New chemical and radiochemical routes to [18F]Rho6G-DEG-F, a delocalized lipophilic cation for myocardial perfusion imaging with PET.
New chemical and radiochemical syntheses are described for the preparation of [18F]Rho6G-DEG-F, an 18F-labeled analogue of the fluoresecent dye rhodamine 6G, which has shown promise as myocardidal perfusion imaging agent. Tosylated precursors of [18F]Rho6G-DEG-F amenable to 18F-labeling were obtained either through a two-step synthesis from rhodamine 6G lactone (33% yield), or in one step from rhodamine 575 (64% yield), then purified by preparative C18 chromatography. Manual synthesis of [18F]Rho6G-DEG-F was achieved in a single radiochemical step from either the tosylate salt or the tosylate/formate double salt in DMSO under standard nucleophillic aliphatic 18F-fluorination conditions (K[18F]F/K2CO3/Kryptofix 2.2.2.). Incorporation of the [18F]F- was found to be satisfactory (≥34% by TLC), despite the protic character of the precursor molecules. [18F]Rho6G-DEG-F was manually synthesized in final decay-corrected radiochemical yields of 11-26% (tosylate salt) and 9-21% (tosylate/formate double salt). The protocol was transferred to an automated synthesis unit, where the product was obtained in 3-9% radiochemical yield (n=3) decay corrected to start-of-synthesis, >99% radiochemical purity, and a molar activity of 122-267 GBq/μmol (3.3-7.2 Ci/μmol).


NTP Toxicology and Carcinogenesis Studies of 3-Chloro-2-methylpropene (Technical grade containing 5% dimethylvinyl chloride) (CAS No. 563-47-3) in F344/N Rats and B6C3F1 Mice (Gavage Studies).
Toxicology and carcinogenesis studies of technical-grade 3-chloro-2-methylpropene (containing 5% dimethylvinyl chloride), a widely used insecticide and a chemical intermediate, were performed on F344/N rats and B6C3F1 mice. In the 13-week studies, 50%-100% mortality occurred in groups of male and female rats receiving 400 mg/kg, male rats receiving 300 mg/kg, and male and female mice receiving 500-1,250 mg/kg. Inflammation and necrosis of the liver were seen in rats and mice, and necrosis of cortical tubules of the kidney was seen in mice. Based on these observations, groups of 50 male and 50 female rats were administered 3-chloro-2-methylpropene in corn oil by gavage at doses of 0, 75, or 150 mg/kg body weight, 5 days per week for 103 weeks, and groups of 50 male and 50 female mice received 3-chloro-2-methylpropene at 0, 100, or 200 mg/kg on the same schedule. In the 2-year studies, the mean body weight of high dose male rats was consistently 10%-15% lower than that of the vehicle control group, and late in the study there was a marginal reduction in survival of high dose male rats. Mean body weights and survival in low dose male rats and in both dosed groups of female rats were comparable to those of their vehicle control groups. Mean body weights of high dose male mice and of both dosed groups of female mice were slightly (5%-9%) lower than those of the vehicle controls, whereas survival in both male and female mice was not affected by 3-chloro-2-methylpropene administration. Dose-related increases in the incidence of forestomach inflammation were observed in male and female mice (male: vehicle control, 0/49; low dose, 9/49; high dose, 7/49; female: vehicle control, 2/50; low dose, 3/48; high dose, 9/44). Increased incidences of forestomach basal cell hyperplasia were observed in rats and mice of each sex. 3-Chloro-2-methylpropene induced forestomach squamous cell papillomas and squamous cell carcinomas in rats and mice as shown in the table. Invasion or metastasis of the squamous cell carcinomas to other organs was observed in two low dose male, three high dose male, and one high dose female mice. Renal tubular cell adenocarcinomas (1/49), renal transitional cell carcinomas (1/49), and transitional cell papillomas (1/49) of the urinary bladder were observed in high dose male rats, and renal tubular cell adenomas (1/50) and renal tubular cell adenocarcinomas (1/50) were seen in low dose male rats. These urinary tract neoplasms were not observed in vehicle controls. The incidences of inflammation of the nasal cavity and of nephropathy/nephrosis were greater in the two dosed groups than in the vehicle control groups of rats and mice of each sex. Negative trends or lower incidences of pheochromocytomas of the adrenal gland and C-cell adenomas or carcinomas (combined) of the thyroid gland were observed in dosed male rats. Negative trends were observed in the incidences of hepatocellular adenomas or carcinomas (combined) in dosed male mice and of hemangiomas or hemangiosarcomas (combined) in dosed female mice. 3-Chloro-2-methylpropene was weakly mutagenic in Salmonella typhimurium strain TA1537 with 10% rat liver S9; results in strain TA100 with 10% Syrian hamster liver S9 or with 10% or 30% rat liver S9 were judged equivocal. Mutagenicity tests with S. typhimurium strains TA1535 and TA98 were negative with or without metabolic activation. 3-Chloro-2-methylpropene was mutagenic in the mouse lymphoma L5178/TK+/- forward mutation assay without exogenous metabolic activation. Cytogenetics tests with cultured Chinese hamster ovary cells were positive for induction of chromosomal aberrations and sister-chromatid exchanges (SCE's) in the absence of rat liver S9. With metabolic activation, SCE levels remained significantly elevated, but the number of chromosomal aberrations was reduced. An audit of the experimental data was conducted for these 2-year carcinogenesis studies on 3-chloro-2-methylpropene. No data discrepancies were found that influenced the final interpretations. Under the conditions of these 2-year gavage studies, d the final interpretations. Under the conditions of these 2-year gavage studies, there was clear evidence of carcinogenicity for 3-chloro-2-methylpropene as shown by the increased incidences of squamous cell neoplasms in the forestomach of male and female F344/N rats and of male and female B6C3F1 mice. Synonyms: 2-methallyl chloride; methyl allyl chloride; b-methallyl chloride; pi-chloroisobutylene; isobutenyl chloride; 3-chloro-2-methyl-1-propene; 2-methyl-2-propenyl chloride


Novel insights into the molecular events linking to cell death induced by tetracycline in the amitochondriate protozoan Trichomonas vaginalis.
Trichomonas vaginalis colonizes the human urogenital tract and causes trichomoniasis, the most common nonviral sexually transmitted disease. Currently, 5-nitroimidazoles are the only recommended drugs for treating trichomoniasis. However, increased resistance of the parasite to 5-nitroimidazoles has emerged as a highly problematic public health issue. Hence, it is essential to identify alternative chemotherapeutic agents against refractory trichomoniasis. Tetracycline (TET) is a broad-spectrum antibiotic with activity against several protozoan parasites, but the mode of action of TET in parasites remains poorly understood. The in vitro effect of TET on the growth of T. vaginalis was examined, and the mode of cell death was verified by various apoptosis-related assays. Next-generation sequencing-based RNA sequencing (RNA-seq) was employed to elucidate the transcriptome of T. vaginalis in response to TET. We show that TET has a cytotoxic effect on both metronidazole (MTZ)-sensitive and -resistant T. vaginalis isolates, inducing some features resembling apoptosis. RNA-seq data reveal that TET significantly alters the transcriptome via activation of specific pathways, such as aminoacyl-tRNA synthetases and carbohydrate metabolism. Functional analyses demonstrate that TET disrupts the hydrogenosomal membrane potential and antioxidant system, which concomitantly elicits a metabolic shift toward glycolysis, suggesting that the hydrogenosomal function is impaired and triggers cell death. Collectively, we provide in vitro evidence that TET is a potential alternative therapeutic choice for treating MTZ-resistant T. vaginalis. The in-depth transcriptomic signatures in T. vaginalis upon TET treatment presented here will shed light on the signaling pathways linking to cell death in amitochondriate organisms.


