An intramolecular G-quadruplex structure with mixed parallel/antiparallel G-strands formed in the human BCL-2 promoter region in solution.
We report the first G-quadruplex structure formed in the promoter region of the human bcl-2. Bcl-2 is a potent oncoprotein that functions as an inhibitor of cell apoptosis and has been found to be aberrantly overexpressed in a wide range of human tumors. A highly GC-rich region upstream of the P1 promoter plays an important role in the regulation of the transcriptional activity of the bcl-2 oncogene. The purine-rich strand of this region contains multiple runs of guanines and can form three distinct intramolecular G-quadruplexes in K+-containing solution. Of these, the G-quadruplex formed within the middle four consecutive guanine runs has been shown to be the most stable G-quadruplex structure, while it is also a mixture of loop isomers. The predominant G-quadruplex structure formed in this region was studied by NMR. Our results demonstrate a novel folding of a unique intramolecular G-quadruplex structure with mixed parallel/antiparallel G-strands. This G-quadruplex structure contains three G-tetrads connected with a single-nucleotide double-chain-reversal side loop and two lateral loops. The first three-nucleotide CGC loop in the bcl-2 promoter sequence forms a lateral loop, as opposed to a double-chain-reversal side loop observed in a similar sequence in the c-MYC promoter, which appears to largely determine the overall folding of the bcl-2 G-quadruplex. Furthermore, both the bcl-2 and c-MYC promoter sequences contain the G3NG3 sequence motif, which forms a stable double-chain-reversal, parallel-stranded structural motif. This predominant bcl-2 G-quadruplex represents an attractive novel target for the design of new anticancer drugs that specifically modulate bcl-2 gene expression.


Superhydrophobic Copper Surfaces with Anticorrosion Properties Fabricated by Solventless CVD Methods.
Due to continuous miniaturization and increasing number of electrical components in electronics, copper interconnections have become critical for the design of 3D integrated circuits. However, corrosion attack on the copper metal can affect the electronic performance of the material. Superhydrophobic coatings are a commonly used strategy to prevent this undesired effect. In this work, a solventless two-steps process was developed to fabricate superhydrophobic copper surfaces using chemical vapor deposition (CVD) methods. The superhydrophobic state was achieved through the design of a hierarchical structure, combining micro-/nanoscale domains. In the first step, O2- and Ar-plasma etchings were performed on the copper substrate to generate microroughness. Afterward, a conformal copolymer, 1H,1H,2H,2H-perfluorodecyl acrylate-ethylene glycol diacrylate [p(PFDA-co-EGDA)], was deposited on top of the metal via initiated CVD (iCVD) to lower the surface energy of the surface. The copolymer topography exhibited a very characteristic and unique nanoworm-like structure. The combination of the nanofeatures of the polymer with the microroughness of the copper led to achievement of the superhydrophobic state. AFM, SEM, and XPS were used to characterize the evolution in topography and chemical composition during the CVD processes. The modified copper showed water contact angles as high as 163° and hysteresis as low as 1°. The coating withstood exposure to aggressive media for extended periods of time. Tafel analysis was used to compare the corrosion rates between bare and modified copper. Results indicated that iCVD-coated copper corrodes 3 orders of magnitude slower than untreated copper. The surface modification process yielded repeatable and robust superhydrophobic coatings with remarkable anticorrosion properties.


Space toxicology: protecting human health during space operations.
Space toxicology is a unique and targeted discipline for spaceflight, space habitation, and occupation of celestial bodies including planets, moons, and asteroids. Astronaut explorers face distinctive health challenges and limited resources for rescue and medical care during space operation. A central goal of space toxicology is to protect the health of the astronaut by assessing potential chemical exposures during spaceflight and setting safe limits that will protect the astronaut against chemical exposures while in a physiologically altered state. In order to maintain sustained occupation in space on the International Space Station (ISS), toxicological risks must be assessed and managed within the context of isolation, continuous exposures, reuse of air and water, limited rescue options, and the need to use highly toxic compounds for propulsion and other purposes. As we begin to explore other celestial bodies, in situ toxicological risks, such as inhalation of reactive mineral dusts, must also be managed.


Sodium current kinetics in cat atrial myocytes.
1. Na+ current kinetics were studied in isolated atrial myocytes from the adult cat using the single suction-pipette voltage-clamp technique. 2. Current-voltage and conductance-voltage relationships were similar to those described in other cardiac myocyte preparations. 3. Analysis of Na+ current decay using single-pulse, double-pulse and tail current measurements were in agreement and demonstrate a second-order process of current decay. 4. Voltage dependence of steady-state inactivation curves was not symmetrical, having an inflexion at about -90 mV. These results suggest more than a single inactivation process for Na+ channel in the negative potential region. 5. Recovery of Na+ current from inactivation had a sigmoid time course: an initial slow component (delay) followed by a fast and then a second slow component. Increasing the pre-pulse duration slowed the time course of recovery. 6. Taken together, the results were consistent with the presence of multiple inactivated states for the atrial myocyte Na+ channel.


DNA polymerase photoprobe 2-[(4-azidophenacyl)thio]-2'-deoxyadenosine 5'-triphosphate labels an Escherichia coli DNA polymerase I Klenow fragment substrate binding site.
The nucleotide photoprobe 2-[(4-azidophenacyl)thio]-2'-deoxyadenosine 5'-triphosphate (1) was evaluated as a photoaffinity label of the DNA polymerase I Klenow fragment. Photolabel [3H]-1 covalently labeled the Klenow fragment with photolysis at 300 nm, reaching saturation at an approximate 1:1 mole ratio at 5.7 microM and with an EC50 (the effective concentration at 50% maximum photoincorporation) of about 0.74 microM. Saturating concentrations of poly(dA).(T)10 protect the Klenow fragment from [3H]-1 photoincorporation, and TTP at a concentration approximately equal to its KD for the free enzyme form shifts the dose-response curve for photoincorporation of [3H]-1 into the Klenow fragment by a factor of 2, indicating a competitive relationship between TTP and 1. Additionally, the photoincorporation of [3H]-1 into the Klenow fragment has an absolute requirement for magnesium, with no significant photoincorporation observed at concentrations of 1 up to 10 microM in the absence of magnesium. These results demonstrate that, as designed, photoprobe 1 binds to both the dNTP and a portion of the template-primer binding sites on the Klenow fragment. Photoaffinity labeling of the Klenow fragment by 1 yielded a single radiolabeled tryptic fragment which was isolated by HPLC; sequence analysis identified Asp732 in the peptide fragment Asp732-Ile733-His734-Arg735 as the site of covalent modification. Molecular modeling and complementary NMR analysis of the conformation of 1 indicated preferred C3'-exo and C2'-exo-C3'-endo symmetrical twist furanose ring puckers, with a high antibase conformation and a +sc C-5 torsional angle. Docking studies using Asp732 as an anchor point for the azide alpha-nitrogen on the photolabel indicate that the dNTP binding site is at the edge of the DNA binding cleft opposite the exonuclease site and that the template binding site includes helix O in the finger motif of the Klenow fragment.


Current State of Evidence: Influence of Nutritional and Nutrigenetic Factors on Immunity in the COVID-19 Pandemic Framework.
The pandemic caused by the new coronavirus has caused shock waves in many countries, producing a global health crisis worldwide. Lack of knowledge of the biological mechanisms of viruses, plus the absence of effective treatments against the disease (COVID-19) and/or vaccines have pulled factors that can compromise the proper functioning of the immune system to fight against infectious diseases into the spotlight. The optimal status of specific nutrients is considered crucial to keeping immune components within their normal activity, helping to avoid and overcome infections. Specifically, the European Food Safety Authority (EFSA) evaluated and deems six vitamins (D, A, C, Folate, B6, B12) and four minerals (zinc, iron, copper and selenium) to be essential for the normal functioning of the immune system, due to the scientific evidence collected so far. In this report, an update on the evidence of the contribution of nutritional factors as immune-enhancing aspects, factors that could reduce their bioavailability, and the role of the optimal status of these nutrients within the COVID-19 pandemic context was carried out. First, a non-systematic review of the current state of knowledge regarding the impact of an optimal nutritional status of these nutrients on the proper functioning of the immune system as well as their potential role in COVID-19 prevention/treatment was carried out by searching for available scientific evidence in PubMed and LitCovid databases. Second, a compilation from published sources and an analysis of nutritional data from 10 European countries was performed, and the relationship between country nutritional status and epidemiological COVID-19 data (available in the Worldometers database) was evaluated following an ecological study design. Furthermore, the potential effect of genetics was considered through the selection of genetic variants previously identified in Genome-Wide Association studies (GWAs) as influencing the nutritional status of these 10 considered nutrients. Therefore, access to genetic information in accessible databases (1000genomes, by Ensembl) of individuals from European populations enabled an approximation that countries might present a greater risk of suboptimal status of the nutrients studied. Results from the review approach show the importance of maintaining a correct nutritional status of these 10 nutrients analyzed for the health of the immune system, highlighting the importance of Vitamin D and iron in the context of COVID-19. Besides, the ecological study demonstrates that intake levels of relevant micronutrients-especially Vitamins D, C, B12, and iron-are inversely associated with higher COVID-19 incidence and/or mortality, particularly in populations genetically predisposed to show lower micronutrient status. In conclusion, nutrigenetic data provided by joint assessment of 10 essential nutrients for the functioning of the immune system and of the genetic factors that can limit their bioavailability can be a fundamental tool to help strengthen the immune system of individuals and prepare populations to fight against infectious diseases such as COVID-19.


