High-density lipoproteins for the systemic delivery of short interfering RNA.
RNA interference (RNAi) is a powerful mechanism for gene silencing with the potential to greatly impact the development of new therapies for many human diseases. Short interfering RNAs (siRNAs) may be the ideal molecules for therapeutic RNAi. However, therapeutic siRNAs face significant challenges that must be overcome prior to widespread clinical use. Many efforts have been made to overcome the hurdles associated with systemic administration of siRNA; however, current approaches are still limited. As such, there is an urgent need to develop new strategies for siRNA delivery that have the potential to impact a broad spectrum of systemic diseases. This review focuses on the promise of siRNA therapies and highlights current siRNA delivery methods. With an eye toward new strategies, this review first introduces high-density lipoprotein (HDL) and describes its natural biological functions, and then transitions into how HDLs may provide significant opportunities as next-generation siRNA delivery vehicles. Importantly, this review describes how synthetic HDLs leverage the natural ability of HDL to stabilize and deliver siRNAs. HDLs are natural nanoparticles that are critical to understanding the systemic delivery of therapeutic nucleic acids, like siRNA. Methods to synthesize biomimetic HDLs are being explored, and data demonstrate that this type of delivery vehicle may be highly beneficial for targeted and efficacious systemic delivery of siRNAs.


Neostigmine to relieve a suspected colonic pseudo-obstruction in a burn patient: a case-based review of the literature.
Neostigmine is one of the treatment options for colonic pseudo-obstruction in the medical patient. However, experience in using neostigmine for this indication in burn patients has not been reported in the literature. We will present a case of a woman who developed colonic pseudo-obstruction during her hospital stay. When conservative management failed, neostigmine was administered with no adverse effects and resolution of the pseudo-obstruction. We will review the literature regarding the pathophysiology and treatment options for acute colonic pseudo-obstruction in burn patients. A 27-year-old woman with 35% total body surface area deep-partial and full-thickness flame burns. On hospital day 17, she developed a nonobstructive ileus. She failed conservative medical therapy. After consultation with colleagues in trauma surgery and a review of the literature (MeSH/PubMed/NLM), the decision was made to try neostigmine therapy rather than a surgical/procedural option such as colonoscopy. The patient was moved to the intensive care unit and 2 mg of neostigmine was administered intravenously over 4 minutes. After 30 minutes, all abdominal examination findings had returned to baseline. No significant adverse effects were noted, and she did not redevelop abdominal distension afterward. This case report provides an alternative treatment modality in which neostigmine was used successfully in a burn patient after conservative medical treatment had failed. The authors believe that neostigmine may be a viable alternative to decompressive colonoscopy in burn patients for whom mechanical obstruction is properly excluded.


A novel mimetic enzymatic fluorescence immunoassay for hepatitis B surface antigen by using a thermal phase separating polymer.
Iron tetrasulfonatophthalocyanine (FeTSPc), a peroxidase mimic, was used as a labeling reagent and poly(N-isopropylacrylamide) (PNIP) as the separation support of the immune complex for the mimetic-enzymatic immunoassay of hepatitis B surface antigen (HBsAg). PNIP was precipitated from aqueous solution when the ambient temperature was higher than its lower critical solution temperature of 31 degrees C. In a sandwich immunoassay, the antigen (HBsAg) first reacted with mouse anti-human HBsAg antibody immobilized on PNIP (PNIP-antibody) and then further reacted with FeTSPc-labeled mouse anti-HBsAg antibody (antibody-FeTSPc) at room temperature in a homogeneous format. After changing the temperature to separate the PNIP-antibody-HBsAg-antibody-FeTSPc conjugate moiety, it was re-dissolved and determined by coupling with the fluorogenic reaction of hydrogen peroxide and p-hydroxyphenylpropionic acid. The sensitivity of this method (3 ng mL-1) was close to that of the traditional ELISA using the same reactants. However, the assay was much faster (the assay time decreased from 100-120 to 45 min). This method was applied to determine HBsAg in human serum with satisfactory results.


[Cognitive-behavioural guidance interventions in adolescents with attention deficit hyperactivity disorder].
Attention deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder that may become manifest at any time in life. In 50-70% of children diagnosed with the disorder it presents in adolescence. Young people with ADHD have high rates of comorbidity with other psychiatric disorders and a high degree of functional compromise. To review the literature on cognitive-behavioural interventions that have been applied to the treatment of ADHD in adolescence. The studies that have been conducted on psychological treatment to date were reviewed, and the interventions were classified into: psychosocial treatments, mindfulness therapy and cognitive-behavioural treatment (individual and in groups). The only study on cognitive-behavioural therapy for adolescents with ADHD is also reviewed, as well as a new intervention protocol for application to groups designed at the Hospital Universitari Vall d'Hebron. Although there has been a recent increase in the number of publications dealing with the psychological treatment of ADHD in adolescents, there is a need for a greater development of intervention protocols and studies on their efficacy/effectiveness.


Design of a prospective, multicenter, global, cohort study of electromagnetic navigation bronchoscopy.
Electromagnetic navigation bronchoscopy (ENB) procedures allow physicians to access peripheral lung lesions beyond the reach of conventional bronchoscopy. However, published research is primarily limited to small, single-center studies using previous-generation ENB software. The impact of user experience, patient factors, and lesion/procedural characteristics remains largely unexplored in a large, multicenter study. NAVIGATE (Clinical Evaluation of superDimension™ Navigation System for Electromagnetic Navigation Bronchoscopy) is a prospective, multicenter, global, cohort study. The study aims to enroll up to 2,500 consecutive subjects presenting for evaluation of lung lesions utilizing the ENB procedure at up to 75 clinical sites in the United States, Europe, and Asia. Subjects will be assessed at baseline, at the time of procedure, and at 1, 12, and 24 months post-procedure. The pre-test probability of malignancy will be determined for peripheral lung nodules. Endpoints include procedure-related adverse events, including pneumothorax, bronchopulmonary hemorrhage, and respiratory failure, as well as quality of life, and subject satisfaction. Diagnostic yield and accuracy, repeat biopsy rate, tissue adequacy for genetic testing, and stage at diagnosis will be reported for biopsy procedures. Complementary technologies, such as fluoroscopy and endobronchial ultrasound, will be explored. Success rates of fiducial marker placement, dye marking, and lymph node biopsies will be captured when applicable. Subgroup analyses based on geography, demographics, investigator experience, and lesion and procedure characteristics are planned. Study enrollment began in April 2015. As of February 19, 2016, 500 subjects had been enrolled at 23 clinical sites with enrollment ongoing. NAVIGATE will be the largest prospective, multicenter clinical study on ENB procedures to date and will provide real-world experience data on the utility of the ENB procedure in a broad range of clinical scenarios. ClinicalTrials.gov NCT02410837 . Registered 31 March 2015.


Phylogeographic characterization of Burkholderia pseudomallei isolated from Bangladesh.
Burkholderia pseudomallei possesses a diverse set of genes which encode a vast array of biological functions reflecting its clinical, ecological and phenotypic diversity. Strain variation is linked to geographic location as well as pattern of land uses. This soil-dwelling Gram-negative pathogen causes melioidosis, a tropical disease endemic in northern Australia and Southeast Asian regions including Bangladesh. Phylogeographic analyses of B. pseudomallei isolates by molecular typing techniques could be used to examine the diversity of this organism as well as to track melioidosis epidemics. In this study, 22 B. pseudomallei isolates, of which 20 clinical and two soil isolates were analyzed, utilizing Real-time PCR assay and multilocus sequence typing (MLST). The sequences were then submitted to PubMLST database for analysis and construction of phylogenetic tree. A total of 12 different sequence types (STs) that includes four novel STs were identified for the first time. Strains having STs 1005, 1007 and 56 were the most widespread STs frequently isolated in Bangladesh. ST 1005, ST 56, ST 1007 and ST 211 have been detected not only in Bangladesh but are also present in many Southeast Asian countries. ST 1005 was detected in both soil and clinical samples of Gazipur. Most prevalent, ST 56 has been previously reported from Myanmar, Thailand, Cambodia and Vietnam, confirming the persistence of the genotype over the entire continent. Further large-scale study is necessary to find out the magnitude of the infection and its different reservoirs in the environment along with phylogeographic association.


[Anticancer drug-induced nephrotoxicity].
Nephrotoxicity is an inherent adverse effect of certain anticancer drugs. Anti neoplasic drugs have a narrow therapeutic index and the amount of drug necessary to produce a significant reduction in tumour burden usually produces significant nephrotoxicity. The dosage used in clinical trials represents often the maximum tolerated doses determined during phase I drug evaluation. Greater toxicity is acceptable during curative therapy than during palliative therapy. But cancer patients often exhibit excretory reduced organ function. Modulation of pharmacokinetics and pharmacodynamics of these drugs in cancer patients is therefore necessary in order to improve tolerance. Patients with malignancies are particularly vulnerable to development of renal abnormalities. Conversely, patients with renal abnormalities who have undergone kidney transplantation are at high risk for malignancy. Clinical syndromes of renal involvement are diverse and sometimes insidious. Despite the recent advances in understanding the mechanism of anticancer drug nephrotoxicity, prevention still relies on drug dosage decrease and active screening for renal abnormalities as part of the usual biological work up in patients treated with anticancer drugs.


[Comparative frequency of phenotypic resemblance of intrafamilial and interfamilial pairs of patients with generalized and focal epilepsy].
The correlation of the frequency of intra-paired phenotypical resemblance of intrafamilial (parents-children - 46 pairs, sibses - 26 pairs) and interfamilial (48 pairs) epilepsy according to the complex of constitutional and clinical features is studied. The intra-paired resemblance of the onset (homochronicity) and clinic (homotypia) of intrafamilial (even in different generations) epilepsy was discovered more often (41.3-88.5%) than interfamilial epilepsy (18.7-33.3%). The complete intra-paired phenotypical resemblance is the most characteristic for generalized, than focal intrafamiliar epilepsy. The comparison of the frequency of intra-paired resemblance according to constitutional features and the medium influence showed that the phenotypical manifestation of the epileptical genotype depends on the complex constellation of hereditary and medium factors being subjected to considerable modification by exogenes. However the clear difference of the frequency of intra-paired clinical resemblance of intra- and interfamilial epilepsy in considerable degree was caused rather by constitutional pecularities than by exogenes influence and may testify to possible hereditary heterogeneity of epilepsy.