Teaching "Shock Pathophysiology" by Flipped Classroom.
To assess usefulness of flipped style of teaching conducted as small-group format in Cardiovascular and Respiration module for Year-I undergraduate medical students at Aga Khan University. The study was planned and conducted over a period of eight months from March to October 2017 including the time taken for planning, mock run, execution followed by analysis and dissemination. It was carried out at the Aga Khan University Medical College, Karachi. Pre and post test scores of students after flipped class room sessions was compared. Moreover, perception of students was assessed on Likert scale (0-4) by a pretested validated questionnaire. The mean pre-test scores of the students was 4.86 ± 0.91 which improved to 6.09 ± 0.81 (p = 0.021) after attending the flipped class session. Students approved that the frame work helped to promote their learning motivation and engagement with improvement in understanding of the course materials and enhancement of learning during Face to Face activity. The flipped classroom approach showed promise in teaching and learning of 'Pathophysiology of Shock' by clinical scenarios in small group discussions. Implementation of flipped class room activity on a wider scale however needs careful selection of course objectives and logistics.


How to get between the sheets: a review of recent works on the electrochemical exfoliation of graphene materials from bulk graphite.
Since the beginning of the 'graphene era' post-2004, there has been significant interest in developing a high purity, high yield, and scalable fabrication route toward graphene materials for both primary research purposes and industrial production. One suitable approach to graphene production lies in the realm of electrochemical exfoliation, in which a potential difference is applied between a graphite anode/cathode in the presence of an electrolyte-containing medium. Herein we review various works on the electrochemical fabrication of graphene materials specifically through the use of electrochemical intercalation and exfoliation of a graphite source electrode, focusing on the quality and purity of products formed. We categorise the most significant works in terms of anodic and cathodic control, highlighting the merits of the respective approaches, as well as indicating the challenges associated with both procedures.


Decomposing variation in male reproductive success: age-specific variances and covariances through extra-pair and within-pair reproduction.
Age-specific variances and covariances in reproductive success shape the total variance in lifetime reproductive success (LRS), age-specific opportunities for selection, and population demographic variance and effective size. Age-specific (co)variances in reproductive success achieved through different reproductive routes must therefore be quantified to predict population, phenotypic and evolutionary dynamics in age-structured populations. While numerous studies have quantified age-specific variation in mean reproductive success, age-specific variances and covariances in reproductive success, and the contributions of different reproductive routes to these (co)variances, have not been comprehensively quantified in natural populations. We applied 'additive' and 'independent' methods of variance decomposition to complete data describing apparent (social) and realised (genetic) age-specific reproductive success across 11 cohorts of socially monogamous but genetically polygynandrous song sparrows (Melospiza melodia). We thereby quantified age-specific (co)variances in male within-pair and extra-pair reproductive success (WPRS and EPRS) and the contributions of these (co)variances to the total variances in age-specific reproductive success and LRS. 'Additive' decomposition showed that within-age and among-age (co)variances in WPRS across males aged 2-4 years contributed most to the total variance in LRS. Age-specific (co)variances in EPRS contributed relatively little. However, extra-pair reproduction altered age-specific variances in reproductive success relative to the social mating system, and hence altered the relative contributions of age-specific reproductive success to the total variance in LRS. 'Independent' decomposition showed that the (co)variances in age-specific WPRS, EPRS and total reproductive success, and the resulting opportunities for selection, varied substantially across males that survived to each age. Furthermore, extra-pair reproduction increased the variance in age-specific reproductive success relative to the social mating system to a degree that increased across successive age classes. This comprehensive decomposition of the total variances in age-specific reproductive success and LRS into age-specific (co)variances attributable to two reproductive routes showed that within-age and among-age covariances contributed substantially to the total variance and that extra-pair reproduction can alter the (co)variance structure of age-specific reproductive success. Such covariances and impacts should consequently be integrated into theoretical assessments of demographic and evolutionary processes in age-structured populations.


The role of 1-D finite size Heisenberg chains in increasing the metal to insulator transition temperature in hole rich VO2.
VO2 samples are grown with different oxygen concentrations leading to different monoclinic, M1, and triclinic, T, insulating phases which undergo a first order metal to insulator transition (MIT) followed by a structural phase transition (SPT) to the rutile tetragonal phase. The metal insulator transition temperature (Tc) was found to be increased with increasing native defects. Vanadium vacancy (VV) is envisaged to create local strains in the lattice which prevents twisting of the V-V dimers promoting metastable monoclinic, M2 and T phases at intermediate temperatures. It is argued that MIT is driven by strong electronic correlation. The low temperature insulating phase can be considered as a collection of one-dimensional (1-D) half-filled bands, which undergo a Mott transition to 1-D infinitely long Heisenberg spin ½ chains leading to structural distortion due to spin-phonon coupling. The presence of VV creates localized holes (d0) in the nearest neighbor, thereby fragmenting the spin ½ chains at the nanoscale, which in turn increases the Tc value more than that of an infinitely long one. The Tc value scales inversely with the average size of the fragmented Heisenberg spin ½ chains following a critical exponent of ⅔, which is exactly the same as predicted theoretically for the Heisenberg spin ½ chain at the nanoscale undergoing SPT (spin-Peierls transition). Thus, the observation of MIT and SPT at the same time in VO2 can be explained from our phenomenological model of reduced 1-D Heisenberg spin ½ chains. The reported increase (decrease) in the Tc value of VO2 by doping with metals having valency less (more) than four can also be understood easily with our unified model, for the first time, considering finite size scaling of Heisenberg chains.


Physical Demand but Not Dexterity Is Associated with Motor Flexibility during Rapid Reaching in Healthy Young Adults.
Healthy humans are able to place light and heavy objects in small and large target locations with remarkable accuracy. Here we examine how dexterity demand and physical demand affect flexibility in joint coordination and end-effector kinematics when healthy young adults perform an upper extremity reaching task. We manipulated dexterity demand by changing target size and physical demand by increasing external resistance to reaching. Uncontrolled manifold analysis was used to decompose variability in joint coordination patterns into variability stabilizing the end-effector and variability de-stabilizing the end-effector during reaching. Our results demonstrate a proportional increase in stabilizing and de-stabilizing variability without a change in the ratio of the two variability components as physical demands increase. We interpret this finding in the context of previous studies showing that sensorimotor noise increases with increasing physical demands. We propose that the larger de-stabilizing variability as a function of physical demand originated from larger sensorimotor noise in the neuromuscular system. The larger stabilizing variability with larger physical demands is a strategy employed by the neuromuscular system to counter the de-stabilizing variability so that performance stability is maintained. Our findings have practical implications for improving the effectiveness of movement therapy in a wide range of patient groups, maintaining upper extremity function in old adults, and for maximizing athletic performance.