Adult neural precursor cells from the subventricular zone contribute significantly to oligodendrocyte regeneration and remyelination.
Parenchymal oligodendrocyte progenitor cells (pOPCs) are considered the principal cell type responsible for oligodendrogenesis and remyelinaton in demyelinating diseases. Recent studies have demonstrated that neural precursor cells (NPCs) from the adult subventricular zone (SVZ) can also generate new oligodendrocytes after demyelination. However, the relative contribution of NPCs versus pOPCs to remyelination is unknown. We used in vivo genetic fate mapping to assess the behavior of each progenitor type within the corpus callosi (CCs) of mice subjected to cuprizone-induced demyelination. Nestin-CreER(T2) and Pdgfra-CreER(T2) transgenic mice were crossed with fluorescent Cre reporter strains to map the fate of NPCs and pOPCs respectively. In cuprizone-challenged mice, substantial numbers of NPCs migrated into the demyelinated CC and contributed to oligodendrogenesis. This capacity was most prominent in rostral regions adjacent to the SVZ where NPC-derived oligodendrocytes significantly outnumbered those generated from pOPCs. Sixty-two percent of all nodes of Ranvier in this region were flanked by at least one paranode generated from an NPC-derived oligodendrocyte. Remarkably, g-ratios (ratio of the axon diameter to the diameter of the axon plus myelin sheath) of myelinated axons in regions subject to significant NPC-derived remyelination were equivalent to those of unchallenged controls, and immunoelectron microscopy revealed that NPC-derived myelin was significantly thicker than that generated by pOPCs, regardless of axonal caliber. We also demonstrate that a reduced efficiency of remyelination in the caudal CC was associated with long-term impairment in the maturation of oligodendrogenic NPCs but only transient delay in pOPC differentiation. Collectively, our data define a major distinct role for NPCs in remyelination, identifying them as a key target for enhancing myelin repair in demyelinating diseases.


Spontaneous atraumatic anterior subluxation of the sternoclavicular joint.
We reviewed the cases of thirty-seven young patients, ten to thirty-six years old, who had had spontaneous atraumatic anterior subluxation of the sternoclavicular joint. Sudden atraumatic subluxation is quite alarming to the patient and the physician. Twenty-nine of the thirty-seven patients were treated non-operatively by observation and rehabilitative measures. Eight patients had been treated elsewhere with attempted operative reconstruction of the sternoclavicular joint or with resection arthroplasty. After an average follow-up of eight years, the twenty-nine patients who were treated non-operatively (group I) had excellent results, with no limitations of activity or changes in life-style. The eight patients who were treated operatively (group II) had numerous problems, including noticeable scars, persistent instability, pain, or limitation of activity that resulted in an alteration in lifestyle. Three of the eight patients had had a resection of the medial portion of the clavicle. Two of them had a second resection and the third patient had a third resection. A thoracic-outlet syndrome developed in this patient and the clavicle had to be totally resected to obtain relief. Since spontaneous atraumatic anterior subluxation of the sternoclavicular joint has a benign natural course, it should not be treated with operative stabilization of the joint. Instead, a conservative approach that includes education and reassurance of the patient will result in an unaltered lifestyle with no limitation of activity and little or no discomfort.


Tropical forests and the global carbon cycle: impacts of atmospheric carbon dioxide, climate change and rate of deforestation.
The remaining carbon stocks in wet tropical forests are currently at risk because of anthropogenic deforestation, but also because of the possibility of release driven by climate change. To identify the relative roles of CO2 increase, changing temperature and rainfall, and deforestation in the future, and the magnitude of their impact on atmospheric CO2 concentrations, we have applied a dynamic global vegetation model, using multiple scenarios of tropical deforestation (extrapolated from two estimates of current rates) and multiple scenarios of changing climate (derived from four independent offline general circulation model simulations). Results show that deforestation will probably produce large losses of carbon, despite the uncertainty about the deforestation rates. Some climate models produce additional large fluxes due to increased drought stress caused by rising temperature and decreasing rainfall. One climate model, however, produces an additional carbon sink. Taken together, our estimates of additional carbon emissions during the twenty-first century, for all climate and deforestation scenarios, range from 101 to 367 Gt C, resulting in CO2 concentration increases above background values between 29 and 129 p.p.m. An evaluation of the method indicates that better estimates of tropical carbon sources and sinks require improved assessments of current and future deforestation, and more consistent precipitation scenarios from climate models. Notwithstanding the uncertainties, continued tropical deforestation will most certainly play a very large role in the build-up of future greenhouse gas concentrations.


Telaprevir for retreatment of HCV infection.
Up to 60% of patients with hepatitis C virus (HCV) genotype 1 infection do not have a sustained virologic response to therapy with peginterferon alfa plus ribavirin. In this randomized, phase 3 trial, we evaluated the addition of telaprevir to peginterferon alfa-2a plus ribavirin in patients with HCV genotype 1 infection who had no response or a partial response to previous therapy or who had a relapse after an initial response. A total of 663 patients were assigned to one of three groups: the T12PR48 group, which received telaprevir for 12 weeks and peginterferon plus ribavirin for a total of 48 weeks; the lead-in T12PR48 group, which received 4 weeks of peginterferon plus ribavirin followed by 12 weeks of telaprevir and peginterferon plus ribavirin for a total of 48 weeks; and the control group (PR48), which received peginterferon plus ribavirin for 48 weeks. The primary end point was the rate of sustained virologic response, which was defined as undetectable HCV RNA 24 weeks after the last planned dose of a study drug. Rates of sustained virologic response were significantly higher in the two telaprevir groups than in the control group among patients who had a previous relapse (83% in the T12PR48 group, 88% in the lead-in T12PR48 group, and 24% in the PR48 group), a partial response (59%, 54%, and 15%, respectively), and no response (29%, 33%, and 5%, respectively) (P<0.001 for all comparisons). Grade 3 adverse events (mainly anemia, neutropenia, and leukopenia) were more frequent in the telaprevir groups than in the control group (37% vs. 22%). Telaprevir combined with peginterferon plus ribavirin significantly improved rates of sustained virologic response in patients with previously treated HCV infection, regardless of whether there was a lead-in phase. (Funded by Tibotec and Vertex Pharmaceuticals; REALIZE ClinicalTrials.gov number, NCT00703118.).


Tumor volume and lymphovascular space invasion as a prognostic factor in early invasive adenocarcinoma of the cervix.
The aim of this study was to investigate the risk and recurrence of early invasive adenocarcinoma of the cervix, and to determine whether non-radical methods of management could be performed. The medical and histopathological records of 50 patients with early invasive adenocarcinoma of the cervix treated at Keio University Hospital between 1993 and 2005 were reviewed, and compared with the literature. The median follow-up period was 64.3 months. The depth of stromal invasion was ≤3 mm in 33 cases and >3 mm, but ≤5 mm in 17 cases. The horizontal spread was ≤7 mm in 25 cases and >7 mm in 25 cases. One of the 33 cases that had tumor volumes of ≤500 mm(3), and three of the 17 cases with tumor volumes of >500 mm(3) were positive for lymph node metastasis. When our data were combined with previously reported results, statistically significant differences were observed between the tumor volume and the frequency of pelvic lymph node metastasis/the rate of recurrence (p<0.0001). The frequency of pelvic lymph node metastases was significantly higher in the lymphovascular space invasion (LVSI)-positive group than in the LVSI-negative group (p=0.02). No adnexal metastasis or parametrial involvement was noted. Assessment of the depth of stromal invasion, tumor volume, and LVSI is critical for selecting an appropriate therapeutic modality. Non-radical methods of management are considered suitable for patients with LVSI-negative adenocarcinoma of the cervix exhibiting a stromal invasion depth of ≤5 mm and a tumor volume of ≤500 mm(3).


The Autonomic Progress Bar Motivates Treatment Completion for Patients of Stimulant Use Disorder and Cannabis Use Disorder.
Background: The intrinsic motivation behind the "need to complete" is more influential than external incentives. We introduced a novel progress-bar tool to motivate the completion of programs designed to treat stimulant and cannabis use disorders. We further examined the effectiveness of the progress bar's scoring approach in forecasting consistently negative urine tests. Methods: This study's participants included 568 patients with stimulant, amphetamine-type, and cannabis use disorders who were undergoing 12-month mandatory treatment programs at Taichung Veterans General Hospital in Taiwan. Patients were given scores of 1, -1, or 0 depending on whether they received negative, positive, or missing urinalysis reports, respectively. The autonomic progress bar generated weekly score totals. At the group level, scorei donated scores from all patients for a given week (i denoted the week). Scorei was standardized to adjusted scorei. We then conducted Autoregressive Integrated Moving Average (ARIMA) Model of time-series analyses for the adjusted scorei. Results: A total of 312 patients maintained treatment progress over the 12-month program. The autonomic score calculator totaled the shared achievements of these patients. The coefficients of the lag variables for mean (p), lag variables for residual error term (q), and number of orders for ensuring stationary (d) were estimated at p = 3, d = 4, and q = 7 for the first half of the treatment program, and were estimated at p = 2, d = 2, and q = 3 for the second half. Both models were stationary and tested as fit for prediction (p < 0.05). Sharply raised adjusted scores were predicted during the high-demand treatment phase. Discussion: This study's novel progress-bar tool effectively motivated treatment completion. It was also effective in forecasting continually negative urine tests. The tool's free open-source code makes it easy to implement among many substance-treatment services.


