Optimization of multi-electrode implant configurations and programming for the delivery of non-ablative electric fields in intratumoral modulation therapy.
Application of low intensity electric fields to interfere with tumor growth is being increasingly recognized as a promising new cancer treatment modality. Intratumoral modulation therapy (IMT) is a developing technology that uses multiple electrodes implanted within or adjacent tumor regions to deliver electric fields to treat cancer. In this study, the determination of optimal IMT parameters was cast as a mathematical optimization problem, and electrode configurations, programming, optimization, and maximum treatable tumor size were evaluated in the simplest and easiest to understand spherical tumor model. The establishment of electrode placement and programming rules to maximize electric field tumor coverage designed specifically for IMT is the first step in developing an effective IMT treatment planning system. Finite element method electric field computer simulations for tumor models with 2 to 7 implanted electrodes were performed to quantify the electric field over time with various parameters, including number of electrodes (2 to 7), number of contacts per electrode (1 to 3), location within tumor volume, and input waveform with relative phase shift between 0 and 2π radians. Homogeneous tissue specific conductivity and dielectric values were assigned to the spherical tumor and surrounding tissue volume. In order to achieve the goal of covering the tumor volume with a uniform threshold of 1 V/cm electric field, a custom least square objective function was used to maximize the tumor volume covered by 1 V/cm time averaged field, while maximizing the electric field in voxels receiving less than this threshold. An additional term in the objective function was investigated with a weighted tissue sparing term, to minimize the field to surrounding tissues. The positions of the electrodes were also optimized to maximize target coverage with the fewest number of electrodes. The complexity of this optimization problem including its non-convexity, the presence of many local minima, and the computational load associated with these stochastic based optimizations led to the use of a custom pattern search algorithm. Optimization parameters were bounded between 0 and 2π radians for phase shift, and anywhere within the tumor volume for location. The robustness of the pattern search method was then evaluated with 50 random initial parameter values. The optimization algorithm was successfully implemented, and for 2 to 4 electrodes, equally spaced relative phase shifts and electrodes placed equidistant from each other was optimal. For 5 electrodes, up to 2.5 cm diameter tumors with 2.0 V, and 4.1 cm with 4.0 V could be treated with the optimal configuration of a centrally placed electrode and 4 surrounding electrodes. The use of 7 electrodes allow for 3.4 cm diameter coverage at 2.0 V and 5.5 cm at 4.0 V. The evaluation of the optimization method using 50 random initial parameter values found the method to be robust in finding the optimal solution. This study has established a robust optimization method for temporally optimizing electric field tumor coverage for IMT, with the adaptability to optimize a variety of parameters including geometrical and relative phase shift configurations.


Ameliorative effects of hempseed (Cannabis sativa) against hypercholesterolemia associated cardiovascular changes.
Hypercholesterolemia (HC) is a major risk factor for cardiovascular (CV) diseases, that are the major cause of mortality worldwide. Free radicals mediated oxidative stress is a critical player in HC-associated pathophysiological insults including atherosclerosis. Unwanted side effects associated with statins, COX-2 inhibitors, and other synthetic drugs limit their use. Thus, modulation of oxidative stress during HC using green pharmaceuticals seems an appropriate approach against deleterious CV consequences without noticeable side-effect. In this regard, owing to an abundance of proteins, fiber and optimal ratios of omega 6 PUFA: omega-3 PUFA in Hempseed (HS), we aim to exploit its anti-inflammatory and antioxidant properties to ameliorate HC- associated CV effects. Comparing the antioxidant capacity of protein and lipid fractions of HS using ABTS and DPPH assays, HS was supplemented to high-fat diets (HFD) induced hypercholesterolemic wistar rats. After treatment schedules, lipid profiles, histological and ultrastructural investigations, gene and protein expressions of inflammatory markers, markers of oxidative stress were studied and correlated with biophysical parameters such as ECG and impedance/conductance across the aorta. HS demonstrating in vitro free radical scavenging activity, ameliorated the signs of HC as seen with improved lipid profiles, aortic tissue damage and ECG patterns compared to HFD groups. HS administration also relieved the COX-2 mediated inflammation, which correlated well with the improved redox status in the tissue. Current study evidently demonstrates that the anti-hypercholesterolemic effects of HS are mediated through redox-sensitive modulation of inflammatory pathways.


Inter-domain communication of human cystathionine β-synthase: structural basis of S-adenosyl-L-methionine activation.
Cystathionine β-synthase (CBS) is a key enzyme in sulfur metabolism, and its inherited deficiency causes homocystinuria. Mammalian CBS is modulated by the binding of S-adenosyl-l-methionine (AdoMet) to its regulatory domain, which activates its catalytic domain. To investigate the underlying mechanism, we performed x-ray crystallography, mutagenesis, and mass spectrometry (MS) on human CBS. The 1.7 Å structure of a AdoMet-bound CBS regulatory domain shows one AdoMet molecule per monomer, at the interface between two constituent modules (CBS-1, CBS-2). AdoMet binding is accompanied by a reorientation between the two modules, relative to the AdoMet-free basal state, to form interactions with AdoMet via residues verified by mutagenesis to be important for AdoMet binding (Phe(443), Asp(444), Gln(445), and Asp(538)) and for AdoMet-driven inter-domain communication (Phe(443), Asp(538)). The observed structural change is further supported by ion mobility MS, showing that as-purified CBS exists in two conformational populations, which converged to one in the presence of AdoMet. We therefore propose that AdoMet-induced conformational change alters the interface and arrangement between the catalytic and regulatory domains within the CBS oligomer, thereby increasing the accessibility of the enzyme active site for catalysis.


Identification of a minimal transforming domain of p53: negative dominance through abrogation of sequence-specific DNA binding.
Mutations in the p53 gene are most frequent in cancer. Many p53 mutants possess transforming activity in vitro. In cells transformed by such mutants, the mutant protein is oligomerized with endogenous cell p53. To determine the relevance of oligomerization for transformation, miniproteins containing C-terminal portions of p53 were generated. These miniproteins, although carrying no point mutation, transformed at least as efficiently as full-length mutant p53. Transforming activity was coupled with the ability to oligomerize with wild-type p53, as well as with the ability to abrogate sequence-specific DNA binding by coexpressed wild-type p53. These findings suggest that p53-mediated transformation may operate through a dominant negative mechanism, involving the generation of DNA binding-incompetent oligomers.


Simplified dietary acute tryptophan depletion: effects of a novel amino acid mixture on the neurochemistry of C57BL/6J mice.
Diet and nutrition can impact on the biological processes underpinning neuropsychiatric disorders. Amino acid (AA) mixtures lacking a specific neurotransmitter precursor can change the levels of brain serotonin (5-HT) or dopamine (DA) in the central nervous system. The availability of these substances within the brain is determined by the blood-brain barrier (BBB) that restricts the access of peripheral AA into the brain. AA mixtures lacking tryptophan (TRP) compete with endogenous TRP for uptake into the brain across the BBB, which in turn leads to a decrease in central nervous 5-HT synthesis. The present study compared the effects of a simplified acute tryptophan depletion (SATD) mixture in mice on blood and brain serotonergic and dopaminergic metabolites to those of a commonly used acute tryptophan depletion mixture (ATD Moja-De) and its TRP-balanced control (BAL). The SATD formula is composed of only three large neutral AAs: phenylalanine (PHE), leucine (LEU), and isoleucine (ILE). BAL, ATD Moja-De, or SATD formulas were delivered to adult male C57BL/6J mice by gavage. TRP, monoamines, and their metabolites were quantified in blood and brain regions (hippocampus, frontal cortex, amygdala, caudate putamen, and nucleus accumbens). Both ATD Moja-De and SATD significantly decreased levels of serum and brain TRP, as well as brain 5-HIAA and 5-HT compared with BAL. SATD reduced HVA levels in caudate but did not alter total DA levels or DOPAC. SATD decreased TRP and serotonergic metabolites comparably to ATD Moja-De administration. A simplified and more palatable combination of AAs can manipulate serotonergic function and might be useful to reveal underlying monoamine-related mechanisms contributing to different neuropsychiatric disorders.


Is malarial placental infection related to peripheral infection at any time of pregnancy?
Placental infection measured by placental smear at delivery is a standard indicator, widely used to characterize malaria infection in pregnant women. However, a single measure can hardly reflect the entire history of infection during pregnancy. To investigate the relation between this indicator and peripheral infection during pregnancy, we used data collected in a randomized trial of malaria prophylaxis in 928 pregnant women in Burkina Faso, 1987-1988, during which repeated measures of peripheral infection were taken. We analyzed placental infection using a logistic model, with two methods for handling missing data. Peripheral infection during two periods of pregnancy was significantly related to placental infection at delivery, before the fifth month: OR = 2.9 [1.3; 6.3]; after 7 months: OR = 4.9 [2.7; 8.8]). Therefore, an early peripheral infection may persist throughout gestation, and placental infection is a good indicator of the women's parasitological status during pregnancy.


Reduced level of glutamic acid decarboxylase-67 kDa in the prefrontal cortex in major depression.
Accumulating evidence suggests dysfunction of the gamma-aminobutyric acid (GABA) system in major depressive disorder (MDD). Neuroimaging studies consistently report reductions of cortical GABA in depressed patients. Our post-mortem analyses demonstrate a reduction in the density and size of GABAergic interneurons in the dorsolateral prefrontal cortex (DLPFC) in MDD. The goal of this study was to test whether the level of glutamic acid decarboxylase (GAD), the GABA synthesizing enzyme, will also be reduced in the same cortical region in MDD. Levels of GAD-65 and GAD-67 proteins were investigated by Western blotting in samples from the DLPFC (BA 9) in 13 medication-free subjects with MDD, and 13 psychiatrically healthy controls. The overall amount of GAD-67 was significantly reduced (-34%) in depressed subjects compared to matched controls. Since recent neuroimaging studies have demonstrated that antidepressants modulate GABA levels, additional experiments were performed to examine the levels of GAD in eight depressed subjects treated with antidepressant medications. Levels of GAD-67 were unchanged in these depressed subjects compared to their respective controls (n=8). The overall amounts of GAD-65 were similar in depressed subjects compared to matched controls, regardless of antidepressant medication. Reduced levels of GAD-67, which is localized to somata of GABA neurons, further support our observation of a decreased density of GABAergic neurons in the PFC in depression. It is likely that a decrease in GAD-67 accounts for the reduction in GABA levels revealed by neuroimaging studies. Moreover, our data support previous neuroimaging observations that antidepressant medication normalizes GABA deficits in depression.


