Polysulfone/pyrene membranes: a new microwell assay platform for bioapplications.
The use of PSU-Py prepared by click chemistry as a platform in membrane-bottom microwell plates for oxidase and hydrolase/oxidase-based enzyme assays is studied. For the GOx assay, the postulated fluorescence mechanism is based on the consumption of glucose by dissolved oxygen and GOx in the microwell plates covered with the PSU-Py membrane. For the AG-GOx assay, maltose is used as AG substrate and hydrolyzed to glucose which is then oxidized by the GOx activity. It is shown that the PSU-Py membrane acts as a fluorescence indicator of the enzymatic reactions, and both GOx and AG/GOx enzyme assays are successfully applied for glucose, maltose and acorbose analysis in the range 0.125-2.0 × 10(-3) M glucose, 0.05-0.5 × 10(-3) M maltose, and 0.0125-0.1 mg · mL(-1) acorbose, respectively.


Mass-gathering health research foundational theory: part 1 - population models for mass gatherings.
The science underpinning the study of mass-gathering health (MGH) is developing rapidly. Current knowledge fails to adequately inform the understanding of the science of mass gatherings (MGs) because of the lack of theory development and adequate conceptual analysis. Defining populations of interest in the context of MGs is required to permit meaningful comparison and meta-analysis between events. Process A critique of existing definitions and descriptions of MGs was undertaken. Analyzing gaps in current knowledge, the authors sought to delineate the populations affected by MGs, employing a consensus approach to formulating a population model. The proposed conceptual model evolved through face-to-face group meetings, structured break out sessions, asynchronous collaboration, and virtual international meetings. Findings and Interpretation Reporting on the incidence of health conditions at specific MGs, and comparing those rates between and across events, requires a common understanding of the denominators, or the total populations in question. There are many, nested populations to consider within a MG, such as the population of patients, the population of medical services providers, the population of attendees/audience/participants, the crew, contractors, staff, and volunteers, as well as the population of the host community affected by, but not necessarily attending, the event. A pictorial representation of a basic population model was generated, followed by a more complex representation, capturing a global-health perspective, as well as academically- and operationally-relevant divisions in MG populations. Consistent definitions of MG populations will support more rigorous data collection. This, in turn, will support meta-analysis and pooling of data sources internationally, creating a foundation for risk assessment as well as illness and injury prediction modeling. Ultimately, more rigorous data collection will support methodology for evaluating health promotion, harm reduction, and clinical-response interventions at MGs. Delineating MG populations progresses the current body of knowledge of MGs and informs the understanding of the full scope of their health effects.


The long-term impact of physical and emotional trauma: the station nightclub fire.
Survivors of physical and emotional trauma experience enduring occupational, psychological and quality of life impairments. Examining survivors from a large fire provides a unique opportunity to distinguish the impact of physical and emotional trauma on long-term outcomes. The objective is to detail the multi-dimensional long-term effects of a large fire on its survivor population and assess differences in outcomes between survivors with and without physical injury. This is a survey-based cross-sectional study of survivors of The Station fire on February 20, 2003. The relationships between functional outcomes and physical injury were evaluated with multivariate regression models adjusted for pre-injury characteristics and post-injury outcomes. Outcome measures include quality of life (Burn Specific Health Scale-Brief), employment (time off work), post-traumatic stress symptoms (Impact of Event Scale-Revised) and depression symptoms (Beck Depression Inventory). 104 fire survivors completed the survey; 47% experienced a burn injury. There was a 42% to 72% response rate range. Although depression and quality of life were associated with burn injury in univariate analyses (p<0.05), adjusted analyses showed no significant relationship between burn injury and these outcomes (p = 0.91; p = .51). Post-traumatic stress symptoms were not associated with burn injury in the univariate (p = 0.13) or adjusted analyses (p = 0.79). Time off work was the only outcome in which physical injury remained significant in the multivariate analysis (p = 0.03). Survivors of this large fire experienced significant life disruption, including occupational, psychological and quality of life sequelae. The findings suggest that quality of life, depression and post-traumatic stress outcomes are related to emotional trauma, not physical injury. However, physical injury is correlated with employment outcomes. The long-term impact of this traumatic event underscores the importance of longitudinal and mental health care for trauma survivors, with attention to those with and without physical injuries.


Combined effects of MC4R and FTO common genetic variants on obesity in European general populations.
Genome-wide association scans recently identified common polymorphisms, in intron 1 of FTO and 188 kb downstream MC4R, that modulate body mass index (BMI) and associate with increased risk of obesity. Although their individual contribution to obesity phenotype is modest, their combined effects and their interactions with environmental factors remained to be evaluated in large general populations from birth to adulthood. In the present study, we analyzed independent and combined effects of the FTO rs1421085 and MC4R rs17782313 risk alleles on BMI, fat mass, prevalence and incidence of obesity and subsequent type 2 diabetes (T2D) as well as their interactions with physical activity levels and gender in two European prospective population-based cohorts of 4,762 Finnish adolescents (NFBC 1986) and 3,167 French adults (D.E.S.I.R.). Compared to participants carrying neither FTO nor MC4R risk allele (20-24% of the populations), subjects with three or four risk alleles (7-10% of the populations) had a 3-fold increased susceptibility of developing obesity during childhood. In adults, their combined effects were more modest (approximately 1.8-fold increased risk) and associated with a 1.27% increase in fat mass (P = 0.001). Prospectively, we demonstrated that each FTO and MC4R risk allele increased obesity and T2D incidences by 24% (P = 0.02) and 21% (P = 0.02), respectively. However, the effect on T2D disappeared after adjustment for BMI. The Z-BMI and ponderal index of newborns homozygous for the rs1421085 C allele were 0.1 units (P = 0.02) and 0.27 g/cm(3) (P = 0.005) higher, respectively, than in those without FTO risk allele. The MC4R rs17782313 C allele was more associated with obesity and fat mass deposition in males than in females (P = 0.003 and P = 0.03, respectively) and low physical activity accentuated the effect of the FTO polymorphism on BMI increase and obesity prevalence (P = 0.008 and P = 0.01, respectively). In European general populations, the combined effects of common polymorphisms in FTO and MC4R are therefore additive, predictive of obesity and T2D, and may be influenced by interactions with physical activity levels and gender, respectively.


Segregation by onset asynchrony.
We describe a simple psychophysical paradigm for studying figure-ground segregation by onset asynchrony. Two pseudorandom arrays of Gabor patches are displayed, to left and right of fixation. Within one array, a subset of elements form a figure, such as a randomly curving path, that can only be reliably detected when their onset is not synchronized with that of the background elements. Several findings are reported. First, for most participants, segregation required an onset asynchrony of 20-40 ms. Second, detection was no better when the figure was presented first, and thus by itself, than when the background elements were presented first, even though in the latter case the figure could not be detected in either of the two successive displays alone. Third, asynchrony segregated subsets of randomly oriented elements equally well. Fourth, asynchronous onsets aligned with the path could be discriminated from those lying on the path but not aligned with it. Fifth, both transient and sustained neural activity contribute to detection. We argue that these findings are compatible with neural signaling by synchronized rate codes. Finally, schizophrenic disorganization is associated with reduced sensitivity. Thus, in addition to bearing upon basic theoretical issues, this paradigm may have clinical utility.


Changes in body core and body surface temperatures during prolonged swimming in water of 10°C-a case report.
This case report describes an experienced open-water ultra-endurance athlete swimming in water of 9.9°C for 6 h and 2 min. Before the swim, anthropometric characteristics such as body mass, body height, skinfold thicknesses, and body fat were determined. During and after the swim, body core (rectum) and body surface (forearm and calf) temperatures were continuously recorded. The swimmer (53 years old, 110.5 kg body mass, 1.76 m body height, 34.9% body fat, and a body mass index of 35.7 kg/m2) achieved a total distance of 15 km while swimming at a mean speed of 2.48 km/h, equal to 0.69 m/s, in water of 9.9°C. Body core temperature was at 37.8°C before the swim, increased to a maximum of 38.1°C after approximately 20 min of swimming, and then decreased continuously to 36.3°C upon finishing the swim. The lowest body core temperature was 36.0°C between 35 and 60 min after finishing the swim. Sixty minutes after the swim, the body core temperature continuously rose to 36.5°C where it remained. At the forearm, the temperature dropped to 19.6°C after approximately 36 min of swimming and decreased to 19.4°C by the end of the swim. The lowest temperature at the forearm was 17.6°C measured at approximately 47 min before the athlete stopped swimming. At the calf, the temperature dropped to 13.0°C after approximately 24 min of swimming and decreased to 11.9°C at the end of the swim. The lowest temperature measured at the calf was 11.1°C approximately 108 min after the start. In both the forearm and the calf, the skin temperature continuously increased after the swim. This case report shows that (1) it is possible to swim for 6 h in water of 9.9°C and that (2) the athlete did not suffer from hypothermia under these circumstances. The high body mass index, high body fat, previous experience, and specific preparation of the swimmer are the most probable explanations for these findings.


Homeostatic sleep regulation in habitual short sleepers and long sleepers.
Homeostatic sleep regulation in habitual short sleepers (sleep episode < 6 h, n = 9) and long sleepers (> 9 h, n = 7) was investigated by studying their sleep structure and sleep electroencephalogram (EEG) during baseline conditions and after prolonging their habitual waking time by 24 h. In each sleep episode, total sleep time was > 3 h longer in the long sleepers than in the short sleepers. Sleep deprivation decreased sleep latency and rapid eye movement (REM) density in REM sleep more in long sleepers than in short sleepers. The enhancement of EEG slow-wave activity (SWA; spectral power density in the 0.75-4.5 Hz range) in non-REM sleep after sleep loss was larger in long sleepers (47%) than in short sleepers (19%). This difference in the SWA response was predicted by the two-process model of sleep regulation on the basis of the different sleep durations. The results indicate that short sleepers live under a higher "non-REM sleep pressure" than long sleepers. However, the two groups do not differ with respect to the homeostatic sleep regulatory mechanisms.