Narrowband Blue and Red LED Supplements Impact Key Flavor Volatiles in Hydroponically Grown Basil Across Growing Seasons.
The use of light-emitting diodes (LEDs) in commercial greenhouse production is rapidly increasing because of technological advancements, increased spectral control, and improved energy efficiency. Research is needed to determine the value and efficacy of LEDs in comparison to traditional lighting systems. The objective of this study was to establish the impact of narrowband blue (B) and red (R) LED lighting ratios on flavor volatiles in hydroponic basil (Ocimum basilicum var. "Genovese") in comparison to a non-supplemented natural light (NL) control and traditional high-pressure sodium (HPS) lighting. "Genovese" basil was chosen because of its high market value and demand among professional chefs. Emphasis was placed on investigating concentrations of important flavor volatiles in response to specific ratios of narrowband B/R LED supplemental lighting (SL) and growing season. A total of eight treatments were used: one non-supplemented NL control, one HPS treatment, and six LED treatments (peaked at 447 nm/627 nm, ±20 nm) with progressive B/R ratios (10B/90R, 20B/80R, 30B/70R, 40B/60R, 50B/50R, and 60B/40R). Each SL treatment provided 8.64 mol ⋅ m-2 ⋅ d-1 (100 μmol ⋅ m-2 ⋅ s-1, 24 h ⋅ d-1). The daily light integral (DLI) of the NL control averaged 9.5 mol ⋅ m-2 ⋅ d-1 during the growth period (ranging from 4 to 18 mol ⋅ m-2 ⋅ d-1). Relative humidity averaged 50%, with day/night temperatures averaging 27.4°C/21.8°C, respectively. Basil plants were harvested 45 days after seeding, and volatile organic compound profiles were obtained by gas chromatography-mass spectrometry. Total terpenoid concentrations were dramatically increased during winter months under LED treatments, but still showed significant impacts during seasons with sufficient DLI and spectral quality. Many key flavor volatile concentrations varied significantly among lighting treatments and growing season. However, the concentrations of some compounds, such as methyl eugenol, were three to four times higher in the control and decreased significantly for basil grown under SL treatments. Maximum concentrations for each compound varied among lighting treatments, but most monoterpenes and diterpenes evaluated were highest under 20B/80R to 50B/50R. This study shows that supplemental narrowband light treatments from LED sources may be used to manipulate secondary metabolic resource allocation. The application of narrowband LED SL has great potential for improving overall flavor quality of basil and other high-value specialty herbs.


Pore Breathing of Metal-Organic Frameworks by Environmental Transmission Electron Microscopy.
Metal-organic frameworks (MOFs) have emerged as a versatile platform for the rational design of multifunctional materials, combining large specific surface areas with flexible, periodic frameworks that can undergo reversible structural transitions, or "breathing", upon temperature and pressure changes, and through gas adsorption/desorption processes. Although MOF breathing can be inferred from the analysis of adsorption isotherms, direct observation of the structural transitions has been lacking, and the underlying processes of framework reorganization in individual MOF nanocrystals is largely unknown. In this study, we describe the characterization and elucidation of these processes through the combination of in situ environmental transmission electron microscopy (ETEM) and computer simulations. This combined approach enables the direct monitoring of the breathing behavior of individual MIL-53(Cr) nanocrystals upon reversible water adsorption and temperature changes. The ability to characterize structural changes in single nanocrystals and extract lattice level information through in silico correlation provides fundamental insights into the relationship between pore size/shape and host-guest interactions.


Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies.
Mitochondria play a pivotal role in energy metabolism, programmed cell death and oxidative stress. Mutated mitochondrial DNA in diseased cells compromises the structure of key enzyme complexes and, therefore, mitochondrial function, which leads to a myriad of health-related conditions such as cancer, neurodegenerative diseases, diabetes and aging. Early detection of mitochondrial and metabolic anomalies is an essential step towards effective diagnoses and therapeutic intervention. Reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) play important roles in a wide range of cellular oxidation-reduction reactions. Importantly, NADH and FAD are naturally fluorescent, which allows noninvasive imaging of metabolic activities of living cells and tissues. Furthermore, NADH and FAD autofluorescence, which can be excited using distinct wavelengths for complementary imaging methods and is sensitive to protein binding and local environment. This article highlights recent developments concerning intracellular NADH and FAD as potential biomarkers for metabolic and mitochondrial activities.


Thermally denaturing high-performance liquid chromatography analysis of primase activity.
Prokaryotic primase, a DNA-dependent RNA polymerase, is a target of interest for the development of novel antibiotics. A new assay was developed to evaluate the inhibition of primase activity while avoiding the limitations of existing assays that require the incorporation of radiolabeled nucleotides into the growing primer followed by electrophoretic separation and autoradiography or scintillation counting. These existing technologies are either time consuming or unable to give detailed information on the kinetics, size, and nature of the primers synthesized. To address these issues in a nonradioactive manner, a thermally denaturing high-performance liquid chromatography (HPLC) assay was developed that was able to (1) measure the two modes of primase activity (de novo and overlong primer synthesis), (2) quantitate de novo primer synthesis kinetics yielding a rate constant of 0.00251 s(-1), and (3) determine that dNTPs inhibited primase activity with an IC50 of 9.5 microM. In addition, the differential elution properties of short DNA and RNA oligonucleotides on an alkylated nonporous polystyrene-divinylbenzene copolymer microsphere bead column were determined. The thermally denaturing HPLC assay provides rapid quantitative analysis of primase function and qualitative analysis of activity with regard to the nature of the primers synthesized.


The role of GABA in modulating the Xenopus electroretinogram.
We have recorded the electroretinogram (ERG) from the superfused eyecup of the Xenopus retina in order to assess the effects of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), and its agonists and antagonists, on individual ERG components. We found that GABA (0.5-10 mM) reduced the amplitudes of both the b- and d-waves of the Xenopus ERG. The GABA uptake blocker nipecotic acid (1 mM) had similar effects on b- and d-waves. GABA at 5 mM and 10 mM also caused an increase in the a-wave. The GABA antagonist picrotoxin (0.1-2 mM) and the GABA/a antagonist bicuculline (0.2 mM) both increased the amplitude of the b- and d-waves of the ERG. The GABA/b agonist baclofen (0.3 mM) reduced the amplitude of the ERG b-wave, enhanced the amplitude of the a-wave, and slightly reduced the amplitude and increased the peak time of the d-wave. The GABA/b antagonists phaclofen and saclofen had no reliable effects on the Xenopus ERG. Glutamate analogs known to affect specific types of retinal neurons were applied to modify the retinal circuitry and then the effects of GABA and its antagonists were examined under these modified conditions. 2-amino-4-phosphonobutyric acid (APB) increased the d-wave, and blocked the b-wave and the effect of GABA on the ERG, but not the antagonist-induced increase in the d-wave. KYN blocked the antagonist-induced increase in the b-wave, while GABA increases the amplitude of the b-wave if the d-wave has been removed by prior superfusion with kynurenic acid (KYN). N-methyl-DL-aspartate (NMDLA), which acts only in the proximal retina, reduced the amplitude of the ERG and blocked the effect of GABA and the antagonist-induced increase in ERG b- and d-waves amplitude. These results suggest that GABAergic mechanisms related to both A and B receptor types can influence the amplitude and light sensitivity of all the components of the Xenopus ERG. Since GABA is found in greatest abundance in the proximal retina, and B type of receptors are present almost exclusively there, the data suggests that most of the effects of GABA agonists and antagonists observed are dependent on proximal retinal mechanisms, and that there are separate mechanisms in the proximal retina related to the b- and the d-waves.


The murine coronavirus mouse hepatitis virus strain A59 from persistently infected murine cells exhibits an extended host range.
In murine 17 Cl 1 cells persistently infected with murine coronavirus mouse hepatitis virus strain A59 (MHV-A59), expression of the virus receptor glycoprotein MHVR was markedly reduced (S. G. Sawicki, J. H. Lu, and K. V. Holmes, J. Virol. 69:5535-5543, 1995). Virus isolated from passage 600 of the persistently infected cells made smaller plaques on 17 Cl 1 cells than did MHV-A59. Unlike the parental MHV-A59, this variant virus also infected the BHK-21 (BHK) line of hamster cells. Virus plaque purified on BHK cells (MHV/BHK) grew more slowly in murine cells than did MHV-A59, and the rate of viral RNA synthesis was lower and the development of the viral nucleocapsid (N) protein was slower than those of MHV-A59. MHV/BHK was 100-fold more resistant to neutralization with the purified soluble recombinant MHV receptor glycoprotein (sMHVR) than was MHV-A59. Pretreatment of 17 Cl 1 cells with anti-MHVR monoclonal antibody CC1 protected the cells from infection with MHV-A59 but only partially protected them from infection with MHV/BHK. Thus, although MHV/BHK could still utilize MHVR as a receptor, its interactions with the receptor were significantly different from those of MHV-A59. To determine whether a hemagglutinin esterase (HE) glycoprotein that could bind the virions to 9-O-acetylated neuraminic acid moieties on the cell surface was expressed by MHV/BHK, an in situ esterase assay was used. No expression of HE activity was detected in 17 Cl 1 cells infected with MHV/BHK, suggesting that this virus, like MHV-A59, bound to cell membranes via its S glycoprotein. MHV/BHK was able to infect cell lines from many mammalian species, including murine (17 Cl 1), hamster (BHK), feline (Fcwf), bovine (MDBK), rat (RIE), monkey (Vero), and human (L132 and HeLa) cell lines. MHV/BHK could not infect dog kidney (MDCK I) or swine testis (ST) cell lines. Thus, in persistently infected murine cell lines that express very low levels of virus receptor MHVR and which also have and may express alternative virus receptors of lesser efficiency, there is a strong selective advantage for virus with altered interactions with receptor (D. S. Chen, M. Asanaka, F. S. Chen, J. E. Shively, and M. M. C. Lai, J. Virol. 71:1688-1691, 1997; D. S. Chen, M. Asanaka, K. Yokomori, F.-I. Wang, S. B. Hwang, H.-P. Li, and M. M. C. Lai, Proc. Natl. Acad. Sci. USA 92:12095-12099, 1995; P. Nedellec, G. S. Dveksler, E. Daniels, C. Turbide, B. Chow, A. A. Basile, K. V. Holmes, and N. Beauchemin, J. Virol. 68:4525-4537, 1994). Possibly, in coronavirus-infected animals, replication of the virus in tissues that express low levels of receptor might also select viruses with altered receptor recognition and extended host range.


