EPIGIST: An observational real-life study on patients with metastatic gastrointestinal stromal tumors receiving imatinib.
Gastrointestinal stromal tumors (GISTs) are rare, but represent the most common mesenchymal neoplasms of the gastrointestinal tract. EPIdemiology GIST, is an observational multicenter longitudinal follow-up cohort study reporting the prescribing patterns of imatinib in patients with GIST and the impact of the treatment in a real-world (standard clinical) setting. Eligible patients had a confirmed diagnosis of unresectable or metastatic KIT-positive GIST and started treatment with imatinib for the first time between May 24, 2002, and June 30, 2010. During routine visits, annual collection of clinical characteristics was requested, i.e., age, GIST stage at diagnosis, history, imatinib treatment duration and dosage, adherence, and concomitant medications. Survival outcomes were estimated using the Kaplan-Meier method. Other data were analyzed using descriptive statistics. Of 151 patients enrolled, imatinib was initiated for 126 patients before enrollment and for 25 patients on the day of enrollment or soon after. The patient characteristics were similar to those in published prospective trials. The estimated 1-, 2-, 3-, and 4-year overall survival rates were 90.4% (95% confidence interval [CI; 84.8%-94.0%]), 84.7% (95% CI [78.1%-89.4%]), 73.0% (95% CI [65.0%-79.4%]), and 60.7% (95% CI [51.4%-68.8%]), respectively. The most common adverse events (AEs) were diarrhea (39%), asthenia (39%), eyelid or periorbital edema (32%), abdominal pain (23%), and anemia (21%). Eight of 126 serious AEs were possibly related to the treatment as assessed by investigators. Study results showed that patients in real-life populations are generally treated in accordance with national and international clinical recommendations and have outcomes comparable to those of patients in clinical trials.


Purification, cloning, and primary structure of an enantiomer-selective amidase from Brevibacterium sp. strain R312: structural evidence for genetic coupling with nitrile hydratase.
An enantiomer-selective amidase active on several 2-aryl and 2-aryloxy propionamides was identified and purified from Brevibacterium sp. strain R312. Oligonucleotide probes were designed from limited peptide sequence information and were used to clone the corresponding gene, named amdA. Highly significant homologies were found at the amino acid level between the deduced sequence of the enantiomer-selective amidase and the sequences of known amidases such as indoleacetamide hydrolases from Pseudomonas syringae and Agrobacterium tumefaciens and acetamidase from Aspergillus nidulans. Moreover, amdA is found in the same orientation and only 73 bp upstream from the gene coding for nitrile hydratase, strongly suggesting that both genes are part of the same operon. Our results also showed that Rhodococcus sp. strain N-774 and Brevibacterium sp. strain R312 are probably identical, or at least very similar, microorganisms. The characterized amidase is an apparent homodimer of Mr 2 x 54,671 which exhibited under our conditions a specific activity of about 13 to 17 mumol of 2-(4-hydroxyphenoxy)propionic R acid formed per min per mg of enzyme from the racemic amide. Large amounts of an active recombinant enzyme could be produced in Escherichia coli at 30 degrees C under the control of an E. coli promoter and ribosome-binding site.


Interfacial Aβ fibril formation is modulated by the disorder-order state of the lipids: The concept of the physical environment as amyloid inductor in biomembranes.
The behavior of amphiphilic molecules such as lipids, peptides and their mixtures at the air/water interface allow us to evaluate and visualize the arrangement formed in a confined and controlled surface area. We have studied the surface properties of the zwitterionic DPPC lipid and Aβ(1-40) amyloid peptide in mixed films at different temperatures (from 15 to 40 °C). In this range of temperature the surface properties of pure Aβ(1-40) peptide remained unchanged, whereas DPPC undergoes its characteristic liquid-expanded → liquid-condensed bidimensional phase transition that depends on the temperature and lateral pressure. This particular property of DPPC makes it possible to dynamically study the influence of the lipid phase state on amyloid structure formation at the interface in a continuous, isothermal and abrupt change on the environmental condition. As the mixed film is compressed the fibril-like structure of Aβ(1-40) is triggered specifically in the liquid-expanded region, independently of temperature, and it is selectively excluded from the well-visible liquid condensed domains of DPPC. The Aβ amyloid fibers were visualized by using BAM and AFM and they were Thio T positive. In mixed DPPC/Aβ(1-40) films the condensed domains (in between 11 mN/m to 20 mN/m) become irregular probably due to the fibril-like structures is imposing additional lateral stress sequestering lipid molecules in the surrounding liquid-expanded phase to self-organize into amyloids.


Quantitative analysis for delta 9-THC, 11-hydroxy-delta 9-THC, and 9-carboxy-delta 9-THC in plasma using GC/CI-MS.
A method for quantitative measurement of delta 9-THC concentrations in plasma has been developed and applied to the analysis of more than 2000 samples over the past 4 years. The method includes addition of deuterium-labeled delta 9-THC to the plasma for use as the internal standard, a simple and relatively rapid solvent extraction procedure, formation of the trimethylsilyl derivative, and quantitation by selected ion monitoring using ammonia chemical ionization. A similar procedure has also been developed for simultaneous analysis of delta 9-THC and two of its major metabolites in plasma. The latter procedure requires use of a glass capillary column for assays of plasma samples in which the delta 9-THC concentration is below 10 ng/ml. Sensitivities for both methods permit quantitation of THC and its metabolites at concentrations as low as 0.2 ng in 1-ml plasma samples.


Characterization of the M2e antibody response following highly pathogenic H5N1 avian influenza virus infection and reliability of M2e ELISA for identifying infected among vaccinated chickens.
A surveillance method able to differentiate between vaccinated and infected poultry is required for those countries that practice vaccination against highly pathogenic avian influenza H5N1. The external domain of the M2 protein (M2e) of influenza virus is a potentially useful differentiating-infected from vaccinated animals (DIVA) antigen but little is known about the M2e antibody response and factors influencing its detection. In this study, the M2e antibody response was characterized in layer birds vaccinated and challenged with an Indonesian H5N1 virus isolate, using a single M2e peptide or four-branched multiple antigenic peptide form of M2e (MAP-M2e) as antigens in two separate ELISAs. Anti-M2e antibodies were absent in chicks with high level of maternal haemagglutination inhibition antibodies and also in all layers vaccinated once, twice or three times with an inactivated commercial H5N1 vaccine. In contrast, anti-M2e antibodies were detected in vaccinated layers challenged with H5N1 virus and their presence was associated with virus isolation and an increase in haemagglutination inhibition titres. The number of birds that developed M2e antibodies, as well as the strength and duration of the M2e antibody response were strongly influenced by the length of the interval between vaccination and challenge. Birds challenged at six weeks after vaccination all developed M2e antibodies by 14 days that lasted until at least 56 days after infection. In birds challenged at two weeks after vaccination, only a proportion of birds developed M2e antibodies by 14 days that lasted only until 28 days post-infection. Both single M2e peptide and MAP-M2e ELISAs had high diagnostic specificity but the diagnostic sensitivity of MAP-M2e ELISA was significantly higher and more effective in detecting M2e antibody in immune and infected birds. The results show that MAP-M2e ELISA would be useful for surveillance in countries using vaccination to control highly pathogenic avian influenza H5N1.


Enhancing anti-AML activity of venetoclax by isoflavone ME-344 through suppression of OXPHOS and/or purine biosynthesis in vitro.
Venetoclax (VEN), in combination with low dose cytarabine (AraC) or a hypomethylating agent, is FDA approved to treat acute myeloid leukemia (AML) in patients who are over the age of 75 or cannot tolerate standard chemotherapy. Despite high response rates to these therapies, most patients succumb to the disease due to relapse and/or drug resistance, providing an unmet clinical need for novel therapies to improve AML patient survival. ME-344 is a potent isoflavone with demonstrated inhibitory activity toward oxidative phosphorylation (OXPHOS) and clinical activity in solid tumors. Given that OXPHOS inhibition enhances VEN antileukemic activity against AML, we hypothesized that ME-344 could enhance the anti-AML activity of VEN. Here we report that ME-344 enhanced VEN to target AML cell lines and primary patient samples while sparing normal hematopoietic cells. Cooperative suppression of OXPHOS was detected in a subset of AML cell lines and primary patient samples. Metabolomics analysis revealed a significant reduction of purine biosynthesis metabolites by ME-344. Further, lometrexol, a purine biosynthesis inhibitor, synergistically enhanced VEN-induced apoptosis in AML cell lines. Interestingly, AML cells with acquired AraC resistance showed significantly increased purine biosynthesis metabolites and sensitivities to ME-344. Furthermore, synergy between ME-344 and VEN was preserved in these AraC-resistant AML cells. In vivo studies revealed significantly prolonged survival upon combination therapy of ME-344 and VEN in NSGS mice bearing parental or AraC-resistant MV4-11 leukemia compared to the vehicle control. This study demonstrates that ME-344 enhances VEN antileukemic activity against preclinical models of AML by suppressing OXPHOS and/or purine biosynthesis.


Nonparametric estimation of the mean function for recurrent event data with missing event category.
Recurrent event data frequently arise in longitudinal studies when study subjects possibly experience more than one event during the observation period. Often, such recurrent events can be categorized. However, part of the categorization may be missing due to technical difficulties. If the event types are missing completely at random, then a complete case analysis may provide consistent estimates of regression parameters in certain regression models, but estimates of the baseline event rates are generally biased. Previous work on nonparametric estimation of these rates has utilized parametric missingness models. In this paper, we develop fully nonparametric methods in which the missingness mechanism is completely unspecified. Consistency and asymptotic normality of the nonparametric estimators of the mean event functions accommodate nonparametric estimators of the event category probabilities, which converge more slowly than the parametric rate. Plug-in variance estimators are provided and perform well in simulation studies, where complete case estimators may exhibit large biases and parametric estimators generally have a larger mean squared error when the model is misspecified. The proposed methods are applied to data from a cystic fibrosis registry.