Clinical implications of distinct metastasizing preferences of different melanoma subtypes.
The incidence and mortality of malignant melanoma have been rising during the past decades, the latter being due to the high invasion capacity and the metastatic potential of melanoma cells to distant organs. We investigated the distribution pattern of melanoma metastases taking into account different clinicopathological subtypes of melanoma. We studied 310 stage IV (AJCC 2009) melanoma patients retrospectively with regard to potential correlations between frequency and occurrence of metastasis and the genetic background and pathological/clinical melanoma subtypes. For all patients, the time to distant metastasis (TTDM) and the distribution patterns of metastases were analyzed and correlated to the median survival time. Superficially Spreading (SSM) and Nodular melanomas (NMM) spread to the brain more frequently than Acrolentiginous (ALM) and Mucosal (MM) melanomas (p = 0.0012). The preference to affect the skeleton was significantly higher for ALM and MM in comparison to SSM and NMM (p = 0.0049). Lentigo maligna (LMM) tumors showed a significantly lower metastatic spread to distant lymph nodes (p = 0.0159). BRAF mutant versus wildtype tumors showed no significant differences concerning localization of metastasis but patients with BRAF mutant tumors were significantly younger at primary diagnosis and had a significantly shorter stage IV survival (p = 0.0106). This study shows a clear distinction of melanoma subtypes with regard to metastasizing preferences. Further knowledge about melanoma subtype specific characteristics, including molecular markers predictive of homing preferences, may help to understand and manage this heterogeneous disease in terms of prognosis and follow-up procedures.


Machine learning identifies a compact gene set for monitoring the circadian clock in human blood.
The circadian clock and the daily rhythms it produces are crucial for human health, but are often disrupted by the modern environment. At the same time, circadian rhythms may influence the efficacy and toxicity of therapeutics and the metabolic response to food intake. Developing treatments for circadian dysfunction, as well as optimizing the daily timing of treatments for other health conditions, will require a simple and accurate method to monitor the molecular state of the circadian clock. Here we used a recently developed method called ZeitZeiger to predict circadian time (CT, time of day according to the circadian clock) from genome-wide gene expression in human blood. In cross-validation on 498 samples from 60 individuals across three publicly available datasets, ZeitZeiger predicted CT in single samples with a median absolute error of 2.1 h. The predictor trained on all 498 samples used 15 genes, only two of which are part of the core circadian clock. By then applying ZeitZeiger to 475 additional samples from the same three datasets, we quantified how the circadian clock in the blood was affected by various perturbations to the sleep-wake and light-dark cycles. Finally, we extended ZeitZeiger (1) to handle intra-individual variation by making predictions based on multiple samples taken a known time apart, and (2) to handle inter-individual variation by personalizing predictions based on samples from the respective individual. Each of these strategies improved prediction of CT by ~20%. Our results are an important step towards precision circadian medicine. In addition, our generalizable extensions to ZeitZeiger may be applicable to the growing number of biological datasets that contain multiple observations per individual.


Transforming growth factor beta 2 and heme oxygenase 1 genes are risk factors for the cerebral malaria syndrome in Angolan children.
Cerebral malaria (CM) represents a severe outcome of the Plasmodium falciparum infection. Recent genetic studies have correlated human genes with severe malaria susceptibility, but there is little data on genetic variants that increase the risk of developing specific malaria clinical complications. Nevertheless, susceptibility to experimental CM in the mouse has been linked to host genes including Transforming Growth Factor Beta 2 (TGFB2) and Heme oxygenase-1 (HMOX1). Here, we tested whether those genes were governing the risk of progressing to CM in patients with severe malaria syndromes. We report that the clinical outcome of P. falciparum infection in a cohort of Angolan children (n = 430) correlated with nine TGFB2 SNPs that modify the risk of progression to CM as compared to other severe forms of malaria. This genetic effect was explained by two haplotypes harboring the CM-associated SNPs (Pcorrec. = 0.035 and 0.036). In addition, one HMOX1 haplotype composed of five CM-associated SNPs increased the risk of developing the CM syndrome (Pcorrec. = 0.002) and was under-transmitted to children with uncomplicated malaria (P = 0.036). Notably, the HMOX1-associated haplotype conferred increased HMOX1 mRNA expression in peripheral blood cells of CM patients (P = 0.012). These results represent the first report on CM genetic risk factors in Angolan children and suggest the novel hypothesis that genetic variants of the TGFB2 and HMOX1 genes may contribute to confer a specific risk of developing the CM syndrome in patients with severe P. falciparum malaria. This work may provide motivation for future studies aiming to replicate our findings in larger populations and to confirm a role for these genes in determining the clinical course of malaria.


Improved oxytetracycline production in Streptomyces rimosus M4018 by metabolic engineering of the G6PDH gene in the pentose phosphate pathway.
The aromatic polyketide antibiotic, oxytetracycline (OTC), is produced by Streptomyces rimosus as an important secondary metabolite. High level production of antibiotics in Streptomycetes requires precursors and cofactors which are derived from primary metabolism; therefore it is exigent to engineer the primary metabolism. This has been demonstrated by targeting a key enzyme in the oxidative pentose phosphate pathway (PPP) and nicotinamide adenine dinucleotide phosphate (NADPH) generation, glucose-6-phosphate dehydrogenase (G6PDH), which is encoded by zwf1 and zwf2. Disruption of zwf1 or zwf2 resulted in a higher production of OTC. The disrupted strain had an increased carbon flux through glycolysis and a decreased carbon flux through PPP, as measured by the enzyme activities of G6PDH and phosphoglucose isomerase (PGI), and by the levels of ATP, which establishes G6PDH as a key player in determining carbon flux distribution. The increased production of OTC appeared to be largely due to the generation of more malonyl-CoA, one of the OTC precursors, as observed in the disrupted mutants. We have studied the effect of zwf modification on metabolite levels, gene expression, and secondary metabolite production to gain greater insight into flux distribution and the link between the fluxes in the primary and secondary metabolisms.


Evolution of microbial virulence: the benefits of stress.
Although genome sequencing of microbial pathogens has shed light on the evolution of virulence, the drivers of the gain and loss of genes and of pathogenicity islands (gene clusters), which contribute to the emergence of new disease outbreaks, are unclear. Recent experiments with the bean pathogen Pseudomonas syringae pv. phaseolicola illustrate how exposure to resistance mechanisms acts as the driving force for genome reorganization. Here we argue that the antimicrobial conditions generated by host defences can accelerate the generation of genome rearrangements that provide selective advantages to the invading microbe. Similar exposure to environmental stress outside the host could also drive the horizontal gene transfer that has led to the evolution of pathogenicity towards both animals and plants.


The person, the soul, and genetic engineering.
Argument about the ethical possibility of the therapeutic use of embryonic stem cells depends critically on the evaluation of the moral status of the very early embryo. Some assert that at the blastocyst stage it is only potentially human, not yet possessing the full ethical status of personhood, while others assert that from its formation the embryo possesses all the moral rights of a human person. It is shown that a decision on this issue is closely related to how human nature is to be understood. The idea of a person as a dual combination of body and spirit correlates naturally with the assertion of absolute personhood from conception, while an idea of human psychosomatic unity encourages a development picture in which the embryo only grows gradually into personhood. The latter view is seen to be encouraged by new advances in science which emphasise the importance of the concept of information in the discussion of complex systems. Other ethical issues related to human genetics are also briefly reviewed.


Resetting of the arterial baroreflex increases orthostatic sympathetic activation and prevents postural hypotension in rabbits.
Since humans are under ceaseless orthostatic stress, the mechanism to maintain arterial pressure (AP) under orthostatic stress against gravitational fluid shift is of great importance. We hypothesized that (1) orthostatic stress resets the arterial baroreflex control of sympathetic nerve activity (SNA) to a higher SNA, and (2) resetting of the arterial baroreflex contributes to preventing postural hypotension. Renal SNA and AP were recorded in eight anaesthetized, vagotomized and aortic-denervated rabbits. Isolated intracarotid sinus pressure (CSP) was increased stepwise from 40 to 160 mmHg with increments of 20 mmHg (60 s for each CSP level) while the animal was placed supine and at 60 deg upright tilt. Upright tilt shifted the CSP-SNA relationship (the baroreflex neural arc) to a higher SNA, shifted the SNA-AP relationship (the baroreflex peripheral arc) to a lower AP, and consequently moved the operating point to marked high SNA while maintaining AP. A simulation study suggests that resetting in the neural arc would double the orthostatic activation of SNA and increase the operating AP in upright tilt by 10 mmHg, compared with the absence of resetting. In addition, upright tilt did not change the CSP-AP relationship (the baroreflex total arc). A simulation study suggests that although a downward shift of the peripheral arc could shift the total arc downward, resetting in the neural arc would compensate this fall and prevent the total arc from shifting downward to a lower AP. In conclusion, upright tilt increases SNA by resetting the baroreflex neural arc. This resetting may compensate for the reduced pressor responses to SNA in the peripheral cardiovascular system and contribute to preventing postural hypotension.


Boltzmann machines reduction by high-order decimation.
Decimation is a common technique in statistical physics that is used in the context of Boltzmann machines (BMs) to drastically reduce the computational cost at the learning stage. Decimation allows to analytically evaluate quantities that should otherwise be statistically estimated by means of Monte Carlo (MC) simulations. However, in its original formulation, this method could only be applied to restricted topologies corresponding to sparsely connected neural networks. In this brief, we present a generalization of the decimation process and prove that it can be used on any BM, regardless of its topology and connectivity. We solve the Monk problem with this algorithm and show that it performs as well as the best classification methods currently available.


Sexual agreement classifications for gay and bisexual men and implications for harm reduction HIV prevention.
HIV prevention educators frequently encourage gay and bisexual men (GBM) to negotiate condom use prior to sexual engagement. Identifying groups of GBM based on their presexual agreements can aid efforts to tailor interventions. Using cross-sectional data from 1,188 GBM who reported having sex with a nonprimary sex partner in the 90 days prior to the survey, the authors identified presexual agreement patterns. For both HIV-positive and HIV-negative GBM, two groups existed: men agreeing to no anal sex and men agreeing to anal sex with condoms. Among HIV-negative participants, there was a group agreeing to anal sex without specifying condom use. Among HIV-positive participants, there was a group agreeing to anal sex without condoms; this was the largest group of HIV-positive participants. Despite a presexual agreement to use condoms, substance use during sex increased the risk of unprotected anal sex. Suggestions for applying harm reduction to HIV prevention based on agreement classifications are discussed.