Viral determinants of human papillomavirus persistence following loop electrical excision procedure treatment for cervical intraepithelial neoplasia grade 2 or 3.
Persistent infection with carcinogenic human papillomavirus (HPV) causes cervical precancer (cervical intraepithelial neoplasia grade 2+) which, in the United States, is commonly treated using the loop electrical excision procedure (LEEP). Data from Atypical Squamous Cells of Undetermined Significance-Low-Grade Squamous Intraepithelial Lesion Triage Study were used to evaluate HPV persistence and reappearance after LEEP. Cervical specimens, collected before LEEP and at 6-month study visits, were tested by L1-PCR for detection of >or=27 HPV types. HPV persistence, defined as the same HPV type being present before and 6 months after LEEP, was evaluated by: (a) genotype, (b) carcinogenicity, and (c) phylogenetic species. HPV infections that cleared post-LEEP (the complement of persistence) were followed for reappearance of the same type. HPV infections (n = 1,130) were detected among 481 women who underwent LEEP. Overall, 20.4% [95% confidence interval (95% CI), 18.2-22.9%] of infections persisted. Assessment of heterogeneity within the three categorizations of HPV showed that phylogenetic species best fit the data. Persistence was significantly greater by HPV types in the alpha3 species [all are noncarcinogenic; 40.9% (95% CI, 31.8-50.4%)] compared with HPV types in the alpha9 (HPV16 and related types) and alpha7 species (HPV18 and related types; 17.6% and 17.9%, respectively; P < 0.001 for both). HPV reappeared in 7.8% (95% CI, 6.1-9.9%) of infections that cleared after LEEP. Infections in the alpha3 species (22.6%) were the most likely to reappear compared with HPV types in the alpha9 (7.5%) and alpha7 (6.8%) species. Patterns of HPV persistence and reappearance following LEEP were better explained by phylogenetic rather than standard classifications. HPV types likely to persist after LEEP as well as those likely to repopulate the cervical/vaginal epithelium were those in the alpha3 species, which are in effect not treated, but are not associated with cervical cancer and are unlikely to cause disease.


Body weight and body composition in old age and their relationship with frailty.
Aging is associated with various changes in body composition, including changes in weight, loss of muscle mass, and increase in fat mass. This article describes the role of body weight and body composition, and their changes, in the risk of frailty in old age. Based on current literature, observational studies on obesity and high waist circumference show most convincing results for an association with frailty. The independent role of muscle mass and muscle fat infiltration remains unclear, mainly due to a lack of studies and a lack of accurate measurement of body composition by computed tomography or MRI. Weight loss and exercise training intervention studies can be of benefit to frail older adults. Obesity and high waist circumference may be important determinants of frailty in old age, whereas the role of muscle mass and muscle fat infiltration is still unclear. More prospective studies that will specifically focus on frailty as an outcome measure are needed to identify specific body composition components as potential targets for the prevention of frailty in old age.


Textile integrated sensors and actuators for near-infrared spectroscopy.
Being the closest layer to our body, textiles provide an ideal platform for integrating sensors and actuators to monitor physiological signals. We used a woven textile to integrate photodiodes and light emitting diodes. LEDs and photodiodes enable near-infrared spectroscopy (NIRS) systems to monitor arterial oxygen saturation and oxygenated and deoxygenated hemoglobin in human tissue. Photodiodes and LEDs are mounted on flexible plastic strips with widths of 4 mm and 2 mm, respectively. The strips are woven during the textile fabrication process in weft direction and interconnected with copper wires with a diameter of 71 μm in warp direction. The sensor textile is applied to measure the pulse waves in the fingertip and the changes in oxygenated and deoxygenated hemoglobin during a venous occlusion at the calf. The system has a signal-to-noise ratio of more than 70 dB and a system drift of 0.37% ± 0.48%. The presented work demonstrates the feasibility of integrating photodiodes and LEDs into woven textiles, a step towards wearable health monitoring devices.


The logic of Ménage à Trois.
Recent studies of socially monogamous species have shown that in many cases females do not copulate exclusively with their pair mates, but are also receptive to other males. The explanation usually given for unfaithful female behavior is that most females are unable to bond with a male they would prefer as genetic father to their offspring. To secure male assistance the female therefore pairs with an available male but also copulates with males of supposedly higher genetic quality. Here we offer an alternative evolutionary explanation for female infidelity, which does not rely upon this 'Good Genes hypothesis of female choice. We show with a simple model that in an evolutionary game between three players, a male, a female and a male lover, solutions exist in which the female can secure more assistance from her mate by being receptive to other males. We conclude that female sexuality can have a decisive role in regulating social behaviour, in which the fertile female is the driving force.


An analysis of enzyme kinetics data for mitochondrial DNA strand termination by nucleoside reverse transcription inhibitors.
Nucleoside analogs used in antiretroviral treatment have been associated with mitochondrial toxicity. The polymerase-gamma hypothesis states that this toxicity stems from the analogs' inhibition of the mitochondrial DNA polymerase (polymerase-gamma) leading to mitochondrial DNA (mtDNA) depletion. We have constructed a computational model of the interaction of polymerase-gamma with activated nucleoside and nucleotide analog drugs, based on experimentally measured reaction rates and base excision rates, together with the mtDNA genome size, the human mtDNA sequence, and mitochondrial dNTP concentrations. The model predicts an approximately 1000-fold difference in the activated drug concentration required for a 50% probability of mtDNA strand termination between the activated di-deoxy analogs d4T, ddC, and ddI (activated to ddA) and the activated forms of the analogs 3TC, TDF, AZT, FTC, and ABC. These predictions are supported by experimental and clinical data showing significantly greater mtDNA depletion in cell culture and patient samples caused by the di-deoxy analog drugs. For zidovudine (AZT) we calculated a very low mtDNA replication termination probability, in contrast to its reported mitochondrial toxicity in vitro and clinically. Therefore AZT mitochondrial toxicity is likely due to a mechanism that does not involve strand termination of mtDNA replication.


Contributions and challenges of cross-national comparative research in migration, ethnicity and health: insights from a preliminary study of maternal health in Germany, Canada and the UK.
Public health researchers are increasingly encouraged to establish international collaborations and to undertake cross-national comparative studies. To-date relatively few such studies have addressed migration, ethnicity and health, but their number is growing. While it is clear that divergent approaches to such comparative research are emerging, public health researchers have not so far given considered attention to the opportunities and challenges presented by such work. This paper contributes to this debate by drawing on the experience of a recent study focused on maternal health in Canada, Germany and the UK. The paper highlights various ways in which cross-national comparative research can potentially enhance the rigour and utility of research into migration, ethnicity and health, including by: forcing researchers to engage in both ideological and methodological critical reflexivity; raising awareness of the socially and historically embedded nature of concepts, methods and generated 'knowledge'; increasing appreciation of the need to situate analyses of health within the wider socio-political setting; helping researchers (and research users) to see familiar issues from new perspectives and find innovative solutions; encouraging researchers to move beyond fixed 'groups' and 'categories' to look at processes of identification, inclusion and exclusion; promoting a multi-level analysis of local, national and global influences on migrant/minority health; and enabling conceptual and methodological development through the exchange of ideas and experience between diverse research teams. At the same time, the paper alerts researchers to potential downsides, including: significant challenges to developing conceptual frameworks that are meaningful across contexts; a tendency to reify concepts and essentialise migrant/minority 'groups' in an effort to harmonize across countries; a danger that analyses are superficial, being restricted to independent country descriptions rather than generating integrated insights; difficulties of balancing the need for meaningful findings at country level and more holistic products; and increased logistical complexity and costs. In view of these pros and cons, the paper encourages researchers to reflect more on the rationale for, feasibility and likely contribution of proposed cross-national comparative research that engages with migration, ethnicity and health and suggests some principles that could support such reflection.


The role of spatial attention in the processing of facial expression: an ERP study of rapid brain responses to six basic emotions.
To investigate the time course of emotional expression processing, we recorded ERP responses to stimulus arrays containing neutral versus angry, disgusted, fearful, happy, sad, or surprised faces. In one half of the experiment, the task was to discriminate emotional and neutral facial expressions. Here, an enhanced early frontocentral positivity was elicited in response to emotional as opposed to neutral faces, followed by a broadly distributed positivity and an enhanced negativity at lateral posterior sites. These emotional expression effects were very similar for all six basic emotional expressions. In the other half of the experiment, attention was directed away from the faces toward a demanding perceptual discrimination task. Under these conditions, emotional expression effects were completely eliminated, demonstrating that brain processes involved in the detection and analysis of facial expression require focal attention. The face-specific N170 component was unaffected by any emotional expression, supporting the hypothesis that structural encoding and expression analysis are independent processes.


Differential impact of tetratricopeptide repeat proteins on the steroid hormone receptors.
Tetratricopeptide repeat (TPR) motif containing co-chaperones of the chaperone Hsp90 are considered control modules that govern activity and specificity of this central folding platform. Steroid receptors are paradigm clients of Hsp90. The influence of some TPR proteins on selected receptors has been described, but a comprehensive analysis of the effects of TPR proteins on all steroid receptors has not been accomplished yet. We compared the influence of the TPR proteins FK506 binding proteins 51 and 52, protein phosphatase-5, C-terminus of Hsp70 interacting protein, cyclophillin 40, hepatitis-virus-B X-associated protein-2, and tetratricopeptide repeat protein-2 on all six steroid hormone receptors in a homogeneous mammalian cell system. To be able to assess each cofactor's effect on the transcriptional activity of on each steroid receptor we employed transient transfection in a reporter gene assay. In addition, we evaluated the interactions of the TPR proteins with the receptors and components of the Hsp90 chaperone heterocomplex by coimmunoprecipitation. In the functional assays, corticosteroid and progesterone receptors displayed the most sensitive and distinct reaction to the TPR proteins. Androgen receptor's activity was moderately impaired by most cofactors, whereas the Estrogen receptors' activity was impaired by most cofactors only to a minor degree. Second, interaction studies revealed that the strongly receptor-interacting co-chaperones were all among the inhibitory proteins. Intriguingly, the TPR-proteins also differentially co-precipitated the heterochaperone complex components Hsp90, Hsp70, and p23, pointing to differences in their modes of action. The results of this comprehensive study provide important insight into chaperoning of diverse client proteins via the combinatorial action of (co)-chaperones. The differential effects of the TPR proteins on steroid receptors bear on all physiological processes related to steroid hormone activity.