Prevalence and factors associated with wound colonization by Staphylococcus spp. and Staphylococcus aureus in hospitalized patients in inland northeastern Brazil: a cross-sectional study.
Infections by Staphylococcus spp. are often associated with wounds, especially in hospitalized patients. Wounds may be the source of bacteria causing cross-contamination, and are a risk factor for methicillin-resistant Staphylococcus aureus (MRSA) infection. The aim of this study was to investigate the prevalence of wound colonization by Staphylococcus spp., especially S. aureus and MRSA, in hospitalized patients, and to identify the factors associated with such colonization. This cross-sectional study enrolled patients with wounds who were hospitalized in a remote and underdeveloped inland region of northeastern Brazil with extreme poverty. Samples were collected using sterile swabs with 0.85% saline solution, and coagulase-negative Staphylococcus spp., S. aureus, and MRSA were identified using standard laboratory procedures. Data regarding the sociodemographic characteristics, antibiotic use, and comorbidities of the patients were collected using the medical records and a questionnaire. A total of 125 wounds were analyzed. The patients had a mean age of 63.88 years and a mean 3.84 years of school education. Eighty-one wounds (64.80%) were colonized by Staphylococcus spp. Twenty-five wounds (20%) were colonized by S. aureus, 32% of which were colonized by MRSA. Wound colonization by Staphylococcus spp. was associated with pneumonia or other respiratory disease (p = 0.03). Wound colonization by S. aureus was associated with nasal colonization by S. aureus (p < 0.001), fewer days of prior antibiotic use (p = 0.04), admission to a medical ward (p = 0.02), and age >65 years (p = 0.05). Among patients with wound colonization by MRSA, 37.50% had a history of prior antibiotic use, 75% had two or more comorbidities, 25% had cancer or diabetes, 50% had cardiovascular disease, and 50% died. Wounds can be the source of Staphylococcus spp. infection, and high proportions of wounds are colonized by S. aureus and MRSA. Nasal colonization by S. aureus may be a source for wound colonization by S. aureus, illustrating the importance of preventing cross-contamination in hospital environments, especially among elderly patients. Wounds should be carefully managed to prevent microbial spread, thereby assisting patient recovery and reducing healthcare costs.


New evidence on the systematic and phylogenetic position of Parides burchellanus (Lepidoptera: Papilionidae).
Parides burchellanus is considered a rare and threatened swallowtail species restricted to central Brazil. It shows considerable morphological similarity to Parides panthonus jaguarae, with which it shares both geographical range and larval host plant. At present, P. burchellanus and P. panthonus are believed to be distinct species, based on minor differences in male genitalia. In this study, the phylogenetic and systematic position of P. burchellanus in relation to three subspecies of P. panthonus (P. p. jaguarae, P. p. lysimachus and P. p. aglaope) was evaluated using molecular evidence: the complete sequence of the mtDNA genes COI and COII and of the nuclear gene EF-1α (c. 3300 bp). In addition, the informativeness of the 'barcode' region next to the 5' end of COI (c. 650 bp) was evaluated for delimiting these taxa. Individual analysis by neighbour-joining, using Kimura 2-parameter distance model, and by maximum parsimony showed that P. p. jaguarae + P. p. lysimachus + P. p. aglaope + P. burchellanus form a strongly supported monophyletic clade, and all molecular regions consistently recovered P. p. jaguarae and P. burchellanus as sister species. The genetic divergence among the subspecies of P. panthonus and P. burchellanus is equivalent to the divergence among conspecifics of other species of Parides, and smaller than the interspecific divergence among different sister species of this genus. The results support the proposal that P. p. jaguarae and P. burchellanus are likely to be synonymous, and suggest that P. burchellanus can be considered conspecific with P. panthonus. The taxonomic classification of P. burchellanus should be revised on the basis of the molecular data.


Spinal Tap Needle Technique for Creating Apical Plug with Mineral Trioxide Aggregate in Immature Permanent Teeth.
This report evaluates the apical plug formation in two cases with necrotic immature maxillary central incisors in 11 and 12 years old female patients by the use of spinal tap needle to create apical plug-in single increment without any residual mineral trioxide aggregate (MTA) attachment to the wall of the root canal. The shape of the immature root canal resembles a truncated cone. To achieve a truncated cone with MTA on the apical 4 mm in order to plug the open apex, the volume of material that will be required can be calculated by taking spinal tap needle (BD Spinal, Spain) equal to the last file used, in a way that the MTA extruded is equal to the amount required to shape the apical plug in a single increment. A uniform void-free apical plug was made in single increments by using this technique. This spinal tap needle is very economical, available in various sizes corresponding to endodontic K files, so spinal tap needle method is a viable method to create uniform void-free accurate apical plug in one increment without the wastage of material and time preventing the material to extrude beyond. Yadav P, Virdi MS. Spinal Tap Needle Technique for Creating Apical Plug with Mineral Trioxide Aggregate in Immature Permanent Teeth. Int J Clin Pediatr Dent 2022;15(3):380-384.


Low-carbohydrate diets and prevalence, incidence and progression of coronary artery calcium in the Multi-Ethnic Study of Atherosclerosis (MESA).
The evidence linking low-carbohydrate diets (LCD) to CVD is controversial, and results from epidemiological studies are inconsistent. We aimed to assess the relationship between LCD patterns and coronary artery Ca (CAC) scores from computed tomography in the Multi-Ethnic Study of Atherosclerosis cohort. Our sample included 5614 men and women free of clinical CVD at baseline (2000-2002), who had a FFQ, a baseline measure and ≥1 measure of CAC during follow-up. We excluded those with implausible energy intake or daily physical activity. The overall, animal-based and plant-based LCD scores were calculated based on intakes of macronutrients. Relative risk regression and robust regression models were used to examine the cross-sectional and longitudinal relationship between LCD score quintile and CAC outcomes, after adjustment for multiple cardiovascular risk factors. The mean age of participants was 63 years. The median intakes of total carbohydrate, fat and protein were 53·7, 30·5 and 15·6 % energy/d, respectively. Among 2892 participants with zero CAC scores at baseline, 264 developed positive scores during 2·4-year follow-up (11-59 months). Among those with positive scores at baseline, the median increase in CAC was 47 units over the course of follow-up. The overall, the animal-based and the plant-based LCD scores were not associated with CAC prevalence, incidence and progression. In conclusion, diets low in carbohydrate and high in fat and/or protein, regardless of the sources of protein and fat, were not associated with higher levels of CAC, a validated predictor of cardiovascular events, in this large multi-ethnic cohort.


A computerized method for measuring respiratory mechanics during mechanical ventilation.
In order to monitor respiratory mechanics in anesthetized ventilated subjects, an automated method was developed and tested on a physical model. The model was composed of an endotracheal tube (curvilinear resistance) coupled to a rigid air-filled box (elastance). Theoretical resistance was determined during steady-state flow experiments and theoretical elastance was estimated from the dimensions of the box. The physical model was connected to a volume-cycled ventilator. Pressure (P) and air flow (V) were measured at the outlet of the tube, and the time integral V of V was calculated. Elastance (E) and curvilinear resistance (R + K/V/) were identified by multiple linear regression analysis, for each ventilatory cycle, according to equation P = EV + RV + K/V/V. When linear regression analysis of P on V, V and /V/V was performed over the entire ventilatory cycle, E was found equal to its estimate, whereas R and K appeared different from their theoretical values. In order to improve determination of R and K, resistive pressure (Pr = P - EV) was calculated using the previously obtained value of E, and multiple linear regression of Pr on V and /V/V was performed over different fractions of the ventilatory cycle. When determined over the expiratory phase corresponding to decreasing flows, R and K were found close to their expected values. Such a method to calculate elastance and curvilinear resistance should prove convenient and efficient in measuring respiratory mechanics during mechanical ventilation.


Cryptochrome 1a of tomato mediates long-distance signaling of soil water deficit.
Although the blue light photoreceptors cryptochromes mediate the expression of genes related to reactive oxygen species, whether cryptochrome 1a (cry1a) regulates local and long-distance signaling of water deficit in tomato (Solanum lycopersicum L.) is unknown. Thus the cry1a tomato mutant and its wild-type (WT) were reciprocally grafted (WT/WT; cry1a/cry1a; WT/cry1a; cry1a/WT; as scion/rootstock) or grown on their own roots (WT and cry1a) under irrigated and water deficit conditions. Plant growth, pigmentation, oxidative stress, water relations, stomatal characteristics and leaf gas exchange were measured. WT and cry1a plants grew similarly under irrigated conditions, whereas cry1a plants had less root biomass and length and higher tissue malondialdehyde concentrations under water deficit. Despite greater oxidative stress, cry1a maintained chlorophyll and carotenoid concentrations in drying soil. Lower stomatal density of cry1a likely increased its leaf relative water content (RWC). In grafted plants, scion genotype largely determined shoot and root biomass accumulation irrespective of water deficit. In chimeric plants grown in drying soil, cry1a rootstocks increased RWC while WT rootstocks maintained photosynthesis of cry1a scions. Manipulating tomato CRY1a may enhance plant drought tolerance by altering leaf pigmentation and gas exchange during soil drying via local and long-distance effects.


Directed evolution of metabolic pathways in microbial populations. I. Modification of the acid phosphatase pH optimum in S. cerevisiae.
An experimental system for directing the evolution of enzymes and metabolic pathways in microbial populations is proposed and an initial test of its power is provided.-The test involved an attempt to genetically enhance certain functional properties of the enzyme acid phosphatase in S. cerevisiae by constructing an environment in which the functional changes desired would be "adaptive". Naturally occurring mutations in a population of 10(9) cells were automatically and continuously screened, over 1,000 generations, for their effect on the efficiency (K(m)) and activity of acid phosphatase at pH 6, and for their effect on the efficiency of orthophosphate metabolism.-The first adaptation observed, M1, was due to a single mutational event that effected a 30% increase in the efficiency of orthophosphate metabolism. The second, M2, effected an adaptive shift in the pH optimum of acid phosphatase and an increase in its activity over a wide range of pH values (an increment of 60% at pH 6). M2 was shown to result from a single mutational event in the region of the acid phosphatase structural gene. The third, M3, effected cell clumping, an adaptation to the culture apparatus that had no effect on phosphate metabolism.-The power of this system for directing the evolution of enzymes and of metabolic pathways is discussed in terms of the kinetic properties of the experimental system and in terms of the results obtained.


Midkine stimulates Wilms' tumor cell proliferation via its signaling receptor.
Midkine, but not pleiotrophin, is mitogenic to human Wilms' tumor cells (G401 line) in dose-dependent and time-dependent fashion. Midkine specifically binds to high affinity (Kd = 0.15 +/- 0.02 nM, 210 kDa) and low affinity receptors (Kd = 0.65 +/- 0.07 nM, 75 kDa) on G401 cell surface, that has been confirmed by cross-linking and competition experiments. In addition, midkine stimulates a tyrosine phosphorylation of several proteins with molecular weight about 110-115 kDa, 130-140 kDa and 210 kDa. These data allow us to suggest that a key point in stimulation of G401 cell proliferation is interaction of midkine to its signaling receptor.


