action. 2. There is a reduced hepatic enzyme activity, which ultimately affects clearance rate. 3. The blood-brain-barrier is not fully developed in the preterm, which results in more access of opioids to binding sites in the CNS. 4. Differentiation of opioid-binding sites into mu, delta, and kappa has not reached its peak; thus, higher doses relative to body weight are needed to establish a sufficient deep plane of analgesia. 5. Caudal parts of the CNS, especially the pons-medullar region exhibit an earlier expression of receptors than the rostral parts. Sequelae of such differences are a more pronounced respiratory depression, often due to muscular rigidity, and bradycardia after which a full analgesic effect takes place. Despite such potential drawbacks, opioids are still the best choice as they sufficiently block nociceptive afferent input and when compared to other anesthetics, they show the least cardiovascular changes. One, however, has to bear in mind that dosing is done according to effect and not to body weight while potential side effects are most prominent in the preterm infant.


Data quality control in genetic case-control association studies.
This protocol details the steps for data quality assessment and control that are typically carried out during case-control association studies. The steps described involve the identification and removal of DNA samples and markers that introduce bias. These critical steps are paramount to the success of a case-control study and are necessary before statistically testing for association. We describe how to use PLINK, a tool for handling SNP data, to perform assessments of failure rate per individual and per SNP and to assess the degree of relatedness between individuals. We also detail other quality-control procedures, including the use of SMARTPCA software for the identification of ancestral outliers. These platforms were selected because they are user-friendly, widely used and computationally efficient. Steps needed to detect and establish a disease association using case-control data are not discussed here. Issues concerning study design and marker selection in case-control studies have been discussed in our earlier protocols. This protocol, which is routinely used in our labs, should take approximately 8 h to complete.


Characteristics of DNA-binding proteins determine the biological sensitivity to high-linear energy transfer radiation.
Non-homologous end-joining (NHEJ) and homologous recombination repair (HRR), contribute to repair ionizing radiation (IR)-induced DNA double-strand breaks (DSBs). Mre11 binding to DNA is the first step for activating HRR and Ku binding to DNA is the first step for initiating NHEJ. High-linear energy transfer (LET) IR (such as high energy charged particles) killing more cells at the same dose as compared with low-LET IR (such as X or gamma rays) is due to inefficient NHEJ. However, these phenomena have not been demonstrated at the animal level and the mechanism by which high-LET IR does not affect the efficiency of HRR remains unclear. In this study, we showed that although wild-type and HRR-deficient mice or DT40 cells are more sensitive to high-LET IR than to low-LET IR, NHEJ deficient mice or DT40 cells are equally sensitive to high- and low-LET IR. We also showed that Mre11 and Ku respond differently to shorter DNA fragments in vitro and to the DNA from high-LET irradiated cells in vivo. These findings provide strong evidence that the different DNA DSB binding properties of Mre11 and Ku determine the different efficiencies of HRR and NHEJ to repair high-LET radiation induced DSBs.


Predictors of self and parental vaccination decisions in England during the 2009 H1N1 pandemic: Analysis of the Flu Watch pandemic cohort data.
During the 2009 H1N1 pandemic, UK uptake of the pandemic influenza vaccine was very low. Furthermore, attitudes governing UK vaccination uptake during a pandemic are poorly characterised. To the best of our knowledge, there is no published research explicitly considering predictors of both adult self-vaccination and decisions regarding whether or not to vaccinate one's children among the UK population during the H1N1 pandemic. We therefore aimed to identify predictors of both self-vaccination decisions and parental vaccination decisions using data collected during the H1N1 pandemic as part of the Flu Watch cohort study. Data were analysed separately for 798 adults and 85 children: exploratory factor analysis facilitated reduction of 16 items on attitudes to pandemic vaccine into a smaller number of factors. Single variable analyses with vaccine uptake as the outcome were used to identify variables that were predictive of vaccination in children and adults. Potential predictors were: attitudinal factors created by data reduction, age group, sex, region, deprivation, ethnicity, chronic condition, vocation, healthcare-related occupation and previous influenza vaccination. Consistent with previous literature concerning adult self-vaccination decisions, we found that vaccine efficacy/safety and perceived risk of pandemic influenza were significant predictors of both self-vaccination decisions and parental vaccination decisions. This study provides the first systematic attempt to understand both the predictors of self and parental vaccination uptake among the UK general population during the H1N1 pandemic. Our findings indicate that concerns about vaccine safety, and vaccine effectiveness may be a barrier to increased uptake for both self and parental vaccination.