[Cu(N^N)(P^P)]+ complexes with 2,2':6',2''-terpyridine ligands as the N^N domain.
The first examples of [Cu(N^N)(POP)](+) complexes (POP = bis(2-(diphenylphosphino)phenyl)ether) in which the N^N domain is a 2,2':6',2''-terpyridine (tpy) ligand have been prepared and characterized; N^N = tpy, 5,5''-dimethyl-2,2':6',2''-terpyridine (1), 4′-(4-tolyl)-2,2′:6′,2′′-terpyridine (2), 4′-(4-npropoxyphenyl)-2,2′:6′,2′′-terpyridine (3) and 4′-(4-nbutoxyphenyl)-2,2′:6′,2′′-terpyridine (4). In solution, the tpy domain in each [Cu(N^N)(POP)][PF6] complex is C2-symmetric, consistent with either tridentate coordination or a low energy dynamic process involving bidentate ligands; for [Cu(2)(POP)][PF6] and [Cu(4)(POP)][PF6], the (1)H NMR spectra showed negligible change between 295 and 210 K. The single crystal structures of [Cu(tpy)(POP)][PF6] and [Cu(4)(POP)][PF6] are presented. The asymmetric unit of [Cu(tpy)(POP)][PF6] contains two independent cations; in one the tpy ligand is tridentate and in the other, it is bidentate with the non-coordinated pyridine ring facing the Cu atom (Cu···N = 3.146(1) Å). In contrast, the solid-state structure of [Cu(4)(POP)][PF6] features a [Cu(4)(POP)](+) cation containing a bidentate tpy-domain with the non-coordinated pyridine ring oriented with the N-atom facing away from the Cu atom; this conformation may be associated with inter-cation NHC non-classical hydrogen bonds. The photophysical properties of [Cu(N^N)(POP)][PF6] with N^N = tpy or 1–4 are described. In the solid state at room temperature, the compounds are poorly emissive. In solution, the emission behaviour is consistent with ligand dissociation. This is supported by (1)H and (31)P NMR spectroscopic data which show POP and [Cu(POP)2](+) in solutions of aged samples; mass spectrometric data are consistent with the formation of [Cu(N^N)2](2+) in these samples.


Psychogenic and neural visual-cue response in PD dopamine dysregulation syndrome.
Dopamine dysregulation syndrome (DDS) in Parkinson's disease (PD) patients refers to the compulsive use of dopaminergic replacement therapy and has serious psycho-social consequences. Mechanisms underlying DDS are not clear although has been linked to dysfunctional brain reward networks. With fMRI, we investigate behavioral and neural response to drug-cues in six PD DDS patients and 12 PD control patients in both the ON and OFF medication state. Behavioral measures of liking, wanting and subjectively 'feeling ON medication' were also collected. Behaviorally, PD DDS patients feel less ON and want their drugs more at baseline compared to PD controls. Following drug-cue exposure, PD DDS patients feel significantly more ON medication, which correlates with significant increases in reward related regions. The results demonstrate that exposure to drug-cues increases the subjective feeling of being 'ON' medication which corresponds to dysfunctional activation in reward related regions in PD DDS patients. These findings should be extended in future studies. Visual stimuli being sufficient to elicit behavioral response through neuroadaptations could have direct implications to the management of addictive behavior.


Failure of adrenal corticosterone production in POMC-deficient mice results from lack of integrated effects of POMC peptides on multiple factors.
Production of corticosteroids from the adrenal gland is a multistep process in which corticosterone is enzymatically processed from its precursor cholesterol. The main hormone regulating the production of corticosterone is the proopiomelanocortin (POMC)-derived adrenocorticotropic hormone (ACTH). Adrenals of POMC-deficient (POMC(-/-)) mice do not produce corticosterone either at basal levels or in response to acute stimulation with ACTH. However, pharmacological amounts of ACTH delivered continuously elicit corticosterone production over time. To define the relative effects of ACTH on individual factors involved in corticosterone production, parameters of adrenal cholesterol metabolism and steroidogenesis were examined in POMC(-/-) mice compared with wild-type and ACTH-treated mutant mice. POMC(-/-) adrenals lack cholesterol esters (CE); adrenal CE is restored with ACTH treatment. However, discontinuation of ACTH treatment stops corticosterone production despite the presence of adrenal CE. Failure of corticosterone production by POMC(-/-) adrenals occurs despite the constitutive presence of transcripts of genes required for cholesterol metabolism and steroidogenesis. Levels of key proteins involved in selective cholesterol uptake and steroidogenesis were attenuated; ACTH treatment increased these protein levels, most significantly those of the receptor responsible for selective uptake of CE, scavenger receptor class B, type I (SR-BI). Our studies reveal that failure of corticosterone production of POMC(-/-) adrenal glands and its pharmacological reconstitution by ACTH are not mediated by any one individual protein, but rather as an integrated effect on multiple factors from import of the substrate cholesterol to its conversion to corticosterone.


Validation of the Korean translation of obesity-related problems scale assessing the quality of life in obese Korean.
The objective of this study was to translate the obesity-related problems (OP) scale for Koreans and to validate it for use in Korean populations. Translation and back-translation of the OP scale was performed and a pilot test was conducted. Following this, patients who had received treatment at the Obesity Center of Inha University Hospital were selected for participation in the field test. Cronbach's alpha (α) was used for assessment of the internal consistency of the OP scale. Spearman's correlation coefficients were used to assess the concurrent validity between the OP scale, the EuroQoL-5D (EQ-5D), and the Beck depression inventory (BDI) scale. One-way analysis of variance and t-test were used to assess the factors associated with the OP scale. A total of 67 individuals participated in the field study. The standardized Cronbach's α of the OP was 0.913. A significant negative correlation was observed between the OP scale and the EQ-5D and a positive correlation was observed between the OP scale and the BDI (the correlation coefficient with EQ-5D = -0.316, and the BDI = 0.305, P < 0.05). The results of this study prove that the Korean version of the OP has been translated and adapted correctly in order to meet the standard of its use.


Bone marrow stroma CD40 expression correlates with inflammatory mast cell infiltration and disease progression in splenic marginal zone lymphoma.
Splenic marginal zone lymphoma (SMZL) is a mature B-cell neoplasm characterized by rather indolent clinical course. However, nearly one third of patients experience a rapidly progressive disease with a dismal outcome. Despite the characterization of clone genetics and the recognition of deregulated immunologic stimulation in the pathogenesis of SMZL, little is known about microenvironment dynamics and their potential biological influence on disease outcome. Here we investigate the effect of stroma-intrinsic features on SMZL disease progression by focusing on the microenvironment of the bone marrow (BM), which represents an elective disease localization endorsing diagnostic and prognostic relevance. We show that the quality of the BM stromal meshwork of SMZL infiltrates correlates with time to progression. In particular, we describe the unfavorable prognostic influence of dense CD40 expression by BM stromal cells, which involves the contribution of CD40 ligand (CD40L)-expressing bystander mast cells infiltrating SMZL BM aggregates. The CD40/CD40L-assisted crosstalk between mesenchymal stromal cells and mast cells populating the SMZL microenvironment finds correlation in p53(-/-) mice developing SMZL and contributes to the engendering of detrimental proinflammatory conditions. Our study highlights a dynamic interaction, playing between nonneoplastic elements within the SMZL niche, toward disease progression.


Potentially serious infections in the aging person: diagnosis, treatment and prevention.
Infections in the elderly patient are a challenge, since the classical signs of infection are absent or ill defined. The present paper describes the presentation, diagnosis, clinical manifestations and treatment for a selected group of potential serious infections including influenza, bacterial pneumonia, urinary tract infections as well as infections caused by multiresistant bacteria, like vacomycin-resistant enterococcus and methicillin resistant S. aureus. We conclude with the need for prevention in the older person with the use of vaccines, specifically the influenza and pneumococcal vaccine as well as the prevention of urinary infections. Influenza is a significant cause of morbidity, whose ill effects can be prevented in many older persons with the use of a vaccine. The use in prophylaxis and treatment of antiviral agents like amantadine, rimatadine, and oseltamivir is presented. Bacterial pneumonia is one of the leading causes of death in the USA among the older persons. The emergence of drug resistant Streptococcus pneumoniae leads to the consideration as empiric therapy the newer fluoroquinolones or the use of third or fourth generation cephalosporis. Of importance is the use of pneumococcal vaccine among people age 60 or above. The frequency of urinary tract infections among the elderly is of primary although in many instances important do not require treatment. When infection of the urinary tract is diagnosed, most authors use a fluoroquinolone as empiric theraphy. The emergence of multiresistant bacteria like methicillin resistant S. aureus and or vancomycin resistant enterococci leads to the need to consider new agents like quinipristin-dalfopristin, linezolid and deptomycin in the management of such patients.


Clinical and dopamine transporter imaging characteristics of non-manifest LRRK2 and GBA mutation carriers in the Parkinson's Progression Markers Initiative (PPMI): a cross-sectional study.
The Parkinson's Progression Markers Initiative (PPMI) is an ongoing observational, longitudinal cohort study of participants with Parkinson's disease, healthy controls, and carriers of the most common Parkinson's disease-related genetic mutations, which aims to define biomarkers of Parkinson's disease diagnosis and progression. All participants are assessed annually with a battery of motor and non-motor scales, 123-I Ioflupane dopamine transporter (DAT) imaging, and biological variables. We aimed to examine whether non-manifesting carriers of LRRK2 and GBA mutations have prodromal features of Parkinson's disease that correlate with reduced DAT binding. This cross-sectional analysis is based on assessments done at enrolment in the subset of non-manifesting carriers of LRRK2 and GBA mutations enrolled into the PPMI study from 33 participating sites worldwide. The primary objective was to examine baseline clinical and DAT imaging characteristics in non-manifesting carriers with GBA and LRRK2 mutations compared with healthy controls. DAT deficit was defined as less than 65% of putamen striatal binding ratio expected for the individual's age. We used t tests, χ2 tests, and Fisher's exact tests to compare baseline demographics across groups. An inverse probability weighting method was applied to control for potential confounders such as age and sex. To account for multiple comparisons, we applied a family-wise error rate to each set of analyses. This study is registered with ClinicalTrials.gov, number NCT01141023. Between Jan 1, 2014, and Jan 1, 2019, the study enrolled 208 LRRK2 (93% G2019S) and 184 GBA (96% N370S) non-manifesting carriers. Both groups were similar with respect to mean age, and about 60% were female. Of the 286 (73%) non-manifesting carriers that had DAT imaging results, 18 (11%) LRRK2 and four (3%) GBA non-manifesting carriers had a DAT deficit. Compared with healthy controls, both LRRK2 and GBA non-manifesting carriers had significantly increased mean scores on the Movement Disorders Society Unified Parkinson's Disease Rating Scale (total score 4·6 [SD 4·4] healthy controls vs 8·4 [7·3] LRRK2 vs 9·5 [9·2] GBA, p<0·0001 for both comparisons) and the Scale for Outcomes for PD - autonomic function (5·8 [3·7] vs 8·1 [5·9] and 8·4 [6·0], p<0·0001 for both comparisons). There was no difference in daytime sleepiness, anxiety, depression, impulsive-compulsive disorders, blood pressure, urate, and rapid eye movement (REM) behaviour disorder scores. Hyposmia was significantly more common only in LRRK2 non-manifesting carriers (69 [36%] of 194 healthy controls vs 114 [55%] of 208 LRRK2 non-manifesting carriers; p=0·0003). Finally, GBA but not LRRK2 non-manifesting carriers showed increased DAT striatal binding ratios compared with healthy controls in the caudate (healthy controls 2·98 [SD 0·63] vs GBA 3·26 [0·63]; p<0·0001), putamen (2·15 [0·56] vs 2·48 [0·52]; p<0·0001), and striatum (2·56 [0·57] vs 2·87 [0·55]; p<0·0001). Our data show evidence of subtle motor and non-motor signs of Parkinson's disease in non-manifesting carriers compared with healthy controls that can precede DAT deficit. Longitudinal data will be essential to confirm these findings and define the trajectory and predictors for development of Parkinson's disease. Michael J Fox Foundation for Parkinson's Research.