Attitudes of ward attendants towards mental illness: comparisons and predictors.
In India few studies exist that explore attitudes of mental health professionals, especially psychiatric ward attendants, towards psychiatric illnesses. This study aimed to explore attitudes of psychiatric ward attendants since they work closely with psychiatric patients in the hospital ward, and which may differ given their cultural and social background. Five hundred psychiatric ward attendants were targeted and a 100 sample size chosen based on systematic random sampling. A control group from guardians attending the outpatient department of Central Institute of Psychiatry was selected using the same methods. Attitudes of both groups were assessed using Community Attitudes towards Mental Illness (CAMI) after written informed consent. There were no significant differences noted in Authoritarianism and Benevolence domains of CAMI. However, there were significant differences in the domains of Social Restrictiveness and Community Mental Health Ideology with psychiatric ward attendants scoring higher than the general attendants. Among the socio-demographic variables, there was significant correlation between age, duration of contact and educational background and various domains of CAMI. Psychiatric ward attendants had more positive attitudes than general attendants towards psychiatric illnesses. Socio-demographic variables like older age, higher education and longer duration of contact with the psychiatrically ill predicted more favourable attitudes.


Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli.
The quorum sensing two-component system (TCS) QseBC has been linked to virulence, motility and metabolism regulation in multiple Gram-negative pathogens, including Enterohaemorrhagic Escherichia coli (EHEC), Uropathogenic E. coli (UPEC) and Salmonella enterica. In EHEC, the sensor histidine kinase (HK) QseC detects the quorum sensing signalling molecule AI-3 and also acts as an adrenergic sensor binding host epinephrine and norepinephrine. Downstream changes in gene expression are mediated by phosphorylation of its cognate response regulator (RR) QseB, and 'cross-talks' with non-cognate regulators KdpE and QseF to activate motility and virulence. In UPEC, cross-talk between QseBC and TCS PmrAB is crucial in the regulation and phosphorylation of QseB RR that acts as a repressor of multiple pathways, including motility. Here, we investigated QseBC regulation of motility in the atypical Enteropathogenic E. coli (EPEC) strain O125ac:H6, causative agent of persistent diarrhoea in children, and its possible cross-talk with the KdpDE and PmrAB TCS. We showed that in EPEC QseB acts as a repressor of genes involved in motility, virulence and stress response, and in absence of QseC HK, QseB is likely activated by the non-cognate PmrB HK, similarly to UPEC. We show that in absence of QseC, phosphorylated QseB activates its own expression, and is responsible for the low motility phenotypes seen in a QseC deletion mutant. Furthermore, we showed that KdpD HK regulates motility in an independent manner to QseBC and through a third unidentified party different to its own response regulator KdpE. We showed that PmrAB has a role in iron adaptation independent to QseBC. Finally, we showed that QseB is the responsible for activation of colistin and polymyxin B resistance genes while PmrA RR acts by preventing QseB activation of these resistance genes.


Primary care physician incentives in medical group practices.
Although medical groups are adapting to changes in financing health care, little is known about individual physician incentives in this environment. To describe methods group practices use to compensate primary care physicians in a managed care environment and to examine the association of revenue sources for the group practice from all patients and primary care physician incentives. We surveyed by mail group practice administrators for practices that had at least 200 members continuously enrolled in 1995. Group practices that had contractual arrangements with Blue Cross/Blue Shield of Minnesota. One hundred of 129 group practices returned usable surveys. Most groups had some portion of primary care physicians' compensation at risk, although 17 groups compensated them through fully guaranteed annual salary. Seventy-one groups used productivity, 4 groups used quality of care, 1 group used utilization, and 30 used group financial performance. Factors reported to significantly influence primary care physician compensation included billings or charges, overall group practice performance, and net revenue or profit. Groups that had a higher proportion of income from various types of fee-for-service arrangements used lower proportions of base salary for primary care physician compensation and were more likely to relate physician income to measures of productivity. Substantial variation exists in the types of primary care physician incentives implemented by medical groups. Base salary, individual productivity, and group financial performance were most frequently used to determine compensation. Physician personal financial risk was higher overall in group practices that derived more revenue from fee-for-service contracts.


Severe hydrogen sulfide exposure in a working adolescent.
We describe an occupational exposure to hydrogen sulfide gas in a 16-year-old boy. While cleaning the reoxygenation tank of a fish hatchery, he and an adult supervisor lost consciousness. The adult died, and the adolescent regained consciousness briefly when emergency medical services personnel administered oxygen. At a local emergency department, he was intubated for respiratory distress. He was transferred to a tertiary care facility for additional management and, over the next 2 weeks, had a recovery to normal function. Hydrogen sulfide is a colorless, malodorous gas that results from the decay of organic material. It is a byproduct of industry and agriculture. The mechanism of its toxicity is related primarily to inhibition of oxidative phosphorylation, which causes a decrease in available cellular energy. Although there is some anecdotal evidence to suggest that the early use of hyperbaric oxygen is beneficial, supportive care remains the mainstay of therapy. This report highlights the sources of exposure, management, and need for more stringent application of safety regulations in industries in which adolescents are employed.


Editorial: mRNA Vaccines and Immunotherapy in Oncology: A New Era for Personalized Medicine.
Synthetic mRNA and the expression of therapeutic proteins have accelerated vaccine development to prevent infection and heralds a new era in targeted immunotherapy in oncology. Therapeutic mRNA vaccines rely on available tumor tissue for gene sequencing analysis to compare the patient's normal cellular DNA sequences and those of the tumor. Carrier-based mRNA vaccines for cancer immunotherapy are now in development that use delivery systems based on peptides, lipids, polymers, and cationic nano-emulsions. There have also been recent developments in dendritic cell-based mRNA vaccines. For patients with available tumor tissue samples, it is possible to develop mRNA vaccines that result in the expression of tumor antigens by antigen-presenting cells (APCs), resulting in innate and adaptive immune responses. Ongoing developments in mRNA immunotherapy include modifications in the route of administration and combined delivery of multiple mRNA vaccines with checkpoint inhibitors. This Editorial aims to present a brief overview of how mRNA immunotherapy may change the therapeutic landscape of personalized medicine for patients with solid malignant tumors.


Deformation heterogeneity and texture in surface severe plastic deformation of copper.
Comprehensive understanding of thermomechanical response and microstructure evolution during surface severe plastic deformation (S2PD) is important towards establishing controllable processing frameworks. In this study, the evolution of crystallographic textures during directional surface mechanical attrition treatment on copper was studied and modelled using the visco-plastic self-consistent framework. In situ high-speed imaging and digital image correlation of surface deformation in circular indentation were employed to elucidate mechanics occurring in a unit process deformation and to calibrate texture model parameters. Material response during directional surface mechanical attrition was simulated using a finite-element model coupled with the calibrated texture model. The crystallographic textures developed during S2PD were observed to be similar to those resultant from uniaxial compression. The implications of these results towards facilitating a processing-based framework to predict deformation mechanics and resulting crystallographic texture in S2PD configurations are briefly discussed.


Effects of dietary protease on nitrogen emissions from broiler production: a holistic comparison using Life Cycle Assessment.
The aim of the study was to quantify the effects of the use of a protease Ronozyme® ProAct in broiler feed on the environmental impacts of broiler and broiler feed production chains. This was done by using a Life Cycle Assessment (LCA) modelling approach with data from trials using both standard soya-based broiler diets and reduced-protein diets with added protease. The results for the feed production chain showed that there was a reduction in all environmental impact categories when protease was used in the diets. The biggest reduction occurred in the category of Global Warming Potential, mainly as a result of decreased carbon dioxide emissions from land use changes related to soya production. In the results for the broiler production chain, there were relatively bigger reductions in Eutrophication Potential and especially in Acidification Potential, mainly as a result of reduced feed protein content and subsequent nitrogen emissions from housing and manure management. The use of protease in the broiler diets reduced the environmental impacts of both feed production and broiler production. The latter is mainly through reduced ammonia emissions, which has substantial benefit per se in the poultry industry.


Histological and Histopathological Study of Incus.
Chronic otits media is long standing infection of middle ear cleft which commonly involves bone erosion. Bone destruction seen in unsafe chronic otitis media mainly involves the ossicles, incus being frequently involved ossicle. So, an investigation of the histopathological changes in incus was carried out to report the various histopathological changes occurring in chronic otitis media. Aim of the study is to report the structural changes occuring in incus bone in chronic otitis media. Ten normal incuses and ten pathological incuses which were removed during the surgery for chronic otitis media (both with and without cholesteotoma) were studied histologically, after staining with haematoxylin and eosin. Normal incus showed compact bone pattern of concentric rings, like that of any long bone of body. Pathological incuses of chronic otitis media (both with and without cholestoetoma) showed similar changes, i.e., stratified squamous epithelium, with distorted concentric rings and increased osseous spaces. The study was undertaken, so that the knowledge to histological changes may help the clinicians to take more rational decisions regarding their diagnosis and therapeutic interventions to prevent the changes occurring in the bone in chronic otitis media.


Dihydroartemisinin exerts cytotoxic effects and inhibits hypoxia inducible factor-1alpha activation in C6 glioma cells.
Artemisinin and its analogue dihydroartemisinin exert cytotoxic effects in some kinds of cancer cell lines. Here we determined whether dihydroartemisinin inhibits the growth and induces apoptosis of rat C6 glioma cells. We found dihydroartemisinin (5-25 microM) inhibited the growth and induced apoptosis of C6 cells in a concentration- and time-dependent manner; however, it was much less toxic to rat primary astrocytes. Dihydroartemisinin (5-25 microM) also increased the generation of reactive oxygen species in C6 cells. These effects of dihydroartemisinin were enhanced by ferrous ions (12.5-100 microM) and reduced by the iron chelator deferoxamine (25-200 microM). Immunoblotting analysis revealed that dihydroartemisinin (5-25 microM) significantly reduced hypoxia- and deferoxamine-induced expression of hypoxia inducible factor-1alpha and its target gene protein, vascular endothelial growth factor, in C6 cells. The results showed that dihydroartemisinin exerts a selective cytotoxic effect on C6 cells by increasing the reactive oxygen species and inhibiting hypoxia inducible factor-1alpha activation.