Trends in the knowledge, attitudes and practices of travel risk groups toward prevention of hepatitis B: results from the repeated cross-sectional Dutch Schiphol Airport Survey 2002-2009.
Previous studies investigating the travellers' knowledge, attitudes and practices (KAP) profile indicated an important educational need among those travelling to risk destinations. In the years 2002-2009 an annually repeated cross-sectional questionnaire-based survey was conducted at the Dutch Schiphol Airport with the aim to study trends in KAP of travel risk groups toward prevention of hepatitis B. The frequently encountered risk groups last-minute travellers, solo-travellers, business travellers, travellers visiting friends and relatives (VFR) and elderly travellers were specifically studied. A total of 3045 respondents were included in the survey. Travellers to destinations with a high risk for hepatitis B had significantly less accurate risk perceptions (knowledge) than travellers to low-risk destinations but no differences were observed in past risk-taking attitude. Protection rates against hepatitis B were significantly higher in travellers to high-risk destinations. There was a positive trend over the years in the proportion of travellers to high-risk destinations seeking travel health advice. In accordance with this, trend analyses also indicated rising protection rates against hepatitis B. No significant trends in protection over time were observed for the travel risk groups. The results of this repeated cross-sectional survey suggest an annual 10% increase in protection rates against hepatitis B in Dutch travellers, both to destinations with a high risk and to destinations with a lower risk of hepatitis B, but these trends in protection rates were not observed for the travel risk groups to high-risk destinations. The KAP profile of last-minute travellers and (to a lesser extent) VFRs showed an increased relative risk in hepatitis B, irrespective of the travel destination, underlining the need for specific targeting of these travel risk groups.


Abundance of six tetracycline resistance genes in wastewater lagoons at cattle feedlots with different antibiotic use strategies.
The abundance of six tetracycline resistance genes tet(O), tet(Q), tet(W), tet(M), tet(B) and tet(L), were quantified over time in wastewater lagoons at concentrated animal feeding operations (CAFO) to assess how feedlot operation affects resistance genes in downstream surface waters. Eight lagoons at five cattle feedlots in the Midwestern United States were monitored for 6 months. Resistance and 16S-rRNA gene abundances were quantified using real-time PCR, and physicochemical lagoon conditions, tetracycline levels, and other factors (e.g. feedlot size and weather conditions) were monitored over time. Lagoons were sorted according to antibiotic use practice at each site, and designated as 'no-use', 'mixed-use' or 'high-use' for comparison. High-use lagoons had significantly higher detected resistance gene levels (tet(R); 2.8 x 10(6) copies ml(-1)) relative to no-use lagoons (5.1 x 10(3) copies ml(-1); P < 0.01) and mixed-use lagoons (7.3 x 10(5) copies ml(-1); P = 0.076). Bivariate correlation analysis on pooled data (n = 54) confirmed that tet(R) level strongly correlated with feedlot area (r = 0.67, P < 0.01) and 'total' bacterial 16S-rRNA gene level in each lagoon (r = 0.51, P < 0.01), which are both characteristic of large CAFOs. tet(M) was the most commonly detected gene, both in absolute number and normalized to 16S-rRNA gene level, although tet(O), tet(Q) and tet(W) levels were also high in the mixed and high-use lagoons. Finally, resistance gene levels were highly seasonal with abundances being 10-100 times greater in the autumn versus the summer. Results show that antibiotic use strategy strongly affects both the abundance and seasonal distribution of resistance genes in associated lagoons, which has implications on water quality and feedlot management practices.


Transition between [R]- and [S]-stereoisomers without bond breaking.
The fifty-year old proposal of a nondissociative racemization reaction of a tetracoordinated tetrahedral center from one enantiomer to another via a planar transition state by Hoffmann and coworkers has been explored by many research groups over the past five decades. A number of stable molecules with planar tetracoordinated and higher-coordinated centers have been designed and experimentally realized; however, there has not been a single example of a molecular system that can possibly undergo such racemization. Here we show examples of molecular species that undergo inversion of stereochemistry around tetrahedral centers (Si, Al- and P+) either via a planar transition state or an intermediate state using quantum mechanical, ab initio quasi-classical dynamics calculations, and Born-Oppenheimer molecular dynamics (BOMD) simulations. This work is expected to provide potential leads for future studies on this fundamental phenomenon in chemistry.


Probing activated sludge with oligonucleotides specific for proteobacteria: inadequacy of culture-dependent methods for describing microbial community structure.
Bacterial community structures in activated sludge samples from aeration tanks of a two-stage system with a high-load first stage and a low-load second stage were analyzed with oligonucleotide probes. The probes were complementary to conserved regions of the rRNA of the alpha, beta, and gamma subclasses of proteobacteria and of all bacteria. Group-specific cell counts were determined by in situ hybridization with fluorescent probe derivatives. Contributions of the proteobacterial subclasses to total bacterial rRNA were quantified by dot blot hybridization with digoxigenin-labeled oligonucleotides. The activated sludge samples were dominated by proteobacteria from the alpha, beta, or gamma subclass. These proteobacteria account for about 80% of all active bacteria found in the activated sludge. For both samples the community structures determined with molecular techniques were compared with the composition of the heterotrophic saprophyte flora isolated on nutrient-rich medium. Probes were used to rapidly classify the isolates and to directly monitor population shifts in nutrient-amended, activated sludge samples. The rich medium favored growth of gamma-subclass proteobacteria (e.g., enterobacteria) and selected against beta-subclass proteobacteria. The culture-dependent community structure analysis of activated sludge produced partial and heavily biased results. A more realistic view will be obtained by using in situ techniques.


Elevated MAL expression is accompanied by promoter hypomethylation and platinum resistance in epithelial ovarian cancer.
We previously found that the gene encoding the Myelin and Lymphocyte protein, MAL, was among the most highly expressed genes in serous ovarian cancers from short-term survivors (<3 years) relative to those of long-term survivors (>7 years). In the present study, we have found that this difference in expression is partially attributable to differences in DNA methylation at a specific region within the MAL promoter CpG island. While MAL was largely unmethylated at the transcription start site (Region 1; -48 to +73 bp) in primary serous ovarian cancers, methylation of an upstream region (Region 2; -452 to -266 bp) was inversely correlated with MAL transcription in the primary cancers (R = -0.463) and ovarian cancer cell lines (R = -0.444). Following treatment of the OVCA432 cell line with 5-azacytidine, methylation of Region 2 decreased from 73.3% to 34.7% (p = 0.007) while Region 1 was unaffected. This was accompanied by a 10-fold increase in MAL expression. Since MAL transcripts are elevated in tumors from short-term survivors, all of whom were treated with platinum-based therapy, MAL may have a role in cisplatin response. We therefore determined the 50% growth inhibitory dose of cisplatin in 30 ovarian cancer cell lines and compared this to MAL expression. MAL transcript levels were higher in the resistant ovarian cell lines (p = 0.04). MAL methylation status may therefore serve as a marker of platinum sensitivity while MAL protein may be a target for development of novel therapies aimed at enhancing sensitivity to platinum-based drugs in ovarian cancer.


Specificity of machine, barbell, and water-filled log bench press resistance training on measures of strength.
The purpose of this study was to compare the effects of 10 weeks of resistance training with an isotonic bench press machine and 2 types of free-weight bench press exercises on several measures bench press strength. Specificity was investigated by comparing the ability to transfer strength gained from a type of training that differed from the mode of testing. Forty-nine men participated in the study. The subjects completed a pretest on the machine (MB), barbell (BB), isokinetic (IB), and log (LB) bench press to determine baseline strength and completed 10 weeks of training on the MB, BB, or LB. The 3 groups were tested to see whether differential training effects occurred from pre- to posttest scores on the BB, MB, LB, and peak force on the IB. By multivariate analysis, the trial-by-group interaction was not statistically significant. The multivariate and subsequent univariate analyses of variance tests indicated statistically significant effects from pre- to posttest for peak force on the IB test and the BB, MB, and LB. Correlations among the strength tests were high (0.92 > or = r < or = 0.97) and moderate between the strength tests and IB peak force (0.62 > or = r < pr = 0.83). Mean 3 repetition maximum MB strength was 8% higher than BB strength, which was 3% higher than LB strength, indicating differences in the amount of stabilization required to control the resistance. The findings of this study showed that all 3 training groups significantly improved in strength during short-term training on the MB, BB, and LB. These data lend evidence that improved strength after training on the MB, BB, and LB equally transfers to strength gains on any of the 4 modes of testing. These results should be considered when including similar exercises varying in stability into the training program to improve strength.


Explanatory models of hypertension among Nigerian patients at a University Teaching Hospital.
To elicit the explanatory models (EM) of hypertension among patients in a hospital-based primary care practice in Nigeria. Design. Semi-structured in-depth individual interviews and focus groups were conducted with 62 hypertensive patients. Interviews and focus groups were audiotaped and transcribed verbatim. Data analysis was guided by phenomenology and content analysis using qualitative research software ATLAS.ti 5.0. Patients expressed four categories of EM of hypertension: (1) perceptions of hypertension, (2) consequences, (3) effect on daily life, and (4) perception of treatment. Focus group discussions and individual interviews yielded a wide range of insights into the social and cultural factors influencing patients' beliefs and health behavior. Participants were aware of the risks of hypertension. There was disagreement between participants' own understanding of the serious nature of hypertension, the need for long-term treatment, and the desire to take long-term medication. Participants acknowledged the use of traditional medicine (e.g., teas and herbs) and healers. Different themes emerged for men versus women such that women often focused on family issues while men tended to discuss external stressors stemming from work as a cause of hypertension. Men were concerned with frequent urination, decreased libido, and erectile dysfunction. Knowledge gained will inform development of patient-centered treatment plans and targeted behavioral and educational interventions.