The influence of age and skull conductivity on surface and subdermal bipolar EEG leads.
Bioelectric source measurements are influenced by the measurement location as well as the conductive properties of the tissues. Volume conductor effects such as the poorly conducting bones or the moderately conducting skin are known to affect the measurement precision and accuracy of the surface electroencephalography (EEG) measurements. This paper investigates the influence of age via skull conductivity upon surface and subdermal bipolar EEG measurement sensitivity conducted on two realistic head models from the Visible Human Project. Subdermal electrodes (a.k.a. subcutaneous electrodes) are implanted on the skull beneath the skin, fat, and muscles. We studied the effect of age upon these two electrode types according to the scalp-to-skull conductivity ratios of 5, 8, 15, and 30 : 1. The effects on the measurement sensitivity were studied by means of the half-sensitivity volume (HSV) and the region of interest sensitivity ratio (ROISR). The results indicate that the subdermal implantation notably enhances the precision and accuracy of EEG measurements by a factor of eight compared to the scalp surface measurements. In summary, the evidence indicates that both surface and subdermal EEG measurements benefit better recordings in terms of precision and accuracy on younger patients.


Comparing three approaches in extending biotic ligand models to predict the toxicity of binary metal mixtures (Cu-Ni, Cu-Zn and Cu-Ag) to lettuce (Lactuca sativa L.).
Metals are always found in the environment as mixtures rather than as solitary elements. However, effect models such as biotic ligand models (BLMs) are usually derived for toxicity prediction of single metals. Our study aimed at predicting mixture toxicity of Cu-Ni, Cu-Zn and Cu-Ag combinations to lettuce (Lactucasativa L.) by combining BLMs with three toxicity indexes: the toxic unit, the overall amounts of metal ions bound to the biotic ligands and the toxic equivalency factor. The accumulation of metal ions at the biotic ligands was used to determine the toxic potency of metals alone or in combination. On the basis of parameters derived from toxicity assessment of individual metals, these three extended BLMs appeared to be all acceptable (p<0.0001) in assessing toxicity of diverse metal mixtures. The BLM-based approaches integrated competition between metal ions in assessing mixture toxicity and showed different predictive ability for each metal combination. The outcome of modeling suggested that the combined toxicity depends on the specific components of the metal mixtures. The best developed models assist in identifying the type of underlying toxic mechanisms of diverse metal mixtures in terrestrial plants.


Does training on a virtual reality robotic simulator improve performance on the da Vinci surgical system?
The primary objective of this study is to determine if training on the Mimic dV-Trainer (MdVT) simulator results in improved ability on the da Vinci surgical system (dVSS) using exercises with inanimate objects. Twelve trainees (MdVT group) and 10 residents and one fellow (dVSS group) were recruited for the study. Each participant in the MdVT group completed one session of five exercises on the dVSS that were scored for timing and accuracy, followed by four training sessions on the MdVT, and concluded with a final session on the dVSS in which the initial exercises were repeated. Improvement on the dVSS exercises was compared with dVSS group who completed four to six training sessions using the same exercises on the dVSS without any simulator training. Both groups had similar significant improvements in the Letter Board and String Running exercises for both timing and accuracy. The MdVT group demonstrated significant improvement in the Pattern Cutting and Peg Board times. Only the dVSS group significantly improved in the Knot Tying time and the Peg Board accuracy. Training with the MdVT provided similar improvement on five exercises performed on the dVSS when compared with training on the dVSS alone. The use of this simulator in resident and student training may help bridge the gap between the safe acquisition of surgical skills and effective performance during live robot-assisted surgery.


Influence of infrapatellar and suprapatellar straps on quadriceps muscle activity and onset timing during the body-weight squat.
The use of knee braces for the treatment of patellofemoral pain syndrome (PFPS) is widely documented, yet the mechanism by which such braces alleviate knee pain remains unclear. This study attempted to clarify this issue by simplifying the brace to the level of only straps. The effectiveness of an infrapatellar strap for PFPS remains controversial, and the use of a suprapatellar strap has not yet been studied. Quadriceps muscle activity and onset timing parameters were measured with surface electromyography (EMG) during a body-weight squat in 19 healthy subjects during 4 different knee-strapping conditions (infra, supra, both, and none). No differences in normalized mean or peak EMG activity in any part of the quadriceps were found. The onset timing of the vastus lateralis (VL) was significantly delayed when using an infrapatellar strap (p < 0.05) or both straps (p < 0.05) and marginally delayed when using a suprapatellar strap (p < 0.10) in comparison with the no-strap (control) condition. No differences in the vastus medialis oblique (VMO) onset timing or VMO-VL onset timing difference were found among the strapping conditions, although an improvement in timing was noted with the suprapatellar condition. The results provide novel evidence that the application of an infrapatellar strap, suprapatellar strap, or both straps improves quadriceps muscle timing imbalances by delaying VL onset. Because the largest delay in VL onset occurred when wearing both straps, the combined application of an infrapatellar and suprapatellar strap may be the most beneficial in managing patellofemoral pain. Knee straps, unlike braces, are cost effective, nonrestrictive, and can be universally fitted to any knee and based on the results deserve further study in the patellofemoral pain population.


Safety of laboratory analyzers for infection testing - results of the market surveillance by the BfArM until End 2007.
The European Directive 98/79/EC on in vitro diagnostic medical devices (IVD) stipulates the marketing and post market surveillance of IVD in the European Economic Area. In cases of issues and field corrective actions, the manufacturers have to inform the responsible Competent Authorities (CA). In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD, with a small subset of IVD for immune hematological and infectiological testing as well as tissue typing as specified in Annex II of the Directive, being within the responsibility of the Paul-Ehrlich-Institute (PEI). In this study, all issues regarding laboratory analyzers for infection testing and their consumables, but not reagents, kits and general culture media, reported to the BfArM between begin 1999 and end of 2007 were analyzed in respect to the sources of report, the underlying product failure and the performed corrective actions. Within the observation period a total of 1471 reports for IVD were received of which 73 related to the IVD for infection testing were included in our study. Reports were predominantly received from manufacturers (56) and competent authorities (15). Affected products were most frequently those for immunological analysis (42) whereas those based on culturing techniques (17) and molecular biological techniques (14) played only minor roles. In all these groups, laboratory analyzers (55) were more frequently affected than their consumables (18). Investigations of the manufacturers were able to identify the underlying root causes of product failures in 62 cases (84.9%). In 2 cases (2.7%) the root cause remained unclear and in 9 cases (12.3%) a product failure was excluded or a user error was the underlying cause. Product failures in laboratory analyzers were most frequently caused by software errors (31) and constructional faults (8) whereas the predominant cause of product failure in consumables were errors in production and quality control (8). Manufacturers issued corrective measures in 66 cases (90.4%) from which 49 and 17 were related to laboratory analyzers and their consumables, respectively. Based on the underlying root causes of product failures these were predominantly customer information (48), recalls (40), software-updates (30) and design changes (9) in the product group of laboratory analyzers as well as customer information (16), recalls (12) and modifications of production and quality management (11) in the group of consumables. The results and experiences obtained since 1999 suggest that the system for post marketing surveillance of IVD is an established tool to ensure product safety, even though the current system can be further enhanced.


Skeletal Indicators of Shark Feeding on Human Remains: Evidence from Florida Forensic Anthropology Cases.
This research examines a series of six Florida forensic anthropology cases that exhibit taphonomic evidence of marine deposition and shark-feeding activities. In each case, we analyzed patterns of trauma/damage on the skeletal remains (e.g., sharp-force bone gouges and punctures) and possible mechanisms by which they were inflicted during shark predation/scavenging. In some cases, shark teeth were embedded in the remains; in the absence of this evidence, we measured interdental distance from defects in the bone to estimate shark body length, as well as to draw inferences about the potential species responsible. We discuss similarities and differences among the cases and make comparisons to literature documenting diagnostic shark-inflicted damage to human remains from nearby regions. We find that the majority of cases potentially involve bull or tiger sharks scavenging the remains of previously deceased, adult male individuals. This scavenging results in a distinctive taphonomic signature including incised gouges in cortical bone.


Pressure-induced hydrogen localization coupled to a semiconductor-insulator transition in a hydrogen-bonded molecular conductor.
Purely organic crystals, κ-X3(Cat-EDT-TTF)2 [X = H or D, Cat-EDT-TTF = catechol-fused tetrathiafulvalene], are a new type of molecular conductor with hydrogen dynamics. In this work, hydrostatic pressure effects on these materials were investigated in terms of the electrical resistivity and crystal structure. The results indicate that the pressure induces and promotes hydrogen (deuterium) localization in the hydrogen bond, in contrast to the case of the conventional hydrogen-bonded materials (where pressure prevents hydrogen localization), and consequently leads to a significant change in the electrical conducting properties (i.e., the occurrence of a semiconductor-insulator transition). Therefore, we have successfully found a new type of pressure-induced phase transition where the cooperation of the hydrogen dynamics and π-electron interactions plays a crucial role.