Highly efficient photocatalytic hydrogen evolution of graphene/YInO3 nanocomposites under visible light irradiation.
Visible-light-driven hydrogen evolution with high efficiency is important in the current photocatalysis research. Here we report for the first time the design and synthesis of a new graphene-semiconductor nanocomposite consisting of YInO3 nanoparticles and two-dimensional graphene sheets as efficient photocatalysts for hydrogen evolution under visible light irradiation. The graphene/YInO3 nanocomposites were synthesized using a facile solvothermal method in which the formation of graphene and the deposition of YInO3 nanoparticles on the graphene sheets can be achieved simultaneously. The addition of graphene as a cocatalyst can narrow the band gap of YInO3 to visible photon energy and prolong the separation and lifetime of electron-hole pairs by the chemical bonding between YInO3 and graphene. The photocatalytic reaction with this nanocomposite reaches a high H2 evolution rate of 400.4 μmol h(-1) g(-1) when the content of graphene is 0.5 wt%, over 127 and 3.7 times higher than that of pure YInO3 and Pt/YInO3, respectively. This work can provide an effective approach to the fabrication of graphene-based photocatalysts with high performance in the field of energy conversion.


Cumulative Risk and Continuity in Nonparental Care from Infancy to Early Adolescence.
Variations in amounts of nonparental care across infancy, preschool, early elementary school, and early adolescence were examined in a longitudinal sample (N = 438). Of interest was (a) continuity in use of the different arrangements, (b) whether the arrangements were additively and cumulatively associated with children's externalizing behavior problems, and (c) whether predictive relations were accounted for by social-ecological (socioeconomic status, mothers' employment status, marital status) and social-experiential (parenting quality, exposure to aggressive peers) factors. Correlations among overall amounts of care provided little evidence of cross-time continuity. Consistent with the cumulative risk perspective, Grade 1 self-care and Grade 6 unsupervised peer contact incrementally predicted Grade 6 externalizing problems. Most of the predictive associations were accounted for by family background and social relationship factors.


Informed consent procedures with cognitively impaired patients: A review of ethics and best practices.
The objectives of this article are to discuss ethical issues of informed consent in cognitively impaired patients and review considerations for capacity determination. We will also discuss how to evaluate capacity, determine competence, and obtain informed consent when a patient is deemed incompetent. This review emphasizes how to carry out informed consent procedures when capacity is questionable and discusses measures supported for use when determining cognitively impaired patients' ability to consent. Information was gathered from medical and psychological codes of ethics, peer-reviewed journals, published guidelines from health-care organizations (e.g., American Medical Association), and scholarly books. Google Scholar and PsycINFO were searched for articles related to 'informed consent' and 'cognitive impairment' published in English between 1975 and 2014. Relevant sources referenced in retrieved publications were subsequently searched and reviewed. We selected 49 sources generated by our search. Sources were included in our review if they presented information related to at least one of our focus areas. These areas included: review of informed consent ethics and procedures, review of cognitive impairment evaluations, recommendations for measuring cognitive capacity, and alternative forms of informed consent. Patients' cognitive impairments can hinder the ability of patients to understand treatment options. Evaluating the capacity of patients with cognitive impairment to understand treatment options is vital for valid informed consent and should be guided by best practices. Thus, proper identification of patients with questionable capacity, capacity evaluation, and determination of competence, as well as reliance upon appropriate alternative consent procedures, are paramount.


Enhanced resistance to Ca2+-induced mitochondrial permeability transition in the long-lived red-footed tortoise Chelonoidis carbonaria.
The interaction between supraphysiological cytosolic Ca2+ levels and mitochondrial redox imbalance mediates the mitochondrial permeability transition (MPT). The MPT is involved in cell death, diseases and aging. This study compared the liver mitochondrial Ca2+ retention capacity and oxygen consumption in the long-lived red-footed tortoise (Chelonoidis carbonaria) with those in the rat as a reference standard. Mitochondrial Ca2+ retention capacity, a quantitative measure of MPT sensitivity, was remarkably higher in tortoises than in rats. This difference was minimized in the presence of the MPT inhibitors ADP and cyclosporine A. However, the Ca2+ retention capacities of tortoise and rat liver mitochondria were similar when both MPT inhibitors were present simultaneously. NADH-linked phosphorylating respiration rates of tortoise liver mitochondria represented only 30% of the maximal electron transport system capacity, indicating a limitation imposed by the phosphorylation system. These results suggested underlying differences in putative MPT structural components [e.g. ATP synthase, adenine nucleotide translocase (ANT) and cyclophilin D] between tortoises and rats. Indeed, in tortoise mitochondria, titrations of inhibitors of the oxidative phosphorylation components revealed a higher limitation of ANT. Furthermore, cyclophilin D activity was approximately 70% lower in tortoises than in rats. Investigation of critical properties of mitochondrial redox control that affect MPT demonstrated that tortoise and rat liver mitochondria exhibited similar rates of H2O2 release and glutathione redox status. Overall, our findings suggest that constraints imposed by ANT and cyclophilin D, putative components or regulators of the MPT pore, are associated with the enhanced resistance to Ca2+-induced MPT in tortoises.


Electrochemical performance of diamond thin-film electrodes from different commercial sources.
The electrochemical properties of two commercial (Condias, Sumitomo) boron-doped diamond thin-film electrodes were compared with those of two types of boron-doped diamond thin film deposited in our laboratory (microcrystalline, nanocrystalline). Scanning electron microscopy and Raman spectroscopy were used to characterize the electrode morphology and microstructure, respectively. Cyclic voltammetry was used to study the electrochemical response, with five different redox systems serving as probes (Fe(CN)(6)(3)(-)(/4)(-), Ru(NH(3))(6)(3+/)(2+), IrCl(6)(2)(-)(/3)(-), 4-methylcatechol, Fe(3+/2+)). The response for the different systems was quite reproducibile from electrode type to type and from film to film for electrodes of the same type. For all five redox systems, the forward reaction peak current varied linearly with the scan rate(1/2) (nu), indicative of electrode reaction kinetics controlled by mass transport (semi-infinite linear diffusion) of the reactant. Apparent heterogeneous electron-transfer rate constants, k degrees (app), for all five redox systems were determined from deltaE(p)-nu experimental data, according to the method described by Nicholson (Nicholson, R. S. Anal. Chem. 1965, 37, 1351.). The rate constants were also verified through digital simulation (DigiSim 3.03) of the voltammetric i-E curves at different scan rates. Good fits between the experimental and simulated voltammograms were found for scan rates up to 50 V/s. k degrees (app) values of 0.05-0.5 cm/s were observed for Fe(CN)(6)(3)(-)(/4)(-), Ru(NH(3))(6)(3+/2+), and IrCl(6)(2)(-)(/3)(-) without any extensive electrode pretreatment (e.g., polishing). Lower k degrees (app) values of 10(-)(4)-10(-)(6) cm/s were found for 4-methylcatechol and Fe(3+/2+). The voltammetric responses for Fe(CN)(6)(3)(-)(/4)(-) and Ru(NH(3))(6)(3+/2+) were also examined at all four electrode types at two different solution pH (1.90, 7.35). Since the hydrogen-terminated diamond surfaces contain few, if any, ionizable carbon-oxygen functionalities (e.g., carboxylic acid, pK(a) approximately 4.5), the deltaE(p), i(p)(ox), and i(p)(red) values for the two systems were, for the most part, unaffected by the solution pH. This is in contrast to the typical behavior of oxygenated, sp(2) carbon electrodes, such as glassy carbon.