A Tractable Method for Measuring Nanomaterial Risk Using Bayesian Networks.
While control banding has been identified as a suitable framework for the evaluation and the determination of potential human health risks associated with exposure to nanomaterials (NMs), the approach currently lacks any implementation that enjoys widespread support. Large inconsistencies in characterisation data, toxicological measurements and exposure scenarios make it difficult to map and compare the risk associated with NMs based on physicochemical data, concentration and exposure route. Here we demonstrate the use of Bayesian networks as a reliable tool for NM risk estimation. This tool is tractable, accessible and scalable. Most importantly, it captures a broad span of data types, from complete, high quality data sets through to data sets with missing data and/or values with a relatively high spread of probability distribution. The tool is able to learn iteratively in order to further refine forecasts as the quality of data available improves. We demonstrate how this risk measurement approach works on NMs with varying degrees of risk potential, namely, carbon nanotubes, silver and titanium dioxide. The results afford even non-experts an accurate picture of the occupational risk probabilities associated with these NMs and, in doing so, demonstrated how NM risk can be evaluated into a tractable, quantitative risk comparator.


Learning from directed evolution: Thermus aquaticus DNA polymerase mutants with translesion synthesis activity.
DNA is being constantly damaged by endo- and exogenous agents such as reactive oxygen species, chemicals, radioactivity, and ultraviolet radiation. Additionally, DNA is inherently labile, and this can result in, for example, the spontaneous hydrolysis of the glycosidic bond that connects the sugar and the nucleobase moieties in DNA; this results in abasic sites. It has long been obscure how cells achieve DNA synthesis past these lesions, and only recently has it been discovered that several specialized DNA polymerases are involved in translesion synthesis. The underlying mechanisms that render one DNA polymerase competent in translesion synthesis while another DNA polymerase fails are still indistinct. Recently two variants of Taq DNA polymerase that exhibited higher lesion bypass ability than the wild-type enzyme were identified by directed-evolution approaches. Strikingly, in both approaches it was independently found that substitution of a single nonpolar amino acid side chain by a cationic side chain increases the capability of translesion synthesis. Here, we combined both mutations in a single enzyme. We found that the KlenTaq DNA polymerase that bore both mutations superseded the wild-type as well as the respective single mutants in translesion-bypass proficiency. Further insights in the molecular basis of the detected gain of translesion-synthesis function were obtained by structural studies of DNA polymerase variants caught in processing canonical and damaged substrates. We found that increased positive charge of the surface potential in the area proximal to the negatively charged substrates promotes translesion synthesis by KlenTaq DNA polymerase, an enzyme that has very limited naturally evolved capability to perform translesion synthesis. Since expanded positively charged surface potential areas are also found in naturally evolved translesion DNA polymerases, our results underscore the impact of charge on the proficiency of naturally evolved translesion DNA polymerases.


A Multi-Institutional Feasibility Study on the Use of Automated Screening Systems for Quality Control Rescreening of Cervical Cytology.
To evaluate the efficacy of the automated screening system FocalPoint for cervical cytology quality control (QC) rescreening. False-negative rates (FNRs) were evaluated by a multi-institutional retrospective study. Cervical cytology slides that had already been reported as negative for intraepithelial lesion or malignancy (NILM) were chosen arbitrarily for FocalPoint rescreening. Slides stratified into the highest 15% probability of being abnormal were rescreened by a cytotechnologist. The slides that were abnormal were reevaluated by a cytopathologist to be false negatives. Rescreening of 12,000 slides, i.e. 9,000 conventional slides and 3,000 liquid-based cytology (LBC) slides, was performed; 9,826 (7,393 conventional and 2,433 LBC) were satisfactory for FocalPoint (2,174 were determined unsatisfactory) and those within the highest 15% of probability (1,496, i.e. 1,123 conventional and 373 LBC) were rescreened. As a result, 117 (96 conventional and 21 LBC) were determined as abnormal (other than NILM) and the FNR was 1.19%. Among these 117 slides, 40 (35 conventional and 5 LBC) were determined as high-grade squamous intraepithelial lesion and greater (HSIL+). Of 117 (1.19%) abnormal slides detected, 40 (0.41%) were determined to be HSIL+. This result suggests that FocalPoint is effective for QC rescreening of cervical cytology.


Characterization of an EcoR124I restriction-modification enzyme produced from a deleted form of the DNA-binding subunit, which results in a novel DNA specificity.
We purified and characterized both the methyltransferase and the endonuclease containing the HsdS delta 50 subunit (type I restriction endonucleases are composed of three subunits--HsdR required for restriction, HsdM required for methylation and HsdS responsible for DNA recognition) produced from the deletion mutation hsdS delta 50 of the type IC R-M system EcoR 124I; this mutant subunit lacks the C-terminal 163 residues of HsdS and produces a novel DNA specificity. Analysis of the purified HsDs delta 50 subunit indicated that during purification it is subject to partial proteolysis resulting in removal of approximately 1 kDa of the polypeptide at the C-terminus. This proteolysis prevented the purification of further deletion mutants, which were determined as having a novel DNA specificity in vivo. After biochemical characterization of the mutant DNA methyltransferase (MTase) and restriction endonuclease we found only one difference comparing with the wild-type enzyme--a significantly higher binding affinity of the MTase for the two substrates of hemimethylated and fully methylated DNA. This indicates that MTase delta 50 is less able to discriminate the methylation status of the DNA during its binding. However, the mutant MTase still preferred hemimethylated DNA as the substrate for methylation. We fused the hsdM and hsdS delta 50 genes and showed that the HsdM-HsdS delta 50 fusion protein is capable of dimerization confirming the model for assembly of this deletion mutant.


Autism Symptoms in Fragile X Syndrome.
Fragile X syndrome (FXS) is recognized as the most common genetic cause of intellectual disability and autism spectrum disorder (ASD). Although symptoms of ASD are frequently observed in patients with FXS, researchers have not yet clearly determined whether the symptoms in patients with FXS differ from the symptoms in patients without ASD or nonsyndromic ASD. Behavioral similarities and differences between FXS and ASD are important to improve our understanding of the causes and correlations of ASD with FXS. Based on the evidence presented in this review, individuals with FXS and comorbid ASD have more severe behavioral problems than individuals with FXS alone. However, patients with FXS and comorbid ASD exhibit less severe impairments in the social and communication symptoms than patients with nonsyndromic ASD. Individuals with FXS also present with anxiety and seizures in addition to comorbid ASD symptoms, and differences in these conditions are noted in patients with FXS and ASD. This review also discusses the role of fragile X mental retardation 1 protein (FMRP) in FXS and ASD phenotypes.


Drastic influence of the half-bowtie resonances on the focusing and collimating capabilities of 2-D extended hemielliptical and hemispherical dielectric lenses.
The interplay between the optical focusing and wavelength-scale resonant features of extended hemielliptical (EHE) and extended hemispherical (EHS) lenses is studied in the two-dimensional (2-D) formulation using highly accurate in-house software based on the Muller boundary integral equations. The influence of the half-bowtie (HBT) resonances on the focusing and collimating capabilities of medium-size EHE and EHS lenses made of silicon is characterized as a function of lens parameters and excitation conditions. As a result, factors determining the parasitic impacts of the HBT resonances on the performance of integrated lens antennas are highlighted.


Isolation and characterization of a ntrC mutant of Bradyrhizobium (Parasponia) sp. ANU289.
A mutant of Bradyrhizobium (Parasponia) sp. ANU289 affected in the regulation of nitrogen metabolism was isolated. The mutant, designated ANU293, was unable to induce ammonium transport (Amt), nitrate reductase (NR) or glutamine synthetase II (GSII) activities under conditions that induce these activities in the wild-type. However, glutamine synthetase I (GSI), which is expressed constitutively in the wild-type, was present at normal levels in the mutant. The mutant also retained the ability to fix nitrogen in vitro and in planta, although nodule development on siratro (Macroptilium atropurpureum) was retarded. Southern blot analysis showed that ntrC, the product of which is involved in regulation of nitrogen metabolism, is the site of pSUP1021 insertion in ANU293. These results indicate that the transcriptional activator NtrC is required for the expression of Amt, NR and GSII, but not GSI or nitrogenase in Bradyrhizobium (Parasponia) sp. ANU289.


A single intratracheal instillation of single-walled carbon nanotubes induced early lung fibrosis and subchronic tissue damage in mice.
Large amounts of nanomaterials may reach both the natural and occupational environments. This represents a potential health hazard. People have forecasted that CNTs may lead to the toxicity such as mesothelioma and fibrosis like asbestos. To identify dominant immune responses induced by SWCNTs, we investigated the composition of bronchioalveolar lavage (BAL) cells, the secretion of cytokine and collagen, histopathology, protein expression, and cell phenotypes over time after a single administration of single-walled carbon nanotubes (SWCNT). In our results, the number of total cells and macrophages remained at the up-regulated level until Day 28, neutrophils rapidly increased at Day 1, and lymphocytes increased from Day 7. In the BAL fluid, pro-inflammatory cytokines rapidly increased at Day 1 and remained at an up-regulated level throughout the experimental period. IL-12 and IL-10 rapidly increased at Day 1 after administration and remained at a similar level until Day 28. IFN-γ and IL-4 reached the maximum at Day 1, and IL-5, TGF-β, and collagen reached the maximum at Day 7. IL-13 and IL-17 increased in a time-dependent manner. The distribution of B cells and cytotoxic T cells markedly increased at Days 7 and 14, and fibrotic lesions were histopathologically observed at Days 7 and 14. The expressions of caspase-3, p53, COL1A1, COX-2, iNOS, MMP-9, and MMP-2 were also markedly increased at Days 7 and 14. In addition, the expression of mesothelin, iNOS, MMP-9, and p53 was up-regulated until Day 28. Based on these findings, we suggest that a single intratracheal instillation of SWCNTs may induce early lung fibrosis and subchronic tissue damage.