High efficiency coherent beam combining of semiconductor optical amplifiers.
We demonstrate 40 W coherently combined output power in a single diffraction-limited beam from a one-dimensional 47-element array of angled-facet slab-coupled optical waveguide amplifiers at 1064 nm. The output from each emitter was collimated and overlapped onto a diffractive optical element combiner using a common transform lens. Phase locking was achieved via active feedback on each amplifier's drive current to maximize the power in the combined beam. The combining efficiency at all current levels was nearly constant at 87%.


[In search of results of technical cooperation].
For the Pan American Health Organization (PAHO), technical cooperation (TC) is the process by which the PAHO Member States work with the Organization, as equal partners, to identify and reach their own health goals and to promote self-sufficiency in health development, through programs that respond to those countries' needs and national priorities. Since 1978, PAHO has used the American Regional Planning, Programming, Monitoring, and Evaluation System (AMPES) to establish management procedures and to facilitate decision-making in health TC. As part of AMPES, PAHO uses a "logical approach to project management" to structure the work program of the Organization and to identify the expected results from TC activities and TC resource investments. This project management approach, which replaces the "functional approaches" system used to date, also helps establish a causative relationship between the programmed activities and the results that PAHO expects, and between the activities and the hoped-for outcomes in the countries. As part of an ongoing process of rethinking international health TC, several years ago PAHO began a four-phase study on the usefulness and validity of functional approaches and on the need to propose new ones or to modify existing ones. The results of the initial phase showed it was difficult to classify the activities because the functional-approaches categories were not mutually exclusive and the TC activities were complex. Further, the expected results did not specify the product for which the PAHO Secretariat was accountable within a certain time frame nor the Secretariat's level of responsibility. Thus, a new and more flexible classification of expected results was proposed, with the following categories: cooperation networks and alliances; surveillance and information systems; standards and guidelines; research and evaluation studies; plans, projects, and policies; methods, models, and technologies; training programs; promotional campaigns and advocacy; and direct support. In the second phase of the study, it was concluded that the proposed classification system made it possible to more precisely identify the products of PAHO technical cooperation projects, the Organization's degree of responsibility, and the bases for estimating needed resources. The new system could also facilitate monitoring and evaluation. In addition, the third phase of analyzing the functional approaches has begun. Its objective is to evaluate the effect of technical cooperation based on the changes incorporated in the programming of activities.


Implementing Yoga Therapy Adapted for Older Veterans Who Are Cancer Survivors.
This goal of this paper is to describe the reach, application, and effectiveness of an 8-week yoga therapy protocol with older cancer survivors within a Veterans Health Administration setting. To document the reach of this intervention, recruitment efforts, attendance, and practice rates were tracked. To explore the application of the protocol to this population, physical therapy pre-assessment and observations by the yoga therapist were recorded to ascertain necessary pose modifications. Effectiveness was measured through pre- and post-course structured interviews, tracking self-reported symptoms of combat-related posttraumatic stress disorder, depression, anxiety, fatigue, insomnia, and pain. Regarding reach, 15% of eligible veterans (n = 14) enrolled, participated in 3-16 classes (M±SD = 11.64±3.39), and practiced at home for 0-56 days (M±SD = 26.36±17.87). Participants were primarily Caucasian (n = 13), male (n = 13), ranged in age from 55 to 78 years (M±SD = 65.64±5.15), and had multiple medical problems. During application, substantial individualized modifications to the yoga therapy protocol were necessary. Effectiveness of the intervention was mixed. During post-course interviews, participants reported a variety of qualitative benefits. Notably, the majority of participants reported that breathing and relaxation techniques were the most useful to learn. Group comparisons of mean pre- and post-course scores on standardized measures showed no significant differences. A minority of older veterans express an interest in yoga, but those who do have high rates of class attendance and home practice. Careful physical pre-assessment and attentive therapists are required to undertake the adaptations required by participants with multiple comorbidities. The effectiveness of yoga in this setting requires additional study.