Potent inhibition of anoxic depolarization by the sodium channel blocker dibucaine.
Recurring waves of peri-infarct depolarizations (PIDs) propagate across gray matter in the hours and days following stroke, expanding the primary site of injury. Ischemic depolarization (termed anoxic depolarization or AD in live brain slices) is PID-like but immediately arises in the more metabolically compromised ischemic core. This causes dramatic neuronal and astrocyte swelling and dendritic beading with spine loss within minutes, resulting in acute cell death. AD is evoked in rodent neocortical slices by suppressing the Na(+)/K(+)-ATPase pump with either oxygen/glucose deprivation (OGD) or exposure to ouabain. The process driving AD and PIDs remains poorly understood. Here we show that dibucaine is a potent drug inhibiting AD because of its high binding affinity to the Na(+) channel. Field recording reveals that, when superfused with ouabain (5 min), neocortical slices pretreated with 1 μM dibucaine for 45 min display either no AD or delayed AD onset compared with untreated controls. If ouabain exposure is extended to 10 min, 1 μM dibucaine is still able to delay AD onset by ∼ 60%. Likewise, it delays OGD-evoked AD onset by ∼ 54% but does not depress action potentials (APs) or evoked orthodromic field potentials. Increasing dibucaine to 10 μM inhibits AP firing, gradually putting the slice into a stasis that inhibits AD onset but also renders the slice functionally quiescent. Two-photon microscopy reveals that 10 μM dibucaine pretreatment prevents or helps reverse ouabain-induced structural neuronal damage. Although the therapeutic range of dibucaine is quite narrow, dibucaine-like drugs could prove therapeutically useful in inhibiting PIDs and their resultant neuronal damage.


Remarkable influence of microwave heating on Morita-baylis-Hillman reaction in PEG-200.
Morita Baylis Hillman (MBH) reaction is used to introduce carbon-carbon or carbon-heteroatom bond in a molecule. The major drawback of this reaction is the relatively low product yield and long reaction time. Though notable changes have been made to improve the reaction rate and yield of MBH adduct by various groups, a reliable synthetic procedure under ambient temperature in presence moisture and air is remain unsolved. Continuing the effort to improve the rate and yield, we report here an eco-friendly and cost-effective method to generate MBH adducts. Non-volatile polyethylene glycol-200 is used as reusable solvents and the reaction was carried out under the influence of microwave energy. Microwave irradiation have a remarkable influence on PEG suspended 4-Diazabicyclo [2.2.2] octane (DABCO) catalysed MBH reaction between aldehydes and ethyl acrylate. Molecular weight of the PEG is found to have a significant influence on the reaction yield. PEG-200 was the most efficient solvent and in combination with DABCO, the medium can be recycled upto three more runs. This reaction condition is successfully applied to obtain MBH adduct of five different aldehydes in very short time with excellent yield and the required catalyst concentration was very low compared to standard MBH reaction. Since the MBH adduct is an important reactive intermediates for many complex organic syntheses, this approach can be successfully utilised as an alternative to existing reaction conditions. A new method was developed to improve the reaction rate and yield of MBH reaction The PEG 200-DABCO combination provides a sustainable, non-volatile, recyclable and environment friendly solvent medium to produce MBH adducts. This medium in combination with microwave energy proved to be very effective to introduce a new carbon-carbon or a carbon-heteroatom bond in a molecule.


Development of Circadian Sleep Regulation in the Rat: A Longitudinal Study Under Constant Conditions.
To better understand the development of sleep, we characterized the development of circadian rhythms in sleep and wakefulness in the artificially-reared, isolated rat pup using an experimental design that minimized the effects of maternal separation. Neonatal rats were reared in constant conditions (dim red light) while electroencephalographic and electromyographic signals were continuously recorded for up to 3 weeks. This time period spanned the preweaned and weaned ages. The distribution of sleep-wake states was analyzed to estimate the emergence of circadian rhythms. Overt ~24-hour rhythms in time spent awake and asleep appear by postnatal day (P)17. A marked bi-modal sleep-wake pattern was also observed, evidenced by the appearance of a pronounced ~12-hour component in the periodogram over the subsequent 3 days (P17-P21). This suggested the presence of two ~24-hour components consistent with the dual-oscillator concept. During this 3-day time window, waking bouts became longer resulting in a repartition of the duration of intervals without non-rapid-eye movement (NREM) sleep into short (<30 minutes) and longer inter-NREM sleep episodes. These longer waking bouts did not immediately result in an increase in NREM sleep delta (0.5-4.0 Hz) power, which is an index of sleep homeostasis in adult mammals. The sleep homeostatic response did not fully mature until P25. These results demonstrate that the maturation of circadian organization of sleep-wake behavior precedes the expression of mature sleep homeostasis.


Biodegradation during contaminant transport in porous media: 4. Impact of microbial lag and bacterial cell growth.
Miscible-displacement experiments were conducted to examine the impact of microbial lag and bacterial cell growth on the transport of salicylate, a model hydrocarbon compound. The impacts of these processes were examined separately, as well as jointly, to determine their relative effects on biodegradation dynamics. For each experiment, a column was packed with porous medium that was first inoculated with bacteria that contained the NAH plasmid encoding genes for the degradation of naphthalene and salicylate, and then subjected to a step input of salicylate solution. The transport behavior of salicylate was non-steady for all cases examined, and was clearly influenced by a delay (lag) in the onset of biodegradation. This microbial lag, which was consistent with the results of batch experiments, is attributed to the induction and synthesis of the enzymes required for biodegradation of salicylate. The effect of microbial lag on salicylate transport was eliminated by exposing the column to two successive pulses of salicylate, thereby allowing the cells to acclimate to the carbon source during the first pulse. Elimination of microbial lag effects allowed the impact of bacterial growth on salicylate transport to be quantified, which was accomplished by determining a cell mass balance. Conversely, the impact of microbial lag was further investigated by performing a similar double-pulse experiment under no-growth conditions. Significant cell elution was observed and quantified for all conditions/systems. The results of these experiments allowed us to differentiate the effects associated with microbial lag and growth, two coupled processes whose impacts on the biodegradation and transport of contaminants can be difficult to distinguish.