Left-hemisphere activation and deficient response modulation in psychopaths.
Psychopathic offenders have difficulty processing contextual or secondary cues once they have initiated goal-directed behavior or allocated attention to a primary task. To test the hypothesis that this deficit in response modulation is specific to conditions in which psychopaths' left-hemisphere resources are engaged, we administered a serial recall task to 21 incarcerated psychopaths and 21 control subjects. Subjects were instructed to memorize eight words that were presented one at a time, each in one of the four corners of the visual display. Subjects' primary task was to recall the words in serial order. Then, without forewarning, they were asked to recall the words' locations. As predicted, psychopaths performed as well as control subjects in recalling words from the left and right spatial fields, but recalled significantly fewer locations from the right spatial field. Thus, psychopaths' deficient response modulation was specific to conditions in which their left-hemisphere resources were actively engaged.


Widening rural-urban disparities in life expectancy, U.S., 1969-2009.
There is limited research on rural-urban disparities in U.S. life expectancy. This study examined trends in rural-urban disparities in life expectancy at birth in the U.S. between 1969 and 2009. The 1969-2009 U.S. county-level mortality data linked to a rural-urban continuum measure were analyzed. Life expectancies were calculated by age, gender, and race for 3-year time periods between 1969 and 2004 and for 2005-2009 using standard life-table methodology. Differences in life expectancy were decomposed by age and cause of death. Life expectancy was inversely related to levels of rurality. In 2005-2009, those in large metropolitan areas had a life expectancy of 79.1 years, compared with 76.9 years in small urban towns and 76.7 years in rural areas. When stratified by gender, race, and income, life expectancy ranged from 67.7 years among poor black men in nonmetropolitan areas to 89.6 among poor Asian/Pacific Islander women in metropolitan areas. Rural-urban disparities widened over time. In 1969-1971, life expectancy was 0.4 years longer in metropolitan than in nonmetropolitan areas (70.9 vs 70.5 years). By 2005-2009, the life expectancy difference had increased to 2.0 years (78.8 vs 76.8 years). The rural poor and rural blacks currently experience survival probabilities that urban rich and urban whites enjoyed 4 decades earlier. Causes of death contributing most to the increasing rural-urban disparity and lower life expectancy in rural areas include heart disease, unintentional injuries, COPD, lung cancer, stroke, suicide, and diabetes. Between 1969 and 2009, residents in metropolitan areas experienced larger gains in life expectancy than those in nonmetropolitan areas, contributing to the widening gap.


Apolipoprotein E distribution among different plaque types in Alzheimer's disease: implications for its role in plaque progression.
We sought to determine the pattern of ApoE immunoreactivity in mesial temporal lobe tissue from 12 Alzheimer patients, age 66-88, and to determine the distribution of this immunoreactivity among different plaque types representing hypothesized stages of plaque evolution. In these patients, the cortical area of ApoE immunoreactivity was 30% that of beta-amyloid. Only 6% of diffuse non-neuritic amyloid deposits were even weakly ApoE immunoreactive (ApoE+). This is in contrast to our previous demonstration that microglia overexpressing interleukin-1 (IL-1) are present in most diffuse non-neuritic deposits. Eighty-three per cent of diffuse neuritic plaques and 86% of dense-core neuritic plaques were highly ApoE+, consistent with IL-1-induced astrocyte activation and synthesis of ApoE resulting in the appearance of ApoE immunoreactivity in neuritic plaques. Dense-core non-neuritic ('burned out') plaques were only rarely (6%) ApoE+. These results, together with the known trophic and toxic effects of ApoE on neurites, suggest that plaque-associated ApoE contributes to the formation of overgrown degenerating (dystrophic) neurites in plaques. However, the fact that some neuritic plaques are not ApoE+ suggests contributions by additional trophic and toxic factors. Our results are also consistent with a role for ApoE in the condensation of diffuse amyloid deposits into a beta-pleated-sheet form that occurs concomitant with dystrophic neurite formation in the neuritic beta-amyloid plaques of Alzheimer's disease.


A radioisotopic method for the measurement of free fatty acid turnover and adrenoceptor response in small fragments of human adipose tissue.
New techniques for measuring rates of triglyceride hydrolysis and the re-esterification of resultant free fatty acids (FFA) in small fragments of human adipose tissue are described. Buffered medium containing [14C]-glucose and [3H]-palmitate of high specific activity is used to label newly synthesized acylglycerides in incubated tissue. 14C is incorporated only into the glyceride 'back bone' of acylglycerides, and the rate of incorporation may be used as a measure of the rate of synthesis of each glyceride molecular species. Due to mixing of exogenous [3H]-palmitate with endogenously released FFA, the actual specific activity of FFA esterified to [14C]-glycerides is always less than that of the [3H]-palmitate in the medium. The degree of dilution of [3H]-palmitate with unlabelled FFA is directly proportional to the rate of lipolysis. Thus, determination of the 14C/3H ratio in newly synthesized triglycerides permits the assessment of adrenoceptor dose-response characteristics (lipolysis) of unweighed fragments of adipose tissue. If rates of 14C-tri- and diglyceride accumulation are measured in tissue fragments whose cellularity and rate of glycerol release has been determined, the fraction of newly lipolyzed FFA which are re-esterified can be calculated. This 'cycle' of re-esterification controls the ratio of FFA: glycerol leaving the adipocyte. These techniques can be applied to serial subcutaneous adipose tissue samples from awake subjects in various clinical and experimental states.


Correlation of Strain Elastography with Conventional Sonography and FNAC/Biopsy.
Elastography is a new promising technique that can be especially helpful when used as an adjunct to conventional B-mode ultrasound in evaluating breast lesions. To evaluate the diagnostic performance of four interpretation criteria for elastography and to compare the diagnostic performance of sonoelastography with that of conventional sonography in characterising breast lesions as benign or malignant with FNAC/biopsy correlation. One hundred breast lesions were prospectively evaluated by ultrasound as well as by strain elastography followed by FNAC/ biopsy correlation. The criteria used were Elastography Score, Strain Ratio, Distance Ratio and Area Ratio. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value were calculated for each modality. The best cut-off point was calculated for each of the interpretation criteria using the MedCalc version 10.1 software. The diagnostic performance of the interpretation criteria was compared with that of conventional sonography by the area under the receiver operating characteristic curve using SPSS software. The elastography score was found to have the best performance among the 4 criteria used with a sensitivity and specificity of 100% and 82.66%, respectively at the best cut-off point between 3 and 4. At a best cut-off point for conventional sonography between BI-RADS categories 4A and 4B, the sensitivity and specificity were found to be 96% and 92% respectively. The area under the curve value was slightly greater for conventional sonography (0.980) than for the elastography score (0.913) using receiver operating characteristic curve analysis. While all interpretation criteria were able to differentiate benign and malignant lesions with statistical significance, the elastography score was found to be the most accurate. While conventional ultrasound remains the primary modality for the characterization of breast masses, elastography was found to have a role in low suspicion lesions (BI-RADS 3 and 4A) where it's greater specificity could justify avoiding unnecessary biopsy.


Determinants of out-of-district health facility bypassing in East Java, Indonesia.
Several large-scale reforms, including policies aimed at achieving universal health coverage, have been implemented to overcome health disparities in Indonesia. However, access to health services remains unequal. Many people 'bypass' health services in their home district to access health services in neighbouring districts, even though their health insurance does not cover such services. This study aims to identify the factors that are associated with this out-of-district bypassing behaviour. We surveyed 500 respondents living in the outermost districts of East Java province. We used data on education, income, district, age, gender, household size, district accessibility, insurance coverage status and satisfaction with health facilities in the home district and logistic regression analysis to model the predictors of out-of-district health facility bypassing. The most important predictors of the bypassing behaviour were education and poor access to health facilities in the home district. Open-ended data also found that the most important reason for seeking care in another district was mostly geographic. In contrast, health insurance coverage does not appear to be a significant predictor. Education and geographic factors are the main predictors of out-of-district bypassing behaviour, which appears to be how border communities express their health facility preferences. Local and central governments should continue their work to reduce inequality in access to health facilities in Indonesia's geographically challenged districts.


Using sequence similarity networks for visualization of relationships across diverse protein superfamilies.
The dramatic increase in heterogeneous types of biological data--in particular, the abundance of new protein sequences--requires fast and user-friendly methods for organizing this information in a way that enables functional inference. The most widely used strategy to link sequence or structure to function, homology-based function prediction, relies on the fundamental assumption that sequence or structural similarity implies functional similarity. New tools that extend this approach are still urgently needed to associate sequence data with biological information in ways that accommodate the real complexity of the problem, while being accessible to experimental as well as computational biologists. To address this, we have examined the application of sequence similarity networks for visualizing functional trends across protein superfamilies from the context of sequence similarity. Using three large groups of homologous proteins of varying types of structural and functional diversity--GPCRs and kinases from humans, and the crotonase superfamily of enzymes--we show that overlaying networks with orthogonal information is a powerful approach for observing functional themes and revealing outliers. In comparison to other primary methods, networks provide both a good representation of group-wise sequence similarity relationships and a strong visual and quantitative correlation with phylogenetic trees, while enabling analysis and visualization of much larger sets of sequences than trees or multiple sequence alignments can easily accommodate. We also define important limitations and caveats in the application of these networks. As a broadly accessible and effective tool for the exploration of protein superfamilies, sequence similarity networks show great potential for generating testable hypotheses about protein structure-function relationships.


Nodal-dependent Cripto signaling promotes cardiomyogenesis and redirects the neural fate of embryonic stem cells.
The molecular mechanisms controlling inductive events leading to the specification and terminal differentiation of cardiomyocytes are still largely unknown. We have investigated the role of Cripto, an EGF-CFC factor, in the earliest stages of cardiomyogenesis. We find that both the timing of initiation and the duration of Cripto signaling are crucial for priming differentiation of embryonic stem (ES) cells into cardiomyocytes, indicating that Cripto acts early to determine the cardiac fate. Furthermore, we show that failure to activate Cripto signaling in this early window of time results in a direct conversion of ES cells into a neural fate. Moreover, the induction of Cripto activates the Smad2 pathway, and overexpression of activated forms of type I receptor ActRIB compensates for the lack of Cripto signaling in promoting cardiomyogenesis. Finally, we show that Nodal antagonists inhibit Cripto-regulated cardiomyocyte induction and differentiation in ES cells. All together our findings provide evidence for a novel role of the Nodal/Cripto/Alk4 pathway in this process.