Persistent beneficial impact of H-reflex conditioning in spinal cord-injured rats.
Operant conditioning of a spinal cord reflex can improve locomotion in rats and humans with incomplete spinal cord injury. This study examined the persistence of its beneficial effects. In rats in which a right lateral column contusion injury had produced asymmetric locomotion, up-conditioning of the right soleus H-reflex eliminated the asymmetry while down-conditioning had no effect. After the 50-day conditioning period ended, the H-reflex was monitored for 100 [±9 (SD)] (range 79-108) more days and locomotion was then reevaluated. After conditioning ended in up-conditioned rats, the H-reflex continued to increase, and locomotion continued to improve. In down-conditioned rats, the H-reflex decrease gradually disappeared after conditioning ended, and locomotion at the end of data collection remained as impaired as it had been before and immediately after down-conditioning. The persistence (and further progression) of H-reflex increase but not H-reflex decrease in these spinal cord-injured rats is consistent with the fact that up-conditioning improved their locomotion while down-conditioning did not. That is, even after up-conditioning ended, the up-conditioned H-reflex pathway remained adaptive because it improved locomotion. The persistence and further enhancement of the locomotor improvement indicates that spinal reflex conditioning protocols might supplement current therapies and enhance neurorehabilitation. They may be especially useful when significant spinal cord regeneration becomes possible and precise methods for retraining the regenerated spinal cord are needed.


Moving mirrors: a high-density EEG study investigating the effect of camera movements on motor cortex activation during action observation.
Action execution-perception links (mirror mechanism) have been repeatedly suggested to play crucial roles in social cognition. Remarkably, the designs of most studies exploring this topic so far excluded even the simplest traces of social interaction, such as a movement of the observer toward another individual. This study introduces a new design by investigating the effects of camera movements, possibly simulating the observer's own approaching movement toward the scene. We conducted a combined high-density EEG and behavioral study investigating motor cortex activation during action observation measured by event-related desynchronization and resynchronization (ERD/ERS) of the mu rhythm. Stimuli were videos showing a goal-related hand action filmed while using the camera in four different ways: filming from a fixed position, zooming in on the scene, approaching the scene by means of a dolly, and approaching the scene by means of a steadycam. Results demonstrated a consistently stronger ERD of the mu rhythm for videos that were filmed while approaching the scene with a steadycam. Furthermore, videos in which the zoom was applied reliably demonstrated a stronger rebound. A rating task showed that videos in which the camera approached the scene were felt as more involving and the steadycam was most able to produce a visual experience close to the one of a human approaching the scene. These results suggest that filming technique predicts time course specifics of ERD/ERS during action observation with only videos simulating the natural vision of a walking human observer eliciting a stronger ERD than videos filmed from a fixed position. This demonstrates the utility of ecologically designed studies for exploring social cognition.


Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype in vitro: implications for cell-based transplantation therapy.
Because mesenchymal stem cells can differentiate into chondrocyte-like cells, we ask the question, can mesenchymal stem cells commit to the nucleus pulposus phenotype? Back pain, a significant source of morbidity in our society, is linked to degenerative changes of the intervertebral disc. Absence of suitable graft tissue limits therapeutic approaches for repair of disc tissue. For this reason, there is considerable interest in developing "tissue engineering" strategies for the regeneration of the nucleus pulposus. Rat mesenchymal stem cells were immobilized in 3-dimensional alginate hydrogels and cultured in a medium containing transforming growth factor-beta1 under hypoxia (2% O2) and normoxia (20% O2). Mesenchymal stem cells were examined by confocal microscopy to evaluate their viability and metabolic status after labeling with Celltracker green, a thiol sensitive dye, and Mitotracker red, a dye sensitive to the mitochondrial membrane potential. Flow cytometry, semiquantitative reverse transcription polymerase chain reaction and Western blot analysis were carried out to evaluate phenotypic and biosynthetic activities and the signaling pathways involved in the differentiation process. Under hypoxic conditions, mesenchymal stem cells formed large aggregates and exhibited positive Celltracker and Mitotracker signals. Glucose transporter-3, matrix metalloproteinase-2, collagen type II and type XI, and aggrecan mRNA and protein expression was upregulated, whereas there was no change in the levels of decorin, biglycan, fibromodulin, and lumican. Hypoxia maintained the expression of CD44 (hyaluronan receptor), ALCAM (CD166), and endoglin (transforming growth factor-beta receptor). Likewise, expression of beta3 and alpha2 integrin was upregulated. Transforming growth factor-beta treatment increased MAPK activity and Sox-9, aggrecan, and collagen type II gene expression. Basal levels of the phosphorylated MAPK isoform ERK1/2, but not p38, were higher under hypoxic conditions than normoxia, and its activation was further augmented by treatment of cells with transforming growth factor-beta. In hypoxia, transforming growth factor-beta sustained phosphorylated p38 expression for an extended time period. Pharmacological inhibition of ERK1/2 and p38 enzymatic activity resulted in a decrease in Sox-9, aggrecan, and collagen type II mRNA levels. Our results indicate that hypoxia and transforming growth factor-beta drive mesenchymal stem cell differentiation towards a phenotype consistent with that of the nucleus pulposus. Measurement of selected signaling molecules and response to specific inhibitors suggest involvement of MAPK signaling pathways. It is concluded that mesenchymal stem cells could be used to repopulate the damaged or degenerate intervertebral disc.


Economic evaluation of desirudin vs heparin in deep vein thrombosis prevention after hip replacement surgery.
Deep vein thrombosis is a serious complication of orthopaedic surgery and can lead to pulmonary embolism and long term post-thrombotic syndrome. A simulation model based on both epidemiological data and data from clinical trials was used to compare the long term cost effectiveness of standard prophylaxis with subcutaneous unfractionated heparin with that of desirudin (recombinant hirudin), in patients undergoing elective hip replacement. The analysis, which was performed before the price of desirudin was set, showed that prophylactic treatment with desirudin saved 4.5 life-years per 100 patients treated, compared with unfractionated heparin. Desirudin prophylaxis was dominant up to a total drug cost of 4400 Swedish kronor (SEK) per treatment. The results were robust against changes in the parameters used in several sensitivity analyses. This study showed that prophylactic therapy with desirudin compared with unfractionated heparin was more cost effective and potentially cost saving under a wide range of assumptions concerning the future price of desirudin in preventing deep vein thrombosis following elective hip surgery.


DSDBASE: a consortium of native and modelled disulphide bonds in proteins.
DSDBASE is a database of disulphide bonds in proteins, which provides information on native disulphides and those that are stereochemically possible between pairs of residues for all known protein structural entries. The modelling of disulphides has been performed, using MODIP, by the identification of residue pairs that can strainlessly accommodate a covalent cross-link. We also assess the stereochemical quality of the covalent cross-link and grade them appropriately. One of the potential uses of the database is to design site-directed mutants in order to enhance the thermal stability of a protein. The proposed sites of mutations can be viewed specifically with respect to active sites of enzymes and across physiological dimers. The occurrence of native and modelled disulphides increases the dimensions of the database enormously. This database can also be employed for proposing three-dimensional models of disulphide-rich short polypeptides. The database can be accessed from http://www.ncbs.res.in/ approximately faculty/mini/dsdbase/dsdbase.html. Supplementary information can be accessed from http://www.ncbs.res.in/ approximately faculty/mini/dsdbase/nar/suppl.htm.


Thyronamines are substrates for human liver sulfotransferases.
Sulfotransferases (SULTs) catalyze the sulfation of many endogenous compounds that include monoamine neurotransmitters, such as dopamine (DA), and thyroid hormones (iodothyronines). Decarboxylation of iodothyronines results in formation of thyronamines. In the mouse, thyronamines act rapidly in a nongenomic fashion to initiate hypothermia and decrease cardiac output and heart rate. These effects are attenuated after 1-4 h, and metabolism of thyronamines via sulfation may be a mechanism for termination of thyronamine action. We carried out this study to test thyronamine (T0AM), 3-iodothyronamine (T1AM), 3,5-diiodothyronamine (T2AM), and 3,5,3'-triiodothyronamine (T3AM) as substrates for human liver and cDNA-expressed SULT activities. We characterized several biochemical properties of SULTs using the thyronamines that acted as substrates for SULT activities in a human liver high-speed supernatant pool (n=3). T1AM led to the highest SULT activity. Activities with T0AM and T3AM were 10-fold lower, and there was no detectable activity with T2AM. Thyronamines were then tested as substrates with eight cDNA-expressed SULTs (1A1, 1A2, 1A3, 1C2, 1E1, 2A1, 2B1a, and 2B1b). Expressed SULT1A3 had the greatest activity with T0AM, T1AM, and T3AM, whereas SULT1A1 showed similar activity only with T3AM. Expressed SULT1E1 had low activity with each substrate. T1AM, the most active thyronamine pharmacologically, was associated with the greatest SULT activity of the thyronamines tested in the liver pool and in both the expressed SULT1A3 and SULT1E1 preparations. Our results support the conclusion that sulfation contributes to the metabolism of thyronamines in human liver and that SULT activities may regulate the physiological effects of endogenous thyronamines.


Genetic applications of an inverse polymerase chain reaction.
A method is presented for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. The method uses the polymerase chain reaction (PCR), but it has the primers oriented in the reverse direction of the usual orientation. The template for the reverse primers is a restriction fragment that has been ligated upon itself to form a circle. This procedure of inverse PCR (IPCR) has many applications in molecular genetics, for example, the amplification and identification of sequences flanking transposable elements. In this paper we show the feasibility of IPCR by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.