DOS, a novel pleckstrin homology domain-containing protein required for signal transduction between sevenless and Ras1 in Drosophila.
The specification of the R7 photoreceptor cell in the developing eye of Drosophila is dependent upon activation of the Sevenless (SEV) receptor tyrosine kinase. By screening for mutations that suppress signaling via a constitutively activated SEV protein, we have identified a novel gene, daughter of sevenless (dos). DOS is required not only for signal transduction via SEV but also in other receptor tyrosine kinase signaling pathways throughout development. The presence of an amino-terminally located pleckstrin homology domain and many potential tyrosine phosphorylation sites suggests that DOS functions as an adaptor protein able to interact with multiple signaling molecules. Our genetic analysis demonstrates that DOS functions upstream of Ras1 and defines a signaling pathway that is independent of direct binding of the DRK SH2/SH3 adaptor protein to the SEV receptor tyrosine kinase.


The effect of environmental change on vascular plant and cryptogam communities from the Falkland Islands and the Maritime Antarctic.
Antarctic terrestrial vegetation is subject to one of the most extreme climates on Earth. Currently, parts of Antarctica are one of the fastest warming regions on the planet. During 3 growing seasons, we investigated the effect of experimental warming on the diversity and abundance of coastal plant communities in the Maritime Antarctic region (cryptogams only) and the Falkland Islands (vascular plants only). We compared communities from the Falkland Islands (51 degrees S, mean annual temperature 7.9 degrees C), with those of Signy Island (60 degrees S, -2.1 degrees C) and Anchorage Island (67 degrees S, -2.6 degrees C), and experimental temperature manipulations at each of the three islands using Open Top Chambers (OTCs). Despite the strong difference in plant growth form dominance between the Falkland Islands and the Maritime Antarctic, communities across the gradient did not differ in total diversity and species number. During the summer months, the experimental temperature increase at 5 cm height in the vegetation was similar between the locations (0.7 degrees C across the study). In general, the response to this experimental warming was low. Total lichen cover showed a non-significant decreasing trend at Signy Island (p < 0.06). In the grass community at the Falkland Islands total vegetation cover decreased more in the OTCs than in adjacent control plots, and two species disappeared within the OTCs after only two years. This was most likely a combined consequence of a previous dry summer and the increase in temperature caused by the OTCs. These results suggest that small temperature increases may rapidly lead to decreased soil moisture, resulting in more stressful conditions for plants. The more open plant communities (grass and lichen) appeared more negatively affected by such changes than dense communities (dwarf shrub and moss).


Comparison of cytokine levels in depressed, manic and euthymic patients with bipolar disorder.
The neurobiology of bipolar disorder is not completely understood. Cytokines have received increasing attention as potential mediators of the interaction with immune, neuroendocrine system and specific pathways involved in mood, energy, and activity control. Previous reports have suggested the association of mania and bipolar depression with a proinflammatory state. However, they did not compare cytokine levels in all phases of bipolar disorder. Sixty-one bipolar patients were recruited for assessment of serum cytokine levels. Of these, 14 were in euthymic state, 23 and 24 were in manic and depressive episodes, respectively. A healthy comparison group included 25 healthy volunteers. Cytokines involved in Th1/Th2 balance, such as TNF-alpha, IL-2, IL-4, IL-6, IL-10, IFN-gamma, were examined by flow cytometry. During mania, proinflammatory cytokines, IL-2, IL-4 and IL-6, were increased in comparison with healthy subjects. Patients in depressive episode showed only increased IL-6 levels. There were no significant differences in cytokine levels between patients in remission and healthy subjects, except for IL-4. Mood symptoms showed a positive correlation with IL-6 and IL-2. These findings suggest that mania, and to a less extent, depression are associated with a proinflammatory state. These changes seem to be related to mood state, as changes in cytokine profile were more pronounced during acute episodes than in euthymia. This study provides further support to investigate the immune system as a target for future treatment development.


Cloning and sequencing of a dehalogenase gene encoding an enzyme with hydrolase activity involved in the degradation of gamma-hexachlorocyclohexane in Pseudomonas paucimobilis.
In Pseudomonas paucimobilis UT26, gamma-hexachlorocyclohexane (gamma-HCH) is converted by two steps of dehydrochlorination to a chemically unstable intermediate, 1,3,4,6-tetrachloro-1,4-cyclohexadiene (1,4-TCDN), which is then metabolized to 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-DDOL) by two steps of hydrolytic dehalogenation via the chemically unstable intermediate 2,4,5-trichloro-2,5-cyclohexadiene-1-ol (2,4,5-DNOL). To clone a gene encoding the enzyme responsible for the conversion of the chemically unstable intermediates 1,4-TCDN and 2,4,5-DNOL, a genomic library of P. paucimobilis UT26 was constructed in Pseudomonas putida PpY101LA into which the linA gene had been introduced by Tn5. An 8-kb BglII fragment from one of the cosmid clones, which could convert gamma-HCH to 2,5-DDOL, was subcloned, and subsequent deletion analyses revealed that a ca. 1.1-kb region was responsible for the activity. Nucleotide sequence analysis revealed an open reading frame (designated the linB gene) of 885 bp within the region. The deduced amino acid sequence of LinB showed significant similarity to hydrolytic dehalogenase, DhlA (D. B. Janssen, F. Pries, J. van der Ploeg, B. Kazemier, P. Terpstra, and B. Witholt, J. Bacteriol. 171:6791-6799, 1989). The protein product of the linB gene was 32 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Not only 1-chlorobutane but also 1-chlorodecane (C10) and 2-chlorobutane, which are poor substrates for other dehalogenases, were good substrates for LinB, suggesting that LinB may be a member of haloalkane dehalogenases with broad-range specificity for substrates.


Novel PRKD gene rearrangements and variant fusions in cribriform adenocarcinoma of salivary gland origin.
Polymorphous low-grade adenocarcinoma (PLGA) and cribriform adenocarcinoma of minor salivary gland (CAMSG) are low-grade carcinomas arising most often in oral cavity and oropharynx, respectively. Controversy exists as to whether these tumors represent separate entities or variants of one spectrum, as they appear to have significant overlap, but also clinicopathologic differences. As many salivary carcinomas harbor recurrent translocations, paired-end RNA sequencing and FusionSeq data analysis was applied for novel fusion discovery on two CAMSGs and two PLGAs. Validated rearrangements were then screened by fluorescence in situ hybridization (FISH) in 60 cases. Histologic classification was performed without knowledge of fusion status and included: 21 CAMSG, 18 classic PLGA, and 21 with "mixed/indeterminate" features. The RNAseq of 2 CAMSGs showed ARID1A-PRKD1 and DDX3X-PRKD1 fusions, respectively, while no fusion candidates were identified in two PLGAs. FISH for PRKD1 rearrangements identified 11 additional cases (22%), two more showing ARID1A-PRKD1 fusions. As PRKD2 and PRKD3 share similar functions with PRKD1 in the diacylglycerol and protein kinase C signal transduction pathway, we expanded the investigation for these genes by FISH. Six additional cases each showed PRKD2 and PRKD3 rearrangements. Of the 26 (43%) fusion-positive tumors, there were 16 (80%) CAMSGs and 9 (45%) indeterminate cases. A PRKD2 rearrangement was detected in one PLGA (6%). We describe novel and recurrent gene rearrangements in PRKD1-3 primarily in CAMSG, suggesting a possible pathogenetic dichotomy from "classic" PLGA. However, the presence of similar genetic findings in half of the indeterminate cases and a single PLGA suggests a possible shared pathogenesis for these tumor types.