The toxicology and carcinogenesis study of phenolphthalein (CAS No. 77-09-8) in genetically modified haploinsufficient p16(Ink4a)/p19(Arf) mice (feed study).
Phenolphthalein was commonly used as a laxative for most of the 20th century. The use of phenolphthalein in laxatives has decreased since 1997 when the United States Food and Drug Administration (FDA) proposed to withdraw its classification as an over-the-counter drug (21 CFR, Part 310). Phenolphthalein has been previously evaluated in 2-year carcinogenicity studies by the National Toxicology Program (1996). The major route of human exposure to phenolphthalein is via ingestion, dermal contact, and inhalation of contaminated air originating from process units manufacturing the compound. In this study, the carcinogenic effects of phenolphthalein were studied in the haploinsufficient p16(Ink4a)/p19(Arf) mouse model as an ongoing goal of the NTP is to seek model systems for toxicology and carcinogenesis studies, especially those that can provide mechanistic information relative to understanding an agent's mode of action. Male and female haploinsufficient p16(Ink4a)/p19(Arf) mice were exposed to phenolphthalein (greater than 97% pure) in feed for 27 weeks. Genetic toxicology studies were conducted in mouse peripheral blood erythrocytes. 27-WEEK STUDY IN MICE: Groups of 15 male and 15 female mice were exposed to 0, 200, 375, 750, 3,000, or 12,000 ppm phenolphthalein (equivalent to average daily doses of approximately 35, 65, 135, 540, and 2,170 mg phenolphthalein/kg body weight to males and 50, 90, 170, 680, 2,770 mg/kg to females) in feed for 27 weeks. Survival of all exposed groups of male and female mice was similar to that of the control groups. Mean body weights of males in the 12,000 ppm group were less than those of the control group after week 11. No differences in feed consumption were noted between exposed and control groups. Atypical hyperplasia of the thymus, a premalignant change of chemically induced thymic lymphoma, occurred in exposed males and females, and the incidence was significantly increased in 12,000 ppm females. Atrophy of the seminiferous tubules in the testis, hyperplasia of the testicular interstitial (Leydig) cells, and epididymal hypospermia occurred in most 3,000 and 12,000 ppm males. Additionally, the left and right testis weights, the left epididymis weights, sperm motility, the numbers of spermatid heads per testis, and sperm heads per cauda and per gram cauda epididymis were generally significantly less in 3,000 and 12,000 ppm males than in the control group. The incidences of nephropathy were significantly increased in 3,000 and 12,000 ppm males; incidences of hypertrophy of renal tubules were significantly increased in males receiving 750 ppm or greater. Hematopoietic cell proliferation of the spleen occurred in all 12,000 ppm males, and the incidences of this lesion were significantly increased in 375, 750, and 12,000 ppm females. The frequency of micronucleated erythrocytes was assessed at four time points during the 27-week study in male and female haploinsufficient p16(Ink4a)/p19(Arf) mice. Significant concentration-related increases in micronucleated cells were observed at all time points in male and female mice. Under the conditions of this 27-week feed study, there was no evidence of carcinogenic activity of phenolphthalein in male or female haploinsufficient p16(Ink4a)/p19(Arf) mice exposed to 200, 375, 750, 3,000, or 12,000 ppm. Because this is a new model, there is uncertainty whether the study possessed sufficient sensitivity to detect a carcinogenic effect. Phenolphthalein induced atypical hyperplasia, a preneoplastic lesion of the thymus, in male and female mice, hematopoietic cell proliferation of the spleen in male and female mice, and toxicity to the kidney and reproductive system in male mice.


Prevalence and socioeconomic characteristics of alcohol disorders among men and women in the Eastern Cape Province, South Africa.
There is growing concern about alcohol problems in low- and middle-income countries. More research is required, particularly among the younger generation. The aim of this study was to investigate the prevalence of alcohol disorders and associated socioeconomic characteristics among young men and women living in the Eastern Cape Province, South Africa. This was a cross-sectional population-based study of 977 participants (52% male and 48% female) aged 18-40, the majority of whom lived in low-income areas. Data collection was carried out in 2012 by trained fieldworkers. The Mini International Neuropsychiatric Interview (DSM-IV) was used to investigate the prevalence of alcohol dependence (increased tolerance to alcohol, failed attempt to cut down, risk of physical and mental effects) and alcohol abuse (harmful use, consistent intoxication, risk behaviour, physically hazardous, social problems). A high 12-month prevalence of alcohol dependence was found (26.5% in total; 39.0% among men and 19.1% among women) as well as of alcohol abuse (9% in total; 19.0% among men and 6.0% among women). Few socioeconomic differences emerged among the men, except older men (OR 1.94, CI 1.11-3.42) and those supported by social grants (OR 2.28, CI 1.06-4.93), who presented higher odd ratios for alcohol dependence than the reference groups. Among the women, more differences emerged: women who were widowed/single (OR 2.35, CI 1.20-4.62), had no education (OR 3.41, CI 1.04-11.21), had a low income (OR 3.26, CI 1.55-6.80) and had no social support from friends when ill presented higher odd ratios (OR 1.73, CI 1.07-2.80). In the adjusted model, marital status and low income remained statistically significant. With regard to alcohol abuse, fewer socioeconomic differences emerged. Interventions need to address the early onset of alcohol misuse in order to meet both current needs and long-standing mental and physical illness.


The Early Cambrian tommotiid Micrina, a sessile bivalved stem group brachiopod.
The tannuolinid Micrina belongs to the tommotiids-a common and widely distributed, but poorly understood, group of Early Cambrian fossil metazoans with multiple external organophosphatic sclerites. Recent findings of sessile articulated tommotiid scleritomes indicate that previous reconstructions of tommotiids as slug-like bilaterians with a dorsal cover of sclerites require detailed re-evaluation. Comparative ultrastructural work has already indicated that the tommotiids might be a sister group to the Brachiopoda, with Micrina representing the most derived and brachiopod-like bimembrate tommotiid. Here we further develop and strengthen this controversial phylogenetic model with a new reconstruction of Micrina, where the two types of sclerites--mitral and sellate--belong to a near bilaterally symmetrical bivalved sessile organism. This new scleritome configuration was tested by recreating an articulated bivalved Micrina from isolated mitral and sellate sclerites; both sclerites have muscles that would have enabled movement of the sclerites. The mitral and sellate sclerites of Micrina are considered to be homologous with the ventral and dorsal valves, respectively, of organophosphatic linguliform brachiopods, indicating that a simple type of filter-feeding within an enclosed bivalved shell had started to evolve in derived tannuolinids. The new reconstruction also indicates that the phylogenetic range of 'bivalved', sessile lophophorates is larger than previously suspected.


In ovo electroporation of miRNA-based-plasmids to investigate gene function in the developing neural tube.
When studying gene function in vivo during development, gene expression has to be controlled in a precise temporal and spatial manner. Technologies based on RNA interference (RNAi) are well suited for such studies, as they allow for the efficient silencing of a gene of interest. In contrast to challenging and laborious approaches in mammalian systems, the use of RNAi in combination with oviparous animal models allows temporal control of gene silencing in a fast and precise manner. We have developed approaches using RNAi in the chicken embryo to analyze gene function during neural tube development. Here we describe the construction of plasmids that direct the expression of one or two artificial microRNAs (miRNAs) to knock down expression of endogenous protein/s of interest upon electroporation into the spinal cord. The miRNA cassette is directly linked to a fluorescent protein reporter, for the direct visualization of transfected cells. The transcripts are under the control of different promoters/enhancers which drive expression in genetically defined cell subpopulations in the neural tube. Mixing multiple RNAi vectors allows combinatorial knockdowns of two or more genes in different cell types of the spinal cord, thus permitting the analysis of complex cellular and molecular interactions in a fast and precise manner. The technique that we describe can easily be applied to other cell types in the neural tube, or even adapted to other organisms in developmental studies.


Storage of Renewable Energy by Reduction of CO2 with Hydrogen.
The main difference between the past energy economy during the industrialization period which was mainly based on mining of fossil fuels, e.g. coal, oil and methane and the future energy economy based on renewable energy is the requirement for storage of the energy fluxes. Renewable energy, except biomass, appears in time- and location-dependent energy fluxes as heat or electricity upon conversion. Storage and transport of energy requires a high energy density and has to be realized in a closed materials cycle. The hydrogen cycle, i.e. production of hydrogen from water by renewable energy, storage and use of hydrogen in fuel cells, combustion engines or turbines, is a closed cycle. However, the hydrogen density in a storage system is limited to 20 mass% and 150 kg/m(3) which limits the energy density to about half of the energy density in fossil fuels. Introducing CO(2) into the cycle and storing hydrogen by the reduction of CO(2) to hydrocarbons allows renewable energy to be converted into synthetic fuels with the same energy density as fossil fuels. The resulting cycle is a closed cycle (CO(2) neutral) if CO(2) is extracted from the atmosphere. Today's technology allows CO(2) to be reduced either by the Sabatier reaction to methane, by the reversed water gas shift reaction to CO and further reduction of CO by the Fischer-Tropsch synthesis (FTS) to hydrocarbons or over methanol to gasoline. The overall process can only be realized on a very large scale, because the large number of by-products of FTS requires the use of a refinery. Therefore, a well-controlled reaction to a specific product is required for the efficient conversion of renewable energy (electricity) into an easy to store liquid hydrocarbon (fuel). In order to realize a closed hydrocarbon cycle the two major challenges are to extract CO(2) from the atmosphere close to the thermodynamic limit and to reduce CO(2) with hydrogen in a controlled reaction to a specific hydrocarbon. Nanomaterials with nanopores and the unique surface structures of metallic clusters offer new opportunities for the production of synthetic fuels.


Diuretic-gene interaction and the risk of myocardial infarction and stroke.
This study investigates whether the interaction between diuretics and alpha-adducin (ADD1) G460W or G-protein beta3-subunit (GNB3) rs2301339 polymorphism modifies the risk of myocardial infarction (MI) or stroke. Data were used from the Rotterdam Study. The drug-gene interaction was determined with a Cox proportional hazard model with adjustment for each drug class as time-dependent covariates. The risk of MI in current users of low-ceiling diuretics with one or two copies of the ADD1 W-allele (hazard ration (HR)=0.92) was similar compared to the expected joint effect of the W-allele and low-ceiling diuretics on a multiplicative scale (1.04 x 0.90=0.94) (synergy index (SI):0.99; 95% confidence interval (CI): 0.43-2.27). No drug-gene interaction was found on the risk of stroke (SI:0.66; 95% CI:0.43-1.27). In addition, a trend towards an interaction was found between current use and the GNB3 rs230119 G/A polymorphism on the risk of MI (SI: 0.51; 95% CI: 0.23-1.15), whereas no interaction on the risk of stroke was found (SI: 0.84; 95% CI: 0.46-1.56).


A balance between positive and negative signals in cytotoxic lymphocytes regulates the polarization of lipid rafts during the development of cell-mediated killing.
Plasma membrane microdomains containing sphingolipids and cholesterol (lipid rafts) are enriched in signaling molecules. The cross-linking of certain types of cell surface receptors initiates the redistribution of these lipid rafts, resulting in the formation of signaling complexes. However, little is known about the regulation of the initial raft redistribution and whether negative regulatory signaling pathways target this phase of cellular activation. We used natural killer (NK) cells as a model to investigate the regulation of raft redistribution, as both positive and negative signals have been implicated in the development of their cellular function. Here we show that after NK cells form conjugates with sensitive tumor cells, rafts become polarized to the site of target recognition. This redistribution of lipid rafts requires the activation of both Src and Syk family protein tyrosine kinases. In contrast, engagement of major histocompatibility complex (MHC)-recognizing killer cell inhibitory receptors (KIRs) on NK cells by resistant, MHC-bearing tumor targets blocks raft redistribution. This inhibition is dependent on the catalytic activity of KIR-associated SHP-1, a Src homology 2 (SH2) domain containing tyrosine phosphatase. These results suggest that the influence of integrated positive and negative signals on raft redistribution critically influences the development of cell-mediated cytotoxicity.