Patient perspectives on barriers and enablers to the use and effectiveness of de-escalation techniques for the management of violence and aggression in mental health settings.
Investigate patient perspectives on barriers and enablers to the use and effectiveness of de-escalation techniques for aggression in mental health settings. De-escalation techniques are the recommended first-line intervention for the management of aggression in mental health settings internationally, yet use of higher risk restrictive practices persists. This indicates de-escalation techniques are not used at optimum frequency and/or there are important factors limiting their use and effect. Descriptive qualitative research using semi-structured interviews and Framework Analysis. Inpatient interviews (N = 26) exploring staff, patient and environmental factors influencing the use and effectiveness of staff de-escalation were conducted mid-2014. Three service user researchers led analysis. Data were synthesized in three deductive themes relating to staff, patient and environmental influences on the use and effectiveness of de-escalation techniques. The dominant view was that restrictive practices, rather than de-escalation techniques, are used in response to escalating patient behaviour. Under-use of de-escalation techniques was attributed to: lack of staff reflection on culture and practice and a need to retain control/dominance over patients. Ward rules, patient factors and a lack of staff respect for patients diluted their effectiveness. Participants identified a systematic process of de-escalation, rule subversion, reduced social distance and staff authenticity as enablers of effective de-escalation. This study investigated patient perspectives on staff, patient and environmental influences on the use and effectiveness of de-escalation techniques. Our framework of barriers and enablers provides indicators of organizational/behaviour change targets for interventions seeking to reduce violence and restrictive practices through enhanced de-escalation techniques.


Absence of aquaporin-4 expression in lesions of neuromyelitis optica but increased expression in multiple sclerosis lesions and normal-appearing white matter.
Aquaporin-4 (AQP4) has recently been implicated in the pathogenesis of neuromyelitis optica (NMO) where it has been identified as the first defined autoantigen pertinent to an inflammatory demyelinating disorder of the human CNS. Furthermore, a recent case report has shown a lack of AQP4 expression in the spinal cord lesions of NMO. However, the pattern of AQP4 expression in multiple sclerosis (MS) tissues has not been well-defined. In the present investigation we have confirmed a lack of expression of AQP4 in optic and spinal cord lesions in NMO which contrasted sharply with the increased levels of AQP4 expression seen in MS lesions. Furthermore a detailed immunohistochemical and semi-quantitative analysis is used to describe the expression pattern of AQP4 on well-characterized tissue microarray samples of MS and control white matter. Anatomically AQP4 was more highly expressed in all categories of MS tissue compared to normal control tissues with the most abundant expression in active lesions. Within active lesions AQP4 expression was significantly correlated with expression of the pro-inflammatory cytokine osteopontin. At the cellular level dual-labeling immunofluoresence demonstrated that increased expression of AQP4 was most pronounced at the astrocytic endfeet but was also associated with the cell bodies of astrocytes in the tissue parenchyma. The finding of increased AQP4 expression in MS lesions in contrast to the lack of expression in NMO lesions may suggest different mechanisms of initiation and progression between the two disease states.


Persistence of antibody in healthcare workers vaccinated against hepatitis B.
A cross-sectional serological survey was undertaken in 82 randomly selected high-risk healthcare workers previously vaccinated with the hepatitis B vaccine. The study design allowed for the identification and testing for hepatitis B surface antibody in equal numbers of employees in six-month intervals up to five years after vaccination. The results showed a consistent decline in antibody level with time, and an increasing proportion of participants with antibody levels below the commonly accepted protective level of greater than or equal to 10 S/N (sample counts/negative control counts) radioimmunoassay units. The percent of vaccinees whose S/N ratio was greater than 10 fell from 10% at one year, to 25% at two years, to over 50% at four years. Based on these figures, as well as the financial costs of hepatitis B in employees and the predictability of booster immunization, it was possible to assess the cost benefit of a hospital policy for reimmunization. At our institution, a strategy of revaccination at fixed intervals could save up to $200,000 over a ten-year period if revaccination was not offered and vaccine efficacy declined. The information obtained should help determine the need for revaccination and the advantages and disadvantages of alternative revaccination strategies.


Actions of adenosine on nitro blue tetrazolium deposition and surface pH during intestinal reperfusion injury.
Mesenteric arteries supplying an intestinal segment were occluded for 5 minutes and then released. During reperfusion, two series of measurements were made with various substances topically applied to the extraluminal surface. In the first series, reduced nitro blue tetrazolium (NBT) was extracted from tissue and measured spectrophotometrically, as an index of oxidative damage. In the second series, mucosal and serosal surface pH was measured as an index of the functional ability to maintain ion gradients. In control conditions, NBT deposition averaged 55-63 micrograms/g tissue. After 60 and 120 minutes of reperfusion, NBT was elevated to 446-479 micrograms/g, which was approximately half as large as the NBT increment (846 micrograms/g) produced by a 15-minute application of xanthine plus xanthine oxidase to well-perfused tissue. As expected, NBT levels were significantly lower (299 micrograms/g) in tissue that was continuously suffused with superoxide dismutase (SOD) plus catalase (CAT) before occlusion and during reperfusion. Similar NBT levels (274 micrograms/g) were observed after reperfusion in animals that were fed a diet supplemented with the antioxidant vitamin E for 4-6 weeks. These observations affirm that some, but not all, NBT deposition after reperfusion can be attributed to oxyradicals. However, with exogenous adenosine (ADO) applied for the first 30 minutes after occlusion, NBT was elevated to 174 micrograms/g after 60 minutes, which was only half as large as the increment with SOD plus CAT, even though those substances were continuously applied. The opposite effect was produced by an ADO receptor antagonist, 8-phenyltheophylline; NBT was increased to 516 micrograms/g.(ABSTRACT TRUNCATED AT 250 WORDS)


Regulation of fibrinolysis by platelet-released plasminogen activator inhibitor 1: light scattering and ultrastructural examination of lysis of a model platelet-fibrin thrombus.
We have investigated the role of plasminogen activator inhibitor 1 (PAI-1) in the regulation of fibrinolysis using a model thrombus composed of thrombin-stimulated platelets, fibrin(ogen), plasminogen, and recombinant tissue-type plasminogen activator. Laser light scattering kinetic measurements showed that clot lysis was significantly delayed both by thrombin-stimulated platelets and their cell-free releasate. This delay in lysis was almost fully reversed by the addition of a PAI-1-specific monoclonal antibody that blocks the ability of PAI-1 to inhibit plasminogen activators. Lysis half-times exhibited a linear dependence on the concentration of PAI-1 antigen present, as determined by enzyme-linked immunosorbent assay (ELISA). Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting confirmed the presence of PAI-1 antigen in the platelet releasates. Scanning electron micrographs of the model thrombus components sampled late in lysis showed considerable unproteolyzed fibrin still attached to platelets. Immunogold cytochemistry detected large amounts of PAI-1 antigen in the partially lysed platelet-fibrin thrombi. This PAI-1 appeared to be bound to the fibrin network rather than to the platelet surface itself. We conclude that the residual clots observed late in lysis represent platelet-associated fibrin to which platelet-released PAI-1 has bound, rendering it less susceptible to degradation.


Metallothionein genes: no association with Crohn's disease in a New Zealand population.
Metallothioneins (MTs) are excellent candidate genes for Inflammatory Bowel Disease (IBD) and have previously been shown to have altered expression in both animal and human studies of IBD. This is the first study to examine genetic variants within the MT genes and aims to determine whether such genetic variants have an important role in this disease. 28 tag SNPs in genes MT1 (subtypes A, B, E, F, G, H, M, X), MT2, MT3 and MT4 were selected for genotyping in a well-characterized New Zealand dataset consisting of 406 patients with Crohn's Disease and 638 controls. We did not find any evidence of association for MT genetic variation with CD. The lack of association indicates that genetic variants in the MT genes do not play a significant role in predisposing to CD in the New Zealand population.


Metabolic response to epigallocatechin-3-gallate in relapsing-remitting multiple sclerosis: a randomized clinical trial.
Muscle weakness and fatigue are common symptoms in multiple sclerosis (MS). Green tea catechins such as (-)epigallocatechin-3-gallate (EGCG) are known to improve energy metabolism at rest and during exercise. We tested the hypothesis that EGCG improves energy metabolism and substrate utilization in patients with MS. Eighteen patients (8 men) with relapsing-remitting MS (expanded disability status scale score <4.5, all receiving glatiramer acetate) participated in this randomized, double-blind, placebo-controlled, crossover trial at a clinical research center. All patients received EGCG (600 mg/d) and placebo over 12 wk (4-wk washout in between). After each intervention, fasting and postprandial energy expenditure (EE), as well as fat oxidation (FAOx) and carbohydrate oxidation (CHOx) rates, were measured either at rest or during 40 min of exercise (0.5 W/kg). At rest, blood samples and microdialysates from adipose tissue and skeletal muscle were also taken. At rest, postprandial EE and CHOx, as well as adipose tissue perfusion and glucose supply, were significantly lower in men but higher in women receiving EGCG compared with placebo. During exercise, postprandial EE was lower after EGCG than after placebo, indicating an increased working efficiency (men > women). After placebo, exercise EE was mainly fueled by FAOx in both men and women. After EGCG, there was a shift to a higher and more stable CHOx during exercise in men but not in women. Our data indicate that EGCG given to patients with MS over 12 wk improves muscle metabolism during moderate exercise to a greater extent in men than in women, possibly because of sex-specific effects on autonomic and endocrine control.


Cytopathic effect of lentogenic strains of Newcastle disease virus in cultured chicken muscle cells.
The ability of lentogenic strains of Newcastle disease virus (NDV) to inhibit the development of embryonic chicken muscle cells in vitro is described. In contrast to the uninfected control, which displayed a thick network of branched multinucleated muscle fibers, cultures infected with NDV exhibited a severely reduced potential to differentiate. The few differentiated muscle fibers that formed in the presence of NDV were poorly developed, displayed an attenuated morphology, and detached from the substrate 2 to 3 days before uninfected cultures showed signs of cell aging. When NDV-infected cultures were stained before muscle fiber detachment with a fluorescent-labeled NDV-specific antibody, the infected muscle fibers reacted strongly relative to neighboring myoblasts and fibroblasts. All six strains of NDV induced a similar inhibitory effect, and no strain-specific differences were detectable. These results indicate that differentiating muscle fibers are highly susceptible to infection by NDV in vitro and that this tissue exhibits an early NDV-induced cytopathic effect.