Deregulation of hyaluronan synthesis, degradation and binding promotes breast cancer.
Clinical and experimental data indicate that hyaluronan accumulates in breast cancer compared with normal breast epithelium, which correlates to poor prognosis. In this review, we discuss the expression of genes encoding enzymes that synthesize or degrade hyaluronan, i.e. hyaluronan synthases and hyaluronidases or bind hyaluronan, i.e. CD44 and receptor for hyaluronan-mediated motility (RHAMM, also designated as HMMR or CD168), in relation to breast cancer progression. Hyaluronan and hyaluronan receptors have multi-faceted roles in signalling events in breast cancer. A better understanding of the molecular mechanisms underlying these signalling pathways is highly warranted and may lead to improvement of cancer treatment.


Limited proteolysis and 'in vitro' mutagenesis of bovine brain inositol monophosphatase identifies an N-terminal region important for activity.
Bovine brain inositol monophosphatase is rapidly cleaved by endoprotease lys-C at a single site in the absence of SDS. Further sites are revealed only after prolonged incubation with high concentrations of protease. The initial cleavage occurs near one end of the enzyme, generating an N-terminally-derived 36-residue peptide, which is blocked, and a large 28 kDa fragment bearing a free N-terminus. The start sequence of this fragment was found to be Xaa-Ser-Pro-Ala-Asp-Leu-Val, consistent with the cDNA sequence, and Lys-36-Ser-37 was identified as the cleavage site. The activity of the cleaved enzyme was markedly decreased to 3% of that of the native enzyme, although its dimeric structure was preserved. The 36-residue peptide was not covalently associated with the large fragment after proteolytic cleavage, although the possibility of non-covalent association could not be excluded. Finally, the epitope for the inhibitory monoclonal antibody G-2A4 [Gee, Howell, Ryan & Ragan (1989) Biochem J. 264. 793-798] was found to lie proximal to the endoprotease lys-C cleavage site. In vitro mutagenesis further mapped the epitope for monoclonal antibody G-2A4 to residues around Cys-8 of the enzyme. These results suggest that the N-terminal region of the enzyme is important for activity.


Screening of blood donations for HIV-1 antibody: 1985-1991.
One hundred and ninety HIV-1 antibody positive donors have been detected, out of 14.85 million blood donations screened in the United Kingdom and the Isle of Man, since the start of testing in October 1985 to the end of March 1991. There were 145 men and 45 women, with an overall seropositivity rate of 0.001%. Records for new donors (ie, those donating for the first time) have been kept since February 1986 and 79 out of 1.9 million donations have been seropositive (58 men and 21 women); a rate of 0.004%. One hundred and forty-one (74.2%) of the 190 positive donors were found to have been exposed to a high risk of HIV infection. Twenty-four (13%) denied any exposure other than heterosexual intercourse with partners who were not considered to be at high risk. In six cases the partners were from countries where the main route of transmission is heterosexual. Seven donors (4%) attributed infection to some other cause. Eighteen (9.5%) have been lost to follow-up, are still being investigated or have not yet been interviewed. Combined tests which screen for both HIV-1 and HIV-2 antibodies were introduced in June 1990. The last 2.55 million donations (including 370,000 new donors) have been tested with these kits. One HIV-2 antibody positive donation had been confirmed in a new donor by the end of March 1991.(ABSTRACT TRUNCATED AT 250 WORDS)


Signal and noise extraction from analog memory elements for neuromorphic computing.
Dense crossbar arrays of non-volatile memory (NVM) can potentially enable massively parallel and highly energy-efficient neuromorphic computing systems. The key requirements for the NVM elements are continuous (analog-like) conductance tuning capability and switching symmetry with acceptable noise levels. However, most NVM devices show non-linear and asymmetric switching behaviors. Such non-linear behaviors render separation of signal and noise extremely difficult with conventional characterization techniques. In this study, we establish a practical methodology based on Gaussian process regression to address this issue. The methodology is agnostic to switching mechanisms and applicable to various NVM devices. We show tradeoff between switching symmetry and signal-to-noise ratio for HfO2-based resistive random access memory. Then, we characterize 1000 phase-change memory devices based on Ge2Sb2Te5 and separate total variability into device-to-device variability and inherent randomness from individual devices. These results highlight the usefulness of our methodology to realize ideal NVM devices for neuromorphic computing.


Optical location of centroids of nonoverlapping objects.
We present an optical technique for finding the centroids of nonoverlapping objects in a scene, thus locating the objects and preserving the underlying advantage of matched filtering approaches to pattern recognition. One is then free to extract any feature desired at these centroid locations rather than restricted to the matched filter test statistic. Furthermore, this allows general feature extraction avoiding prior scene segmentation into individual objects. The technique can also be used for tracking the motion of rigid or nonrigid objects. It consists of cross-correlating the input f(x,y) with a windowed version of the function x + iy and detecting the zeros of the magnitude of the resulting correlation. At these points the x and y first moments vanish. The window is selected based on the size and separation of the objects in a scene. Experimental verification as well as restrictions are also presented.


Do Family Responsibilities and a Clinical Versus Research Faculty Position Affect Satisfaction with Career and Work-Life Balance for Medical School Faculty?
Balancing career and family obligations poses challenges to medical school faculty and contributes to dissatisfaction and attrition from academics. We examined the relationship between family setting and responsibilities, rank, and career and work-life satisfaction for faculty in a large U.S. medical school. Baseline faculty surveys were analyzed from the first year of a 4-year National Institutes of Health-funded study to evaluate awareness, knowledge, attitudes, and use of family friendly policies and career satisfaction. The study focus was on the impact of family responsibilities and characteristics of the faculty position (rank, clinical vs. nonclinical, and academic series) in multivariate comparisons between primary predictors and outcomes of interest. Both clinical and family responsibilities for children under 18 play a major and interacting role in satisfaction with career and work-life balance. Clinical faculty respondents without children at home reported significantly greater career satisfaction and better work-life balance than their nonclinical counterparts. Nonclinical faculty respondents with children reported greater satisfaction and better balance than counterparts without family responsibilities. However, the advantage in career satisfaction and work-life balance for clinical faculty respondents disappeared for those with responsibility for young children. No gender-based differences were noted in the results or across faculty rank for respondents; however, for women, reaching associate professor resulted in greater career satisfaction. This study suggests that both work-related factors and family responsibilities influence satisfaction with career and work-life balance, but the predictors appear to interact in complex and nuanced ways. Further research is needed to delineate more clearly these interactions and to explore other factors that may play important additional roles.


Comparison of amikacin lung delivery between AKITA® and eFlow rapid® nebulizers in healthy controls and patients with CF: A randomized cross-over trial.
Nebulization plays a key role in the treatment of cystic fibrosis. The Favorite function couple to jet nebulizers (AKITA®) emerged recently. The aim of this study was to assess the efficiency of the lung delivery by the AKITA® by comparing the urinary concentration of amikacin after nebulization with the AKITA® and the eFlow rapid®, in healthy subjects and patients with CF (PwCF). The two samples (healthy subjects and PwCF) were randomized (cross-over 1:1) for two nebulizations (500 mg of amikacin diluted in 4 mL of normal saline solution), with the AKITA® and with the eFlow rapid®. The primary endpoint was the amount of urinary excretion of amikacin over 24 h. The constant of elimination (Ke) was calculated based on the maximal cumulative urinary amikacin excretion plotted over time. The total amount of urinary amikacin excretion was greater when AKITA® was used in PwCF (11.7 mg (8.2-14.1) vs 6.1 mg (3.7-13.3); p = 0.02) but not different in healthy subjects (14.5 mg (11.7-18.5) vs 12.4 mg (8.0-17.1); p = 0.12). The duration of the nebulization was always shorter with eFlow rapid® than with AKITA® (PwCF: 6.5 ± 0.6 min vs 9.2 ± 1.8 min; p = 0.001 - Healthy: 4.7 ± 1.3 min vs 9.7 ± 1.6 min; p = 0.03). The constant of elimination was similar between the two modalities in CF subjects (0.153 (0.071-0.205) vs 0.149 (0.041-0.182); p = 0.26) and in healthy subjects (0.166 (0.130-0.218) vs 0.167 (0.119-0.210), p = 0.25). the Favorite inhalation is better to deliver a specific amount of drug than a mesh nebulizer (eFlow rapid®) in PwCF but not in healthy subjects.


U.S. HIV Incidence and Transmission Goals, 2020 and 2025.
The recently updated U.S. National HIV/AIDS Strategy sets key HIV prevention and care targets for 2020, but the trajectory of the epidemic remains unclear. Authors modeled HIV incidence, prevalence, and mortality for the U.S. over 10 years to determine whether an ambitious trajectory toward "ending AIDS" by 2025 would be achievable. Authors utilized recently published 2010-2013 Centers for Disease Control and Prevention surveillance data to model HIV incidence, prevalence, and mortality. Authors applied a 90/90/90 framework (90% awareness of serostatus, 90% of diagnosed individuals in care, and 90% of individuals on antiretroviral therapy virally suppressed) by 2020 and 95/95/95 by 2025 to assess the feasibility of meeting epidemiologic targets. Analyses were conducted in 2016. With a goal of reducing infections to 21,000 new HIV infections in 2020, authors project a transmission rate of 1.74, 12,571 deaths, and a total of 1,205,515 people living with HIV. By 2025, with a target of 12,000 new HIV infections (a 69% decrease in HIV incidence), authors project a transmission rate of 0.98, 12,522 deaths, and a total of 1,220,615 people living with HIV. With a 90/90/90 framework by 2020 and a 95/95/95 framework by 2025, these epidemiologic targets would be feasible. Key programmatic milestones provide an ambitious, but important, pathway to reduce U.S. HIV incidence below 12,000 new infections by 2025. HIV incidence would decrease below mortality in 2025, marking a transition toward ending the HIV/AIDS epidemic. Such goals will require a sustained and intensified national commitment over the next decade.