Major intrinsic proteins and arsenic transport in plants: new players and their potential role.
Arsenic (As) is a toxic and highly abundant metalloid that endangers human health through drinking water and the food chain. The most common forms of As in the environment re arsenate [As(V)] and arsenite [As(III)]. As(V) is a nonfunctional phosphate analog that enters the food chain via plant phosphate transporters. Recently, evidence was provided that uptake of As(III)--the second most abundant As species in soils--is mediated by plant nodulin26-like intrinsic proteins (NIPs), a subfamily of plant major intrinsic proteins (MIPs). Specific NIPs are also essential for the uptake of the metalloids boron and silicon and aquaglyceroporins from microbes and mammals were shown to be the major routes of As uptake. Therefore As(III) transport through MIPs is a conserved and ancient feature. In this chapter we summarize the current view on As transport in plants and address the potential physiological significance of As(III) transport through NIPs.


Improving our understanding of multi-tasking in healthcare: Drawing together the cognitive psychology and healthcare literature.
Multi-tasking is an important skill for clinical work which has received limited research attention. Its impacts on clinical work are poorly understood. In contrast, there is substantial multi-tasking research in cognitive psychology, driver distraction, and human-computer interaction. This review synthesises evidence of the extent and impacts of multi-tasking on efficiency and task performance from health and non-healthcare literature, to compare and contrast approaches, identify implications for clinical work, and to develop an evidence-informed framework for guiding the measurement of multi-tasking in future healthcare studies. The results showed healthcare studies using direct observation have focused on descriptive studies to quantify concurrent multi-tasking and its frequency in different contexts, with limited study of impact. In comparison, non-healthcare studies have applied predominantly experimental and simulation designs, focusing on interleaved and concurrent multi-tasking, and testing theories of the mechanisms by which multi-tasking impacts task efficiency and performance. We propose a framework to guide the measurement of multi-tasking in clinical settings that draws together lessons from these siloed research efforts.


Identification of mutations in the WT1 gene in tumours from patients with the WAGR syndrome.
Individuals with constitutional, heterozygous deletions of chromosome region 11p13 are predisposed to the development of Wilms' tumour, indicating the site of the tumour predisposition gene. The WT1 gene is a candidate for this cancer predisposition gene. If this gene is truly involved in tumorigenesis it would be expected to be mutant in tumour tissue from patients with 11p13 deletions. We have used single-stranded conformation polymorphism (SSCP) and polymerase chain reaction sequencing to test this hypothesis in an exon-by-exon analysis of the gene. Four tumours were analysed, two of which were from unilaterally affected individuals and two from a bilaterally affected patient. SSCP analysis identified mutations in the two unilateral tumours which, on sequencing, were shown to involve a 10-bp insertion in exon 7 and a single base pair change in exon 8. Both mutations result in the generation of premature stop codons and the predicted proteins would lack part of the zinc finger motif. Despite complete sequencing of the WT1 gene in both of the bilateral tumours, no mutations were identified. These results possibly suggest that WT1 may not be involved in tumorigenesis in all tumours. All four tumours retained heterozygosity in the 11p15 region, making it unlikely that a second recessive oncogene in this region was involved in tumorigenicity.


Protocol endomyocardial biopsy beyond 6 months-It is time to move on.
The optimal duration and frequency of routine surveillance endomyocardial biopsy (EMB) have been questioned in the current era of heart transplantation (HT), where the advances in immunosuppression and donor selection strategies have led to a decline in acute allograft rejection. We investigated the utility of routine EMB beyond 6 months post-HT. A single-center retrospective review was performed on 2963 EMBs from 220 HT recipients over 10 years. Each EMB was categorized into protocol or symptom-triggered biopsy and reviewed for rejection. Heart transplant recipients with ≥2 known risk factors for rejection were designated as an elevated risk group. The majority of rejections occurred within 3 months following HT. The yield of routine protocol EMBs was significantly lower than symptom-triggered EMBs, not only during the first 6 months post-HT (1.6% vs. 33.3%, P < .0001), but more so during the 6-12 months (0.1% vs 83.0%, P < .0001). A similar pattern was observed in heart transplant recipients at both elevated and standard risk for rejection. In conclusion, EMB was found to be a low-yield screening modality for rejection beyond 6 months post-HT.


Patient treatment preferences in localized prostate carcinoma: The influence of emotion, misconception, and anecdote.
Multiple therapeutic options exist for localized prostate carcinoma, without conclusive evidence to guide the choice of treatment. Thus, treatment should reflect trade-offs between the probability of curing disease and the desire to avoid treatment-associated side effects. Factors that actually influence patient treatment preferences are poorly understood. We reviewed medical records and carried out in-depth, semistructured interviews of 20 men with newly-diagnosed, clinically-localized prostate carcinoma in a Veterans Affairs Hospital following their first consultations with urologists and before treatments were initiated. Six to eight months after treatment, we carried out follow-up interviews. Interviews explored beliefs and attitudes about prostate cancer and treatment options, emotional reactions to the diagnosis, treatment preferences, information sources, and perceptions of interactions with urologists. Patient treatment preferences were not based on careful assessments of numerical risks for various clinical outcomes. Instead, feelings of fear and uncertainty contributed to a desire for rapid treatment, and specific preferences were profoundly influenced by misconceptions, especially about prostatectomy, and by anecdotes about the experiences of others with cancer. Few patients wanted to seek second opinions. Most patients received treatments that matched their initial preferences. Afterwards, they justified their choices in terms of the same misconceptions and anecdotal influences invoked during treatment deliberation. For men with localized prostate carcinoma, the treatment decision-making process would benefit from interventions that moderate feelings of fear and a desire for rapid treatment, dispel common and powerful misconceptions about prostate cancer and its therapies, and help patients avoid over-reliance on anecdotes.


Inhibition of H1N1 influenza A virus growth and induction of inflammatory mediators by the isoquinoline alkaloid berberine and extracts of goldenseal (Hydrastis canadensis).
In this study we tested whether the isoquinoline alkaloid berberine can inhibit the growth of influenza A. Our experiments showed strong inhibition of the growth of H1N1 influenza A strains PR/8/34 or WS/33 in RAW 264.7 macrophage-like cells, A549 human lung epithelial-derived cells and murine bone marrow derived macrophages, but not MDCK canine kidney cells. Studies of the mechanism underlying this effect suggest that berberine acts post-translationally to inhibit virus protein trafficking/maturation which in turn inhibits virus growth. Berberine was also evaluated for its ability to inhibit production of TNF-α and PGE(2) from A/PR/8/34 infected-RAW 264.7 cells. Our studies revealed strong inhibition of production of both mediators and suggest that this effect is distinct from the anti-viral effect. Finally, we asked whether berberine-containing ethanol extracts of goldenseal also inhibit the growth of influenza A and production of inflammatory mediators. We found strong effectiveness at high concentrations, although upon dilution extracts were somewhat less effective than purified berberine. Taken together, our results suggest that berberine may indeed be useful for the treatment of infections with influenza A.


Effects of the difference between lumbar lordosis in the supine and standing positions on the clinical outcomes of decompression surgery for lumbar spinal stenosis.
The authors sought to evaluate the relationship between the difference in lumbar lordosis (DiLL) in the preoperative supine and standing positions and spinal sagittal alignment in patients with lumbar spinal stenosis (LSS) and to determine whether this difference affects the clinical outcome of laminectomy. Sixty patients who underwent single-level unilateral laminectomy for bilateral decompression of LSS were evaluated. Spinopelvic parameters in the supine and standing positions were measured preoperatively and at 3 months and 2 years postoperatively. DiLL between the supine and standing positions was determined as follows: DiLL = supine LL - standing LL. On the basis of this determination patients were then categorized into DiLL(+) and DiLL(-) groups. The relationship between DiLL and preoperative spinopelvic parameters was evaluated using Pearson's correlation coefficient. In addition, clinical outcomes such as visual analog scale (VAS) and Oswestry Disability Index (ODI) scores between the two groups were measured, and their relationship to DiLL was evaluated using two-group comparison and multivariate analysis. There were 31 patients in the DiLL(+) group and 29 in the DiLL(-) group. DiLL was not associated with supine LL but was strongly correlated with standing LL and pelvic incidence (PI) - LL (PI - LL). In the preoperative spinopelvic alignment, LL and SS in the standing position were significantly smaller in the DiLL(+) group than in the DiLL(-) group, and PI - LL was significantly higher in the DiLL(+) group than in the DiLL(-) group. There was no difference in the clinical outcomes 3 months postoperatively, but low-back pain, especially in the sitting position, was significantly higher in the DiLL(+) group 2 years postoperatively. DiLL was associated with low-back pain in the sitting position, which was likely to persist in the DiLL(+) group postoperatively. We evaluated the relationship between DiLL and spinal sagittal alignment and the influence of DiLL on postoperative outcomes in patients with LSS. DiLL was strongly correlated with PI - LL, and in the DiLL(+) group, postoperative low-back pain relapsed. DiLL can be useful as a new spinal alignment evaluation method that supports the conventional spinal sagittal alignment evaluation.


Translocation of a UV-damaged DNA binding protein into a tight association with chromatin after treatment of mammalian cells with UV light.
A UV-damaged DNA binding protein (UV-DDB) is the major source of UV-damaged DNA binding activity in mammalian cell extracts. This activity is defective in at least some xeroderma pigmentosum group E (XP-E) patients; microinjection of the UV-DDB protein into their fibroblasts corrects nucleotide excision repair (NER). In an in vitro reconstituted NER system, small amounts of UV-DDB stimulate repair synthesis a few fold. After exposure to UV, mammalian cells show an early dose-dependent inhibition of the extractable UV-DDB activity; this inhibition may reflect a tight association of the binding protein with UV-damaged genomic DNA. To investigate the dynamics and location of UV-DDB with respect to damaged chromatin in vivo, we utilized nuclear fractionation and specific antibodies and detected translocation of the p127 component of UV-DDB from a loose to a tight association with chromatinized DNA immediately after UV treatment. A similar redistribution was found for other NER proteins, i.e. XPA, RP-A and PCNA, suggesting their tighter association with genomic DNA after UV. These studies revealed a specific protein-protein interaction between UV-DDB/p127 and RP-A that appears to enhance binding of both proteins to UV-damaged DNA in vitro, providing evidence for the involvement of UV-DDB in the damage-recognition step of NER. Moreover, the kinetics of the reappearance of extractable UV-DDB activity after UV treatment of human cells with differing repair capacities positively correlate with the cell's capacity to repair 6-4 pyrimidine dimers (6-4 PD) in the whole genome, a result consistent with an in vivo role for UV-DDB in recognizing this type of UV lesion.