Exosomes as divine messengers: are they the Hermes of modern molecular oncology?
Exosomes are cell-derived vesicles that convey key elements with the potential to modulate intercellular communication. They are known to be secreted from all types of cells, and are crucial messengers that can regulate cellular processes by 'trafficking' molecules from cells of one tissue to another. The exosomal content has been shown to be broad, composed of different types of cytokines, growth factors, proteins, or nucleic acids. Besides messenger RNA (mRNA) they can also contain noncoding transcripts such as microRNAs (miRNAs), which are small endogenous cellular regulators of protein expression. In diseases such as cancer, exosomes can facilitate tumor progression by altering their vesicular content and supplying the tumor niche with molecules that favor the progression of oncogenic processes such as proliferation, invasion and metastasis, or even drug resistance. The packaging of their molecular content is known to be tissue specific, a fact that makes them interesting tools in clinical diagnostics and ideal candidates for biomarkers. In the current report, we describe the main properties of exosomes and explain their involvement in processes such as cell differentiation and cell death. Furthermore, we emphasize the need of developing patient-targeted treatments by applying the conceptualization of exosomal-derived miRNA-based therapeutics.


Gold standards in pharmacovigilance: the use of definitive anecdotal reports of adverse drug reactions as pure gold and high-grade ore.
Anecdotal reports of adverse drug reactions are generally regarded as being of poor evidential quality. This is especially relevant for postmarketing drug safety surveillance, which relies heavily on spontaneous anecdotal reports. The numerous limitations of spontaneous reports cannot be overemphasised, but there is another side to the story: these datasets also contain anecdotal reports that can be considered to describe definitive adverse reactions, without the need for further formal verification. We have previously defined four categories of such adverse reactions: (i) extracellular or intracellular tissue deposition of the drug or a metabolite; (ii) a specific anatomical location or pattern of injury; (iii) physiological dysfunction or direct tissue damage demonstrable by physicochemical testing; and (iv) infection, as a result of the administration of an infective agent as the therapeutic substance or because of demonstrable contamination. In this article, we discuss the implications of these definitive ('between-the-eyes') adverse effects for pharmacovigilance.


Cerebrospinal fluid {beta}-amyloid 42 and tau proteins as biomarkers of Alzheimer-type pathologic changes in the brain.
There is a clear need to develop an objective diagnostic test for Alzheimer disease (AD). Changes in the levels of cerebrospinal fluid (CSF) tau protein and beta-amyloid 42 (Abeta42) peptide in patients with AD have been well documented, but the relationship between these biomarkers and neuropathologic changes in the brain is not established. To study the relationship between antemortem CSF biomarker levels and Alzheimer-type neuropathologic changes in the brain. Cross-sectional study to correlate levels of CSF Abeta42, total tau, and phosphorylated tau protein with neuropathologic changes in the brain. Academic research. Patients The study included 123 patients (79 with clinically diagnosed AD, 29 with other dementia, and 15 with other neurologic disease). All underwent clinical evaluation and provided antemortem lumbar CSF samples, and neuropathologic data were collected from September 11, 1990, to March 13, 2003, in the Department of Neuroscience and Neurology, University of Kuopio, Kuopio, Finland. Levels of CSF Abeta42, total tau, and phosphorylated tau protein were measured using standard commercial immunoassays. Neuropathologic evaluations included the classic silver impregnation method and immunohistochemistry for Abeta, hyperphosphorylated tau, and alpha-synuclein. Cerebrospinal fluid Abeta42 and tau protein levels were related to amyloid load and the presence of neurofibrillary pathologic abnormalities in the brain. Cerebrospinal fluid Abeta42 level correlated inversely with total Abeta load in the brain, and CSF tau level correlated with results of immunohistochemistry for hyperphosphorylated tau and with the presence of neocortical neurofibrillary tangles. In multivariate logistic regression analysis, the number of neuritic plaques in the brain remained a significant predictor of decreased CSF Abeta42 level and of increased CSF tau level. Based on the ratio of phosphorylated tau level to Abeta42 level, sensitivity was 91.6%, and specificity was 85.7%, with an overall accuracy of 90.2% for the presence of pathologic neuritic plaque in the brain. Cerebrospinal fluid Abeta42 and tau proteins are biomarkers of AD-associated pathologic changes in the brain. The combination of abnormally low CSF Abeta42 level and abnormally high CSF tau level predicted the presence of AD pathologic features with high accuracy. This combination assay may be helpful in diagnosing the presence of AD pathologic changes in the brain.


Age-related sex differences in body condition and telomere dynamics of red-sided garter snakes.
Life-history strategies vary dramatically between the sexes, which may drive divergence in sex-specific senescence and mortality rates. Telomeres are tandem nucleotide repeats that protect the ends of chromosomes from erosion during cell division. Telomeres have been implicated in senescence and mortality because they tend to shorten with stress, growth and age. We investigated age-specific telomere length in female and male red-sided garter snakes, Thamnophis sirtalis parietalis We hypothesized that age-specific telomere length would differ between males and females given their divergent reproductive strategies. Male garter snakes emerge from hibernation with high levels of corticosterone, which facilitates energy mobilization to fuel mate-searching, courtship and mating behaviours during a two to four week aphagous breeding period at the den site. Conversely, females remain at the dens for only about 4 days and seem to invest more energy in growth and cellular maintenance, as they usually reproduce biennially. As male investment in reproduction involves a yearly bout of physiologically stressful activities, while females prioritize self-maintenance, we predicted male snakes would experience more age-specific telomere loss than females. We investigated this prediction using skeletochronology to determine the ages of individuals and qPCR to determine telomere length in a cross-sectional study. For both sexes, telomere length was positively related to body condition. Telomere length decreased with age in male garter snakes, but remained stable in female snakes. There was no correlation between telomere length and growth in either sex, suggesting that our results are a consequence of divergent selection on life histories of males and females. Different selection on the sexes may be the physiological consequence of the sexual dimorphism and mating system dynamics displayed by this species.


How particle-particle and liquid-particle interactions govern the fate of evaporating liquid marbles.
Liquid marbles refer to droplets that are covered with a layer of non-wetting particles. They are observed in nature and have practical significance. These squishy objects bounce, coalesce, break, inflate, and deflate while the liquid does not touch the substrate underneath. Despite the considerable cross-disciplinary interest and value of the research on liquid marbles, a unified framework for describing the mechanics of deflating liquid marbles-as the liquid evaporates-is unavailable. For instance, analytical approaches for modeling the evaporation of liquid marbles exploit empirical parameters that are not based on liquid-particle and particle-particle interactions. Here, we have combined complementary experiments and theory to fill this gap. To unentangle the contributions of particle size, roughness, friction, and chemical make-up, we investigated the evaporation of liquid marbles formed with particles of sizes varying over 7 nm-300 μm and chemical compositions ranging from hydrophilic to superhydrophobic. We demonstrate that the potential final states of evaporating liquid marbles are characterized by one of the following: (I) constant surface area, (II) particle ejection, or (III) multilayering. Based on these insights, we developed an evaporation model for liquid marbles that takes into account their time-dependent shape evolution. The model fits are in excellent agreement with our experimental results. Furthermore, this model and the general framework can provide mechanistic insights into extant literature on the evaporation of liquid marbles. Altogether, these findings advance our fundamental understanding of liquid marbles and should contribute to the rational development of technologies.


The mesoderm determinant snail collaborates with related zinc-finger proteins to control Drosophila neurogenesis.
The Snail protein functions as a transcriptional regulator to establish early mesodermal cell fate. Later, in germ band-extended embryos, Snail is also expressed in most neuroblasts. Here we present evidence that this expression of Snail is required for central nervous system (CNS) development. The neural function of snail is masked by two closely linked genes, escargot and worniu. Both Escargot and Worniu contain zinc-finger domains that are highly homologous to that of Snail. Although not affecting expression of early neuroblast markers, the deletion of the region containing all three genes correlates with loss of expression of CNS determinants including fushi tarazu, pdm-2 and even-skipped. Transgenic expression of each of the three Snail family proteins can rescue efficiently the fushi tarazu defects, and partially the pdm-2 and even-skipped CNS patterns. These results demonstrate that the Snail family proteins have essential functions during embryonic CNS development, around the time of ganglion mother cell formation.


Implementing one-shot multiple-FID acquisition into homonuclear and heteronuclear NMR experiments.
Multiple-FID acquisition (MFA) within the same scan is applied to acquire simultaneously multiple 2D spectra from a single NMR experiment. A discussion on the incorporation of the MFA strategy in several homonuclear and heteronuclear 2D pulse sequences is presented. As a proof of concept, a set of novel COSY, TOCSY and HMBC experiments are reported as a time-efficient solution in small-molecule NMR spectroscopy.


What have we learnt from SARS?
With outbreaks of infectious disease emerging from animal sources, we have learnt to expect the unexpected. We were, and are, expecting a new influenza A pandemic, but no one predicted the emergence of an unknown coronavirus (CoV) as a deadly human pathogen. Thanks to the preparedness of the international network of influenza researchers and laboratories, the cause of severe acute respiratory syndrome (SARS) was rapidly identified, but there is no complacency over the global or local management of the epidemic in terms of public health logistics. The human population was lucky that only a small proportion of infected persons proved to be highly infectious to others, and that they did not become so before they felt ill. These were the features that helped to make the outbreak containable. The next outbreak of another kind of transmissible disease may well be quite different.


A basic study on molecular hydrogen (H2) inhalation in acute cerebral ischemia patients for safety check with physiological parameters and measurement of blood H2 level.
In animal experiments, use of molecular hydrogen ( H2) has been regarded as quite safe and effective, showing benefits in multiple pathological conditions such as ischemia-reperfusion injury of the brain, heart, kidney and transplanted tissues, traumatic and surgical injury of the brain and spinal cord, inflammation of intestine and lung , degenerative striatonigral tissue and also in many other situations. However, since cerebral ischemia patients are in old age group, the safety information needs to be confirmed. For the feasibility of H2 treatment in these patients, delivery of H2 by inhalation method needs to be checked for consistency. Hydrogen concentration (HC) in the arterial and venous blood was measured by gas chromatography on 3 patients, before, during and after 4% (case 1) and 3% (case2,3) H2 gas inhalation with simultaneous monitoring of physiological parameters. For a consistency study, HC in the venous blood of 10 patients were obtained on multiple occasions at the end of 30-min H2 inhalation treatment. The HC gradually reached a plateau level in 20 min after H2 inhalation in the blood, which was equivalent to the level reported by animal experiments. The HC rapidly decreased to 10% of the plateau level in about 6 min and 18 min in arterial and venous blood, respectively after H2 inhalation was discontinued. Physiological parameters on these 3 patients were essentially unchanged by use of hydrogen. The consistency study of 10 patients showed the HC at the end of 30-min inhalation treatment was quite variable but the inconsistency improved with more attention and encouragement. H2 inhalation of at least 3% concentration for 30 min delivered enough HC, equivalent to the animal experiment levels, in the blood without compromising the safety. However, the consistency of H2 delivery by inhalation needs to be improved.