Degradation and formation of wood odorant β-cyclocitral during permanganate oxidation.
The effect of permanganate oxidation on the formation and degradation of wood odorant β-cyclocitral in water was investigated. Oxidation experiments were conducted for β-cyclocitral prepared from pure chemicals and extracted from Microcystis aeruginosa. The data were simulated with appropriate kinetic rate models. The formation and degradation of β-cyclocitral during the oxidation of β-carotene were also monitored and modeled. The degradation of β-cyclocitral prepared from pure chemicals followed second-order kinetics with a rate constant of 91.7 ± 2.4M(-1)s(-1), and that of the β-cyclocitral precursor, β-carotene, followed first-order kinetics with a rate constant of 0.0054 ± 0.0004 min(-1). During the oxidation of β-carotene, β-cyclocitral was produced. The formation and degradation can both be simulated by first-order kinetics with respect to β-carotene and β-cyclocitral concentrations, respectively. The degradation rate of β-cyclocitral produced from β-carotene was found to be much slower than that for β-cyclocitral obtained from pure chemicals, very likely due to a mass transfer limitation. The kinetic models were further employed to simulate the oxidation of β-cyclocitral in the presence of β-carotene and for β-cyclocitral extracted from M. aeruginosa, respectively. The models well predict/fit the experimental data, with rate constants similar to other experiments, indicating that the models may be used for simulating the formation and degradation of β-cyclocitral in water treatment systems.


A recommendation to perform a blood culture before the administration of intravenous antibiotics increased the detection of Staphylococcus aureus bacteremia.
In 2004, the Surviving Sepsis Campaign was launched to increase awareness and improve the outcome of severe sepsis. Accordingly, in Jönköping County, Sweden, a strong recommendation to perform a blood culture before the start of intravenous antibiotic treatment was introduced in 2007. Moreover, a reminder was included in the laboratory report to consult an infectious disease specialist when Staphylococcus aureus was isolated from a blood culture. Retrospectively, patients with at least one blood culture growing S. aureus during 2002 through 2003 (pre intervention n = 58) or during 2008 through 2009 (post intervention n = 100) were included. Medical records were evaluated regarding clinical data and outcome. Blood culture isolates were characterized by antibiotic susceptibility testing (AST) and S. aureus protein A (spa) gene typing. The annual incidence of S. aureus bacteremia (SAB) increased from 28 per 100,000 inhabitants at the pre intervention period to 45 per 100,000 at the post intervention period (p = 0.046). During post intervention, the SAB incidence was significantly higher in men (p = 0.009). The mortality rate during hospital stay was 14 % during pre intervention and 18 % during post intervention (p = 0.47). The most common spa types were t012 and t084. The Surviving Sepsis Campaign resulted in an increased number of detected cases of SAB. The mortality rate was the same before and after the intervention, and no spa type correlated to certain clinical manifestations or mortality.


Clusters of high abundance of plants detected from local indicators of spatial association (LISA) in a semi-deciduous tropical forest.
Plants are rarely randomly distributed across communities, and patchiness is a common spatial pattern in most tropical forests. Clusters of high density of plant individuals are related to internal and external forces, as well as to historical events. The detection of aggregated patterns of plant individuals allows for a better understanding of the internal and external factors that guide the distribution of species. The aim of this research was to detect and characterize clusters of high abundance of plants and species richness in semi-deciduous forests in the Dominican Republic. For this, we collected vegetation data from 575 quadrats in 23 transects (2300 m2 in total) within the Ocoa river basin. Using local Moran's I statistics, we isolated 18 quadrats of high density of individuals. We show that density of individuals can be 2.5 times larger on average than in non-aggregated quadrats, and can reach higher values for shrubs species as well as for palms and vines species. In addition, we found that shrub species are the most abundant group in aggregated quadrats, and density of tree species is significantly smaller than that of shrub species. High density quadrats are predominantly occupied by shrubs, palms and vines, following patterns of species composition and lithology. Detecting clusters of high density of individuals could help in the efficient assessment of richness in semi-deciduous tropical forests, and may support new conservation practices for this valuable but threatened ecosystem.


The effects of external potassium and long duration voltage conditioning on the amplitude of sodium currents in the giant axon of the squid, Loligo pealei.
Giant axons were voltage-clamped in solutions of constant sodium concentration (230 mM) and variable potassium concentrations (from 0 to 210 mM). The values of the peak initial transient current, I(p), were measured as a function of conditioning prepulse duration over the range from less than 1 msec to over 3 min. Prepulse amplitudes were varied from E(m) = -20 mv to E(m) = -160 mv. The attenuation of the I(p) values in high [K(o)] was found to vary as a function of time when long duration conditioning potentials were applied. In both high and low [K(o)], I(p) values which had reached a quasi-steady-state level within a few milliseconds following a few milliseconds of hyperpolarization were found to increase following longer hyperpolarization. A second plateau was reached with a time constant of about 100-500 msec and a third with a time constant in the range of 30 to 200 sec. The intermediate quasi-steady-state level was absent in K-free ASW solutions. Sodium inactivation curves, normalized to I(pmax) values obtained at either the first or second plateaus, were significantly different in different [K(o)]. The inactivation curves, however, tended to superpose after about 1 min of hyperpolarizing conditioning. The time courses and magnitudes of the intermediate and very slow sodium conductance restorations induced by long hyperpolarizing pulses are in agreement with those predicted from the calculated rates and magnitudes of [K(+)] depletion in the space between the axolemma and the Schwann layer.


Predictive factors of central lymph node metastasis in papillary thyroid carcinoma.
The aim of this study was to evaluate the correlation between central lymph node (CLN) metastasis and clinicopathologic characteristics of papillary thyroid cancer (PTC). In addition, we investigated the incidence and risk factors for contralateral CLN metastasis in unilateral PTC. This study suggests the appropriate surgical extent for CLN dissection. A prospective study of 500 patients with PTC who underwent total thyroidectomy and prophylactic bilateral CLN dissection was conducted. Of 500 patients, 255 had CLN metastases. The rate of CLN metastasis was considerably higher in cases of younger patients (<45 years old) (P < 0.001; odds ratio [OR], 2.357) and of a maximal tumor size greater than 1 cm (P < 0.001; OR, 3.165). Ipsilateral CLN metastasis was detected in 83.1% of cases (133/160) of unilateral PTC, only contralateral CLN metastases in 3.7% of cases (6/160), and bilateral CLN metastases in 13.1% of cases (21/160). The rate of contralateral CLN metastasis was considerably higher in cases of PTC with a large tumor size (≥1 cm) (P = 0.019; OR, 4.440) and with ipsilateral CLN metastasis (P = 0.047; OR, 2.613). Younger age (<45 years old) and maximal tumor size greater than 1 cm were independent risk factors for CLN metastasis. Maximal tumor size greater than 1 cm and presence of ipsilateral CLN macrometastasis were independent risk factors for contralateral CLN metastasis. Therefore, both CLN dissections should be considered for unilateral PTC with a maximal tumor size greater than 1 cm or presence of ipsilateral CLN macrometastasis.


Health policy in South Australia 2003-10: primary health care workforce perceptions of the impact of policy change on health promotion.
This paper examines recent Australian health reform policies and considers how the primary health care (PHC) workforce experiences subsequent change and perceives its impact on health promotion practice. Health policy documents were analysed to determine their intended impact on health promotion. Interviews were conducted with 39 respondents from four State-funded PHC services to gain their perceptions of the impact of policy change on health promotion. There have been a plethora of policy and strategy documents over the last decade relevant to PHC, and these suggest an intention to strengthen health promotion. However, respondents report that changes to the role and focus of PHC services have led to fewer opportunities for health promotion. Services are struggling to engage in health promotion activity, while funding and policy directions are prioritised to targeted, individual behaviour change. The experience of PHC workforce respondents in South Australia suggests that, despite policy intentions, health promotion practice is much reduced. Our research suggests that rigorous evaluation of health sector reforms should be undertaken to assess both intended and unintended outcomes in terms of service quality and delivery. SO WHAT? Health promoters are experiencing a contradictory policy and practice environment, and this research should assist health promoters in advocating for more government accountability in the implementation of policies in order to advance comprehensive PHC.


Electrical properties of cultured human adrenocorticotropin-secreting adenoma cells: effects of high K+, corticotropin-releasing factor, and angiotensin II.
ACTH-secreting pituitary adenoma cells were cultured from specimens obtained by transphenoidal hypophysectomy in five patients with Cushing's disease. The majority of adenoma cells (90%) stained specifically with antiserum against human ACTH. The electrophysiological properties and response to hormones of these cells were studied with intracellular recording techniques under current clamp and voltage clamp conditions. Most (80%) of the cells fired action potentials that were Ca2+-dependent inasmuch as they were blocked by Co2+ (5 mM) and by removal of Ca2+ from the medium, but were unaffected by tetrodotoxin (0.3 mM) and by Na+ removal. The cells responded to factors known to stimulate ACTH release, including high K+, CRF, and angiotensin II (AII). High K+ (50 mM) induced a membrane depolarization in association with an increase in conductance. CRF (100 nM) produced a depolarization, a decrease in conductance, an increase in spike firing, and an increase in spike duration. Although AII was inactive in ordinary recordings, in cells loaded with lithium (Li+) to promote the phospholipid-dependent second messenger system, the peptide produced an increase in spike firing and spike duration with no change in membrane potential. The combination of CRF and AII (CRF + AII; 100 nM each) in Li+-loaded cells caused a greater excitatory effect than either peptide alone. Under voltage clamp, the response either to CRF or to CRF + AII could be attributed, at least in part, to the inhibition of a slow, voltage-dependent K+ current that is persistently active at resting potential. These results indicate that modulation of action potential firing may be an early step in the regulation of ACTH release from pituitary cells by known secretagogues. Since action potentials in these cells are associated with Ca2+ entry, the resulting changes in intracellular Ca2+ levels could mediate the effects of the hormones on secretion.


The effect of alcohol on the choline requirement. I. Changes in the rat's liver following prolonged ingestion of alcohol.
Rats maintained for a period of 7 months on a fluid intake of 15 per cent alcohol and a diet marginal in lipotropic activity developed fatty infiltration and mild fibrosis of the liver. Similar changes were observed in pair-fed controls given an isocaloric equivalent of sucrose instead of alcohol, but not in pair-fed controls receiving neither alcohol nor sucrose supplements. To exclude the possibility that the alcohol effect was related to an augmentation of the caloric intake, a third group of controls was given the same amount of alcohol, but a limited number of calories. This was accomplished by subtracting from the basal diet an amount of sucrose equivalent in calories to the alcohol consumed. Under these conditions the hepatic changes following alcohol ingestion appeared to be enhanced. Choline or methionine, on the other hand, abolished the effects of both alcohol and sucrose supplements. There was no increase in fecal nitrogen excretion following alcohol ingestion, and no histological changes were observed in the pancreas. These results are consistent with the hypothesis that alcohol increases the choline requirement of the rat, but do not support the contention that this effect is the consequence of an augmented caloric intake. Further studies are needed to establish conclusively the relationship between alcohol ingestion and the choline requirement, and to elucidate the mechanisms involved.