The genetic epidemiology of rheumatoid arthritis.
This review considers the epidemiological aspects of the genetic investigation of rheumatoid arthritis (RA). Problems both of disease definition and disease heterogeneity render studies difficult to undertake and interpret. The selection of individuals both in population and family studies can explain divergent results. Despite the insights into the immunopathogenesis of RA afforded by the demonstration of an association between the disease and HLA class II genes, the latter have only a limited role in explaining differences, both between individuals and between populations, in susceptibility to RA.


Stromatoxin-sensitive, heteromultimeric Kv2.1/Kv9.3 channels contribute to myogenic control of cerebral arterial diameter.
Cerebral vascular smooth muscle contractility plays a crucial role in controlling arterial diameter and, thereby, blood flow regulation in the brain. A number of K(+) channels have been suggested to contribute to the regulation of diameter by controlling smooth muscle membrane potential (E(m)) and Ca(2+) influx. Previous studies indicate that stromatoxin (ScTx1)-sensitive, Kv2-containing channels contribute to the control of cerebral arterial diameter at 80 mmHg, but their precise role and molecular composition were not determined. Here, we tested if Kv2 subunits associate with 'silent' subunits from the Kv5, Kv6, Kv8 or Kv9 subfamilies to form heterotetrameric channels that contribute to control of diameter of rat middle cerebral arteries (RMCAs) over a range of intraluminal pressure from 10 to 100 mmHg. The predominant mRNAs expressed by RMCAs encode Kv2.1 and Kv9.3 subunits. Co-localization of Kv2.1 and Kv9.3 proteins at the plasma membrane of dissociated single RMCA myocytes was detected by proximity ligation assay. ScTx1-sensitive native current of RMCA myocytes and Kv2.1/Kv9.3 currents exhibited functional identity based on the similarity of their deactivation kinetics and voltage dependence of activation that were distinct from those of homomultimeric Kv2.1 channels. ScTx1 treatment enhanced the myogenic response of pressurized RMCAs between 40 and 100 mmHg, but this toxin also caused constriction between 10 and 40 mmHg that was not previously observed following inhibition of large conductance Ca(2+)-activated K(+) (BK(Ca)) and Kv1 channels. Taken together, this study defines the molecular basis of Kv2-containing channels and contributes to our understanding of the functional significance of their expression in cerebral vasculature. Specifically, our findings provide the first evidence of heteromultimeric Kv2.1/Kv9.3 channel expression in RMCA myocytes and their distinct contribution to control of cerebral arterial diameter over a wider range of E(m) and transmural pressure than Kv1 or BK(Ca) channels owing to their negative range of voltage-dependent activation.


Implications of fibrotic extracellular matrix in diabetic retinopathy.
Fibrosis is an accumulation of extracellular matrix (ECM) proteins and fibers in a disordered fashion, which compromises cell and tissue functions. High glucose-induced fibrosis, a major pathophysiological change of diabetic retinopathy (DR), severely affects vision by compromising the retinal vasculature and ultimately disrupting retinal tissue organization. The retina is a highly vascularized, stratified tissue with multiple cell types organized into distinct layers. Chronically high blood glucose stimulates certain retinal cells to increase production and assembly of ECM proteins resulting in excess ECM deposition primarily in the capillary walls on the basal side of the endothelium. This subendothelial fibrosis of the capillaries is the earliest histological change in the diabetic retina and has been linked to the vascular dysfunction that underlies DR. Proteins that are not normally abundant in the capillary basement membrane (BM) matrix, such as the ECM protein fibronectin, are assembled in significant quantities, disrupting the architecture of the BM and altering its properties. Cell culture models have identified multiple mechanisms through which elevated glucose can stimulate fibronectin matrix assembly, including intracellular signaling pathways, alternative splicing, and non-enzymatic glycation of the ECM. The fibrotic subendothelial matrix alters cell adhesion and supports further accumulation of other ECM proteins leading to disruption of endothelial cell-cell junctions. We review evidence supporting the notion that these molecular changes in the ECM contribute to the pathogenesis of DR, including vascular leakage, loss of endothelial cells and pericytes, changes in blood flow, and neovascularization. We propose that the accumulation of ECM, especially fibronectin matrix, first around the vasculature and later in extravascular locations, plays a critical role in DR and vision loss. Strategies for DR prevention and treatment should consider the ECM a potential therapeutic target.


ApoE Genotype-Dependent Response to Antioxidant and Exercise Interventions on Brain Function.
This study determined whether antioxidant supplementation is a viable complement to exercise regimens in improving cognitive and motor performance in a mouse model of Alzheimer's disease risk. Starting at 12 months of age, separate groups of male and female mice expressing human Apolipoprotein E3 (GFAP-ApoE3) or E4 (GFAP-ApoE4) were fed either a control diet or a diet supplemented with vitamins E and C. The mice were further separated into a sedentary group or a group that followed a daily exercise regimen. After 8 weeks on the treatments, the mice were administered a battery of functional tests including tests to measure reflex and motor, cognitive, and affective function while remaining on their treatment. Subsequently, plasma inflammatory markers and catalase activity in brain regions were measured. Overall, the GFAP-ApoE4 mice exhibited poorer motor function and spatial learning and memory. The treatments improved balance, learning, and cognitive flexibility in the GFAP-ApoE3 mice and overall the GFAP-ApoE4 mice were not responsive. The addition of antioxidants to supplement a training regimen only provided further benefits to the active avoidance task, and there was no antagonistic interaction between the two interventions. These outcomes are indicative that there is a window of opportunity for treatment and that genotype plays an important role in response to interventions.


Multisensory integration with a head-mounted display: role of mental and manual load.
The aim of this study was to replicate the finding that multisensory integration with a head-mounted display (HMD) is particularly difficult when a person is walking and hearing sound from a free-field speaker, and to extend the finding with a response method intended to reduce workload. HMDs can support the information needs of workers whose work requires mobility, but some low-cost solutions for delivering auditory information may be less effective than others. For the study, 24 participants detected whether shapes moving on the HMD screen made a sound appropriate to their forms when they collided with other shapes. Independent variables were self-motion (participants were mobile or seated), sound delivery (free-field speakers or an earpiece), and response method (noting mismatches via a mental count or via a manual clicker). Unexpectedly, overall mismatch task accuracy was worse with the clicker (p = .027) than without. Participants also reported that it was harder to time-share the mismatch task with clicker responses (p = .033). In the clicker condition, self-motion and sound delivery interacted but in the opposite direction to the previous study. The best way of delivering auditory information to mobile workers performing a multisensory integration task with an HMD may depend on whether responding involves mental load or manual load. Broader theories are needed to capture factors influencing performance. Until more powerful theory is developed, designers should perform careful formative and summative tests of whether the activities to be performed by mobile HMD wearers will make some sound delivery solutions less effective than others.


Correlation between pretreatment levels of interferon response genes and clinical responses to an immune response modifier (Imiquimod) in genital warts.
Imiquimod (IQ) has been successfully used in treatment of genital warts. In clinical settings, patients responded well but wart reduction rates varied. Our aim was to find a correlation between clinical responses and pretreatment (constitutive) levels of genes that might be involved in the molecular action of IQ. Since IQ is a cytokine inducer, we analyzed levels of expression of genes of the JAK/STAT signaling pathway and their inhibitors as well as interferon response factors (IRFs) in pretreatment biopsy specimens from complete responders (99 to 100% wart reduction rate) versus incomplete responders (75 to 92% wart reduction rate) by reverse transcription-PCR. We found that mRNA levels of signal transducer and activator of transcription 1 (STAT1) and IRF1 were higher in complete responders than in incomplete responders. Incomplete responders expressed larger amounts of STAT3, IRF2, and protein inhibitor of activated STAT1 (PIAS1) mRNAs compared to complete responders before IQ treatment. We hypothesize that high-level expression of STAT1 and IRF1 is advantageous for a better IQ response. The observed differences in constitutive mRNA levels of these genes may be the consequence of alterations in cellular differentiation and/or variable expression of endogenous interferons. Previous in vitro studies showed that keratinocyte differentiation coordinates the balance between positive and negative signals along the JAK/STAT pathway by regulating the IRF1:IRF2 and STAT1:PIAS1 ratios and thus affecting induction of IQ-inducible genes. Specifically, differentiation supports constitutive expression of STAT1 and IRF1 mRNAs but not expression of IRF2 and PIAS1. Our data are in good agreement with studies that showed the importance of STAT1 in cytokine induction and activation of interferon-responsive genes by IQ.


How is charge transport different in ionic liquids? The effect of high pressure.
Modern ionic liquids (ILs) are considered green solvents for the future applications due to their inherited advantages and remarkable transport properties. One of the ubiquitous properties of ILs is their intrinsic ionic conductivity. However, understanding of the super-Arrhenius behavior of the ionic conductivity process at elevated pressure still remains elusive and crucial in glass science. In this work, we investigate the ion transport properties of 1-butyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]imide: [C4mim][NTf2], 1-butylimidazolium bis[(trifluoromethyl)-sulfonyl]imide: [C4Him][NTf2] and 1-butylimidazolium hydrogen sulfate: [C4Him][HSO4] ILs in the supercooled liquid state using dielectric spectroscopy at ambient and high pressure. We present the experimental data in the dynamic window of the conductivity formalism to examine the charge transport properties. The frequency-dependent ionic conductivity data have been analyzed using the time-temperature superposition principle. In the Arrhenius diagram, the thermal evolution of the dc-conductivity reveals similar temperature dependence for both protic and aprotic ILs thus making it difficult to distinguish the ion dynamics. However, our results demonstrate the key role of high pressure that unambiguously separates the charge transport properties of protic ILs from aprotic ones through the apparent activation volume parameter. We also highlight that the activation volume can be employed to assess the information connecting the ability of ionic systems to form H-bond networks and the impact of proton transfer involved in the conduction process.