The initiation of breast and prostate cancer.
The agents responsible for the initiation of human mammary and prostatic cancers remain unidentified. Population migration studies on breast and prostate cancer risk have revealed that incidence rates in migrants from low-risk to high-risk "Westernized" countries rise over time to match those of the host populations. The parallels suggest that the two diseases may share a common aetiology, with changes in diet, rather than in environment, being responsible for the migration-related increases in cancer incidence. Genotoxins, such as polycyclic aromatic hydrocarbons and heterocyclic aromatic amines, are formed when foodstuffs are cooked at elevated temperatures and can be extracted with solvents: other genotoxins may only be released from cooked proteins when digestion occurs in the gastrointestinal tract. Human mammary and prostatic epithelial cells are known to be capable of metabolically activating members of different classes of chemical carcinogens to DNA-reactive species and, in rodents, five out of six mammary carcinogens can also induce prostatic neoplasms. Genotoxins have been detected in some 40% of breast lipid and milk samples donated by UK-resident women but the agents, currently thought to be of dietary origin, have not been characterized or identified as yet. Reduction mammoplasty and lactation both reduce breast cancer risk and the reduction is proportional either to the amount of tissue removed or to the total duration of lactation. As DNA damage has been detected in otherwise untreated mammary epithelial cells isolated both from breast tissue and from breast milk, we have proposed that reduction mammoplasty and lactation reduce risk through a common mechanism, i.e. the loss of pre-malignant cells. Further research, perhaps aimed particularly at the characterization of all the carcinogens formed when different dietary components are cooked in different ways, should succeed in identifying the agents that initiate breast and prostate cancer.


Characterization of interactions between inclusion membrane proteins from Chlamydia trachomatis.
Chlamydiae are obligate intracellular pathogens of eukaryotes. The bacteria grow in an intracellular vesicle called an inclusion, the membrane of which is heavily modified by chlamydial proteins called Incs (Inclusion membrane proteins). Incs represent 7-10% of the genomes of Chlamydia and, given their localization at the interface between the host and the pathogen, likely play a key role in the development and pathogenesis of the bacterium. However, their functions remain largely unknown. Here, we characterized the interaction properties between various Inc proteins of C. trachomatis, using a bacterial two-hybrid (BACTH) method suitable for detecting interactions between integral membrane proteins. To validate this approach, we first examined the oligomerization properties of the well-characterized IncA protein and showed that both the cytoplasmic domain and the transmembrane region independently contribute to IncA oligomerization. We then analyzed a set of Inc proteins and identified novel interactions between these components. Two small Incs, IncF, and Ct222, were found here to interact with many other Inc proteins and may thus represent interaction nodes within the inclusion membrane. Our data suggest that the Inc proteins may assemble in the membrane of the inclusion to form specific multi-molecular complexes in an hierarchical and temporal manner. These studies will help to better define the putative functions of the Inc proteins in the infectious process of Chlamydia.


Tobramycin resistance of Pseudomonas aeruginosa cells growing as a biofilm on urinary catheter material.
When disks of urinary catheter material were exposed to the flow of artificial urine containing cells of Pseudomonas aeruginosa, a thick adherent biofilm, composed of these bacteria and of their exopolysaccharide products, developed on the latex surface within 8 h. After this colonization, sterile artificial urine containing 1,000 micrograms of tobramycin per ml was flowed past this established biofilm, and a significant proportion of the bacterial cells within the biofilm were found to be still viable after 12 h of exposure to this very high concentration of aminoglycoside antibiotic. Planktonic (floating) cells taken from the test system just before the exposure of the biofilm to the antibiotic were completely killed by 50 micrograms of tobramycin per ml. The MIC of tobramycin for cells taken from the seeding cultures before colonization of the catheter material, and for surviving cells recovered directly from the tobramycin-treated biofilm, was found to be 0.4 micrograms/ml when dispersed cells were assayed by standard methods. These data indicate that growth within thick adherent biofilms confers a measure of tobramycin resistance on cells of P. aeruginosa.


Spontaneous regression of hepatocellular carcinoma in a Caucasian male patient: A case report and review of the literature.
Hepatocellular carcinoma (HCC) is the most common primary hepatic cancer. As it often presents at an advanced stage, it generally has a poor prognosis. Spontaneous regression is a rare finding in HCC, and is often associated with tumour ischemia or a systemic inflammatory response. We herein present the case of a Caucasian patient with spontaneous regression of advanced HCC, not associated with either mechanism. Our patient was a 74-year old Caucasian male who presented with advanced biopsy-proven HCC, with α-fetoprotein levels of >16,600 kU/l. On computed tomography examination, multiple metastases were identified. Due to the poor performance status and as the patient declined treatment, he was referred to a general practitioner for supportive palliative care. Six months later, the patient was alive and feeling well. He had suffered a cerebrovascular accident with ensuing partial hemiparesis, and had been initiated on enalapril, furosemide and curcumin. On re-evaluation, the lung lesions had disappeared, whereas both the liver and peritoneal lesions had decreased in size. Following a review of the literature, the conclusion was that this case of spontaneous regression of advanced HCC was not caused by ischemia, inflammation, or any other previously reported mechanism. Further research is required to elucidate the mechanisms underlying spontaneous regression of HCC.


Symbiotic bacterium modifies aphid body color.
Color variation within populations of the pea aphid influences relative susceptibility to predators and parasites. We have discovered that infection with a facultative endosymbiont of the genus Rickettsiella changes the insects' body color from red to green in natural populations. Approximately 8% of pea aphids collected in Western Europe carried the Rickettsiella infection. The infection increased amounts of blue-green polycyclic quinones, whereas it had less of an effect on yellow-red carotenoid pigments. The effect of the endosymbiont on body color is expected to influence prey-predator interactions, as well as interactions with other endosymbionts.


Children's participation in vaccine research: parents' views.
Vaccine studies that evaluate the persistence of protection following immunisation require subjects to continue participation in a research protocol over many years. As parents' attitudes and opinions may change over time, and with experience of research, it is important to consider the factors influencing parents' decision-making about their child's continued participation in such prolonged vaccine studies. Parental views about participation of their child in a one-year follow-up vaccine study were explored by means of a self-administered questionnaire. Of the 254 eligible parents, 187 took part (74 per cent). Parents who provided consent were more likely to agree that having a home visit to take blood was very helpful (p=0.005) and that information obtained during the earlier part of the study influenced their decision to take part in a follow-up study (p<0.0001). Parents who did not consent to their child's participation were more likely to report the presence of personal reasons as a variable influencing their decision (p<0.0001). The relationship between study staff and parents is the cornerstone for success in performing studies involving vaccines and children. Provision of clear study information (oral and written) and offering the convenience of home visits are important in retaining participants in paediatric vaccine trials.