Zinc deficiency suppresses the development of oral tolerance in rats.
Oral tolerance is a specific immune unresponsiveness to food antigens to prevent hypersensitivity reactions. We investigated whether zinc deficiency affects oral tolerance. Rats were fed a control (C) or zinc-deficient (ZD) diet, or pair-fed (PF) to ZD rats for 28 d. Beginning on d 7, rats were administered ovalbumin (OVA) orally to induce tolerance, or PBS 3 times/wk, and were then immunized by OVA injection. The proliferation of mesenteric lymph node (MLN) and spleen lymphocytes after in vitro OVA stimulation and the delayed-type hypersensitivity were higher in OVA-fed ZD than in OVA-fed C rats and not different between OVA- and PBS-fed ZD rats, indicating a suppression of tolerance. Lymphocyte proliferation did not differ between PF and C rats. Expressions of cytokines involved in oral tolerance, i.e., interleukin (IL)-4, IL-10 and transforming growth factor-beta, were higher in OVA- than in PBS-fed C rats, but not in ZD rats. Apoptosis was higher in OVA- than in PBS-fed C rats but not different between OVA- and PBS-fed ZD rats. Inflammation and ulcerations that were not present in ZD rats on d 7 (ZD(7)) developed in OVA- or PBS-fed ZD rats. Compared with ZD(7) rats, tumor necrosis factor-alpha and cytokine-induced neutrophil chemoattractant were higher in OVA- and PBS-fed ZD rats, whereas interferon-gamma increased only in OVA-fed ZD rats. In conclusion, zinc deficiency suppresses oral tolerance through dysregulation of cytokine expression and lack of antigen-specific clonal deletion. We suggest that abrogation of tolerance may lead to development of mucosal inflammation and damage.


A diagnostic tool for self-poisoned patients: Analysis of gastric content and lavage fluids.
Approximately 500 patients per year are admitted to the emergency department (ED) of the Erasmus University Medical Center presenting with intoxications with medication. For adequate treatment, it is sometimes important to know which drugs in which quantities were ingested. This can require laboratory analysis of blood or urine samples; however, these samples do not provide information about the possible effects that can still be expected. We performed toxicological screening on the gastric content of three patients admitted to our ED in January and February 2018. These patients underwent gastric lavage or received a gastric tube as part of routine care. The gastric fluid was analysed via UPLC-MS/MS using the Waters method for toxicological screening. In all three patients, we successfully determined drugs in the gastric content. In two patients, we identified more different drugs in the gastric content than in blood plasma. In the other patient, admitted approximately six hours after a severe autointoxication with the betablocker metoprolol, we found significant amounts of metoprolol in the gastric content acquired by gastric lavage. We therefore believe that analysis of gastric content after an intoxication can have multiple applications; for example, it may provide information about symptoms of intoxication that can be expected, it may aid patient care and may provide insight in the toxicokinetics of different drugs. In conclusion, we demonstrate that toxicological screening and quantification of drug levels in gastric content is possible and has potential as an adjunct in patient care, but limitations need to be addressed before implementation in clinical practice.


Rational development and characterization of humanized anti-EGFR variant III chimeric antigen receptor T cells for glioblastoma.
Chimeric antigen receptors (CARs) are synthetic molecules designed to redirect T cells to specific antigens. CAR-modified T cells can mediate long-term durable remissions in B cell malignancies, but expanding this platform to solid tumors requires the discovery of surface targets with limited expression in normal tissues. The variant III mutation of the epidermal growth factor receptor (EGFRvIII) results from an in-frame deletion of a portion of the extracellular domain, creating a neoepitope. We chose a vector backbone encoding a second-generation CAR based on efficacy of a murine scFv-based CAR in a xenograft model of glioblastoma. Next, we generated a panel of humanized scFvs and tested their specificity and function as soluble proteins and in the form of CAR-transduced T cells; a low-affinity scFv was selected on the basis of its specificity for EGFRvIII over wild-type EGFR. The lead candidate scFv was tested in vitro for its ability to direct CAR-transduced T cells to specifically lyse, proliferate, and secrete cytokines in response to antigen-bearing targets. We further evaluated the specificity of the lead CAR candidate in vitro against EGFR-expressing keratinocytes and in vivo in a model of mice grafted with normal human skin. EGFRvIII-directed CAR T cells were also able to control tumor growth in xenogeneic subcutaneous and orthotopic models of human EGFRvIII(+) glioblastoma. On the basis of these results, we have designed a phase 1 clinical study of CAR T cells transduced with humanized scFv directed to EGFRvIII in patients with either residual or recurrent glioblastoma (NCT02209376).


Development and validation of a multiplex real-time PCR assay for simultaneous genotyping and human T-lymphotropic virus type 1, 2, and 3 proviral load determination.
The human T-lymphotropic virus (HTLV) proviral load remains the best surrogate marker for disease progression. Real-time PCR techniques have been developed for detection and quantification of cosmopolitan HTLV type 1a (HTLV-1a) and HTLV-2. Since a growing level of diversity in subtypes and genotypes is observed, we developed a multiplex quantitative PCR for simultaneous detection, genotyping, and quantification of proviral loads of HTLV-1, 2, and 3. Our assay uses tax type-specific primers and dually labeled probes and has a dynamic range of 10(5) to 10 HTLV copies. One hundred sixty-three samples were analyzed, among which all of the different subtypes within each HTLV genotype could be detected. The performance of proviral load determination of our multiplex assay was compared with that of a previously published HTLV-1 singleplex quantitative PCR based on SYBR green detection, developed at a different institute. Linear regression analysis showed a statistically significant (P < 0.0001) and strong (r(2) = 0.87) correlation between proviral load values measured with the two distinct real-time PCR assays. In conclusion, our novel assay offers an accurate molecular diagnosis and genotyping, together with the determination of the proviral load of HTLV-infected individuals, in a single amplification reaction. Moreover, our molecular assay could offer an alternative when current available serological assays are insufficient.


Contribution of BK Ca2+-activated K+ channels to auditory neurotransmission in the Guinea pig cochlea.
Large-conductance calcium-activated potassium (BK) channels are known to play a prominent role in the hair cell function of lower vertebrates where these channels determine electrical tuning and regulation of neurotransmitter release. Very little is known, by contrast, about the role of BK channels in the mammalian cochlea. In the current study, we perfused specific toxins in the guinea pig cochlea to characterize the role of BK channels in cochlear neurotransmission. Intracochlear perfusion of charybdotoxin (ChTX) or iberiotoxin (IbTX) reversibly reduced the compound action potential (CAP) of the auditory nerve within minutes. The cochlear microphonics (CM at f1 = 8 kHz and f2 = 9.68 kHz) and their distortion product (DPCM at 2f1-f2) were essentially not affected, suggesting that the BK specific toxins do not alter the active cochlear amplification at the outer hair cells (OHCs). We also tested the effects of these toxins on the whole cell voltage-dependent membrane current of isolated guinea pig inner hair cells (IHCs). ChTX and IbTX reversibly reduced a fast outward current (activating above -40 mV, peaking at 0 mV with a mean activation time constant tau ranging between 0.5 and 1 ms). A similar block of a fast outward current was also observed with the extracellular application of barium ions, which we believe permeate through Ca2+ channels and block BK channels. In situ hybridization of Slo antisense riboprobes and immunocytochemistry demonstrated a strong expression of BK channels in IHCs and spiral ganglion and to a lesser extent in OHCs. Overall, our results clearly revealed the importance of BK channels in mammalian cochlear neurotransmission and demonstrated that at the presynaptic level, fast BK channels are a significant component of the repolarizing current of IHCs.


Significant variations in mortality occur at similarly designated trauma centers.
Mortality rates vary across designated trauma centers (TC), even after controlling for injury severity. Retrospective analysis of state trauma registry data. Designated Level 1 and 2 TCs in 2003 in a large Southwestern state. Adult trauma patients (n = 18,584) treated at 15 designated Level 1 and 2 TCs. Risk-adjusted survival was calculated for each trauma center using logistic regression analysis to adjust for differences in age, sex, race, injury mechanism, and injury severity. The model was developed using half of the study population and validated in the remaining half. It was then applied to the entire study population, with inclusion of TC identification codes. Observed vs Expected survival ratios were then calculated for each TC. Adjusted odds ratios (OR) for survival at each TC were also calculated. Adjusted OR of survival were significantly different from crude OR at 6 of the 14 TCs, underscoring the importance of risk adjustment when performing quality comparisons. One TC performed significantly worse than the others, 8 achieved significantly better survival, and 5 performed the same as the referent. Observed vs Expected ratios demonstrated that one trauma center had significantly worse severity-adjusted outcomes, some were marginal, some performed as well as expected, and none performed better than expectations. Considerable variations in risk-adjusted mortality rates exist across similarly designated TCs. Such variability in outcomes may reflect variations in quality of care, and reasons for this discrepancy should be explored as the next step in the trauma care quality improvement process.