Light-induced retinal injury enhanced neurotrophins secretion and neurotrophic effect of mesenchymal stem cells in vitro.
To investigate neurotrophins expression and neurotrophic effect change in mesenchymal stem cells (MSCs) under different types of stimulation. Rats were exposed in 10,000 lux white light to develop light-induced retinal injury. Supernatants of homogenized retina (SHR), either from normal or light-injured retina, were used to stimulate MSCs. Quantitative real time for polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were conducted for analysis the expression change in basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) in MSCs after stimulation. Conditioned medium from SHR-stimulated MSCs and control MSCs were collected for evaluation their effect on retinal explants. Supernatants of homogenized retina from light-injured rats significantly promoted neurotrophins secretion from MSCs (p<0.01). Conditioned medium from mesenchymal stem cells stimulated by light-injured SHR significantly reduced DNA fragmentation (p<0.01), up-regulated bcl-2 (p<0.01) and down-regulated bax (p<0.01) in retinal explants, displaying enhanced protective effect. Light-induced retinal injury is able to enhance neurotrophins secretion from mesenchymal stem cells and promote the neurotrophic effect of mesenchymal stem cells.


The financial impact of HIV/AIDS on poor households in South Africa.
Rising mortality rates caused by HIV/AIDS in South Africa have substantial and lingering impacts on poor households. This is a descriptive paper using a new dataset of daily income, expenditure and financial transactions collected over a year from a total of 181 poor households in South African rural and urban areas. One of the key pathways through which HIV/AIDS impacts on household wellbeing is through the socioeconomic impacts of death, which this dataset is especially useful in quantifying. The key impacts of death on households are funerals and the loss of income. Funerals often cost up to 7 months of income. Nearly all households in the sample attempt to cover such costs by holding a portfolio of funeral insurance. Despite these efforts to insure against funeral costs, 61% of households are underinsured against the cost of a funeral. Nearly half the sample households are dependent on a regular wage earner, and another quarter are dependent on a grant recipient. Eighty per cent of these households would lose over half of their monthly income should the highest income recipient in the household die. Even by selling liquid assets, only one third of the sample households would be able to maintain their pre-death living standards for a year or more. Death poses substantial and lingering burdens from the funerals that surviving household members need to finance and the ongoing loss of income once brought into the household by the deceased. These costs pose so great a threat to households that they dominate household saving and insurance behavior.


Characteristics of older adults with diabetes: What does the current aged care resident look like?
The prevalence of diabetes in older adults in residential aged care (RAC) is twice that of community dwelling older adults. Older adults with diabetes have been highlighted as being at high risk of frailty and malnutrition, particularly when managed on a therapeutic diet. However, assumptions may be based on clinical presentation of our oldest old which may be at conflict with the clinical presentation of younger older adults. The aim of this retrospective audit was to identify the characteristics of aged care residents with diabetes, their comorbidities and malnutrition risk. Residents' with diabetes paper-based records were audited for demographic data, diet code, medical and medication history, malnutrition risk screening scores and anthropometric data. A total of 295 residents with diabetes records were audited across 13 sites in South Australia. Younger older adults (65-74 years) were generally in a healthy weight range (23-30 kg/m2 ) or obese (>30 kg/m2 ) and had gained weight since admission to RAC; whereas the oldest old were more likely to have lost weight and had a lower body mass index range 22.6-28.9 kg/m2 . An unexpected finding was the variability between RAC sites in dietary management of diabetes and potential inappropriate dietary management of malnutrition, as indicated by their food service diet codes. Findings suggest that younger older adults in RAC are more likely to be overweight which is maintained over a typical length of stay. Study findings indicate that diabetes management is inconsistent and highlights the need for mandated RAC menu guidelines.


Psychiatric disorders as risk factors for the development of type 2 diabetes mellitus: an umbrella review protocol.
Numerous longitudinal studies, systematic reviews and meta-analyses have examined psychiatric disorders as risk factors for the development of type 2 diabetes mellitus. A more comprehensive overview of the area is warranted to summarise current evidence and discuss strengths and weaknesses to guide future research. The aim of this umbrella review is to determine whether and to what extent different psychiatric disorders are associated with the development of type 2 diabetes mellitus. Furthermore, the umbrella review also assesses the evidence on potential mediating mechanisms. The present umbrella review will consist of a comprehensive systematic search of published systematic reviews and meta-analyses of observational longitudinal studies investigating whether a psychiatric disorder is associated with the risk of developing type 2 diabetes. PubMed, Embase, PsychINFO and the Cochrane Database of Systematic Reviews will be searched, and the results will be screened for inclusion by two independent reviewers. Furthermore, the reference lists of included publications will be manually searched. Two independent reviewers will extract data and assess the methodological quality in the included systematic reviews and meta-analyses. Evidence on potential mediating mechanisms included in the systematic reviews and meta-analyses will also be reviewed. The implications of the overview will be discussed in light of the quality of the included studies, and suggestions for clinical practice and future research will be made. Ethical approval is not required for this umbrella review. Our review will be submitted for publication in a peer-reviewed international journal using open access option if available. The results will also be disseminated at international conferences. CRD42018096362.


Genetics of arrhythmia: disease pathways beyond ion channels.
Diseases of the electrical conduction system that lead to irregularities in cardiac rhythm can have morbid and often lethal clinical outcomes. Linkage analysis has been the principal tool used to discover the genetic mutations responsible for Mendelian arrhythmic disease. Although the majority of arrhythmias can be accounted for by mutations in genes encoding ion channels, linkage analysis has also uncovered the role of other gene families such as those encoding members of the desmosome. With a list of candidates in mind, mutational analysis has helped confirm the suspicion that proteins found in caveolae or gap junctions also play a role in arrhythmogenesis. Atrial fibrillation and sudden cardiac death are relatively common arrhythmias that may be caused by multiple factors including common genetic variants. Genome-wide association studies are already revealing the important and poorly understood role of intergenic regions in atrial fibrillation. Despite the great advancements that have been made in our understanding of the genetics of these diseases, we are still far from able to routinely use genomic data to make clinical management decisions. There remain several hurdles in the study of genetics of arrhythmia, including the costs of genotyping, the need to find large affected families for linkage analysis, or to recruit large numbers of patients for genome-wide studies. Novel techniques that incorporate epigenetic information, such as known gene-gene interactions, biologic pathways, and experimental gene expression, will need to be developed to better interpret the large amount of genetic data that can now be generated. The study of arrhythmia genetics will continue to elucidate the pathophysiology of disease, help identify novel therapies, and ultimately allow us to deliver the individualized medical therapy that has long been anticipated.


A serological survey of African horse sickness in Botswana.
A retrospective serological survey of African horse sickness (AHS) in Botswana covering a 10-year period (1995-2004) is reported. The survey involved horses showing clinical symptoms of the disease; the horses had not been vaccinated against AHS. Over the period surveyed, serological evidence suggestive of infection with AHS virus (AHSV) was found in 99 clinical cases out of which 41.4% (41/99) cases were found during the 1st half (1995-1999) and 58.6 % (58/99) cases were found in the 2nd half of the survey period (2000-2004). These serological findings are discussed in relation to AHSV serotypes isolated from diseased horses in Botswana before and during the period of this serological survey.


Effect of epidermal growth factor receptor expression level on survival in patients with epithelial ovarian cancer.
Several lines of laboratory evidence support the epidermal growth factor receptor (EGFR) as an adverse prognostic indicator in ovarian cancers. However, different methods of immunohistochemical assessment have yielded conflicting results. Here, we sought to determine the prognostic value of EGFR in ovarian cancer using a novel method of compartmentalized in situ protein analysis. A tissue array composed of 150 advanced-stage ovarian cancers uniformly treated, with surgical debulking followed by platinum-paclitaxel combination chemotherapy, was constructed. For evaluation of EGFR protein expression, we used an immunofluorescence-based method of automated in situ quantitative measurement of protein analysis (AQUA). Mean follow-up time for the entire cohort was 34.4 months. Eighty-one of 150 cases had sufficient tissue for AQUA analysis. High tumor EGFR expression was associated with poor outcome for overall survival (P=0.0001) and disease-free survival (P=0.0005) at 3 years. In multivariable analysis, adjusting for well-characterized prognostic variables, EGFR expression status was the most significant prognostic factor for disease-free and overall survival. The conflicting results in the literature regarding the prognostic value of EGFR may be due to the technical difficulties inherent in assessing EGFR with immunocytochemistry. In the present study, we show that measurement of EGFR protein levels in ovarian cancer using AQUA is feasible and can give important prognostic information.


My prosthesis as a part of me: a qualitative analysis of living with an osseointegrated prosthetic limb.
Bone-anchored prosthesis is still a rather unusual treatment for patients with limb loss. The aim of this study was to improve our understanding about the experience of living with an osseointegrated prosthesis (OI-prosthesis) compared to one suspended with a socket, through the use of qualitative research methodology. A qualitative phenomenological research method. Thirteen Swedish patients (37-67 years) with unilateral upper or lower limb amputation (10 transfemoral, 2 transhumeral, 1 transradial), who had been using OI-prostheses for 3 to 15 years, were recruited by means of purposive sampling. An audio-taped in-depth interview was performed. The guiding question was 'How do you experience living with your osseointegrated prosthesis compared to your earlier prostheses suspended with sockets?'. The empirical phenomenological psychological method was used for data analysis. The results showed that all participants described living with an OI-prosthesis as a revolutionary change. These experiences were described in terms of three typologies, called 'Practical prosthesis', 'Pretend limb' and 'A part of me'. The most important finding was that the change went beyond the functional improvements, integrating the existential implications in the concept of quality of life. This qualitative in-depth interview study on patients using bone-anchored prosthetic limbs showed that all described a revolutionary change in their lives as amputees and the meaning of that change went beyond the functional improvements, integrating existential implications in the concept of quality of life.