[¿Azitromicina como tratamiento contra Chlamydia trachomatis?]
Azithromycin and doxycycline effectiveness has been demonstrated in the treatment of urogenital chlamydiasis, which has remained unchanged for a long time. Autoinoculation has been proposed as a method of reinfection and persistence of the disease in women and probably also owing to azithromycin pharmacokinetics in this tissue. With the new diagnostic methods and tests of cure, a difference has been demonstrated in favor of doxycycline in the treatment of rectal chlamydiasis Antimicrobial resistance has not played a relevant role since no treatment-resistant strains have been found in vivo. Nevertheless, azithromycin remains a first-choice drug, since it can be administered as a single dose, which favors therapeutic adherence.


Transforming growth factor beta can mediate apoptosis via the expression of TRAIL in human hepatoma cells.
Transforming growth factor beta (TGF-beta) has been shown to induce apoptotic cell death in normal and transformed hepatocytes. However, the exact mechanism through which TGF-beta induces cell death is still unknown. We examined a potential role of various death receptor/ligand systems in TGF-beta-induced apoptosis and identified the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) as a mediator of TGF-beta-induced apoptosis in hepatoma cells. TGF-beta-induced apoptosis is significantly impaired upon blockage of TRAIL. We show that TRAIL is upregulated in hepatoma cells upon treatment with TGF-beta, whereas TRAIL receptor levels remain unchanged. In conclusion, our results provide evidence that the TRAIL system is critically involved in TGF-beta-induced cell death in liver pathology.


Adjuvant chemotherapy for patients with high-grade soft-tissue sarcomas of the extremity.
We have previously reported the results of a randomized trial that demonstrated the survival benefit of adjuvant chemotherapy in the treatment of patients with high-grade extremity sarcomas compared with no chemotherapy. This regimen included doxorubicin, cyclophosphamide, and methotrexate. This report updates and extends our experience. The median follow-up of this trial is now 7.1 years and reveals a 5-year disease-free survival of 75% and 54% for chemotherapy and no chemotherapy groups, respectively (two-sided P [P2] = .037). The 5-year overall survival for patients in this trial was 83% and 60% for the chemotherapy and no chemotherapy groups, respectively, with a trend towards improved survival in the chemotherapy arm (P2 = .124). Because of doxorubicin-induced cardiomyopathy we performed a subsequent randomized trial comparing this high-dose regimen to reduced cumulative doses of doxorubicin and cyclophosphamide without methotrexate. Eighty-eight patients were entered into this trial which has a median follow-up of 4.4 years. The 5-year disease-free and overall survival for patients treated with the reduced doses of chemotherapy was 72% and 75%, respectively, and was not significantly different from the high-dose regimen. No patients developed congestive heart failure on this study. We conclude that adjuvant chemotherapy improves disease-free survival in patients with extremity soft-tissue sarcomas. The overall survival advantage in patients receiving adjuvant chemotherapy in our initial randomized high-dose chemotherapy trial has diminished though it continues to favor the chemotherapy group. A reduced-dose chemotherapy regimen was found to be comparable to the high-dose regimen.


The Benefits of Closed-Loop Transcranial Alternating Current Stimulation on Subjective Sleep Quality.
Poor sleep quality is a common complaint, affecting over one third of people in the United States. While sleep quality is thought to be related to slow-wave sleep (SWS), there has been little investigation to address whether modulating slow-wave oscillations (SWOs) that characterize SWS could impact sleep quality. Here we examined whether closed-loop transcranial alternating current stimulation (CL-tACS) applied during sleep impacts sleep quality and efficiency. CL-tACS was used in 21 participants delivered at the same frequency and in phase with endogenous SWOs during sleep. Sleep quality was assessed in the morning following either verum or sham control stimulation during sleep, with order counterbalanced within participants. Higher sleep quality and efficiency were found after verum stimulation nights compared to control. The largest effects on sleep quality were found immediately following an adaptation night in the laboratory for which sleep quality was reduced. Applying CL-tACS at the same frequency and phase as endogenous SWOs may offer a novel method to improve subjective sleep quality after a night with poor quality sleep. CL-tACS might be helpful for increasing sleep quality and efficiency in otherwise healthy people, and in patients with clinical disorders that involve sleep deficits.


A single dose of sodium nitrate does not improve oral glucose tolerance in patients with type 2 diabetes mellitus.
Dietary nitrate (NO3(-)) supplementation has been proposed as an emerging treatment strategy for type 2 diabetes. We hypothesized that ingestion of a single bolus of dietary NO3(-) ingestion improves oral glucose tolerance in patients with type 2 diabetes. Seventeen men with type 2 diabetes (glycated hemoglobin, 7.3% ± 0.2%) participated in a randomized crossover experiment. The subjects ingested a glucose beverage 2.5 hours after consumption of either sodium NO3(-) (0.15 mmol NaNO3(-) · kg(-1)) or a placebo solution. Venous blood samples were collected before ingestion of the glucose beverage and every 30 minutes thereafter during a 2-hour period to assess postprandial plasma glucose and insulin concentrations. The results show that plasma NO3(-) and nitrite levels were increased after NaNO3(-) as opposed to placebo ingestion (treatment-effect, P = .001). Despite the elevated plasma NO3(-) and nitrite levels, ingestion of NaNO3(-) did not attenuate the postprandial rise in plasma glucose and insulin concentrations (time × treatment interaction, P = .41 for glucose, P = .93 for insulin). Despite the lack of effect on oral glucose tolerance, basal plasma glucose concentrations measured 2.5 hours after NaNO3(-) ingestion were lower when compared with the placebo treatment (7.5 ± 0.4 vs 8.3 ± 0.4 mmol/L, respectively; P = .04). We conclude that ingestion of a single dose of dietary NO3(-) does not improve subsequent oral glucose tolerance in patients with type 2 diabetes.


Phenolic Plant Extracts Induce Sirt1 Activity and Increase Antioxidant Levels in the Rabbit's Heart and Liver.
Several dietary phytochemicals potentially regulate the equilibrium between oxidant and antioxidant species. The aim of this study was to evaluate the effects of Lippia citriodora, Raphanus sativus, and Solanum lycopersicum on blood parameters, oxidative/antioxidant status, and SIRT1 activity in the rabbit's heart and liver. Twenty rabbits were divided into 4 groups of 5 animals each. The control group (CN) received a feed without any additives. One intervention group received a supplement containing verbascoside (VB), another Raphanus sativus extract (RAP), and lastly lycopene (LYC). Oxidant-antioxidant parameters and SIRT1 activity were measured in plasma and in the heart and liver, respectively. The treatment with VB, RAP, and LYC resulted in a marked improvement in the blood lipid and glycaemic profile in respect to CN. VB was the most effective, but all three plant extracts induced a significant reduction in oxidant parameters as well as an increase in antioxidant tissue activity and vitamin A and E levels. SIRT1 activity was significantly increased in both VB and LYC compared to CN, but the increased levels in the VB group were far the highest. The multivariate analysis suggests that the benefits of VB, particularly the antiglycaemic and antioxidant effects, might be mediated by increasing SIRT1 activity.


In vitro transfusion of red blood cells results in decreased cytokine production by human T cells.
Transfusion-related immunomodulation consists of both proinflammatory and anti-inflammatory responses after transfusion of blood products. Stored red blood cells (RBCs) suppress human T-cell proliferation in vitro, but the mechanism remains unknown. We hypothesized that cytokine synthesis by T cells may be inhibited when stored RBCs are present and that suppression between fresh and stored RBCs would be different. Purified human T cells were stimulated to proliferate with anti-CD3/anti-CD28 and then exposed to stored or fresh RBCs. Cells were placed in culture for 5 days. Cell culture supernatants were analyzed for the production of typical T-cell cytokines using multianalyte ELISArray kits. Stimulated T cells proliferated. RBC exposure markedly suppressed this proliferation. Interleukin 10, interleukin 17a, interferon γ, tumor necrosis factor α, and granulocyte macrophage colony-stimulating factor were increased in response to stimulation but depressed in the presence of stored RBCs. The use of fresh RBCs also resulted in depression of these cytokines when compared with stimulated T cells with no RBCs; however, this depression was less pronounced. T-cell activation is associated with both proinflammatory and anti-inflammatory cytokine release, comparable with patterns seen in trauma and acute injury. All of these responses are depressed by an exposure to stored RBCs. Decreased levels of these cytokines after RBC transfusion represents a potential contributor to the immunosuppressive complications seen in trauma patients after transfusion. This provides insight for future mechanistic studies to delineate the role of RBC transfusion in transfusion-related immunomodulation.


Negative Impact of Skeletal Muscle Loss after Systemic Chemotherapy in Patients with Unresectable Colorectal Cancer.
Skeletal muscle depletion (sarcopenia) is closely associated with limited physical ability and high mortality. This study evaluated the prognostic significance of skeletal muscle status before and after chemotherapy in patients with unresectable colorectal cancer (CRC). We conducted a retrospective analysis of 215 consecutive patients with unresectable CRC who underwent systemic chemotherapy. Skeletal muscle cross-sectional area was measured by computed tomography. We evaluated the prognostic value of skeletal muscle mass before chemotherapy and the rate of skeletal muscle change in cross-sectional area after chemotherapy. One-hundred-eighty-two patients met our inclusion criteria. There were no significant differences in progression-free survival (PFS) or overall survival (OS) associated with skeletal muscle mass before chemotherapy. However, 22 patients with skeletal muscle loss (>5%) after chemotherapy showed significantly shorter PFS and OS compared with those without skeletal muscle loss (PFS, log-rank p = 0.029; OS, log-rank p = 0.009). Multivariate Cox regression analysis revealed that skeletal muscle loss after chemotherapy (hazard ratio, 2.079; 95% confidence interval, 1.194-3.619; p = 0.010) was independently associated with OS. Skeletal muscle loss after chemotherapy was an independent, negative prognostic factor in unresectable CRC.