Sacrococcygeal teratoma. Outcome and management. An analysis of 17 cases.
The authors compare their experience of 17 cases of sacrococcygeal teratoma (SCT) with the literature in an attempt to clarify the natural history of this tumor and to identify factors related to its prognosis and management. The obstetrical, neonatal and surgical data were analyzed for 17 cases of SCT observed between July 1985 and December 1998. Three fetuses died in utero or shortly after birth. In the remaining 14, the tumors were removed. Twelve of the infants are currently tumor-free, with good sphincter control and lower-limb function. The remaining two died: one had a malignant tumor, and the other had a recurrence of an embryonal carcinoma. Recurrent tumors (mature histotypes) were also removed from two of the 12 patients who survived. Benign SCTs generally have favorable prognosis. Negative prognostic factors for SCT include solid tumors, those detected early in pregnancy, malignant histotypes, polyhydramnios, placentomegaly, and fetal hydrops.


Dietary fat in breast cancer survival.
Laboratory evidence suggests a plausible role for dietary fat in breast cancer pathophysiology. We conducted a systematic literature review to assess the epidemiological evidence on the impact of total dietary fat and fat subtypes, measured pre- and/or postcancer diagnosis, in relation to breast cancer-specific and all-cause mortality among breast cancer survivors. Studies were included if they were in English, had a sample size ≥200, and presented the hazard ratio/rate ratio for recurrence, disease-specific mortality, or all-cause mortality (n = 18). Although the results are mixed, most studies suggested that higher saturated fat intake prediagnosis was associated with increased risk of breast cancer-specific and all-cause mortality. Postdiagnostic trans fat intake was associated with a 45% and 78% increased risk of all-cause mortality. Higher monounsaturated fat intake before and after diagnosis was generally associated with increased risk of all-cause and breast cancer-specific mortality, albeit the majority of the studies were statistically nonsignificant. Two studies evaluating omega-3 fat intake suggested an inverse association with all-cause mortality. Although there were too few studies on fat subtypes to draw definitive conclusions, high consumption of saturated fat may exert a detrimental effect on breast cancer-specific and all-cause mortality, whereas omega-3 fat may be beneficial. The inconsistent and limited evidence warrants research to assess the impact of consumption of fat subtypes on breast cancer recurrence and mortality.


The Culture of Translational Science Research: Participants' Stories.
We apply a symbolic interactionist framework and a qualitative methodology to the examination of the everyday reality of translational science research (TSR). This is a growing scientific movement that aims to facilitate the efficient application of basic research to clinical service design and delivery. We describe the emerging culture of translational research at a mid-size medical center that received a Clinical and Translational Science Award from the National Institutes of Health. The stories related by scientists, clinicians, and students in interviews indicate that they make sense of the emerging inter- and cross-disciplinary, team-oriented culture of TSR through the refinement and redefinition of the significant symbols that inform their work while they attempt to master translational research by addressing the dilemmas it produces for them and their work. We see the strength, currency, adaptability, and energy of the core self-definition of "scientist" to be significant in shaping the emerging culture of translational research. We conclude by celebrating the value of interpretive ethnography for evaluation research.


Androgens exacerbate motor asymmetry in male rats with unilateral 6-hydroxydopamine lesion.
Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by dopamine neuron loss in the nigrostriatal pathway that shows greater incidence in men than women. The mechanisms underlying this gender bias remain elusive, although one possibility is that androgens may increase dopamine neuronal vulnerability to oxidative stress. Motor impairment can be modeled in rats receiving a unilateral injection of 6-hydroxydopamine (6-OHDA), a neurotoxin producing nigrostriatal degeneration. To investigate the role of androgens in PD, we compared young (2 months) and aged (24 months) male rats receiving gonadectomy (GDX) and their corresponding intact controls. One month after GDX, rats were unilaterally injected with 6-OHDA, and their motor impairment and asymmetry were assessed 2 weeks later using the cylinder test and the amphetamine-induced rotation test. Plasma samples were also collected to assess the concentration of testosterone and advanced oxidation protein products, a product of oxidative stress. GDX decreased lesion-induced asymmetry along with oxidative stress and increased amphetamine-induced rotations. These results show that GDX improves motor behaviors by decreasing motor asymmetry in 6-OHDA-treated rats, an effect that may be ascribed to increased release of striatal dopamine and decreased oxidative stress. Collectively, the data support the hypothesis that androgens may underlie the gender bias observed in PD.


Nucleotide sequence of a cloned structural gene coding for a precursor of pancreatic somatostatin.
We have constructed and cloned, in bacteria, recombinant plasmids containing DNA complementary to mRNA coding for a pancreatic pre-prosomatostatin, a product of the cell-free translation of pancreatic islet mRNAs shown previously by immunoprecipitation to be a precursor of somatostatin. A clone containing a nearly full-length cDNA insert of 550 base pairs was identified and appeared to contain the entire coding sequence for the somatostatin precursor in addition to portions of the 5' and 3' untranslated regions. mRNA coding for the pre-prosomatostatin is 600-630 bases long as determined by agarose gel electrophoresis and hybridization with labeled cDNA. Analyses of the nucleotide sequence of the cDNA revealed a protein of 119 amino acid beginning with methionine followed by a typical leader sequence containing 18 hydrophobic amino acids. The tetradecapeptide somatostatin, identical in sequence to mammalian hypothalamic somatostatin, is located at the carboxy terminus followed immediately by a stop codon. An ARg-Lys sequence immediately preceding the sequence of somatostatin is typical of a prohormone cleavage site. A sequence Ala-Pro-Arg-Glu preceding the Arg-Lys cleavage site is identical to that found in porcine prosomatostatin. The evolutionary conservation of the identical amino acid sequence of the somatostatin tetradecapeptide from fish to mammals is remarkable. In addition, similar conservation, in fish and mammals, of the sequence Ala-Pro-Arg-Glu-Arg-Lys preceding the coding region for somatostatin suggets that this particular sequence may have biologic importance in cellular processing of the somatostatin precursor.


Protective mechanisms of inosine in platelet activation and cerebral ischemic damage.
Inosine is a naturally occurring nucleoside degraded from adenosine. Recent studies have demonstrated that inosine has potent immunomodulatory and neuroprotective effects. In the present study, we further investigated the inhibitory effects of inosine on platelet activation in vitro and in vivo, as well as in attenuating middle cerebral artery occlusion (MCAO)-induced focal cerebral ischemia in rats. Inosine concentration-dependently (0.5 to 6.0 mmol/L) inhibited platelet aggregation stimulated by agonists. Inosine (1.5 and 3.0 mmol/L) inhibited phosphoinositide breakdown, [Ca+2]i, and TxA2 formation in human platelets stimulated by collagen (1 microg/mL). In addition, inosine (1.5 and 3.0 mmol/L) markedly increased levels of cyclic guanylate monophosphate (GMP) and cyclic GMP-induced vasodilator-stimulated phosphoprotein Ser157 phosphorylation. Rapid phosphorylation of a platelet protein of molecular weight 47,000 (P47), a marker of protein kinase C activation, was triggered by collagen (1 microg/mL). This phosphorylation was markedly inhibited by inosine (3.0 mmol/L). Inosine (1.5 and 3.0 mmol/L) markedly reduced hydroxyl radical in collagen (1 microg/mL)-activated platelets. In in vivo studies, inosine (400 mg/kg) significantly prolonged the latency period of inducing platelet plug formation in mesenteric venules of mice, and administration of 2 doses (100 mg/kg) or a single dose (150 mg/kg) of inosine significantly attenuated MCAO-induced focal cerebral ischemia in rats. Platelet aggregation contributes significantly to MCAO-induced focal cerebral ischemia. The most important findings of this study suggest that inosine markedly inhibited platelet activation in vitro and in vivo, as well as cerebral ischemia. Thus, inosine treatment may represent a novel approach to lowering the risk of or improving function in thromboembolic-related disorders and ischemia-reperfusion brain injury.


Loss of function mutations in the gene encoding Omi/HtrA2 in Parkinson's disease.
Recently targeted disruption of Omi/HtrA2 has been found to cause neurodegeneration and a parkinsonian phenotype in mice. Using a candidate gene approach, we performed a mutation screening of the Omi/HtrA2 gene in German Parkinson's disease (PD) patients. In four patients, we identified a novel heterozygous G399S mutation, which was absent in healthy controls. Moreover, we identified a novel A141S polymorphism that was associated with PD (P<0.05). Both mutations resulted in defective activation of the protease activity of Omi/HtrA2. Immunohistochemistry and functional analysis in stably transfected cells revealed that S399 mutant Omi/HtrA2 and to a lesser extent, the risk allele of the A141S polymorphism induced mitochondrial dysfunction associated with altered mitochondrial morphology. Cells overexpressing S399 mutant Omi/HtrA2 were more susceptible to stress-induced cell death than wild-type. On the basis of functional genomics, our results provide a novel link between mitochondrial dysfunction and neurodegeneration in PD.


Effects of Chinese herbal medicine Fuzhisan on autologous neural stem cells in the brain of SAMP-8 mice.
Fuzhisan (FZS), a Chinese herbal complex prescription, has been used in the treatment of Alzheimer's disease (AD) for more than 16 years. However the underlying mechanism remains to be explored. The effects of the aqueous extract of FZS on the cognitive functions of the aged mice and the pharmacological basis for its therapeutic efficacy were investigated. The results showed that FZS improved impaired cognitive ability of aged SAMP-8 mice. FZS (2.4, 4.8 g/kg/d) increased hippocampal neurogenesis and the long-term survival of BrdU-labeled cells without affecting the proportion of BrdU-positive neurons and glial cells. FZS also increased the number of BrdU-positive cells in the subventricular zone (SVZ) of the lateral ventricles of 8-month-old SAMP-8 mice. These studies suggest that FZS upregulates neurogenesis by increasing proliferation of neural progenitor cells and prolonging survival of the newborn cells in the hippocampal DG. FZS may be beneficial for the treatment of senile dementia, especially Alzheimer's disease.