Somatostatin receptors subtypes 2 and 5, dopamine receptor type 2 expression and gsp status as predictors of octreotide LAR responsiveness in acromegaly.
We present two acromegalic patients in which clinical and molecular data are discussed in regard to their ability to predict long term octreotide LAR therapy response. Patient 1: female, 36 years old at diagnosis. Basal GH and IGF-I at diagnosis were 133 ng/mL and 181% above the upper limit of reference values (ULRV), respectively. Growth hormone during acute test with subcutaneous octreotide decreased from 133 to 13 ng/mL. Patient started on primary octreotide LAR therapy (20mg q28 days) and achieved biochemical parameters of disease control after 6 months. Molecular analysis of tumor fragments: gsp +; quantitative analysis of SSTR (somatostatin receptor) and DR (dopamine receptor) mRNA - SSTR2 23954; SSTR5 2407; DR2 total 17016 copies. Patient 2: male, 38 years old at diagnosis. Basal GH and IGF-I at diagnosis were 120 ng/mL and 114% ULRV, respectively. Patient underwent non-curative trans-sphenoidal surgery. Post-operative GH and IGF-I were 112 ng/mL and 137% ULRV, respectively. Growth hormone during acute test with subcutaneous octreotide decreased from 112 to 7 ng/mL. Octreotide LAR therapy (20 mg q28 days) was then initiated. After 6 months of treatment, patient did not attain biochemical control of disease and displayed increased tumor volume. Molecular analysis of tumor fragments: gsp not done; quantitative analysis of SSTR and DR mRNA - SSTR2 416; SSTR5 3767; DR2 total 3439 copies. In conclusion, these two cases illustrate how laboratory data can be conflicting as predictors of octreotide LAR responsiveness and how molecular analysis of tumor fragments can help explain different behaviors in clinically similar patients.


Proteolysis as a regulatory mechanism.
Proteases can play key roles in regulation by controlling the levels of critical components of, for example, signal transduction pathways. Proteolytic processing can remove regulatory proteins when they are not needed, while transforming others from the dormant into the biologically active state. The latter mechanism often involves a subsequent change of cellular localization such as the movement from the membrane to the nucleus. The investigation of these processes has revealed a new type of proteolytic activity, regulated intramembrane proteolysis, and a reversible switch in activity occurring in the HtrA family of serine proteases. The bacterial RseA and the human amyloid precursor processing pathways are used as models to review these novel principles that are evolutionarily conserved and have wide biological implications.


Effects of self-efficacy on oral health behaviours and gingival health in university students aged 18- or 19-years-old.
Although self-efficacy is known to affect various health-related practises, few studies have clearly examined how self-efficacy correlates with oral health behaviors or the oral health condition. We examined the relationship between gingivitis, oral health behaviors and self-efficacy in university students. A total of 2,111 students (1,197 males, 914 females) aged 18 and 19 years were examined. The degree of gingivitis was expressed as the percentage of bleeding on probing (%BOP). Additional information was collected via a questionnaire regarding oral health behaviors (daily frequency of tooth-brushing, use of dental floss and regular check-up). Self-efficacy was assessed using the Self-Efficacy Scale for Self-care (SESS). Path analysis was used to test pathways from self-efficacy to oral health behaviors and %BOP. In the final structural model, self-efficacies were related to each other, and they affected oral health behaviors. Good oral health behaviors reduced dental plaque and calculus, and lower levels of dental plaque and calculus resulted in lower %BOP. Higher self-efficacy correlated with better oral health behaviours and gingival health in university students. Improving self-efficacy may be beneficial for maintaining good gingival health in university students. To prevent gingivitis, the approach of enhancing self-efficacy in university students would be useful.


A dynamic map of antigen recognition by CD4 T cells at the site of Leishmania major infection.
CD4 T helper cells play a central role in the control of infection by intracellular parasites. How efficiently pathogen-specific CD4 T cells detect infected cells in vivo is unclear. Here, we employed intravital two-photon imaging to examine the behavior of pathogen-specific CD4 T cells at the site of Leishmania major infection. While activated CD4 T cells enter the inflamed tissue irrespective of their antigen specificity, pathogen-specific T cells preferentially decelerated and accumulated in infected regions of the dermis. Antigen recognition by CD4 T cells was heterogeneous, involving both stable and dynamic contacts with infected phagocytes. However, not all infected cells induced arrest or deceleration of pathogen-specific T cells, and dense clusters of infected cells were poorly accessible to migrating T cells. Thus, disparities in the dynamics of T cell contacts with infected cells and local variation in T cell access to infected cells are important elements of the host-pathogen interplay.


Separating Business Logic from Medical Knowledge in Digital Clinical Workflows Using Business Process Model and Notation and Arden Syntax.
Evidence-based clinical guidelines have a major positive effect on the physician's decision-making process. Computer-executable clinical guidelines allow for automated guideline marshalling during a clinical diagnostic process, thus improving the decision-making process. Implementation of a digital clinical guideline for the prevention of mother-to-child transmission of hepatitis B as a computerized workflow, thereby separating business logic from medical knowledge and decision-making. We used the Business Process Model and Notation language system Activiti for business logic and workflow modeling. Medical decision-making was performed by an Arden-Syntax-based medical rule engine, which is part of the ARDENSUITE software. We succeeded in creating an electronic clinical workflow for the prevention of mother-to-child transmission of hepatitis B, where institution-specific medical decision-making processes could be adapted without modifying the workflow business logic. Separation of business logic and medical decision-making results in more easily reusable electronic clinical workflows.


Inhibition of plasminogen activator inhibitor-1 restores skeletal muscle regeneration in untreated type 1 diabetic mice.
Type 1 diabetes leads to impairments in growth, function, and regenerative capacity of skeletal muscle; however, the underlying mechanisms have not been clearly defined. With the use of Ins2(WT/C96Y) mice (model of adolescent-onset type 1 diabetes), muscle regeneration was characterized in terms of muscle mass, myofiber size (cross-sectional area), and protein expression. Blood plasma was analyzed for glucose, nonesterified fatty acids, insulin, and plasminogen activator inhibitor-1 (PAI-1). PAI-039, an effective inhibitor of PAI-1, was orally administered to determine if PAI-1 was attenuating muscle regeneration in Ins2(WT/C96Y) mice. Ins2(WT/C96Y) mice exposed to 1 or 8 weeks of untreated type 1 diabetes before chemically induced muscle injury display significant impairments in their regenerative capacity as demonstrated by decreased muscle mass, myofiber cross-sectional area, myogenin, and Myh3 expression. PAI-1, a physiologic inhibitor of the fibrinolytic system and primary contributor to other diabetes complications, was more than twofold increased within 2 weeks of diabetes onset and remained elevated throughout the experimental period. Consistent with increased circulating PAI-1, regenerating muscles of diabetic mice exhibited excessive collagen levels at 5 and 10 days postinjury with concomitant decreases in active urokinase plasminogen activator and matrix metalloproteinase-9. Pharmacologic inhibition of PAI-1 with orally administered PAI-039 rescued the early regenerative impairments in noninsulin-treated Ins2(WT/C96Y) mice. Taken together, these data illustrate that the pharmacologic inhibition of elevated PAI-1 restores the early impairments in skeletal muscle repair observed in type 1 diabetes and suggests that early interventional studies targeting PAI-1 may be warranted to ensure optimal growth and repair in adolescent diabetic skeletal muscle.


Pharmacology of appetite suppression: implication for the treatment of obesity.
Given the current global epidemic of obesity there is a demand for new anti-obesity drugs to overcome the problem. Many pharmacological agents reduce food intake and significantly decrease body mass when administered to animals but affect feeding behaviour in a profoundly different way indicating the variety of biological mechanisms by which such agents act (appetite verses non-appetite). More limited clinical data demonstrates that some of the same drugs produce decreases in food intake and weight loss in humans. A few of these drugs do so by modifying the functioning of the appetite system as measured by subjective changes in feelings of hunger and fullness (indices of satiety). These drugs that modify the daily flux of appetite could be considered as 'appetite suppressants' with clinical potential as anti-obesity agents. Drugs that can be considered suitable candidates for appetite suppressants are agents that enhance peripherally satiety peptide systems (such as CCK, Bombesin/GRP, Enterostatin and GLP-1), alter the CNS levels of various hypothalamic neuropeptides (NPY, Galanin, Orexin, CART and Melanocortins) or monoamine neurotransmitters (such as serotonin, nor-adrenaline and possibly dopamine). Recently, the hormone leptin has become regarded as a key hormonal signal linking adipose tissue status with a number of key central nervous system circuits (NPY, CART, CRF, Melanocortins and possibly Orexins). This tonic system may also provide drug targets for the control of appetite. Any changes induced by a potential appetite suppressant should be considered in terms of the (i) psychological experience and behavioural expression of appetite, (ii) metabolism and peripheral physiology, and (iii) functioning of CNS neural pathways. In humans, such modulation of appetite will involve changes in total caloric consumption, subjective changes in feelings of hunger and fullness, preferences for specific food items, and general macronutrient preferences. These may be expressed behaviourally as changes in meal patterns, snacking behaviour and food choice. Within the next 20 years it is certain that clinicians will have a new range of anti-obesity compounds available to choose from. Such novel compounds may act on a single component of the appetite system or target a combination of these components detailed in this review. Such compounds used in combination with life style changes and dietary intervention may be critical in dealing with the rising world epidemic of obesity.


Choline acetyltransferase-deficient mutants of the nematode Caenorhabditis elegans.
We have identified five independent allelic mutations, defining the gene cha-1, that result in decreased choline acetyltransferase (ChAT) activity in Caenorhabditis elegans. Four of the mutant alleles, when homozygous, lead to ChAT reductions of greater than 98%, as well as recessive phenotypes of uncoordinated behavior, small size, slow growth and resistance to cholinesterase inhibitors. Animals homozygous for the fifth allele retain approximately 10% of the wild-type enzyme level; purified enzyme from this mutant has altered Km values for both choline and acetyl-CoA and is more thermolabile than the wild-type enzyme. These qualitative alterations, together with gene dosage data, argue that cha-1 is the structural gene for ChAT. cha-1 has been mapped to the left arm of linkage group IV and is within 0.02 map unit of the gene unc-17, mutant alleles of which lead to all of the phenotypes of cha-1 mutants except for the ChAT deficiency. Extensive complementation studies of cha-1 and unc-17 alleles reveal a complex complementation pattern, suggesting that both loci may be part of a single complex gene.


Effect of losartan on muscle sympathetic activity and baroreceptor function in systemic hypertension.
Angiotensin II directly stimulates muscle sympathetic nerve activity and facilitates adrenergic sympathetic transmission. The hypotheses that the chronic blockade of angiotensin II receptors (AT(1) type) reduces muscle sympathetic activity and that there is an interaction with baroreceptor function in patients with mild to moderate hypertension were investigated. Muscle sympathetic nerve activity decreased from 51.7 +/- 3.5 to 45.9 +/- 4.2 bursts/min (p = 0.022), and cardiac baroreceptor sensitivity increased from 3.2 +/- 1.3 to 4.9 +/- 1.8 ms/mm Hg (p = 0.007). This study for the first time demonstrates that in hypertensive patients, chronic AT(1) receptor antagonism inhibits muscle sympathetic nerve activity and that baroreceptor function is improved under these conditions.