The role of side chain entropy and mutual information for improving the de novo design of Kemp eliminases KE07 and KE70.
Side chain entropy and mutual entropy information between residue pairs have been calculated for two de novo designed Kemp eliminase enzymes, KE07 and KE70, and for their most improved versions at the end of laboratory directed evolution (LDE). We find that entropy, not just enthalpy, helped to destabilize the preference for the reactant state complex of the designed enzyme as well as favoring stabilization of the transition state complex for the best LDE enzymes. Furthermore, residues with the highest side chain couplings as measured by mutual information, when experimentally mutated, were found to diminish or annihilate catalytic activity, some of which were far from the active site. In summary, our findings demonstrate how side chain fluctuations and their coupling can be an important design feature for de novo enzymes, and furthermore could be utilized in the computational steps in lieu of or in addition to the LDE steps in future enzyme design projects.


Correlation between estimates of tumor perfusion from microbubble contrast-enhanced sonography and dynamic contrast-enhanced magnetic resonance imaging.
We compared measurements of tumor perfusion from microbubble contrast-enhanced sonography (MCES) and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in an animal tumor model. Seven mice were implanted with Lewis lung carcinoma cells on their hind limbs and imaged 14 days later with a Philips 5- to 7-MHz sonography system (Philips Medical Systems, Andover, MA) and a Varian 7.0-T MRI system (Varian, Inc, Palo Alto, CA). For sonographic imaging 100 microL of a perfluoropropane microbubble contrast agent (Definity; Bristol-Myers Squibb Medical Imaging, Billerica, MA) was injected and allowed to reach a pseudo steady state, after which a high-mechanical index pulse was delivered to destroy the microbubbles within the field of view, and the replenishment of the microbubbles was imaged for 30 to 60 seconds. The MRI included acquisition of a T(10) map and 35 serial T(1)-weighted images (repetition time, 100 milliseconds; echo time, 3.1 milliseconds; alpha, 30 degrees ) after the injection of 100 microL of 0.2-mmol/kg gadopentetate dimeglumine (Magnevist; Berlex, Wayne, NJ). Region-of-interest and voxel-by-voxel analyses of both data sets were performed; microbubble contrast-enhanced sonography returned estimates of microvessel cross-sectional area, microbubble velocity, and mean blood flow, whereas DCE-MRI returned estimates of a perfusion-permeability index and the extravascular extracellular volume fraction. Comparing similar regions of tumor tissue seen on sonography and MRI, region-of-interest analyses revealed a strong (r(2) = 0.57) and significant relationship (P < .002) between the estimates of perfusion obtained by the two modalities. Microbubble contrast-enhanced sonography can effectively depict intratumoral heterogeneity in preclinical xenograft models when voxel-by-voxel analysis is performed, and this analysis correlates with similar DCE-MRI measurements.


Minor elevations in troponin I are associated with mortality and adverse cardiac events in patients with atrial fibrillation.
In patients with atrial fibrillation, minor troponin I elevation is regularly detected; however, the prognostic significance of this finding is unknown. We therefore sought to examine the prognostic value of elevated troponin I in patients with atrial fibrillation. A prospective study was conducted analysing all consecutive patients admitted with atrial fibrillation in a 2-year period. Patients with an ST-elevation myocardial infarction (MI) were excluded. Minor troponin elevation was defined as a troponin I level between 0.15 and 0.65 ng/mL, which is still below the 99th percentile of the upper reference limit. A positive troponin I was defined as ≥ 0.65 ng/mL. Study outcomes were all-cause mortality (death), death and myocardial infarction (death/MI), or all major adverse cardiac events (MACE: death, MI, or revascularization). A total of 407 patients were eligible for inclusion. The median duration of follow-up was 688 days. A minor elevation occurred in 81 (20%) patients and 77 (19%) had a positive troponin I. In a multivariate model, minor troponin I elevation and a positive troponin I were independently associated with death [hazard ratio (HR): 2.36, 95% confidence interval (CI): 1.17-4.73 for minor elevation and HR: 3.77, 95% CI: 1.42-10.02 for positive troponin I]. Also, there was an independent correlation between the combined endpoints of death/MI and MACE and both a minor elevation and a positive troponin I. Minor elevations in troponin I on hospital admission are associated with mortality and cardiac events in patients with atrial fibrillation and might be useful for risk stratification.