Effects of reward and behavioral context on neural activity in the primate inferior colliculus.
Neural activity in the inferior colliculus (IC) likely plays an integral role in the processing of various auditory parameters, such as sound location and frequency. However, little is known about the extent to which IC neural activity may be influenced by the context in which sounds are presented. In this study, we examined neural activity of IC neurons in the rhesus monkey during an auditory task in which a sound served as a localization target for a saccade. Correct performance was rewarded, and the magnitude of the reward was varied in some experiments. Neural activity was also assessed during a task in which the monkey maintained fixation of a light while ignoring the sound, as well as when sounds were presented in the absence of any task. We report that neural activity increased late in the trial in the saccade task in 58% of neurons and that the level of activity throughout the trials could be modulated by reward magnitude for many neurons. The late-trial neural activity similarly increased in the fixation task in 39% of the neurons tested for this task but was not observed when sounds were presented in the absence of a behavioral task and reward. Together, these results suggest that a reward-related signal influences neural activity in the IC.


Sensor data fusion for responsive high resolution ultrasonic temperature measurement using piezoelectric transducers.
Ultrasonic temperature measurement allows for responsive measurements across an entire ultrasonic pathway, unlike most conventional temperature sensors that respond to the temperature at the point of their placement only after a notable response time. The high cost of required ultrasonic instrumentation can be reduced substantially by using ultrasonic oscillating temperature sensors (UOTS) consisting of inexpensive narrowband piezo transducers and driving electronics. An UOTS produces sustained oscillations at a frequency that relates to the temperature of the medium between the transducers. The existence of thermal hysteresis in UOTS readings, observed experimentally and apparently related to the fundamental properties of piezoelectric materials, makes conversion of the output frequency readings to the temperature values ambiguous. This makes it complicated to calibrate and use UOTS on their own. In the reported experiment (heating, then naturally cooling of a water vessel equipped with both UOTS and conventional sensors), this hysteresis was solved by fusing UOTS data with conventional temperature sensor readings. As the result, the combination of one UOTS plus one conventional reference sensor allowed improving both the temperature resolution and responsiveness of the latter and ambiguity of the readings of the former. Data fusion effectively led to calibrating the UOTS at every change of the conventional sensor's reading, removing any concerns related to the thermal expansion/contraction of the ultrasonic pathway itself and/or hysteresis of piezoelectric transducers.


A commentary on Marja Jylhä's "What is self-rated health and why does it predict mortality? Towards a unified conceptual model"(69:3, 2009, 307-316).
This is a commentary on Marja Jylhä's paper "What is self-rated health and why does it predict mortality? Toward a unified conceptual model" (2009) published recently in this journal. In this commentary we address two issues that we believe the paper raises. Firstly, we react to the theory proposed by Jylhä about how individuals form an assessment of their own health. Secondly, we discuss what we believe amounts to an existing confusion about what self-rated health can and should be considered to measure. We conclude that the cognitive process that Jylhä introduces in her paper should be expanded to include external and internal states that influence the process illogically and irrationally, as well as a long-term ongoing component reflecting that people are continuously presented with health-relevant information during their lives and continuously evaluate and change their assessment of health according to this information. Further, we conclude that self-rated health should best be viewed as a measure of people's perception of their health rather than a measure of true health.


Thyroid-Related Orbital Decompression Surgery: A Multivariate Analysis of Risk Factors and Outcomes.
Despite the number of publications on orbital decompression surgery for thyroid eye disease, there are few comparative studies and most studies are underpowered. The goal of the study is to use multivariable analysis to identify independent patient and disease-related predictors of response to decompression surgery and of need for secondary decompressions. The authors retrospectively reviewed all patients who underwent transorbital thyroid-related orbital decompression surgery at the Kellogg Eye Center of the University of Michigan between 1999 and 2014. Demographic, medical, and surgical covariates were collected. Decompression techniques included medial, lateral, and balanced decompressions, with or without orbital fat removal. Main outcomes included proptosis reduction and secondary decompressions, both analyzed at the orbital level. Univariate and multivariable analyses (with adjustment for interorbit correlation) were conducted to determine predictors of the outcomes of interest. Mean proptosis reduction was 3.8 ± 2.4 mm (mean ± standard deviation, N = 420 orbits). The secondary decompression rate was 13.8% (82/594). On multivariable mixed linear regression, larger preoperative proptosis (p < 0.0001), balanced decompression (p = 0.0002), thyroid eye disease duration < 4 years (p = 0.0093), and history of orbital radiation (p = 0.0111) were all predictive of greater proptosis reduction. On multivariable survival modeling, factors associated with increased hazard for secondary decompression include younger age (p = 0.0434), larger preoperative proptosis (p = 0.0001), unilateral decompression (p = 0.0272), preoperative steroid treatment (p = 0.0200), and normal thyroid function (p = 0.0148). Factors associated with decreased hazard include adjunctive fat decompression (p = 0.0004), balanced decompression (vs. lateral, p = 0.0039), and African-American ethnicity (p = 0.0076). Despite a diverse study cohort, the authors have identified factors associated with clinically relevant outcomes of decompression surgery for thyroid eye disease, including proptosis reduction and incidence of secondary decompression. Randomized controlled trials of different treatment algorithms for TED are needed to devise optimized guidelines for individualizing surgical care.


Sinus and atrioventricular nodal function: Preoperative and early postoperative assessment in children.
Ninety-three rapid atrial pacing studies were performed in 38 children to compare preoperative and early postoperative function of the sinus and atrioventricular (AV) nodes. The interval between the preoperative and postoperative studies was under 6 months in the majority of patients. Postoperative studies were performed within 48 hours of operation and between 4 and 8 days after operation. Sinus nodal function as measured by sinus nodal recovery time (SNRT) was an unreliable index in determining depression since the number who improved postoperatively (10/55) was nearly equal to the number that worsened (12/55). The majority who had abnormal function postoperatively demonstrated a junctional rather than sinus recovery focus. This finding appears a more definitive and more reproducible indicator of sinus node depression in the postoperative patient. Postoperative AV nodal function was decreased (as measured by the cycle length [CL] at which Wenckebach periodicity occurred) in 15 of 55 studies (27%) of the entire group. There was nearly an equal chance for improvement (24%) in function. This also applied to those patients who had sequential studies. Therefore, this method of assessment for AV nodal function was unreliable, or else the operation did not affect the node significantly. The latter is unlikely in view of late postoperative data. The greatest utility of this test was to determine the capability for AV conduction in certain patients with slow escape rhythms in the absence of surface P waves, and to differentiate complete heart block from AV dissociation when atrial activity was absent. Despite the variability of effects on the sinus and AV nodes in these patients, those who demonstrated depression had a significantly higher incidence of dysrhythmias (80% of patients with sinus nodal depression and 100% of patients with AV nodal depression).


[Correction of schoolchildren's diets with biologically active additives].
The actual nutrition of schoolchildren in the Nizni-Novgorod district is imbalanced due to the deficiency of protein and vitamins and to the high contents of fats and carbohydrates. The authors provide evidence for a combined preparation to correct the children's diets, which incorporates animal protein, biologically active plant additives, and egg-shell calcium. The use of the preparation in combination with liquid bifidumbacterin contributes to increases in morphofunctional parameters in adolescents.


In Vivo Damage of the Head-Neck Junction in Hard-on-Hard Total Hip Replacements: Effect of Femoral Head Size, Metal Combination, and 12/14 Taper Design.
Recently, concerns have been raised about the potential effect of head-neck junction damage products at the local and systemic levels. Factors that may affect this damage process have not been fully established yet. This study investigated the possible correlations among head-neck junction damage level, implant design, material combination, and patient characteristics. Head-neck junctions of 148 retrieved implants were analysed, including both ceramic-on-ceramic (N = 61) and metal-on-metal (N = 87) bearings. In all cases, the male taper was made of titanium alloy. Damage was evaluated using a four-point scoring system based on damage morphology and extension. Patient age at implantation, implantation time, damage risk factor, and serum ion concentration were considered as independent potential predicting variables. The damage risk factor summarises head-neck design characteristics and junction loading condition. Junction damage correlated with both implantation time and damage factor risk when the head was made of ceramic. A poor correlation was found when the head was made of cobalt alloy. The fretting-corrosion phenomenon seemed mainly mechanically regulated, at least when cobalt alloy components were not involved. When a component was made of cobalt alloy, the role of chemical phenomena increased, likely becoming, over implantation time, the damage driving phenomena of highly stressed junctions.


Fatigue in systemic lupus: the role of disease activity and its correlates.
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that leads to a variety of negative health outcomes resulting from inflammation in various organ systems. Although treatment continues to advance, fatigue remains one of the most salient, poorly understood and addressed patient complaints. Understanding the mechanisms of fatigue can help guide the development of interventions to improve health outcomes. The aim of this research was to evaluate the contribution of six variables (disease activity, insomnia, depression, stress, pain and physical health) to fatigue in SLE without concomitant fibromyalgia (FM). A total of 116 ethnically diverse, primarily female participants (91%) with SLE, receiving care at university medical centers, completed assessments of disease activity and quality of life outcomes (FACIT-FT, Insomnia Severity Index, Perceived Stress Scale (PSS-4), Pain Inventory, Depression-PHQ-9, and LupusPRO-physical function). All patients met the American College of Rheumatology classification criteria for SLE and did not have a known diagnosis of FM. Multivariate linear and stepwise regression analyses were conducted with fatigue (FACIT-FT) as the dependent variable, and the above six variables as independent variables. Mean (SD) age was 39.80 (13.87) years; 50% were African American, 21% Caucasian, 13% Hispanic, 9% Asian and 8% other. Mean (SD) FACIT-FT was 20.09 (12.76). Collectively, these six variables explained 57% of the variance in fatigue. In the multivariate model, depression, stress and pain were significantly and independently associated with fatigue, but not disease activity, sleep or physical health. Stress had the largest effect on fatigue (β 0.77, 95% CI 0.17-1.38, p = 0.01), followed by depression (β 0.66, 95% CI 0.21-1.10, p = 0.005). On stepwise regression analysis, only stress, depression and pain were retained in the model, and collectively explained 56% of the variance in fatigue. All three remained independent correlates of fatigue, with the largest contribution being stress (β 0.84, 95% CI 0.27-1.42, p = 0.005), followed by depression (β 0.79, 95% CI 0.44-1.14, p < 0.001) with fatigue. Stress, depression and pain are the largest independent contributors to fatigue among patients with SLE, without concurrent FM. Disease activity, sleep and physical health were not associated with fatigue. The evaluation of stress, depression and pain needs to be incorporated during assessments and clinical trials of individuals with SLE, especially within fatigue. This stress-depression-fatigue model requires further validation in longitudinal studies and clinical trials. Significance and innovation: • Disease activity, sleep, pain, stress, depression, and physical health have been reported individually to be associated with fatigue in lupus. This analysis evaluated the role of each and all of these six variables collectively in fatigue among patients with SLE without a known diagnosis of FM. • Disease activity, sleep and physical health were not significantly related to fatigue, but depression, stress and pain were. • The results emphasize the need to evaluate and treat fatigue in individuals with SLE utilizing a biopsychosocial approach, particularly in the realm of clinical trials. Behavioral medicine interventions are shown to be most effective for the treatment of depression, stress and pain.