Expression of the anti-Mullerian hormone, kisspeptin 1, and kisspeptin 1 receptor in polycystic ovary syndrome and controlled ovarian stimulation rat models.
Polycystic ovary syndrome represents a significant cause of female infertility. The aim of this study was to investigate the expression of anti-Mul-lerian hormone (AMH), kisspeptin 1 (KISS-1), and kisspeptin 1 receptor (KISS1r) in rat models of polycystic ovary syndrome (PCOS) and controlled ovarian stimulation (COS). For this purpose, 28 rats were assigned into four groups. Estrus and Diestrus groups consisted of rats in estrus and diestrus phases, respectively, while COS and PCOS groups consisted of rats with induced COS and PCOS, respectively. The serum AMH, KISS-1, and estradiol levels, and ovarian KISS1r levels were analyzed by enzyme-linked immunosorbent assay. Furthermore, histopathological analysis of the ovary tissue was done and ovarian KISS-1 expression was determined by immunohistochemical assay. The results revealed that ovarian KISS1r levels were higher in the Estrus (1271.43±51.98 pg/mL) and COS (1191.43±85.67 pg/mL) groups, compared to Diestrus and PCOS groups. The highest level of AMH was found in the Estrus group (16.91±2.12 ng/mL). The results indicate that AMH had no effect on the development of COS and PCOS, while KISS-1 was found to affect the development of COS in rats.


The anterolateral approach leads to less disruption of the femoral head-neck blood supply than the posterior approach during hip resurfacing.
In 12 patients, we measured the oxygen concentration in the femoral head-neck junction during hip resurfacing through the anterolateral approach. This was compared with previous measurements made for the posterior approach. For the anterolateral approach, the oxygen concentration was found to be highly dependent upon the position of the leg, which was adjusted during surgery to provide exposure to the acetabulum and femoral head. Gross external rotation of the hip gave a significant decrease in oxygenation of the femoral head. Straightening the limb led to recovery in oxygen concentration, indicating that the blood supply was maintained. The oxygen concentration at the end of the procedure was not significantly different from that at the start. The anterolateral approach appears to produce less disruption to the blood flow in the femoral head-neck junction than the posterior approach for patients undergoing hip resurfacing. This may be reflected subsequently in a lower incidence of fracture of the femoral neck and avascular necrosis.


Anti-Oxidant and Anti-Inflammatory Activity of Ketogenic Diet: New Perspectives for Neuroprotection in Alzheimer's Disease.
The ketogenic diet, originally developed for the treatment of epilepsy in non-responder children, is spreading to be used in the treatment of many diseases, including Alzheimer's disease. The main activity of the ketogenic diet has been related to improved mitochondrial function and decreased oxidative stress. B-Hydroxybutyrate, the most studied ketone body, has been shown to reduce the production of reactive oxygen species (ROS), improving mitochondrial respiration: it stimulates the cellular endogenous antioxidant system with the activation of nuclear factor erythroid-derived 2-related factor 2 (Nrf2), it modulates the ratio between the oxidized and reduced forms of nicotinamide adenine dinucleotide (NAD⁺/NADH) and it increases the efficiency of electron transport chain through the expression of uncoupling proteins. Furthermore, the ketogenic diet performs anti-inflammatory activity by inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) activation and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome as well as inhibiting histone deacetylases (HDACs), improving memory encoding. The underlying mechanisms and the perspectives for the treatment of Alzheimer's disease are discussed.


RELICA: a method for estimating the reliability of independent components.
Independent Component Analysis (ICA) is a widely applied data-driven method for parsing brain and non-brain EEG source signals, mixed by volume conduction to the scalp electrodes, into a set of maximally temporally and often functionally independent components (ICs). Many ICs may be identified with a precise physiological or non-physiological origin. However, this process is hindered by partial instability in ICA results that can arise from noise in the data. Here we propose RELICA (RELiable ICA), a novel method to characterize IC reliability within subjects. RELICA first computes IC "dipolarity" a measure of physiological plausibility, plus a measure of IC consistency across multiple decompositions of bootstrap versions of the input data. RELICA then uses these two measures to visualize and cluster the separated ICs, providing a within-subject measure of IC reliability that does not involve checking for its occurrence across subjects. We demonstrate the use of RELICA on EEG data recorded from 14 subjects performing a working memory experiment and show that many brain and ocular artifact ICs are correctly classified as "stable" (highly repeatable across decompositions of bootstrapped versions of the input data). Many stable ICs appear to originate in the brain, while other stable ICs account for identifiable non-brain processes such as line noise. RELICA might be used with any linear blind source separation algorithm to reduce the risk of basing conclusions on unstable or physiologically un-interpretable component processes.


Developing polarized protein-specific charges for protein dynamics: MD free energy calculation of pKa shifts for Asp26/Asp20 in thioredoxin.
Ab initio quantum mechanical calculation of protein in solution is carried out to generate polarized protein-specific charge(s) (PPC) for molecular dynamics (MD) stimulation of protein. The quantum calculation of protein is made possible by developing a fragment-based quantum chemistry approach in combination with the implicit continuum solvent model. The computed electron density of protein is utilized to derive PPCs that represent the polarized electrostatic state of protein near the native structure. These PPCs are atom-centered like those in the standard force fields and are thus computationally attractive for molecular dynamics simulation of protein. Extensive MD simulations have been carried out to investigate the effect of electronic polarization on the structure and dynamics of thioredoxin. Our study shows that the dynamics of thioredoxin is stabilized by electronic polarization through explicit comparison between MD results using PPC and AMBER charges. In particular, MD free-energy calculation using PPCs accurately reproduced the experimental value of pK(a) shift for ionizable residue Asp(26) buried inside thioredoxin, whereas previous calculations using standard force fields overestimated pK(a) shift by twice as much. Accurate prediction of pK(a) shifts by rigorous MD free energy simulation for ionizable residues buried inside protein has been a significant challenge in computational biology for decades. This study presented strong evidence that electronic polarization of protein plays an important role in protein dynamics.


Modelling mortality and discharge of hospitalized stroke patients using a phase-type recovery model.
We model the length of in-patient hospital stays due to stroke and the mode of discharge using a phase-type stroke recovery model. The model allows for three different types of stroke: haemorrhagic (the most severe, caused by ruptured blood vessels that cause brain bleeding), cerebral infarction (less severe, caused by blood clots) and transient ischemic attack or TIA (the least severe, a mini-stroke caused by a temporary blood clot). A four-phase recovery process is used, where the initial phase depends on the type of stroke, and transition from one phase to the next depends on the age of the patient. There are three differing modes of absorption for this phase-type model: from a typical recovery phase, a patient may die (mode 1), be transferred to a nursing home (mode 2) or be discharged to the individual's usual residence (mode 3). The first recovery phase is characterized by a very high rate of mortality and very low rates of discharge by the other two modes. The next two recovery phases have progressively lower mortality rates and higher mode 2 and 3 discharge rates. The fourth recovery phase is visited only by those who experience a very mild TIA, and they are discharged to home after a short stay. The novelty of our approach to phase representation is two-fold: first, it aligns the phases with labelled diagnosis states, representing stages of illness severity; second, the model allows us to obtain expressions for Key Performance Indicators that are of use to healthcare professionals. This allows us to use a backward estimation process where we leverage the fact that we know the phase of admission (the diagnosis), but not which phases are subsequently entered or when this happens; this strategy improves both computational efficiency and accuracy. The model has clear practical value as it yields length of stay distributions by age and type of stroke, which are useful in resource planning. Also, inclusion of the three modes of discharge permits analyses of outcomes.


Comparison of the tissue receptors for Vibrio cholerae and Escherichia coli enterotoxins by means of gangliosides and natural cholera toxoid.
The in vitro binding properties of enterotoxins of Vibrio cholerae and Escherichia coli to different pure gangliosides and related neutral glycosphin-golipids were analyzed with a sorbent assay utilizing plastic tubes to which the glycolipid substances had been coupled. It was found that the cholera toxin bound to G(M1) ganglioside better than to the other tested substances G(M3), G(M3)-NGN, G(M2), G(D1a), G(D1b), G(T), G(A1), tetrahexoside-GlcNac and globoside. With this assay using G(M1)-coated tubes it is possible to measure cholera toxin even at concentrations below 1 ng/ml. Also enterotoxin of various E. coli strains bound to G(M1), but the affinity was much less than for cholera toxin. The G(M1) ganglioside, in contrast to the other glycosphingolipids, effectively inactivated cholera toxin as determined with the intradermal and the ileal loop assays; approximately equimolar concentrations of the ganglioside in relation to toxin sufficed. Also, the skin and ileal loop activities of E. coli enterotoxins could be inhibited by G(M1); however, several orders more of the ganglioside were required for such inhibition than for inactivation of the cholera toxin, and the differences between G(M1) and the other substances were less pronounced for E. coli toxins. Preincubation of rabbit ileal loops with choleragenoid, a natural toxoid of V. cholerae which has binding properties to the G(M1) ganglioside similar to cholera toxin, made the loops resistant to subsequently added enterotoxin of V. cholerae. The responsiveness to enterotoxin of E. coli was not reduced by this toxoid. A likely interpretation of these data is that the G(M1) ganglioside constitutes or at least contains the structure of functional tissue receptors for the cholera toxin, whereas the weak binding to G(M1) by E. coli enterotoxins is probably a pathogenetically insignificant reflection of structural similarities between these toxins and cholera toxin. Consequently, the cholera toxoid by occupying functional intestinal G(M1) receptors for the cholera toxin could inhibit the ileal response to this toxin, but not the response to E. coli enterotoxin since the intestinal receptors for the latter toxin are not affected by the cholera toxoid.


[Prevalence of metabolic syndrome in Mapuche individuals living in urban and rural environment in Chile].
Metabolic Syndrome (MS) increases the risk of diabetes and mortality associated with cardiovascular disease. However, the prevalence of MS could differ by ethnicity and lifestyle factors. To determine the prevalence of MS in Mapuche individuals living in urban and rural environments in Chile and to investigate whether the prevalence and risk of MS in urban and rural environments differs by sex, age and nutritional status. A total of 1077 Mapuche participants were recruited from urban (MU = 288) and rural (MR = 789) settings. Body mass index, waist circumference and blood pressure were measured. A fasting blood sample was obtained to measure serum glucose, HDL cholesterol and triacylglycerol. The prevalence of MS was determined using the unified IDF and ATP-III criteria. An environment and sex interaction was found for the prevalence of MS (p = 0.042). The prevalence was significantly lower in male MR (13%) compared to other groups (22, 23 and 25% among female MR, female MU and male MU respectively). Also, the prevalence of central obesity and low HDL-cholesterol were significantly lower in male MR. MU are at an increased risk of developing MS compared to MR, with an odds ratio of 1.59 (95% confidence intervals 1.1 to 2.2). This risk increases along with age or body mass index of the population. The adoption of an urbanized lifestyle increases the risk of developing MS in Mapuche individuals. This risk is enhanced by age and nutritional status.