In Vivo Cleavage Map Illuminates the Central Role of RNase E in Coding and Non-coding RNA Pathways.
Understanding RNA processing and turnover requires knowledge of cleavages by major endoribonucleases within a living cell. We have employed TIER-seq (transiently inactivating an endoribonuclease followed by RNA-seq) to profile cleavage products of the essential endoribonuclease RNase E in Salmonella enterica. A dominating cleavage signature is the location of a uridine two nucleotides downstream in a single-stranded segment, which we rationalize structurally as a key recognition determinant that may favor RNase E catalysis. Our results suggest a prominent biogenesis pathway for bacterial regulatory small RNAs whereby RNase E acts together with the RNA chaperone Hfq to liberate stable 3' fragments from various precursor RNAs. Recapitulating this process in vitro, Hfq guides RNase E cleavage of a representative small-RNA precursor for interaction with a mRNA target. In vivo, the processing is required for target regulation. Our findings reveal a general maturation mechanism for a major class of post-transcriptional regulators.


Development and performance of inactivated vaccines against foot and mouth disease.
The historical background of foot and mouth disease (FMD) vaccine production is briefly described. Improvements achieved through the use of monolayer and suspension cultures are outlined. Elements that are crucial in the production of modern vaccines are discussed, such as inactivation of viral antigen, successive concentration and purification of the antigen and the final formulation of the vaccine. Storage of concentrated antigen at ultra-low temperatures creates greater flexibility for the producer and has also enabled national and international organisations to establish vaccine banks. The purification of FMD viral antigens, including the removal of non-structural proteins (NSPs), enables the immune responses of vaccinated animals to be distinguished from the responses of animals infected with live FMD virus. Consequently, the combined use of purified vaccine and tests for the detection of antibodies against NSPs essentially provides a marker system to distinguish between vaccinated animals that subsequently become infected and those that have not. Bearing in mind the good record of modern vaccines in the control of outbreaks and the possibility of screening vaccinated herds for carriers, the author proposes that the OIE reconsider the differences between the requirements for regaining export status following the use of stamping-out as opposed to vaccination in outbreak situations.


Tissue and gene specificity of mouse renin expression.
The Ren-1 locus of mice encodes the protease renin, which with converting enzyme processes angiotensinogen to the potent vasopressor angiotensin II. Some strains of mice appear to carry a duplication of the renin structural gene (Ren-2) near the Ren-1 locus. Strains with the gene duplication can exhibit as much as 100-fold higher levels of submaxillary gland renin compared to strains with a single gene copy. In contrast, kidney renin levels appear to be unaffected by the gene duplication. Sequence analysis of a 319 bp renin cDNA recombinant isolated from a kidney library from the two-gene strain DBA/2Ha corresponds to a transcript of the Ren-1 gene. Moreover, a single base substitution of A for G at residue #996 in the kidney renin mRNA creates a potential glycosylation recognition site that may, in part, account for the differential glycosylation of kidney and submaxillary gland renins. In addition, our tissue surveys indicate that mature mRNAs from the Ren loci are detectable in adrenal gland and testes, as well as sublingual and parotid salivary glands, and reveal length variation for the renin transcripts in at least the submaxillary gland.


PSYCHOLOGY. Estimating the reproducibility of psychological science.
Reproducibility is a defining feature of science, but the extent to which it characterizes current research is unknown. We conducted replications of 100 experimental and correlational studies published in three psychology journals using high-powered designs and original materials when available. Replication effects were half the magnitude of original effects, representing a substantial decline. Ninety-seven percent of original studies had statistically significant results. Thirty-six percent of replications had statistically significant results; 47% of original effect sizes were in the 95% confidence interval of the replication effect size; 39% of effects were subjectively rated to have replicated the original result; and if no bias in original results is assumed, combining original and replication results left 68% with statistically significant effects. Correlational tests suggest that replication success was better predicted by the strength of original evidence than by characteristics of the original and replication teams.


Travel to, and use of, twenty-one Michigan trails.
This study examined trail use among 857 trail users on 21 trails in Michigan from 2008 to 2011 using a valid and reliable intercept survey. Most of the 857 participants traveled to the trail from their home (92.6%), lived within 15 min of the trails (74.8%), and used active transport to travel to the trails 69.7%. The odds of active transport to the trails were greater among those who had not graduated high school (OR=3.49; 95% CI=1.02, 11.99) and high school graduates (OR=7.432; 95% CI=2.02, 27.30) compared to college graduates. Whites and adults also had greater odds of active transport than non-Whites (OR=3.160, 95% CI: 1.65, 6.05), and older adults (OR=1.75; 95% CI: 1.20, 2.54). The majority of respondents (89.7%) reported using trails for recreational purposes. A significantly greater proportion of females (73.3%) compared to males (64.7%) reported using the trail with others. The findings from this study might enable health and parks and recreation professionals to better promote physical activity on trails.


A new approach for monitoring ebolavirus in wild great apes.
Central Africa is a "hotspot" for emerging infectious diseases (EIDs) of global and local importance, and a current outbreak of ebolavirus is affecting multiple countries simultaneously. Ebolavirus is suspected to have caused recent declines in resident great apes. While ebolavirus vaccines have been proposed as an intervention to protect apes, their effectiveness would be improved if we could diagnostically confirm Ebola virus disease (EVD) as the cause of die-offs, establish ebolavirus geographical distribution, identify immunologically naïve populations, and determine whether apes survive virus exposure. Here we report the first successful noninvasive detection of antibodies against Ebola virus (EBOV) from wild ape feces. Using this method, we have been able to identify gorillas with antibodies to EBOV with an overall prevalence rate reaching 10% on average, demonstrating that EBOV exposure or infection is not uniformly lethal in this species. Furthermore, evidence of antibodies was identified in gorillas thought previously to be unexposed to EBOV (protected from exposure by rivers as topological barriers of transmission). Our new approach will contribute to a strategy to protect apes from future EBOV infections by early detection of increased incidence of exposure, by identifying immunologically naïve at-risk populations as potential targets for vaccination, and by providing a means to track vaccine efficacy if such intervention is deemed appropriate. Finally, since human EVD is linked to contact with infected wildlife carcasses, efforts aimed at identifying great ape outbreaks could have a profound impact on public health in local communities, where EBOV causes case-fatality rates of up to 88%.


Gonioscopic features in patients with acute and chronic angle-closure glaucoma.
A number of ocular biometric parameters, iris hiotologic and anatomic characters have been suggested as inciting factors for converting patients with narrow angle to angle-closure glaucoma. This study was conducted to determine if there was any goniscopic difference between patients with acute angle-closure glaucoma (AACG) and chronic angle-closure glaucoma (CACG). The study is a retrospective analysis of the charts of 97 patients with asymmetric CACG and 15 patients with unilateral AACG. The age, sex, type of glaucoma, gonioscopic findings and optic nerve head cup/disc ratio were recorded for all patients. Dynamic gonioscopy and Spaeth's convention were used to grade the drainage angle. The eyes with AACG or more optic nerve damage in CACG groups were considered as involved eye, and the contralateral eyes in the AACG and CACG groups were considered as noninvolved and less-involved, respectively. There was no significant difference between patients with AACG and CACG in terms of age, gender, refraction, and laterality of the involved eyes. In intragroup analysis, no significant difference was observed for distribution of iris attachment, irido-corneal angle, iris configuration, or trabecular pigmentation. In intergroup analysis, the superior iris was attached more anterior in the involved eyes of AACG compared to that in CACG (P=0.007). Moreover, the iris root attachment was also more anterior in both the superior (P=0.001) and inferior (P=0.002) angles of the noninvolved eyes of AACG vs. than those in the less-involved eyes of CACG group. The findings of the study indicate that there is no significant difference between the eyes with AACG or CACG in terms of goniscopic findings. However, the superior iris attachment was located more anterior in eyes with AACG compared to that in eyes with CACG.