Association of Chronic Kidney Disease with Prior Tinnitus: A Case-Control Study.
This population-based, case−control study aims to explore the relationship between prior tinnitus and the occurrence of chronic kidney disease (CKD) using a nationwide, population-based cohort study. We used data from the Taiwan National Health Insurance Research Database to explore the association of CKD with tinnitus. We identified 15,314 patients aged ≥40 years old with a first-time diagnosis of CKD as the cases. We used propensity-scored matching to select 45,942 controls (1:3 ratio). We performed multivariate logistic regression to estimate the odds ratio (OR) of a prior tinnitus diagnosis among the CKD group vs. the control group. Analysis showed that 770 (1.26%) out of the 61,256 sampled patients had previously diagnosed tinnitus. Chi-square testing revealed a significant difference in the rate of previously diagnosed tinnitus between cases and controls (3.86% vs. 0.93%, p < 0.001). Univariate logistic regression analysis showed an OR of prior tinnitus for cases of 10.249 (95% confidence interval (CI): (8.662~12.126)) relative to controls. In adjusted analysis, cases were more likely than controls to have a prior diagnosis of tinnitus (OR = 10.970, 95% CI = 9.255~13.004, p < 0.001) after adjusting for age, sex, monthly income, geographic location, urbanization level, hypertension, diabetes, coronary heart disease, hyperlipidemia, obesity, and autoimmune disease. Our study shows that CKD patients have a higher likelihood of having suffered from tinnitus before CKD was diagnosed, but we have no data suggesting that tinnitus is a predictor of subsequent CKD. Patients diagnosed with tinnitus may benefit from proactive measures to prevent CKD and detect it early through lifestyle modifications and regular renal function examinations, regardless of CKD-related symptoms.


Facile synthesis of Ba(1-x)K(x)Fe2As2 superconductors via hydride route.
We have developed a fast, easy, and scalable synthesis method for Ba(1-x)K(x)Fe2As2 (0 ≤ x ≤ 1) superconductors using hydrides BaH2 and KH as a source of barium and potassium metals. Synthesis from hydrides provides better mixing and easier handling of the starting materials, consequently leading to faster reactions and/or lower synthesis temperatures. The reducing atmosphere provided by the evolved hydrogen facilitates preparation of oxygen-free powders. By a combination of methods we have shown that Ba(1-x)K(x)Fe2As2 obtained via hydride route has the same characteristics as when it is prepared by traditional solid-state synthesis. Refinement from synchrotron powder X-ray diffraction data confirms a linear dependence of unit cell parameters upon K content as well as the tetragonal to orthorhombic transition at low temperatures for compositions with x < 0.2. Magnetic measurements revealed dome-like dependence of superconducting transition temperature Tc upon K content with a maximum of 38 K for x close to 0.4. Electron diffraction and high-resolution high-angle annular dark-field scanning transmission electron microscopy indicates an absence of Ba/K ordering, while local inhomogeneity in the Ba/K distribution takes place at a scale of several angstroms along [110] crystallographic direction.


NuMA phosphorylation by CDK1 couples mitotic progression with cortical dynein function.
Spindle positioning and spindle elongation are critical for proper cell division. In human cells, an evolutionary conserved ternary complex (NuMA/LGN/Gαi) anchors dynein at the cortex during metaphase, thus ensuring correct spindle positioning. Whether this complex contributes to anaphase spindle elongation is not known. More generally, the mechanisms coupling mitotic progression with spindle behaviour remain elusive. Here, we uncover that levels of cortical dynein markedly increase during anaphase in a NuMA-dependent manner. We demonstrate that during metaphase, CDK1-mediated phosphorylation at T2055 negatively regulates NuMA cortical localization and that this phosphorylation is counteracted by PPP2CA phosphatase activity. We establish that this tug of war is essential for proper levels of cortical dynein and thus spindle positioning during metaphase. Moreover, we find that upon CDK1 inactivation in anaphase, the rise in dephosphorylated NuMA at the cell cortex leads to cortical dynein enrichment, and thus to robust spindle elongation. Our findings uncover a mechanism whereby the status of NuMA phosphorylation coordinates mitotic progression with proper spindle function.


Energy-density distribution inside large nonabsorbing spheres by using Mie theory and geometrical optics.
Mie theory and geometrical-optics ray tracing are used to obtain the distribution of electric energy density inside a nonabsorbing micrometer-sized sphere illuminated by a polarized plane wave. The Mie solution shows the multiply reflected geometrical-optics rays inside a sphere having a diameter of ~ 150 free-space wavelengths (size parameter = circumference/wavelength = 500). The geometrical-optics result shows the major features of the Mie solution and provides a physical interpretation of the electromagnetic interactions that result in the observed energy-density distributions. Both solutions show internal on-axis energy-density maxima inside the shadow surface of the sphere. The region of greatest enhanced energy density is approximately one internal wavelength in diameter and approximately twenty internal wavelengths in length.


A randomized trial of direct-to-patient communication to enhance adherence to beta-blocker therapy following myocardial infarction.
Although beta-blockers are routinely prescribed at hospital discharge after myocardial infarction (MI), patients' adherence has been shown to decline substantially over time. We sought to test the hypothesis that a simple, direct-to-patient intervention can improve adherence to beta-blocker therapy following MI. We conducted a cluster randomized controlled trial in 4 geographically dispersed health maintenance organizations testing the hypothesis that a simple direct-to-patient intervention could improve adherence. The study was carried out from June 2004 to March 2005. The primary analyses were based on 836 post-MI patients who were dispensed a beta-blocker prescription after discharge. The intervention consisted of 2 mailings 2 months apart describing the importance of beta-blocker use. The main outcomes were proportion of days covered with beta-blocker therapy and percentage of patients with at least 80% of days covered in the 9 months after the first mailing. Analyses were adjusted for age, sex, total medications dispensed, days between MI and intervention, and intervention site. Over the entire follow-up period, patients in the treatment arm had a mean absolute increase of 4.3% of days covered per month compared with patients in the control arm (a 5.7% relative change from baseline), representing 1.3 extra days (P = .04). Treatment patients were 17% more likely (relative risk, 1.17; 95% confidence interval, 1.02-1.29) to have 80% of days covered. For every 16 patients receiving the intervention, 1 additional patient would become adherent (80% or more days covered per month). A low-cost, easily replicable effort to increase adherence can have a demonstrable impact on beta-blocker adherence following MI. Trial Registration clinicaltrials.gov Identifier: NCT00211172.


Executive function and uptake of 99mTc-exametazime shown by single photon emission tomography after oral idazoxan in probable Alzheimer-type dementia.
Preliminary reports suggest improved executive function in patients with lobar dementia after treatment with single doses of the alpha 2 adrenoceptor antagonist, idazoxan. The potential for use in probable Alzheimer-type dementia prompted the present study. Fifteen patients with probable Alzheimer-type dementia were examined twice with neuropsychological measures and 14 also with single photon emission tomography (SPET) after a single double blind oral administration of 40 mg idazoxan or placebo in a balanced cross-over design. Brain perfusion maps were spatially transformed into standard stereotactic space and compared pixel-by-pixel. A parametric analysis was used to examine the relationship between the drug effect, verbal fluency and brain perfusion. Two to 3 h after idazoxan, measures of reaction time, Stroop test, category fluency and anxiety were unchanged. Verbal fluency (letter) and spatial working memory were impaired and performance on the Tower of London test in a sub-set of patients showed a trend to impairment in the idazoxan condition. Idazoxan produced a modest relative activation in left thalamus and inferior occipital cortex: decreases occurred in inferior anterior cingulate and left insular cortex. There were significant correlations on both days between measures of fluency and brain perfusion in left lateral prefrontal cortex. The reduced performance with idazoxan was directly correlated with reduced perfusion in left lateral prefrontal cortex, supporting an important interaction between drug and task performance. The imaging component of the study therefore suggested that activation of frontal networks is necessary for performing fluency tasks in Alzheimer-type dementia. Brain networks involving prefrontal cortex are the locus for the primary cognitive effects of noradrenergic drugs. The direction of the effect of any dose of agonist or antagonist may depend critically upon the age and pathology of the experimental subjects and the relationship between performance, noradrenergic drive and task difficulty.


The fundamental operating principles of electronic root canal length measurement devices.
It is generally accepted that root canal treatment procedures should be confined within the root canal system. To achieve this objective the canal terminus must be detected accurately during canal preparation and precise control of working length during the process must be maintained. Several techniques have been used for determining the apical canal terminus including electronic methods. However, the fundamental electronic operating principles and classification of the electronic devices used in this method are often unknown and a matter of controversy. The basic assumption with all electronic length measuring devices is that human tissues have certain characteristics that can be modelled by a combination of electrical components. Therefore, by measuring the electrical properties of the model, such as resistance and impedance, it should be possible to detect the canal terminus. The root canal system is surrounded by dentine and cementum that are insulators to electrical current. At the minor apical foramen, however, there is a small hole in which conductive materials within the canal space (tissue, fluid) are electrically connected to the periodontal ligament that is itself a conductor of electric current. Thus, dentine, along with tissue and fluid inside the canal, forms a resistor, the value of which depends on their dimensions, and their inherent resistivity. When an endodontic file penetrates inside the canal and approaches the minor apical foramen, the resistance between the endodontic file and the foramen decreases, because the effective length of the resistive material (dentine, tissue, fluid) decreases. As well as resistive properties, the structure of the tooth root has capacitive characteristics. Therefore, various electronic methods have been developed that use a variety of other principles to detect the canal terminus. Whilst the simplest devices measure resistance, other devices measure impedance using either high frequency, two frequencies, or multiple frequencies. In addition, some systems use low frequency oscillation and/or a voltage gradient method to detect the canal terminus. The aim of this review was to clarify the fundamental operating principles of the different types of electronic systems that claim to measure canal length.