Measurement of glomerular filtration rate in cats: methods and advantages over routine markers of renal function.
Routinely used markers of renal function in clinical practice include urea and creatinine. However, these are insensitive markers, particularly in the early stages of kidney disease. Measurement of glomerular filtration rate (GFR) is regarded as the most sensitive index of functioning renal mass. It may be useful for feline patients in varying clinical scenarios; for example, where a more accurate measurement of renal function may aid diagnosis, to enable response to therapeutic interventions to be more closely monitored, or to evaluate renal function prior to the use of nephrotoxic or renally cleared drugs. Traditional methods of measuring GFR, such as renal clearance or multisample plasma clearance techniques, are generally impractical for clinical use. Limited sampling and single sample plasma clearance methods using the filtration marker iohexol have been validated in cats. These have the advantages of reduced stress to cats associated with repeated sampling and reduced costs of analysis, and therefore offer greater clinical utility. Attempts to develop an estimated GFR (eGFR) formula similar to that used in human patients have been made in cats, although currently an accurate and reliable formula is not available. This review presents the basis for the theoretical understanding and practical measurement of GFR for any veterinary practitioner wishing to obtain a more accurate and sensitive assessment of renal function than routinely used markers provide. The review draws evidence from peer-reviewed publications, the author's PhD thesis and also clinical experience.


Ventricular tachycardia ablation remains treatment of last resort in structural heart disease: argument for earlier intervention.
Despite advances in ablation of ventricular tachycardia (VT), recognized toxicity of amiodarone, and potential harm of implantable cardioverter defibrillator (ICD) shocks, there appears to be reluctance to pursue catheter ablation. We tested the hypothesis that patients with structural heart disease and VT are referred late for ablation and may have worse outcomes as a result. Consecutive patients with VT and structural heart disease referred to a single center, between January 2008 and April 2009 were studied. Patients with prior VT ablations were excluded. Late referrals were defined as those with 2 or more episodes of VT, separated by at least 1 month. Ninety-eight consecutive patients were analyzed. Ninety-six percent of patients had an ICD implanted prior to ablation, 58% were in VT storm and 67% taking ≥400 mg daily of amiodarone or amiodarone intolerant (10%). Thirty-six patients fit the definition of early referral and 62 late. Overall acute procedural success was achieved in 89%. Amiodarone dose decreased from a mean and median of 559 and 400 mg daily preablation to 98 and 0 postablation (P < 0.01). Mean and median VT episodes decreased from 17 and 6 in the month preceding ablation to 1 and 0 in the 6 months following ablation (P < 0.01). In Kaplan-Meier analysis, the early referral group had superior 1-year VT free survival (P = 0.01). VT ablation is frequently reserved for patients receiving recurrent ICD shocks despite high dose amiodarone. Stronger consideration should be given to earlier referral for VT ablation in patients with structural heart disease.


ALK and NSCLC: Targeted therapy with ALK inhibitors.
For many years treatment for advanced or metastatic non-small cell lung cancer (NSCLC) has employed chemotherapy regimens for patient care, with limited effect. Five-year survival rates for these patients are not encouraging. However, for a subgroup of these patients, there have been radical changes over recent years. Our understanding of the basic pathology behind NSCLC at the molecular level has offered up a host of new molecularly targeted therapies, which are revolutionizing this area of cancer care. Results from recent clinical trials provide hope for NSCLC patients harboring oncogenic translocations involving the anaplastic lymphoma kinase (ALK) receptor tyrosine kinase. Just as inhibition of the breakpoint cluster region-ABL complex has changed the face of chronic myeloid leukemia diagnosis, oncogenic ALK fusions offer a step forward in the diagnosis and treatment of ALK-positive NSCLC. This article discusses the current knowledge and potential implications concerning ALK inhibitors and NSCLC.


Genetic variations in a PTEN/AKT/mTOR axis and prostate cancer risk in a Chinese population.
Genetic variations in a PTEN/AKT/mTOR signaling axis may influence cellular functions including cell growth, proliferation and apoptosis, and then increase the individual's risk of cancer. Accordingly, we explore the association between single nucleotide polymorphisms (SNPs) of these genes and prostate cancer (PCa) in our Chinese population. Subjects were recruited from 666 PCa patients and 708 cancer-free controls, and eight SNPs in the PTEN/AKT/mTOR axis were determined by the TaqMan assay. Odds ratios (OR) and 95% confidence intervals (95% CI) were evaluated by logistic regression. We observed significant associations between PCa risk and mTOR rs2295080 [P = 0.027, OR = 0.85, 95%CI = 0.74-0.98], and AKT2 rs7254617 (P = 0.003, OR = 1.35, 95%CI = 1.11-1.64). When estimated these two SNPs together, the combined genotypes with 2-4 risk alleles (rs2295080 T and rs7254617 A alleles) were associated with an increased risk of PCa compared with 0-1 risk alleles, which was more pronounced among subgroups of age >71 years, smokers, drinkers and no family history of cancer. Results of stratified analyses by cliniopathological parameters revealed that the frequencies of the combined genotypes with 2-4 risk alleles in advanced stage were significantly higher than in localized stage (P = 0.022), but there was no significant association in Gleason score and PSA level. Our results indicate, for the first time that the two variants in AKT2 and mTOR, particularly the joint genotypes with 2-4 risk alleles may influence PCa susceptibility and progression in Chinese, and the association appeared to be more strong in the subgroup of smokers and drinkers.


Identification of amino acid residues responsible for differences in substrate specificity and inhibitor sensitivity between two human liver dihydrodiol dehydrogenase isoenzymes by site-directed mutagenesis.
Human liver dihydrodiol dehydrogenase isoenzymes (DD1 and DD2), in which only seven amino acid residues are substituted, differ remarkably in specificity for steroidal substrates and inhibitor sensitivity: DD1 shows 20alpha-hydroxysteroid dehydrogenase activity and sensitivity to 1,10-phenanthroline, whereas DD2 oxidizes 3alpha-hydroxysteroids and is highly inhibited by bile acids. In the present study we performed site-directed mutagenesis of the seven residues (Thr-38, Arg-47, Leu-54, Cys-87, Val-151, Arg-170 and Gln-172) of DD1 to the corresponding residues (Val, His, Val, Ser, Met, His and Leu respectively) of DD2. Of the seven mutations, only the replacement of Leu-54 with Val produced an enzyme that had almost the same properties as DD2. No significant changes were observed in the other mutant enzymes. An additional site-directed mutagenesis of Tyr-55 of DD1 to Phe yielded an inactive protein, suggesting the catalytically important role of this residue. Thus a residue at a position before the catalytic Tyr residue might play a key role in determining the orientation of the substrates and inhibitors.


The effect of the ketogenic diet on hippocampal GluR5 and Glu(6 mRNA expression and Q/R site editing in the kainate-induced epilepsy model.
Treatment with the ketogenic diet has been used to control refractory epilepsy for many years, although its anticonvulsant mechanism is unknown. By modulating synaptic transmission in the hippocampus, kainate receptors and their RNA editing might play a role in the antiseizure action of the treatment. To investigate the potential effect of the ketogenic diet on GluR(5) mRNA and GluR(6) mRNA expression and Q/R site editing, we used the kainate-induced epilepsy model in the present study. Reverse transcription polymerase chain reaction was performed to determine GluR(5) and GluR(6) mRNA expression, and RNA editing was analyzed with the BbvIota restriction enzyme assay. The results demonstrated that expression of GluR(6) mRNA, but not GluR(5) mRNA, was significantly increased after 8 weeks of dietary treatment. Neither the GluR(5) nor the GluR(6) RNA editing rate at the Q/R site was significantly changed by dietary treatment. These data indicate that GluR(6) may be involved in the anticonvulsant mechanism of ketogenic diet treatment.


Identification and characterization of Cryptosporidium parvum Clec, a novel C-type lectin domain-containing mucin-like glycoprotein.
Cryptosporidium species are waterborne apicomplexan parasites that cause diarrheal disease worldwide. Although the mechanisms underlying Cryptosporidium-host cell interactions are not well understood, mucin-like glycoproteins of the parasite are known to mediate attachment and invasion in vitro. We identified C. parvum Clec (CpClec), a novel mucin-like glycoprotein that contains a C-type lectin domain (CTLD) and has orthologs in C. hominis and C. muris. CTLD-containing proteins are ligand-binding proteins that function in adhesion and signaling and are present in a wide range of organisms, from humans to viruses. However, this is the first report of a CTLD-containing protein in protozoa and in Apicomplexa. CpClec is predicted to be a type 1 membrane protein, with a CTLD, an O-glycosylated mucin-like domain, a transmembrane domain, and a cytoplasmic tail containing a YXX sorting motif. The predicted structure of CpClec displays several characteristics of canonical CTLD-containing proteins, including a long loop region hydrophobic core associated with calcium-dependent glycan binding as well as predicted calcium- and glycan-binding sites. CpClec expression during C. parvum infection in vitro is maximal at 48 h postinfection, suggesting that it is developmentally regulated. The 120-kDa mass of native CpClec is greater than predicted, most likely due to O-glycosylation. CpClec is localized to the surface of the apical region and to dense granules of sporozoites and merozoites. Taken together, these findings, along with the known functions of C. parvum mucin-like glycoproteins and of CTLD-containing proteins, strongly implicate a significant role for CpClec in Cryptosporidium-host cell interactions.


Epstein-Barr virus seronegativity is a risk factor for late-onset posttransplant lymphoroliferative disorder in adult renal allograft recipients.
Posttransplant lymphoproliferative disorder (PTLD) remains a difficult management issue; therefore, many studies focus on the identification of risk factors to allow for preventive strategies. We investigated risk factors for PTLD in the adult renal transplant setting. A single-center, matched case-control study design was used. Cases were identified from patients who underwent a first renal transplant between January 1, 1985, and December 1, 2001. Two controls were chosen per case, matched (+/-1 year) by date of transplant and graft survival. Clinical and demographic data were ascertained from medical records. Pretransplant serology for Epstein-Barr virus (EBV) and cytomegalovirus was confirmed on frozen, stored sera. Statistical analysis included univariate and multivariable examination of putative risk factors using conditional logistic regression. Twenty cases of PTLD were identified, an incidence of 2.4%. Median time from transplant to diagnosis was 55 months (range, 3-168 months), with 16 cases of late-onset PTLD (>1 year posttransplant). The only significant risk in univariate analysis was EBV-negative status at transplant (risk ratio 6.0, P=0.03). In multivariable analysis, EBV-negative status remained significant (adjusted risk ratio 8.9, P=0.01). The risk related to EBV status held true when late cases were analyzed separately (adjusted risk ratio 7.1, P=0.03). Pretransplant EBV-seronegative status is a strong risk for development of PTLD in adult renal allograft recipients, even in late disease. These results indicate that primary infection with EBV may have a pathogenic role in some cases of late PTLD.