The Use of Poly-4-Hydroxybutyrate (P4HB) Scaffold in the Ptotic Breast: A Multicenter Clinical Study.
Mastopexy and reduction mammaplasty are often limited by the patient's poor native soft tissue quality, resulting in ptosis recurrence and loss of rejuvenated surgical results. Surgical scaffolds and acellular dermal matrices are used in these procedures to provide physical and mechanical stabilization of weakened or compromised tissue. GalaFLEX scaffold, made from poly-4-hydroxybutyrate (P4HB), is a next-generation product for soft tissue reinforcement that resorbs gradually while aiding tissue regeneration to achieve excellent outcomes. To assess the clinical performance of GalaFLEX scaffold in soft tissue reinforcement during elective mastopexy and reduction mammaplasty. This multicenter, single-arm, observational study assessed product performance and outcomes of GalaFLEX scaffold when used in breast surgery. Outcomes included ptosis correction and maintenance, associated adverse events, patient and surgeon satisfaction, and mammographic and ultrasound imaging evaluation. At 6 centers in the US, 62 of 69 enrolled patients were treated. Of this population, 89.7% had successful ptosis correction and maintenance at 1 year, with high patient and surgeon satisfaction for breast shape, droop/sag of the breast, and maintenance of results at 1 year. There were 5 adverse events deemed related to the device (8.0%), including nerve pain, breast swelling, ptosis, and 2 instances of asymmetry. GalaFLEX scaffold safely and successfully supports and elevates breast tissue in mastopexy and reduction mammaplasty, with maintained support at 1 year. Surgeon and patient satisfaction were high. No mammogram or ultrasound interference was detected.


Hydrodynamics of isotropic and liquid crystalline active polymer solutions.
We describe the large-scale collective behavior of solutions of polar biofilaments and stationary and mobile crosslinkers. Both mobile and stationary crosslinkers induce filament alignment promoting either polar or nematic order. In addition, mobile crosslinkers, such as clusters of motor proteins, exchange forces and torques among the filaments and render the homogeneous states unstable via filament bundling. We start from a Smoluchowski equation for rigid filaments in solutions, where pairwise crosslink-mediated interactions among the filaments yield translational and rotational currents. The large-scale properties of the system are described in terms of continuum equations for filament and motor densities, polarization, and alignment tensor obtained by coarse-graining the Smoluchovski equation. The possible homogeneous and inhomogeneous states of the systems are obtained as stable solutions of the dynamical equations and are characterized in terms of experimentally accessible parameters. We make contact with work by other authors and show that our model allows for an estimate of the various parameters in the hydrodynamic equations in terms of physical properties of the crosslinkers.


Comparison of the primary stability of orthodontic miniscrews after repeated insertion cycles.
To compare the primary stability of miniscrews after repeated cycles of insertion through insertion torque (IT) measurements and resonance frequency analysis (RFA). Sixty titanium miniscrews were divided into two groups according to the insertion protocol: one with predrilled sites and the other self-drilled into porcine iliac crest bone specimens. Each group had three cycles of reinsertion. After each insertion, the IT and RFA were measured. The IT was measured by using a torque meter, and the RFA was measured using the Osstell ISQ device. A total of five miniscrews of each group were selected for sequential assessment of the morphology of their tip and threads using scanning electron microscopy after each insertion cycle. No statistically significant differences were found in the IT values of miniscrews reinserted up to three times in the group with predrilled surgical sites. The IT value increased significantly with the number of reinsertions in the self-drilled group. The RFA value decreased as the number of insertions increased in both groups. Under the conditions of this in vitro study, reinserting miniscrews deteriorates the integrity of their tip and thread. Reinsertion should be discouraged particularly when insertion sites are not predrilled.


In the clinic. Constipation.
This issue provides a clinical overview of constipation, focusing on prevention, diagnosis, treatment, and patient information. The content of In the Clinic is drawn from the clinical information and education resources of the American College of Physicians (ACP), including ACP Smart Medicine and MKSAP (Medical Knowledge and Self-Assessment Program). Annals of Internal Medicine editors develop In the Clinic from these primary sources in collaboration with the ACP's Medical Education and Publishing divisions and with the assistance of science writers and physician writers. Editorial consultants from ACP Smart Medicine and MKSAP provide expert review of the content. Readers who are interested in these primary resources for more detail can consult http://smartmedicine.acponline.org, http://mksap.acponline.org, and other resources referenced in each issue of In the Clinic.


Piloting a needs assessment to guide development of a survivorship program for a community cancer center.
To develop and pilot a survey to assess needs and distress of cancer survivors receiving care in a community cancer center. Descriptive, quantitative. A community cancer center in the southeastern United States partnering with a local college of nursing faculty. Convenience sample of 307 adult cancer survivors. Voluntary completion of a modified survey of needs. Cancer survivor needs and distress according to five subscales (physical effects, social issues, emotional aspects, spiritual issues, and other issues), age, and gender. Patients on average reported experiencing more than 25 of 50 possible survivorship needs. Average distress scores associated with individual needs were low. The most frequently experienced needs were fatigue, fear of recurrence, and sleep disturbance. Middle-aged survivors experienced significantly greater need and distress across all subscales. Need and distress exist among adult cancer survivors receiving treatment and follow-up in community cancer care settings, with the middle-age phase of life creating unique barriers. Survey data may provide documentation of the multidimensional impact of cancer on quality of life and can help direct survivorship program development. Nurses can address a barrier to survivorship care in community care settings by using the Pearlman-Mayo Survey of Needs to assess outcomes relevant to survivors. Partnership between community hospital RNs and college of nursing faculty may create local or regional solutions and serve as useful models for survivorship care.


Kindled seizures result in decreased responsiveness of benzodiazepine receptors to gamma-aminobutyric acid (GABA).
Repeated kindled seizures result in increased numbers of benzodiazepine receptors in fascia dentata membranes. Our previous work localized these receptors to a discrete neuronal population, the dentate granule cells. The present investigation characterizes the regulation of this binding by NaCl, pentobarbital and gamma-aminobutyric acid (GABA). [3H]Flunitrazepam binding in membranes from kindled rats exceeded that from controls by 24%. NaCl increased this binding in membranes prepared from control and kindled animals by roughly equivalent amounts (18 and 14%, respectively). No significant differences were found in the effects of pentobarbital on benzodiazepine binding between kindled and control groups. In contrast, addition of GABA increased benzodiazepine binding by a significantly smaller percentage in the kindled (116 +/- 5%) compared to the control (138 +/- 11%) group. We propose that this altered molecular response to GABA is intimately related to the enhanced GABA-mediated inhibition of granule cells present after kindled seizures.


Transcranial Doppler ultrasonography in the assessment of cerebral circulation arrest: improving sensitivity by transcervical and transorbital carotid insonation and serial examinations.
Transcranial Doppler (TCD) can detect the cerebral circulation arrest (CCA) in brain death. TCD is highly specific, but less sensitive because of false-negatives accounting for up to 10%. The aim of the study was to explore the diagnostic accuracy of TCD and to determine whether it can be augmented by strategies such as the insonation of the extracranial internal carotid artery (ICA) and sequential examinations. Data of 184 patients, who met clinical criteria of brain death, observed from 1998 through 2006, were retrospectively reviewed. The study of cerebral arteries was performed through the transtemporal approach, suboccipital insonation of the vertebro-basilar system, transorbital insonation of the ICA and ophthalmic artery, and transcervical insonation of the extracranial ICA. Repeated exams were performed in cases of persistent diastolic flow. The specificity of the testing was 100%, no false-positive cases were recorded. The sensitivity of conventional TCD examination was 82.1%. The insonation of the extracranial ICA increased sensitivity to 88% allowing the detection of CCA in those patients lacking temporal windows; serial examinations further increased sensitivity to 95.6%. The addition of insonation of the cervical ICA and of the siphon increased sensitivity of TCD. Nevertheless, a CCA flow patterns may appear later on those segments. Serial examinations, may be needed in those cases.


Epidemiology of intussusception before and after rotavirus vaccine introduction in Fiji.
In 2012, Fiji introduced rotavirus vaccine (Rotarix, GSK) into the national immunisation schedule. We describe the intussusception epidemiology prior to rotavirus vaccine, temporal association of intussusception cases to administration of rotavirus vaccine, and estimate the additional number of intussusception cases that may be associated with rotavirus vaccine. A retrospective review of intussusception cases for children aged <24 months old was undertaken between January 2007 and October 2012 pre-vaccine. All admissions and deaths with a discharge diagnosis of intussusception, bowel obstruction, paralytic ileus, or intussusception ICD10-AM codes were extracted from national databases and hospital records. Nationwide active intussusception surveillance was established for three years post-vaccine (2013-2015). There were 24 definite intussusception cases in the pre-rotavirus vaccine period, 96% were confirmed by surgery. The median age was 6.5 months. The incidence rate was 22.2 (95% CI: 13.9-33.7) per 100,000 infants. There were no deaths. Active surveillance identified 25 definite intussusception cases, 96% of which were among children who were age-eligible for rotavirus vaccine. None were potentially vaccine related. We estimated one to five additional cases of intussusception every five years. The incidence of intussusception pre-rotavirus vaccine in Fiji is low. Intussusception associated with rotavirus vaccine is likely a rare event in Fiji.


Patient safety skills in primary care: a national survey of GP educators.
Clinicians have a vital role in promoting patient safety that goes beyond their technical competence. The qualities and attributes of the safe hospital doctor have been explored but similar work within primary care is lacking. Exploring the skills and attributes of a safe GP may help to inform the development of training programmes to promote patient safety within primary care. This study aimed to determine the views of General Practice Educational Supervisors (GPES) regarding the qualities and attributes of a safe General Practitioner (GP) and the perceived trainability of these 'safety skills' and to compare selected results with those generated by a previous study of hospital doctors. This was a two-stage study comprising content validation of a safety skills questionnaire (originally developed for hospital doctors) (Stage 1) and a prospective survey of all GPES in Scotland (n = 691) (Stage 2). Stage 1: The content-validated questionnaire comprised 66 safety skills/attributes across 17 broad categories with an overall content validation index of 0.92. Stage 2: 348 (50%) GPES completed the survey. GPES felt the skills/attributes most important to being a safe GP were honesty (93%), technical clinical skills (89%) and conscientiousness (89%). That deemed least important/relevant to being a safe GP was leadership (36%). This contrasts sharply with the views of hospital doctors in the previous study. GPES felt the most trainable safety skills/attributes were technical skills (93%), situation awareness (75%) and anticipation/preparedness (71%). The least trainable were honesty (35%), humility (33%) and patient awareness/empathy (30%). Additional safety skills identified as relevant to primary care included patient advocacy, negotiation skills, accountability/ownership and clinical intuition ('listening to that worrying little inner voice'). GPES believe a broad range of skills and attributes contribute to being a safe GP. Important but subtle differences exist between what primary care and secondary care doctors perceive as core safety attributes. Educationalists, GPs and patient safety experts should collaborate to develop and implement training in these skills to ensure that current and future GPs possess the necessary competencies to engage and lead in safety improvement efforts.