Nanocarriers in therapy of infectious and inflammatory diseases.
Nanotechnology is a growing science that has applications in various areas of medicine. The composition of nanocarriers for drug delivery is critical to guarantee high therapeutic performance when targeting specific host sites. Applications of nanotechnology are prevalent in the diagnosis and treatment of infectious and inflammatory diseases. This review summarizes recent advancements in the application of nanotechnology to the therapy of infectious and inflammatory diseases. The major focus is on the design and fabrication of various nanomaterials, characteristics and physicochemical properties of drug-loaded nanocarriers, and the use of these nanoscale drug delivery systems in treating infectious and inflammatory diseases, such as AIDS, hepatitis, tuberculosis, melanoma, and representative inflammatory diseases. Clinical trials and future perspective of the use of nanocarriers are also discussed in detail. We hope that such a review will be valuable to researchers who are exploring nanoscale drug delivery systems for the treatment of specific infectious and inflammatory diseases.


A randomized placebo-controlled trial of fluvastatin for prevention of restenosis after successful coronary balloon angioplasty; final results of the fluvastatin angiographic restenosis (FLARE) trial.
The 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors competitively inhibit biosynthesis of mevalonate, a precursor of non-sterol compounds involved in cell proliferation. Experimental evidence suggests that fluvastatin may, independent of any lipid lowering action, exert a greater direct inhibitory effect on proliferating vascular myocytes than other statins. The FLARE (Fluvastatin Angioplasty Restenosis) Trial was conceived to evaluate the ability of fluvastatin 40 mg twice daily to reduce restenosis after successful coronary balloon angioplasty (PTCA). Patients were randomized to either placebo or fluvastatin 40 mg twice daily beginning 2-4 weeks prior to planned PTCA and continuing after a successful PTCA (without the use of a stent), to follow-up angiography at 26+/-2 weeks. Clinical follow-up was completed at 40 weeks. The primary end-point was angiographic restenosis, measured by quantitative coronary angiography at a core laboratory, as the loss in minimal luminal diameter during follow-up. Clinical end-points were death, myocardial infarction, coronary artery bypass graft surgery or re-intervention, up to 40 weeks after PTCA. Of 1054 patients randomized, 526 were allocated to fluvastatin and 528 to placebo. Among these, 409 in the fluvastatin group and 427 in the placebo group were included in the intention-to-treat analysis, having undergone a successful PTCA after a minimum of 2 weeks of pre-treatment. At the time of PTCA, fluvastatin had reduced LDL cholesterol by 37% and this was maintained at 33% at 26 weeks. There was no difference in the primary end-point between the treatment groups (fluvastatin 0.23+/-0.49 mm vs placebo 0.23+/-0.52 mm, P=0.95) or in the angiographic restenosis rate (fluvastatin 28%, placebo 31%, chi-square P=0.42), or in the incidence of the composite clinical end-point at 40 weeks (22.4% vs 23.3%; logrank P=0.74). However, a significantly lower incidence of total death and myocardial infarction was observed in six patients (1.4%) in the fluvastatin group and 17 (4.0%) in the placebo group (log rank P=0.025). Treatment with fluvastatin 80 mg daily did not affect the process of restenosis and is therefore not indicated for this purpose. However, the observed reduction in mortality and myocardial infarction 40 weeks after PTCA in the fluvastatin treated group has not been previously reported with statin therapy. Accordingly, a priori investigation of this finding is indicated and a new clinical trial with this intention is already underway.


Contralateral microemboli following carotid artery stenting in patients with a contralateral internal carotid artery occlusion.
Subclinical microembolization identified on diffusion-weighted magnetic resonance imaging is recognized as an important outcome measure for carotid revascularization procedures. It is generally believed that arch manipulation is the primary reason for developing microemboli in the contralateral hemisphere during carotid artery stenting. However, we identified three patients who developed postprocedure microemboli of the contralateral hemisphere despite a known chronic contralateral internal carotid artery occlusion. Our cases highlight that ipsilateral microemboli may be an underappreciated but an important source of contralateral lesions through patent intracranial collateral pathways.


Developmental expression of FMRP in the astrocyte lineage: implications for fragile X syndrome.
One of the most common causes of mental retardation in humans, Fragile X syndrome, results from the absence of FMRP, the protein product of the FMR1 gene. In the nervous system, expression of FMRP has been thought to be confined mainly to neurons as little research has examined FMRP expression in non-neuronal lineages. We present evidence that, in addition to neuronal expression, FMRP is expressed in developing CNS glial cells in vitro and in vivo. The neurosphere assay was used to establish cultures of stem and progenitor cells from the brains of wildtype and FMRP knockout (B6.129.FMR1/FvBn) mouse pups. When the neurospheres were differentiated in vitro, approximately 50% of the FMRP positive cells also expressed GFAP. Immunocytochemical studies of the embryonic and postnatal mouse brain revealed coexpression of FMRP and GFAP in the developing hippocampus. Prominent coexpression was also observed in ependymal cells surrounding the third ventricle and astrocytes of the glia limitans. No double-labeled cells were evident in the brains of young adult mice. Cells coexpressing FMRP and the oligodendrocyte precursor marker NG2 were also identified in the hippocampus and corpus callosum of the early postnatal brain. Our results suggest that FMRP is expressed in cells of non-neuronal lineage(s) during development. This represents potential involvement of glial cells in the neural development of fragile X syndrome.


Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Mutation rate and the cost of complexity.
Two recent theoretical studies of adaptation suggest that more complex organisms tend to adapt more slowly. Specifically, in Fisher's "geometric" model of a finite population where multiple traits are under optimizing selection, the average progress ensuing from a single mutation decreases as the number of traits increases--the "cost of complexity." Here, I draw on molecular and histological data to assess the extent to which on a large phylogenetic scale, this predicted decrease in the rate of adaptation per mutation is mitigated by an increase in the number of mutations per generation as complexity increases. As an index of complexity for multicellular organisms, I use the number of visibly distinct types of cell in the body. Mutation rate is the product of mutational target size and population mutation rate per unit target. Despite much scatter, genome size appears to be positively correlated with complexity (as indexed by cell-type number), which along with other considerations suggests that mutational target size tends to increase with complexity. In contrast, effective population mutation rate per unit target appears to be negatively correlated with complexity. The net result is that mutation rate probably does tend to increase with complexity, although probably not fast enough to eliminate the cost of complexity.


Neural maps of interaural time and intensity differences in the optic tectum of the barn owl.
This report describes the binaural basis of the auditory space map in the optic tectum of the barn owl (Tyto alba). Single units were recorded extracellularly in ketamine-anesthetized birds. Unit tuning for interaural differences in timing and intensity of wideband noise was measured using digitally synthesized sound presented through earphones. Spatial receptive fields of the same units were measured with a free field sound source. Auditory units in the optic tectum are sharply tuned for both the azimuth and the elevation of a free field sound source. To determine the binaural cues that could be responsible for this spatial tuning, we measured in the ear canals the amplitude and phase spectra produced by a free field noise source and calculated from these measurements the interaural differences in time and intensity associated with each of 178 locations throughout the frontal hemisphere. For all frequencies, interaural time differences (ITDs) varied systematically and most strongly with source azimuth. The pattern of variation of interaural intensity differences (IIDs) depended on frequency. For low frequencies (below 4 kHz) IID varied primarily with source azimuth, whereas for high frequencies (above 5 kHz) IID varied primarily with source elevation. Tectal units were tuned for interaural differences in both time and intensity of dichotic stimuli. Changing either parameter away from the best value for the unit decreased the unit's response. The tuning of units to either parameter was sharp: the width of ITD tuning curves, measured at 50% of the maximum response with IID held constant (50% tuning width), ranged from 18 to 82 microsecs. The 50% tuning widths of IID tuning curves, measured with ITD held constant, ranged from 8 to 37 dB. For most units, tuning for ITD was largely independent of IID, and vice versa. A few units exhibited systematic shifts of the best ITD with changes in IID (or shifts of the best IID with changes in ITD); for these units, a change in the value of one parameter to favor one ear shifted the best value of the other parameter in favor of the same ear, i.e., in the direction opposite to that expected from "time-intensity trading." Overall sound intensity had little or no effect on ITD tuning, but did increase the best IIDs of units tuned to nonzero IIDs. The tuning of units for ITD and IID changed systematically along different dimensions of the optic tectum to create coextensive, independent neurophysiological maps of ITD and IID.(ABSTRACT TRUNCATED AT 400 WORDS)


Discovery of prosimian and afrotherian foamy viruses and potential cross species transmissions amidst stable and ancient mammalian co-evolution.
Foamy viruses (FVs) are a unique subfamily of retroviruses that are widely distributed in mammals. Owing to the availability of sequences from diverse mammals coupled with their pattern of codivergence with their hosts, FVs have one of the best-understood viral evolutionary histories ever documented, estimated to have an ancient origin. Nonetheless, our knowledge of some parts of FV evolution, notably that of prosimian and afrotherian FVs, is far from complete due to the lack of sequence data. Here, we report the complete genome of the first extant prosimian FV (PSFV) isolated from a lorisiforme galago (PSFVgal), and a novel partial endogenous viral element with high sequence similarity to FVs, present in the afrotherian Cape golden mole genome (ChrEFV). We also further characterize a previously discovered endogenous PSFV present in the aye-aye genome (PSFVaye). Using phylogenetic methods and available FV sequence data, we show a deep divergence and stable co-evolution of FVs in eutherian mammals over 100 million years. Nonetheless, we found that the evolutionary histories of bat, aye-aye, and New World monkey FVs conflict with the evolutionary histories of their hosts. By combining sequence analysis and biogeographical knowledge, we propose explanations for these mismatches in FV-host evolutionary history. Our discovery of ChrEFV has expanded the FV host range to cover the whole eutherian clade, and our evolutionary analyses suggest a stable mammalian FV-host co-speciation pattern which extends as deep as the exafroplacentalian basal diversification. Nonetheless, two possible cases of host switching were observed. One was among New World monkey FVs, and the other involves PSFVaye and a bat FV which may involve cross-species transmission at the level of mammalian orders. Our results highlight the value of integrating multiple sources of information to elucidate the evolutionary history of viruses, including continental and geographical histories, ancestral host locations, in addition to the natural history of host and virus.