Is a lower dose of vitamin D supplementation enough to increase 25(OH)D status in a sunny country?
Calcium and vitamin D are essential nutrients for bone metabolism Vitamin D can either be obtained from dietary sources or cutaneous synthesis. The study was conducted in subtropic weather; therefore, some might believe that the levels of solar radiation would be sufficient in this area. To evaluate calcium and vitamin D supplementation in postmenopausal women with osteoporosis living in a sunny country. A 3-month controlled clinical trial with 64 postmenopausal women with osteoporosis, mean age 62 + or - 8 years. They were randomly assigned to either the supplement group, who received 1,200 mg of calcium carbonate and 400 IU (10 microg) of vitamin D(3,) or the control group. Dietary intake assessment was performed, bone mineral density and body composition were measured, and biochemical markers of bone metabolism were analyzed. Considering all participants at baseline, serum vitamin D was under 75 nmol/l in 91.4% of the participants. The concentration of serum 25(OH)D increased significantly (p = 0.023) after 3 months of supplementation from 46.67 + or - 13.97 to 59.47 + or - 17.50 nmol/l. However, the dose given was limited in effect, and 86.2% of the supplement group did not reach optimal levels of 25(OH)D. Parathyroid hormone was elevated in 22.4% of the study group. After the intervention period, mean parathyroid hormone tended to decrease in the supplement group (p = 0.063). The dose given (400 IU/day) was not enough to achieve 25(OH)D concentration, considered optimal for bone health.


Why Use Nutraceutical Strategies for the Irritable Bowel Syndrome?
Irritable bowel syndrome (IBS) is a chronic functional bowel disorder, often stress-related, identified by many abdominal symptoms, the most important of which is chronic visceral abdominal pain. Therefore, IBS commonly impairs the quality of life of patients, moreover, it is frequently linked to depressive and anxiety symptoms. The treatment of IBS primarily focuses on symptoms relief. Unfortunately, up to now, no efficacious therapies have been found. Therefore, it would be important to develop new anti- IBS interventions. The aim of this brief review is to summarize the current evidence of nutraceutical supplementation in IBS treatment, with probiotics, prebiotics, synbiotics, butyrate, palmitoylethanolamide and colostrum. Since nutraceutics are over-the-counter products, the review has the purpose to better inform the medicinal chemist and the practitioner about the possible beneficial mechanisms and the many advantages that these therapies offer. All of these compounds present multiple mechanisms of action, such as restoring the physiological microbiota, potentiating gastrointestinal barrier's function, immunomodulatory, anti-inflammatory and antinociceptive activities. From the literature data, it results that these compounds are not only capable of improving IBS symptomatology, but mainly display an optimal safety and tolerability profile. Although extensive studies must be carried out to reinforce the evidence from the so far limited clinical trials, the supplementation with these compounds may be useful considering the warnings of prescription medicines for special populations of patients, such as elders, youngsters, or patients who need combination therapy. Finally, the nutraceutical approach may improve adherence to treatment, given its better acceptance by the patients compared to pharmacological therapy.


[Genetic etiology of esophageal cancer. V. Fragile sites on chromosome of cultured lymphocytes from members of high risk cancer families in Linxian County].
The chromosome fragile sites of cultured peripheral lymphocytes from 40 members of 4 high risk cancer families and 10 members of 4 low risk cancer families in Linxian County were analysed. The results showed that 46 fragile sites in 7045 lymphocytes expression at 502 times (7.13%) were found in high risk cancer families and 8 fragile sites in 1053 lymphocytes expression at 26 times (2.47%) were found in low risk cancer families. There was a significant difference between the two groups (P less than 0.01). In 46 fragile sites carried by 40 members of high risk cancer families, 27 were common, 5 rare, 12 provisional and 2 new fragile sites. Among them, the fragile sites at 1p22-p36 and 4q21-q31 were detected in members of high risk cancer families and in patients with esophageal cancer, meanwhile, uniform breakpoint in chromosome deletion and rearrangement was also found in 4 esophageal cancer cell lines. Therefore, the author conjectures that these fragile sites at 1p13-p36 and 4q21-q31 may be fragile site-specific for high risk cancer families and patients with esophageal cancer, and they may be breakpoint-specific for esophageal cancer cells. These fragile sites may play an important role in esophageal carcinogenesis in high risk cancer families.


Influence of Different Pretreatments on the Microtensile Bond Strength to Eroded Dentin.
To evaluate the influence of different pretreatments on the microtensile bond strength (µTBS) of an etch-and-rinse adhesive to eroded dentin. Thirty-six human teeth were ground down to their dentin layer and randomly divided into six groups (G1-G6; n = 6), G1 being the control group. Only in the test groups (G2-G6) were samples subjected to erosion using citric acid (pH 2.6) 10 x 2 min per day for five days. Between the erosive attacks, samples were stored in artificial saliva. After pretreatment - none (G1); none (G2); 2% chlorhexidine (30 s) (G3); prolonged primer application (1 min) (G4); roughening with a diamond bur (G5) and 10.5% NaOCl (1 min) (G6) - the adhesive OptiBond FL was applied. After the application of composite, samples were stored in water (7 d) and µTBS was determined. Data were evaluated using one-way ANOVA and Dunnett-T post-hoc test (p < 0.05). Eroded dentin without pretreatment (G2) resulted in significant reduction of µTBS compared with uneroded dentin (G1). µTBS after pretreatment with a diamond bur (G5) or NaOCl (G6) was not significantly different from that of the uneroded control group (G1). µTBS after pretreatment with chlorhexidine (G3) or with prolonged primer application (G4) was significantly lower than in the uneroded control group (G1), and not significantly different from the eroded control group (G2). The present data suggests that µTBS to eroded dentin pretreated with bur abrasion or NaOCl is similar to the µTBS to sound, uneroded dentin.


Characterization and inflammatory response of perivascular-resident macrophage-like melanocytes in the vestibular system.
A large number of perivascular cells expressing both macrophage and melanocyte characteristics (named perivascular-resident macrophage-like melanocytes, PVM/Ms), previously found in the intra-strial fluid-blood barrier, are also found in the blood-labyrinth barrier area of the vestibular system in normal adult cochlea, including in the three ampullae of the semicircular canals (posterior, superior, and horizontal), utricle, and saccule. The cells were identified as PVM/Ms, positive for the macrophage and melanocyte marker proteins F4/80 and GSTα4. Similar to PVM/Ms present in the stria vascularis, the PVM/Ms in the vestibular system are closely associated with microvessels and structurally intertwined with endothelial cells and pericytes, with a density in normal (unstimulated) utricle of 225 ± 43/mm(2); saccule 191 ± 25/mm(2); horizontal ampullae 212 ± 36/mm(2); anterior ampullae 238 ± 36/mm(2); and posterior ampullae 223 ± 64/mm(2). Injection of bacterial lipopolysaccharide into the middle ear through the tympanic membrane causes the PVM/Ms to activate and arrange in an irregular pattern along capillary walls in all regions within a 48-h period. The inflammatory response significantly increases vascular permeability and leakage. The results underscore the morphological complexity of the blood barrier in the vestibular system, with its surrounding basal lamina, pericytes, as well as second line of defense in PVM/Ms. PVM/Ms may be important to maintain blood barrier integrity and initiating local inflammatory response in the vestibular system.


Magnesium, hemostasis, and outcomes in patients with intracerebral hemorrhage.
We tested the hypothesis that admission serum magnesium levels are associated with hematoma volume, hematoma growth, and functional outcomes in patients with intracerebral hemorrhage (ICH). Patients presenting with spontaneous ICH were enrolled in an observational cohort study that prospectively collected demographic, clinical, laboratory, radiographic, and outcome data. We performed univariate and adjusted multivariate analyses to assess for associations between serum magnesium levels and initial hematoma volume, final hematoma volume, and in-hospital hematoma growth as radiographic measures of hemostasis, and functional outcome measured by the modified Rankin Scale (mRS) at 3 months. We included 290 patients for analysis. Admission serum magnesium was 2.0 ± 0.3 mg/dL. Lower admission magnesium levels were associated with larger initial hematoma volumes on univariate (p = 0.02), parsimoniously adjusted (p = 0.002), and fully adjusted models (p = 0.006), as well as greater hematoma growth (p = 0.004, p = 0.005, and p = 0.008, respectively) and larger final hematoma volumes (p = 0.02, p = 0.001, and p = 0.002, respectively). Lower admission magnesium level was associated with worse functional outcomes at 3 months (i.e., higher mRS; odds ratio 0.14, 95% confidence interval 0.03-0.64, p = 0.011) after adjustment for age, admission Glasgow Coma Scale score, initial hematoma volume, time from symptom onset to initial CT, and hematoma growth, with evidence that the effect of magnesium is mediated through hematoma growth. These data support the hypothesis that magnesium exerts a clinically meaningful influence on hemostasis in patients with ICH.


Ceftaroline: a cephalosporin with expanded Gram-positive activity.
Infections caused by resistant Gram-positive pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) are now posing a major health risk for patients in hospital and community settings. There is a need to evaluate new antibiotics that would offer reliable clinical efficacy combined with a favorable safety profile for the treatment of such infections. Ceftaroline is a new member of the cephalosporin class of antibiotics with expanded activity against Gram-positive pathogens such as MRSA, vancomycin-intermediate S. aureus, vancomycin-resistant S. aureus and multidrug-resistant Streptococcus pneumoniae, while retaining good activity against common Gram-negative organisms. Phase II and III studies have shown ceftaroline to be an effective and well-tolerated treatment for complicated skin and skin-structure infections compared with standard therapy. Trials are ongoing in the treatment of community-acquired pneumonia.