Long read transcript profiling of ion channel splice isoforms.
Alternative splicing of RNA transcripts allows a single gene to generate multiple products and is a key means of generating functionally diverse voltage-gated ion channels. Splicing can be regulated according to cell type, cell state, and stage of development to produce a bespoke complement of protein isoforms. Characterizing the identities of full-length transcript isoforms is essential in order to fully understand a gene's expression and function. However, the repertoire of transcript isoforms is not well characterized for most genes. Long read nanopore sequencing allows full-length isoforms to be sequenced, therefore identifying full-length transcripts. Using this approach, we recently discovered that the human CACNA1C gene gives rise to a far greater repertoire of splice isoforms than previously appreciated. Here we provide a detailed overview of the technical approach we used to achieve this. The method described in this chapter combines long read nanopore sequencing with PCR targeting to selectively sequence transcripts of a specific gene of interest.


Comorbidities of psychiatric and headache disorders in Nepal: implications from a nationwide population-based study.
Headache disorders, anxiety and depression - the major disorders of the brain - are highly comorbid in the western world. Whether this is so in South Asia has not been investigated, but the question is of public-health importance to countries in the region. We aimed to investigate associations, and their direction(s), between headache disorders (migraine, tension-type headache [TTH] and headache on ≥15 days/month) and psychiatric manifestations (anxiety, depression and neuroticism), and how these might affect quality of life (QoL). In a nationwide, cross-sectional survey of the adult Nepalese population (N = 2100), trained interviewers applied: 1) a culturally-adapted version of the Headache-Attributed Restriction, Disability, Social Handicap and Impaired Participation (HARDSHIP) questionnaire to diagnose headache disorders; 2) a validated Nepali version of the Hospital Anxiety and Depression Scale (HADS) to detect anxiety (HADS-A), depression (HADS-D) and comorbid anxiety and depression (HADS-cAD); 3) a validated Nepali version of the Eysenck Personality Questionnaire Revised Short Form-Neuroticism (EPQRS-N); and 4) the World Health Organization Quality of Life 8-question scale (WHOQOL-8). Associations with headache types were analysed using logistic regression for psychiatric caseness and linear regression for neuroticism. Adjustments were made for age, gender, household consumption, habitat, altitude and use of alcohol and marijuana. HADS-A was associated with any headache (p = 0.024), most strongly headache on ≥15 days/month (AOR = 3.2) followed by migraine (AOR = 1.7). HADS-cAD was also associated with any headache (p = 0.050, more strongly among females than males [p = 0.047]) and again most strongly with headache on ≥15 days/month (AOR = 2.7), then migraine (AOR = 2.3). Likewise, neuroticism was associated with any headache (p < 0.001), most strongly with headache on ≥15 days/month (B = 1.6), followed by migraine (B = 1.3). No associations were found between HADS-D and any headache type, or between TTH and any psychiatric manifestation. Psychiatric caseness of any sort, when comorbid with migraine or TTH, aggravated the negative impact on QoL (p < 0.001). Headache disorders are highly comorbid with anxiety and show associations with neuroticism in Nepal, with negative consequences for QoL. These findings call for reciprocal awareness, and a holistic coordinated approach to management and in the health service. Care for common headache and common psychiatric disorders should be integrated in primary care.