In-situ characterization of ultrathin nickel silicides using 3D medium-energy ion scattering.
Epitaxial ultrathin films are of utmost importance for state-of-the-art nanoelectronic devices, such as MOSFET transistors and non-volatile memories. At the same time, as the film thickness is reduced to a few nanometers, characterization of the materials is becoming challenging for commonly used methods. In this report, we demonstrate an approach for in-situ characterization of phase transitions of ultrathin nickel silicides using 3D medium-energy ion scattering. The technique provides simultaneously depth-resolved composition and real-space crystallography of the silicide films using a single sample and with a non-invasive probe. We show, for 10 nm Ni films on Si, that their composition follows a normal transition sequence, such as Ni-Ni2Si-NiSi. However, the transition process is significantly different for samples with initial Ni thickness of 3 nm. Depth-resolved crystallography shows that the Ni films transform from an as-deposited disordered layer to an epitaxial silicide layer at the temperature of ~290 °C, significantly lower than previously reported. The high depth resolution of the technique permits us to determine the composition of the ultrathin films to be 38% Ni and 62% Si.


The prospective pioglitazone clinical trial in macrovascular events (PROactive): can pioglitazone reduce cardiovascular events in diabetes? Study design and baseline characteristics of 5238 patients.
The PROspective pioglitAzone Clinical Trial In macroVascular Events (PROactive) assesses the effect of pioglitazone, a peroxisome proliferator-activated receptor agonist, with anti-inflammatory and vascular properties, on the secondary prevention of macrovascular events in type 2 diabetes. PROactive is an on-going randomized, double-blind outcome study in patients with type 2 diabetes managed with diet and/or oral blood glucose-lowering drugs (combination of oral agents with insulin is permitted) who have a history of macrovascular disease. Patients are randomized to receive pioglitazone (forced titration from 15 to 30 to 45 mg, depending on tolerability) or placebo in addition to existing therapy. The primary end point is the time from randomization to occurrence of a new macrovascular event or death. Follow-up is estimated to span 4 years. A total of 5238 patients have been randomized from 19 countries. At entry into the study, patients enrolled are a mean age of 61.8 years, with type 2 diabetes for a mean of 9.5 years; 60.9 and 61.5% are taking metformin or a sulfonylurea, respectively; and 33.6% are using insulin in addition to oral glucose-lowering drugs. The majority of patients are men (66.1%). Patients are required to meet one or more of entry criteria, as follows: >6 months' history of myocardial infarction (46.7%); coronary artery revascularization (30.8%), stroke (18.8%), or acute coronary syndrome for >3 months (13.7%); other evidence of coronary artery disease (48.1%); or peripheral arterial occlusive disease (19.9%). One-half (48.5%) of the patients have two or more of these risk factors. Three-quarters (75.4%) have hypertension, and 58.8% are current or previous smokers. The cohort of patients enrolled in PROactive is a typical type 2 diabetic population at high risk of further macrovascular events. The characteristics of this population are ideal for assessing the ability of pioglitazone to reduce the cardiovascular risk of patients with type 2 diabetes.


Identification and sequencing of a cDNA encoding 6-phosphogluconate dehydrogenase from a fungus, Cunninghamella elegans and expression of the gene in Escherichia coli.
The fungus, Cunninghamella elegans has been widely used in bioremediation and microbial models of mammalian studies in many laboratories. Using the polymerase chain reaction to randomly amplify the insert directly from the single non-blue plaques of a C. elegans cDNA library, then partly sequencing and comparing with GenBank sequences, we have identified a clone which contains C. elegans 6-phosphogluconate dehydrogenase gene. The polymerase chain reaction product was cloned into a plasmid, pGEM-T Easy vector for full insert DNA sequencing. The 6-phosphogluconate dehydrogenase gene (1458 bases) and the deduced protein sequence were determined from the insert DNA sequence. The gene was found by open reading frame analysis and confirmed by the alignment of the deduced protein sequence with other published 6-phosphogluconate dehydrogenase sequences. Several highly conserved regions were found for the 6-phosphogluconate dehydrogenase sequences. The 6-phosphogluconate dehydrogenase gene was subcloned and over-expressed in a plasmid-E. coli system (pQE30). The cell lysate of this clone has a very high 6-phosphogluconate dehydrogenase enzyme activity. Most of the recombinant protein in this system was formed as insoluble inclusion bodies, but soluble in high concentration of urea-buffer. Ni-NTA resin was used to purify the recombinant protein which showed 6-phosphogluconate dehydrogenase enzyme activity. The recombinant protein has a predicted molecular size correlating with that revealed by sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. The C. elegans 6-phosphogluconate dehydrogenase was in a cluster with yeast' 6-phosphogluconate dehydrogenase in the phylogenetic tree. Bacterial 6-phosphogluconate dehydrogenase and higher organisms' 6-phosphogluconate dehydrogenase were found in different clusters.


Lipocalin-type prostaglandin D synthase is not a biomarker of atherosclerotic manifestations.
Over the last decades Lipocalin-type prostaglandin D synthase (L-PGDS), Osteoprotegerin (OPG), Osteopontin (OPN) and Pregnancy associated plasma protein A (PAPP-A) have been reported to be associated with coronary artery disease, and L-PGDS has been proposed as a potential new diagnostic tool in the setting of stable coronary artery disease. We set out to investigate if measurement of concentrations of these biomarkers could be used to differentiate between four groups of individuals with different atherosclerotic manifestations. A total of 120 individuals from four equal gender- and age-matched groups were studied: (i) no previous cardiovascular disease (CVD) and no coronary calcifications [CAC-negative group], (ii) no previous CVD but evidence of severe coronary calcifications [CAC-positive group], (iii) acute coronary syndrome [ACS-group], and (iv) clinical stable patients with CVD, who were referred for cardiovascular surgery [CVD-group]. Concentrations of L-PGDS, OPG, OPN and PAPP-A were analyzed and compared between the four groups. We did not find any significant differences in L-PGDS concentrations between the four groups (p = 0.32). OPG concentrations differed significantly (p = 0.003), with the highest concentration observed in ACS patients. Considering OPN (p = 0.12) and PAPP-A (p = 0.53) their concentrations between groups did not differ significantly. The main message from this study is the observation that L-PGDS based on a single blood test appears to be less valuable than previously proposed in identification of patients with coronary artery disease. However, ACS patients have higher OPG concentrations than patients with different manifestations of stable atherosclerosis. Neither OPN nor PAPP-A concentrations differed between groups.


Cytokine expression in a rat model of Staphylococcus aureus endophthalmitis.
To examine the ability of viable Staphylococcus aureus to induce the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, cytokine-induced neutrophil chemoattractant (CINC), and interferon (IFN)-gamma after intravitreal injection. Experimental rat eyes were injected with a 25-microl volume of approximately 80 colony-forming units of viable S. aureus; control eyes received sterile saline. Eyes were graded daily for signs of clinical inflammation and were removed 6, 24, 48, and 72 hours after injection. One group was prepared for histologic analysis, and vitreous was removed from the other group for cytokine analysis, using standard enzyme-linked immunosorbent assay procedures. TNF-alpha, IL-1beta, CINC, and IFN-gamma were detected in experimental vitreous samples at increased levels that peaked at 24 hours. TNF-alpha, IL-1beta, and CINC declined at 48 hours, but IFN-gamma remained elevated. At 72 hours, levels returned to baseline. Statistically significant elevations of TNF-alpha, IL-1beta, and CINC were detected in experimental samples at 24, but not at 6 and 48 hours compared with levels in saline control samples (P < 0.03). A statistically significant increase in IFN-gamma was detected at 24 and 48 hours compared with control levels (P < 0.03). In experimental animals, clinical inflammation and inflammatory cells peaked at 24 hours, persisted at 48 hours, and began to decline thereafter. Neutrophils were the predominant inflammatory cell detected at 24 (72.3% of cells) and 48 (60.1%) hours. By 72 hours, the total number of inflammatory cells had decreased by 75.0%, and the cellular infiltrate had changed so that neutrophils equaled monocytes-macrophages. S. aureus induced the expression of TNF-alpha, IL-1beta, CINC, and IFN-gamma. The time course of these cytokine levels could account for the clinical inflammatory responses and the entry and decline of vitreous cells in this model of bacterial endophthalmitis.


Increased vesicular and vacuolar transendothelial transport in traumatic human brain oedema. A review.
The endothelial vacuolar and vesicular transports in traumatic human brain oedema have been reviewed and compared with experimental brain oedema in order to establish their role in both oedema formation and oedema resolution. Normal or "non-activated" and "activated" capillaries are found. The activated capillaries showed predominantly an enhanced abluminally orientated vesicular transport by means of small, medium and large uncoated and clathrin coated vesicles, as well as the presence of endothelial tubular structures. Activation of the endothelial nuclear zone is featured by the increased amount of micropinocytotic vesicles. Vesicles internalizing to the hypertrophic Golgi complex, lysosomes and multivesicular bodies are observed. The protein vacuolar transport is predominant in most cortical capillaries. A wide spectrum of endothelial cell mechanisms is observed increasing the vesicular and vacuolar transport, such as deep invaginations of the luminal surface, large coated vesicles, tubular structures, and transient and incomplete transendothelial channels formed either by chained plasmalemmal vesicles or elongated protein-containing vacuoles. Uncoated vesicles are seen surrounding lysosomes. Vesicular transport might be discriminated between abluminally orientated or transendothelial transport (oedema formation) and intraendothelial transport (oedema resolution) directed towards cell lysosomes to be degraded by lysosomal enzymes. The transendothelial passage via large vacuoles is mainly caused by macromolecular protein transport.