A new oxidative stress model, 2,2-azobis(2-amidinopropane) dihydrochloride induces cardiovascular damages in chicken embryo.
It is now well established that the developing embryo is very sensitive to oxidative stress, which is a contributing factor to pregnancy-related disorders. However, little is known about the effects of reactive oxygen species (ROS) on the embryonic cardiovascular system due to a lack of appropriate ROS control method in the placenta. In this study, a small molecule called 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radicals generator, was used to study the effects of oxidative stress on the cardiovascular system during chick embryo development. When nine-day-old (stage HH 35) chick embryos were treated with different concentrations of AAPH inside the air chamber, it was established that the LD50 value for AAPH was 10 µmol/egg. At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos. Impacts of AAPH on younger embryos were also examined and discovered that it inhibited the development of vascular plexus on yolk sac in HH 18 embryos. AAPH also dramatically repressed the development of blood islands in HH 3+ embryos. These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis. Furthermore, we observed heart enlargement in the HH 40 embryo following AAPH treatment, where the left ventricle and interventricular septum were found to be thickened in a dose-dependent manner due to myocardiac cell hypertrophy. In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo. The current study also provided a new developmental model, as an alternative for animal and cell models, for testing small molecules and drugs that have anti-oxidative activities.


Evolution and phylogenetic relationships of chitin synthases from yeasts and fungi.
Chitin, the structural component that provides rigidity to the cell wall of fungi is the product of chitin synthases (Chs). These enzymes are not restricted to fungi, but are amply distributed in four of the five eukaryotic 'crown kingdoms'. Dendrograms obtained by multiple alignment of Chs revealed that fungal enzymes can be classified into two divisions that branch into at least five classes, independent of fungal divergence. In contrast, oomycetes and animals each possess a single family of Chs. These results suggest that Chs originated as a branch of beta-glycosyl-transferases, once the kingdom Plantae split from the evolutionary line of eukaryotes. The existence of a single class of Chs in animals and Stramenopiles, against the multiple families in fungi, reveals that Chs diversification occurred after fungi departed from these kingdoms, but before separation of fungal groups. Accordingly, each fungal taxon contains members with enzymes belonging to different divisions and classes. Multiple alignment revealed the conservation of specific motifs characteristic of class, division and kingdom, but the strict conservation of only three motifs QXXEY, EDRXL and QXRRW, and seven isolated amino acids in the core region of all Chs. Determination of different structural features in this region of Chs brought to light a noticeable conservation of secondary structure in the proteins.


Can Oral Bacteria and Mechanical Fatigue Degrade Zirconia Dental Implants in Vitro?
Zirconia (ZrO2) is an emerging alternative to titanium for dental implant systems due to its material properties including high mechanical strength and chemical stability. However, oral environmental factors such as bacterial adhesion and mechanical fatigue may trigger low-temperature degradation of ZrO2, leading to reduced mechanical strength and potential implant fracture. Although failure modes of ZrO2 in orthopedic applications have been studied, they have yet to be thoroughly investigated in the context of dental implant systems. Thus, the goal of the present study was to assess the surface of ZrO2 dental implants for signs of degradation after exposure to oral bacteria and oral bacteria in combination with mechanical fatigue. ZrO2 dental implants were subjected to 30-day immersion in (i) early or (ii) late colonizing oral bacteria or (iii) were mechanically loaded for 2 × 106 cycles with oral bacteria in circulation. Optical microscopy, Raman microscopy, and X-ray photoelectron spectroscopy (XPS) were used to evaluate the surface morphology, phase composition, and chemical composition, respectively. Post-immersion, all implants exhibited minimal changes in surface features, and all loaded implants survived cyclic fatigue tests. All implants had <1% monoclinic phase at the collar, junction, and screw regions, excluding the screw threads, for which monoclinic phase was significantly higher but <10%. XPS revealed an increase in carbon- and nitrogen-based organic debris on the implants exposed to early colonizers as compared to those immersed in late colonizers or synergistically with mechanical loading. Within the limitations of the present study, ZrO2 is a suitable alternative material for dental implant systems based on its ability to resist both physical and chemical degradation imposed by oral bacteria and applied cyclic loads.


Impact of Intensive Gait Training With and Without Electromechanical Assistance in the Chronic Phase After Stroke-A Multi-Arm Randomized Controlled Trial With a 6 and 12 Months Follow Up.
Introduction: Movement related impairments and limitations in walking are common long-term after stroke. This multi-arm randomized controlled trial explored the impact of training with an electromechanically assisted gait training (EAGT) system, i.e., the Hybrid Assistive Limb® (HAL), when integrated with conventional rehabilitation focused on gait and mobility. Material and Methods: Participants, aged 18-70 years with lower extremity paresis but able to walk with manual support or supervision 1-10 years after stroke, were randomized to (A) HAL-training on a treadmill, combined with conventional rehabilitation interventions (HAL-group), or (B) conventional rehabilitation interventions only (Conventional group), 3 days/week for 6 weeks, or (C) no intervention (Control group). Participants in the Control group were interviewed weekly regarding their scheduled training. Primary outcome was endurance in walking quantified by the 6 Minute Walk Test (6MWT). A rater blinded to treatment allocation performed assessments pre- and post-intervention and at follow-ups at 6 and 12 months. Baseline assessment included the National Institute of Health Stroke Scale (NIHSS) and the Modified Ranking Scale (MRS). Secondary outcomes included the Fugl Meyer Assessment- Lower Extremity, 10 Meter Walk Test, Berg Balance Scale (BBS), Barthel Index (BI) and perceived mobility with the Stroke Impact Scale. Results: A total of 48 participants completed the intervention period. The HAL-group walked twice as far as the Conventional group during the intervention. Post-intervention, both groups exhibited improved 6 MWT results, while the Control group had declined. A significant improvement was only found in the Conventional group and when compared to the Control group (Tukey HSD p = 0.022), and not between the HAL group and Conventional group (Tukey HSD p = 0.258) or the HAL- group and the Control group (Tukey HSD p = 0.447). There was also a significant decline in the Conventional group from post-intervention to 6 months follow up (p = 0.043). The best fitting model to predict outcome included initial balance (BBS), followed by stroke severity (NIHSS), and dependence in activity and participation (BI and MRS). Conclusion: Intensive conventional gait training induced significant improvements long-term after stroke while integrating treadmill based EAGT had no additional value in this study sample. The results may support cost effective evidence-based interventions for gait training long-term after stroke and further development of EAGT. Trial registration: Published on clinicaltrials.gov (NCT02545088) August 24, 2015.


Discrete Optimal Graph Clustering.
Graph-based clustering is one of the major clustering methods. Most of it works in three separate steps: 1) similarity graph construction; 2) clustering label relaxing; and 3) label discretization with k -means (KM). Such common practice has three disadvantages: 1) the predefined similarity graph is often fixed and may not be optimal for the subsequent clustering; 2) the relaxing process of cluster labels may cause significant information loss; and 3) label discretization may deviate from the real clustering result since KM is sensitive to the initialization of cluster centroids. To tackle these problems, in this paper, we propose an effective discrete optimal graph clustering framework. A structured similarity graph that is theoretically optimal for clustering performance is adaptively learned with a guidance of reasonable rank constraints. Besides, to avoid the information loss, we explicitly enforce a discrete transformation on the intermediate continuous label, which derives a tractable optimization problem with a discrete solution. Furthermore, to compensate for the unreliability of the learned labels and enhance the clustering accuracy, we design an adaptive robust module that learns the prediction function for the unseen data based on the learned discrete cluster labels. Finally, an iterative optimization strategy guaranteed with convergence is developed to directly solve the clustering results. Extensive experiments conducted on both real and synthetic datasets demonstrate the superiority of our proposed methods compared with several state-of-the-art clustering approaches.


Extracellular Vesicles in Premature Aging and Diseases in Adulthood Due to Developmental Exposures.
The developmental origins of health and disease (DOHaD) is a paradigm that links prenatal and early life exposures that occur during crucial periods of development to health outcome and risk of disease later in life. Maternal exposures to stress, some psychoactive drugs and alcohol, and environmental chemicals, among others, may result in functional changes in developing fetal tissues, creating a predisposition for disease in the individual as they age. Extracellular vesicles (EVs) may be mediators of both the immediate effects of exposure during development and early childhood as well as the long-term consequences of exposure that lead to increased risk and disease severity later in life. Given the prevalence of diseases with developmental origins, such as cardiovascular disease, neurodegenerative disorders, osteoporosis, metabolic dysfunction, and cancer, it is important to identify persistent mediators of disease risk. In this review, we take this approach, viewing diseases typically associated with aging in light of early life exposures and discuss the potential role of EVs as mediators of lasting consequences.


Super suppressors in Neurospora crassa I. Induction, genetic localization and relationship to a missense suppressor.
Genetic analyses have been made to test the feasibility of using coincident reversions to prototrophy of multiple mutants to select super suppressors (ssu) in Neurospora crassa. Of five double-mutant strains examined, only those mutant combinations in which both members had the properties of nonsense mutations did revert coincidently. Forty-eight genetically purified coincident revertants were crossed to the wild type, and each was shown to contain a suppressor mutation. Five super suppressors were examined more thoroughly. Tetrad and random spore analysis was used to demonstrate that each behaved as a single gene in crosses. Two super suppressors, ssu-1 and ssu-4 were localized respectively on the right and left arm of linkage group 7. Two others, ssu-2 and ssu-3, appear to map on the right arm of linkage group 1. The fifth super suppressor mapped, ssu-7, lies between ad-8 and ylo-1 on linkage group 6. One super suppressor, ssu-1, was interesting because it mapped near the location reported for the suppressor of the missense mutant tryp-3(td201) (Yourno and Suskind 1964a). However, no overlap was found in action spectrum of the two suppressors. Tetrad analysis showed the two suppressors were located about 10 map units apart, the missense suppressor being the more distal to the centromere.


Resonant Rayleigh light scattering of single Au nanoparticles with different sizes and shapes.
Scientific interest in nanotechnology is driven by the unique and novel properties of nanometer-sized metallic materials such as the strong interaction between the conductive electrons of the nanoparticles and the incident light, caused by localized surface plasmon resonances (LSPRs). In this article, we analysed the relationship of the Rayleigh scattering properties of a single Au nanoparticle with its size, shape, and local dielectric environment. We also provided a detailed study on the refractive index sensitivity of three types of differently shaped Au nanoparticles, which were nanospheres, oval-shaped nanoparticles and nanorods. This study helps one to differentiate the Rayleigh light scattering from individual nanoparticles of different sizes and/or shapes and precisely obtain quantitative data as well as the correlated optical spectra of single gold nanoparticles from the inherently inhomogeneous solution of nanoparticles. These results suggest that the shape, size and aspect ratio of Au nanoparticles are important structural factors in determining the resonant Rayleigh light scattering properties of a single Au nanoparticle such as its spectral peak position, scattering-cross-section and refractive index sensitivity, which gives a handle for the choice of gold nanoparticles for the design and fabrication of single nanosensors.


Life without thyroxine to 3,5,3'-triiodothyronine conversion: studies in mice devoid of the 5'-deiodinases.
Considerable indirect evidence suggests that the type 2 deiodinase (D2) generates T(3) from T(4) for local use in specific tissues including pituitary, brown fat, and brain, whereas the type I deiodinase (D1) generates T(3) from T(4) in the thyroid and peripheral tissues primarily for export to plasma. From studies in deiodinase-deficient mice, the importance of the D2 for local T(3) generation has been confirmed. However, the phenotypes of these D1 knockout (KO) and D2KO mice are surprisingly mild and their serum T(3) level, general health, and reproductive capacity are unimpaired. To explore further the importance of 5'deiodination in thyroid hormone economy, we used a mouse devoid of both D1 and D2 activity. In general, the phenotype of the D1/D2KO mouse is the sum of the phenotypes of the D1KO and D2KO mice. It appears healthy and breeds well, and most surprisingly its serum T(3) level is normal. However, impairments in brain gene expression and possibly neurological function are somewhat greater than those seen in the D2KO mouse, and the serum rT(3) level is elevated 6-fold in the D1/D2KO mouse but only 2-fold in the D1KO mouse and not at all in the D2KO mouse. The data suggest that whereas D1 and D2 are not essential for the maintenance of the serum T(3) level, they do serve important roles in thyroid hormone homeostasis, the D2 being critical for local T(3) production and the D1 playing an important role in iodide conservation by serving as a scavenger enzyme in peripheral tissues and the thyroid.


Two novel missense mutations in the myelin protein zero gene causes Charcot-Marie-Tooth type 2 and Déjérine-Sottas syndrome.
The Charcot-Marie-Tooth (CMT) phenotype caused by mutation in the myelin protein zero (MPZ) gene varies considerably, from early onset and severe forms to late onset and milder forms. The mechanism is not well understood. The myelin protein zero (P0) mediates adhesion in the spiral wraps of the Schwann cell's myelin sheath. The crystalline structure of the extracellular domain of the myelin protein zero (P0ex) is known, while the transmembrane and intracellular structure is unknown. One novel missense mutation caused a milder late onset CMT type 2, while the second missense mutation caused a severe early onset phenotype compatible with Déjérine-Sottas syndrome. The phenotypic variation caused by different missense mutations in the MPZ gene is likely caused by different conformational changes of the MPZ protein which affects the functional tetramers. Severe changes of the MPZ protein cause dysfunctional tetramers and predominantly uncompacted myelin, i.e. the severe phenotypes congenital hypomyelinating neuropathy and Déjérine-Sottas syndrome, while milder changes cause the phenotypes CMT type 1 and 2.


The Longue Durée of Black Lives Matter.
Black Lives Matter was first articulated just a few years ago, but it has been the leitmotif of antiracist struggles for generations. The Movement for Black Lives extends the work of previous movements that challenged forms of oppression that act on Black bodies with impunity. It should be understood in the context of Ida B. Wells' anti-lynching campaign, Fannie Lou Hamer's reproductive justice demands, and the Black Panther Party's health activism. The 50th anniversary of the Black Panther Party is an occasion to recall that its work confronted the callous neglect and the corporeal surveillance and abuse of poor Black communities. Similar demands have been the centrifugal force of social movements that for centuries have refused to have Black lives cast beyond the human boundary.


Universal entropy of word ordering across linguistic families.
The language faculty is probably the most distinctive feature of our species, and endows us with a unique ability to exchange highly structured information. In written language, information is encoded by the concatenation of basic symbols under grammatical and semantic constraints. As is also the case in other natural information carriers, the resulting symbolic sequences show a delicate balance between order and disorder. That balance is determined by the interplay between the diversity of symbols and by their specific ordering in the sequences. Here we used entropy to quantify the contribution of different organizational levels to the overall statistical structure of language. We computed a relative entropy measure to quantify the degree of ordering in word sequences from languages belonging to several linguistic families. While a direct estimation of the overall entropy of language yielded values that varied for the different families considered, the relative entropy quantifying word ordering presented an almost constant value for all those families. Our results indicate that despite the differences in the structure and vocabulary of the languages analyzed, the impact of word ordering in the structure of language is a statistical linguistic universal.


Therapeutic self-disclosure in integrative psychotherapy: When is this a clinical error?
Ascending to prominence in virtually all forms of psychotherapy, therapist self-disclosure (TSD) has recently been identified as a primarily integrative intervention (Ziv-Beiman, 2013). In the present article, we discuss various instances in which using TSD in integrative psychotherapy might constitute a clinical error. First, we briefly review extant theory and empirical research on TSD, followed by our preferred version of integrative psychotherapy (i.e., a version of Wachtel's Cyclical Psychodynamics [Wachtel, 1977, 1997, 2014]), which we title cognitive existential psychodynamics. Next, we provide and discuss three examples in which implementing TSD constitutes a clinical error. In essence, we submit that using TSD constitutes an error when patients, constrained by their representational structures (object relations), experience the subjectivity of the other as impinging, and thus propels them to "react" instead of "emerge." (PsycINFO Database Record


Cerebrospinal fluid otorrhea through a congenitally patent fallopian canal.
Cerebrospinal fluid (CSF) otorrhea is a relatively rare entity that may occur either as a spontaneous occurrence or as a result of trauma or surgery. Spontaneous CSF leaks may be found during tympanocentesis, myringotomy, or tube insertion for chronic middle ear effusion. Rapid identification of the problem and timely treatment are required to avoid life-threatening complications such as meningitis. The site of leakage must also be identified so that the disorder can be treated effectively. Computed tomography, magnetic resonance imaging, and radionucleotide localization scanning all play a role in the early identification of the leakage site. A detailed knowledge of possible CSF leakage pathways aids in evaluating imaging studies. We report 2 rare cases of CSF otorrhea through a congenitally patent facial canal and their management.


Neurospora spore killers Sk-2 and Sk-3 suppress meiotic silencing by unpaired DNA.
In Neurospora crassa, pairing of homologous DNA segments is monitored during meiotic prophase I. Any genes not paired with a homolog, as well as any paired homologs of that gene, are silenced during the sexual phase by a mechanism known as meiotic silencing by unpaired DNA (MSUD). Two genes required for MSUD have been described previously: sad-1 (suppressor of ascus dominance), encoding an RNA-directed RNA polymerase, and sad-2, encoding a protein that controls the perinuclear localization of SAD-1. Inactivation of either sad-1 or sad-2 suppresses MSUD. We have now shown that MSUD is also suppressed by either of two Spore killer strains, Sk-2 and Sk-3. These were both known to contain a haplotype segment that behaves as a meiotic drive element in heterozygous crosses of killer x sensitive. Progeny ascospores not carrying the killer element fail to mature and are inviable. Crosses homozygous for either of the killer haplotypes suppress MSUD even though ascospores are not killed. The killer activity maps to the same 30-unit-long region within which recombination is suppressed in killer x sensitive crosses. We suggest that the region contains a suppressor of MSUD.


Effect of human immunodeficiency virus type 1 protein R (vpr) gene expression on basic cellular function of fission yeast Schizosaccharomyces pombe.
The human immunodeficiency virus type 1 (HIV-1) Vpr protein affects cell morphology and prevents proliferation of human cells by induction of cell cycle G2 arrest. In this study, we used the fission yeast Schizosaccharomyces pombe as a model system to investigate the cellular effects of HIV-1 vpr gene expression. The vpr gene was cloned into an inducible fission yeast gene expression vector and expressed in wild-type S. pombe cells, and using these cells, we were able to demonstrate the specific Vpr-induced effects by induction and suppression of vpr gene expression. Induction of HIV-1 vpr gene expression affected S. pombe at the colonial, cellular, and molecular levels. Specifically, Vpr induced small-colony formation, polymorphic cells, growth delay, and cell cycle G2 arrest. Additionally, Vpr-induced G2 arrest appeared to be independent of cell size and morphological changes. The cell cycle G2 arrest correlated with increased phosphorylation of p34cdc2, suggesting negative regulation of mitosis by HIV-1 Vpr. Treatment of Vpr-induced cell with a protein phosphatase inhibitor, okadaic acid, transiently suppressed cell cycle arrest and morphological changes. This observation implicates possible involvement of protein phosphatase(s) in the effects of Vpr. Together, these data showed that the HIV-1 Vpr-induced cellular changes in S. pombe are similar to those observed in human cells. Therefore, the S. pombe system is suited for further investigation of the HIV-1 vpr gene functions.


Antibody responses to Sarcoptes scabiei apolipoprotein in a porcine model: relevance to immunodiagnosis of recent infection.
No commercial immunodiagnostic tests for human scabies are currently available, and existing animal tests are not sufficiently sensitive. The recombinant Sarcoptes scabiei apolipoprotein antigen Sar s 14.3 is a promising immunodiagnostic, eliciting high levels of IgE and IgG in infected people. Limited data are available regarding the temporal development of antibodies to Sar s 14.3, an issue of relevance in terms of immunodiagnosis. We utilised a porcine model to prospectively compare specific antibody responses to a primary infestation by ELISA, to Sar s 14.3 and to S. scabiei whole mite antigen extract (WMA). Differences in the antibody profile between antigens were apparent, with Sar s 14.3 responses detected earlier, and declining significantly after peak infestation compared to WMA. Both antigens resulted in >90% diagnostic sensitivity from weeks 8-16 post infestation. These data provide important information on the temporal development of humoral immune responses in scabies and further supports the development of recombinant antigen based immunodiagnostic tests for recent scabies infestations.


Neurochondrin neurological autoimmunity.
To describe the neurologic spectrum and treatment outcomes for neurochondrin-IgG positive cases identified serologically in the Mayo Clinic Neuroimmunology Laboratory. Archived serum and CSF specimens previously scored positive for IgGs that stained mouse hippocampal tissue in a nonuniform synaptic pattern by immunofluorescence assay (89 among 616,025 screened, 1993-2019) were reevaluated. Antibody characterization experiments revealed specificity for neurochondrin, confirmed by recombinant protein assays. IgG in serum (9) or CSF (4) from 8 patients yielded identical neuron-restricted CNS patterns, most pronounced in hippocampus (stratum lucidum in particular), cerebellum (Purkinje cells and molecular layer), and amygdala. All were neurochondrin-IgG positive. Five were women; median symptom onset age was 43 years (range, 30-69). Of 7 with clinical data, 6 presented with rapidly progressive cerebellar ataxia, brainstem signs, or both; 1 had isolated unexplained psychosis 1 year prior. Five of 6 had cerebellar signs, 4 with additional brainstem symptoms or signs (eye movement abnormalities, 3; dysphagia, 2; nausea and vomiting, 1). One patient with brainstem signs (vocal cord paralysis and VII nerve palsy) had accompanying myelopathy (longitudinally extensive abnormality on MRI; aquaporin-4-IgG and myelin oligodendrocyte glycoprotein-IgG negative). The 7th patient had small fiber neuropathy only. Just 1 of 7 had contemporaneous cancer (uterine). Six patients with ataxia or brainstem signs received immunotherapy, but just 1 remained ambulatory. At last follow-up, 5 had MRI evidence of severe cerebellar atrophy. In our series, neurochondrin autoimmunity was usually accompanied by a nonparaneoplastic rapidly progressive rhombencephalitis with poor neurologic outcomes. Other phenotypes and occasional paraneoplastic causes may occur.


Structural brain differences in alcohol-dependent individuals with and without comorbid substance dependence.
Over 50% of individuals with alcohol use disorders (AUD) also use other substances; brain structural abnormalities observed in alcohol dependent individuals may not be entirely related to alcohol consumption. This MRI study assessed differences in brain regional tissue volumes between short-term abstinent alcohol dependent individuals without (ALC) and with current substance use dependence (polysubstance users, PSU). Nineteen, one-month-abstinent PSU and 40 ALC as well as 27 light-drinkers (LD) were studied on a 1.5 T MR system. Whole brain T1-weighted images were segmented automatically into regional gray matter (GM), white matter (WM), and cerebrospinal fluid (CSF) volumes. MANOVA assessed group differences of intracranial volume-normalized tissue volumes of the frontal, parietal, occipital, and temporal lobes and regional subcortical GM volumes. The volumetric measures were correlated with neurocognitive measures to assess their functional relevance. Despite similar lifetime drinking and smoking histories, PSU had significantly larger normalized WM volumes than ALC in all lobes. PSU also had larger frontal and parietal WM volumes than LD, but smaller temporal GM volumes and smaller lenticular and thalamic nuclei than LD. ALC had smaller frontal, parietal, and temporal GM, thalamic GM and cerebellar volumes than LD. ALC had more sulcal CSF volumes than both PSU and LD. One-month-abstinent ALC and PSU exhibited different patterns of gross brain structural abnormalities. The larger lobar WM volumes in PSU in the absence of widespread GM volume loss contrast with widespread GM atrophy in ALC. These structural differences may demand different treatment approaches to mitigate specific functionally relevant brain abnormalities.


Enhanced Cancer DNA Vaccine via Direct Transfection to Host Dendritic Cells Recruited in Injectable Scaffolds.
Deoxyribonucleic acid (DNA) vaccines are a promising cancer immunotherapy approach. However, effective delivery of DNA to antigen-presenting cells (e.g., dendritic cells (DCs)) for the induction of an adaptive immune response is limited. Conventional DNA delivery via intramuscular, intradermal, and subcutaneous injection by hypodermal needles shows a low potency and immunogenicity. Here, we propose the enhanced cancer DNA vaccine by direct transfection to the high number of DCs recruited into the chemoattractant-loaded injectable mesoporous silica microrods (MSRs). Subcutaneous administration of the MSRs mixed with tumor-antigen coding DNA polyplexes resulted in DC recruitment in the macroporous space of the scaffold formed by the spontaneous assembly of high-aspect-ratio MSRs, thereby allowing for enhanced cellular uptake of antigen-coded DNA by host DCs. The MSR scaffolds delivering the DNA vaccine trigger a more robust DC activation, antigen-specific CD8+ T cell response, and Th1 immune response compared to the bolus DNA vaccine. Additionally, the immunological memory can be induced with a single administration of the vaccine. The combination of the vaccination and antiprogrammed cell death-1 antibody significantly eliminates established lung metastasis. These results indicate that MSRs serve as a powerful platform for DNA vaccine delivery to DCs for effective cancer immunotherapy.


Ecological covariates of Ascaris lumbricoides infection in schoolchildren from rural KwaZulu-Natal, South Africa.
To identify environmental factors that could serve to predict Ascaris lumbricoides infection patterns and thus guide control efforts in the absence of epidemiological information; to assess whether A. lumbricoides infection is positively associated with the soil clay content. Information on A. lumbricoides infection and re-infection in a cohort of primary schoolchildren and interview data on their socioeconomic background and behaviour were combined with environmental data using a geographical information system (GIS). Multivariate models served to explore the covariation of environmental and infection patterns adjusted for possible confounders. Prevalence maps and spatial statistics revealed considerable spatial clustering of infection in the small study area. Logistic multivariate regression models showed strong positive associations of infection with vegetation density measured as the normalized difference vegetation index (NDVI) at baseline [odds ratio (OR) for a 10% increase: 1.82; 95% confidence interval (95% CI): 1.24-2.68; P=0.002] and after re-infection (OR: 2.22; 95% CI: 1.71-2.87; P<0.001). We also found a strong negative association of re-infection with the sun exposure of the soil surface as estimated from digital elevation models (OR: 0.78; 95% CI: 0.88; P<0.001). The soil clay content was only moderately positively associated with infection and re-infection. Socioeconomic and behavioural variables, although correlated with A. lumbricoides infection, did not appear to confound the above associations in the demographically homogeneous study area. Spatial analysis of the model residuals suggested that as the models accounted for most of the spatial pattern, the model standard errors should not be affected by spatial clustering. NDVI seems to have a high potential for the prediction of A. lumbricoides infection as it was strongly associated with infection patterns in the study area. Further advantages are that NDVI information is easy to use, affordable and available with global coverage.


A local agreement pattern measure based on hazard functions for survival outcomes.
Assessing agreement is often of interest in biomedical and clinical research when measurements are obtained on the same subjects by different raters or methods. Most classical agreement methods have been focused on global summary statistics, which cannot be used to describe various local agreement patterns. The objective of this work is to study the local agreement pattern between two continuous measurements subject to censoring. In this article, we propose a new agreement measure based on bivariate hazard functions to characterize the local agreement pattern between two correlated survival outcomes. The proposed measure naturally accommodates censored observations, fully captures the dependence structure between bivariate survival times and provides detailed information on how the strength of agreement evolves over time. We develop a nonparametric estimation method for the proposed local agreement pattern measure and study theoretical properties including strong consistency and asymptotical normality. We then evaluate the performance of the estimator through simulation studies and illustrate the method using a prostate cancer data example.


2,5-Bis[2-({bis-[3-(dimethyl-aza-nium-yl)prop-yl]aza-nium-yl}meth-yl)phen-yl]-1,3,4-oxadiazole hexa-kis-(perchlorate) sesquihydrate.
In the title hydrated salt, C36H66N8O(6+)·6ClO4(-)·1.5H2O, the asymmetric unit consists of a hexa-protonated [H6L](6+) cation, five perchlorate anions in general positions, two on twofold rotation axes (one of which is disordered), and two water mol-ecules of crystallization in general positions, one of them disordered around a twofold crystallographic axis. In the [H6L](6+) cation, two strong intra-molecular N-H⋯N hydrogen bonds occur, involving the N atoms of the oxadiazole ring as acceptors and the closest NH(+) groups of each dipropyl-enetriamine unit. In the crystal, the [H6L](6+) cations form channels along the a-axis direction, in which the perchlorate counter-ions and the water mol-ecules are lodged. The crystal packing features a network of N-H⋯O and O-H⋯O hydrogen bonds involving the NH(+) groups of the [H6L](6+) cation, the perchlorate anions and the water mol-ecules.


Long-term outcomes after transmyocardial revascularization.
Two independent reports documented substantially higher operative mortality associated with transmyocardial revascularization (TMR) when used in isolation than that reported in the premarket clinical trials. To clarify the state of the art, this article assesses temporal trends in the use of TMR, short-term and long-term outcomes, and outcomes stratified by procedure type (TMR only and TMR + coronary artery bypass graft [CABG]) and by the 2 specific TMR devices. The study population included all patients undergoing TMR in isolation or in combination with CABG at 435 cardiothoracic hospitals in the United States participating in the Society of Thoracic Surgeons (STS) Adult Cardiac Surgery Database (ACSD) from January 2000 through November 2006 (n = 15,386). Analysis of long-term outcomes was accomplished through linkage to Medicare claims data. Short-term and long-term (7 years) adverse outcomes were assessed and compared between the 2 TMR device types. The use of TMR in conjunction with CABG surgery is increasing. This study showed modest differences in short-term morbidity and mortality between the 2 devices. In combination with CABG, after risk adjustment, patients treated with the holmium:YAG laser (experienced a higher rate of operative mortality (3.5% vs 2.5%; adjusted hazard ratio 1.39, 95% confidence level 1.03 to 1.87) but no difference in the composite short-term rate of major morbidity or mortality, compared with the Heart Laser CO2 transmyocardial revascularization system (PLC Medical Systems, Inc, Milford, MA). However, there were no clinically meaningful differences in long-term results. Modest differences in short-term morbidity and mortality between the 2 devices suggest the usefulness of further research.


Tetrachloroaurate (III)-induced oxidation increases non-thermal plasma-induced oxidative stress.
Non-thermal plasma (NTP) devices have been explored for medical applications. NTP devices discharge electrons, positive ions, ultraviolet (UV), reactive oxygen species (ROS) and reactive nitrogen species (RNS), such as the hydroxyl radical (•OH), singlet oxygen (1O2), superoxide (O2•-), hydrogen peroxide (H2O2), ozone, and nitric oxide, at near-physiological temperature. At preclinical stages or in human clinical trials, NTP promotes blood coagulation, eradication of bacterial, viral, and biofilm-related infections, wound healing, and cancer cell death. Here, we observed that ferric, vanadium, and gold(III) ions significantly elevated lipid peroxidation, which was measured by 2-thiobarbituric acid-reactive substances (TBARS) in combination with NTP exposure. Using 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (M4PO) as a spin probe in electron paramagnetic resonance (EPR), we observed that tetrachloroaurate (III) yielded an M4PO-X spin adduct. Tetrachloroaurate-induced oxidation was attenuated efficiently by reduced (GSH) and oxidized glutathione (GSSG), while glycine (Gly), and L-glutamate (Glu), components of GSH, were ineffective. Furthermore, GSH and GSSG efficiently suppressed tetrachloroaurate-induced lipid peroxidation, while Gly and Glu were ineffective in suppressing TBARS elevation. These results indicate that tetrachloroaurate-induced oxidation is attenuated by GSH as well as GSSG. Further studies are warranted to elucidate the redox reactions between metal ions and biomolecules to advance the clinical application of NTP.