The association between self-reported lack of sleep, low vitality and impaired glucose tolerance: a Swedish cross-sectional study.
The increased incidence of impaired glucose tolerance (IGT), are serious public health issues, and several studies link sleeping disorders with increased risk of developing type 2 diabetes, impaired glucose tolerance and insulin resistance (IR). This study explore how self-reported lack of sleep and low vitality, are associated with IGT in a representative Swedish population. A cross-sectional survey conducted in two municipalities in South-western Sweden. Participants aged 30-75 were randomly selected from the population in strata by sex and age. Altogether, 2,816 participants were surveyed with a participation rates at 76%. Participants with normal glucose tolerance (n=2,314), and those with IGT (n=213) were retained for analyses. The participants answered a questionnaire before the oral glucose tolerance test (OGTT). Associations for questions concerning sleeping disorders, vitality and IGT were analysed using logistic regression and were expressed as odds ratios (OR) with 95% CI. In men a statistically significant age-adjusted association was found between self-reported lack of sleep and IGT: OR 2.4 (95% CI: 1.1-5.4). It did not weaken after further adjustment for body mass index (BMI), smoking, education, and leisure time physical activity 2.3 (1.0-5.5, p=0.044). No such associations were found in females. Corresponding age-adjusted associations between low vitality and IGT in both men 2.8 (1.3-5.8), and women 2.0 (1.2-3.4) were successively lost with increasing adjustment. Insufficient sleep seems independently associated with IGT in men, while low vitality was not independently associated with IGT neither in men nor women, when multiple confounders are considered. IGT should be considered in patients presenting these symptoms, and underlying mechanisms further explored.


Profiling of a prescription drug library for potential renal drug-drug interactions mediated by the organic cation transporter 2.
Drug-drug interactions (DDIs) are major causes of serious adverse drug reactions. Most DDIs have a pharmacokinetic basis in which one drug reduces the elimination of a second drug, leading to potentially toxic drug levels. As a major organ of drug elimination, the kidney represents an important site for DDIs. Here, we screened a prescription drug library against the renal organic cation transporter OCT2/SLC22A2, which mediates the first step in the renal secretion of many cationic drugs. Of the 910 compounds screened, 244 inhibited OCT2. Computational analyses revealed key properties of inhibitors versus noninhibitors, which included overall molecular charge. Four of six potential clinical inhibitors were transporter-selective in follow-up screens against additional transporters: OCT1/SLC22A1, MATE1/SLC47A1, and MATE2-K/SLC47A2. Two compounds showed different kinetics of interaction with the common polymorphism OCT2-A270S, suggesting a role of genetics in modulating renal DDIs.


Evaluation of 3 different registration techniques in image-guided bimaxillary surgery.
Perioperative navigation is an upcoming tool in orthognathic surgery. This study aimed to access the feasibility of the technique and to evaluate the success rate of 3 different registration methods--facial surface registration, anatomic landmark-based registration, and template-based registration. The BrainLab navigation system (BrainLab AG, Feldkirchen, Germany) was used as an additional precision tool for 85 patients who underwent bimaxillary orthognathic surgery from February 2010 to June 2012. Eighteen cases of facial surface-based registration, 63 cases of anatomic landmark-based registration, and 8 cases of template-based registration were analyzed. The overall success rate of facial surface-based registration was 39%, which was significant lower than template-based (100%, P = 0.013) and anatomic landmark-based registration (95%, P < 0.0001). In all cases with successful registration, the further procedure of surgical navigation was performed. The concept of navigation of the maxilla during bimaxillary orthognathic surgery has been proved to be feasible. The registration process is the critical point regarding success of intraoperative navigation. Anatomic landmark-based registration is a reliable technique for image-guided bimaxillary surgery. In contrast, facial surface-based registration is highly unreliable.


Imaging of haemodialysis-associated amyloidosis with 123I-serum amyloid P component.
Long-term haemodialysis is frequently complicated by amyloid deposition in which the fibrils consist of beta 2-microglobulin. Dialysis-related amyloid disease causes extensive morbidity and has been associated with deaths in some cases. All amyloid deposits contain amyloid P component that is derived from the normal circulating protein, serum amyloid P component (SAP). We have used scintigraphic imaging after injection of 123I-labelled SAP to assess the distribution of amyloidosis in 38 patients receiving long-term haemodialysis for end-stage renal failure. There was focal localisation of tracer at all sites where histological examination confirmed amyloid deposition. Splenic uptake was seen in 12 patients, indicating splenic amyloidosis, but there was no evidence of other visceral involvement. 6 control subjects who had been dialysed for under 1.5 years showed no localisation of tracer, nor was there any uptake of 123I-labelled human serum albumin in 3 long-term dialysis patients with histologically confirmed amyloidosis and positive 123I-SAP images. Negative scans were also obtained in 5 patients who had been transplanted 0.8-2.4 years previously, despite past evidence of dialysis arthropathy (5) and histologically proven amyloidosis (4). 123I-SAP scintigraphy may be helpful as a non-invasive method for both the diagnosis and monitoring of dialysis-associated amyloidosis.


Granulocyte-colony stimulating factor and umbilical cord blood cell transplantation: Synergistic therapies for the treatment of traumatic brain injury.
Traumatic brain injury (TBI) is now characterized as a progressive, degenerative disease and continues to stand as a prevalent cause of death and disability. The pathophysiology of TBI is complex, with a variety of secondary cell death pathways occurring which may persist chronically following the initial cerebral insult. Current therapeutic options for TBI are minimal, with surgical intervention or rehabilitation therapy existing as the only viable treatments. Considering the success of stem-cell therapies in various other neurological diseases, their use has been proposed as a potential potent therapy for patients suffering TBI. Moreover, stem cells are highly amenable to adjunctive use with other therapies, providing an opportunity to overcome the inherent limitations of using a single therapeutic agent. Our research has verified this additive potential by demonstrating the efficacy of co-delivering human umbilical cord blood (hUCB) cells with granulocyte-colony stimulating factor (G-CSF) in a murine model of TBI, providing encouraging results which support the potential of this approach to treat patients suffering from TBI. These findings justify ongoing research toward uncovering the mechanisms which underlie the functional improvements exhibited by hUCB + G-CSF combination therapy, thereby facilitating its safe and effect transition into the clinic. This paper is a review article. Referred literature in this paper has been listed in the reference section. The datasets supporting the conclusions of this article are available online by searching various databases, including PubMed. Some original points in this article come from the laboratory practice in our research center and the authors' experiences.


Development of an assessment tool to measure the influence of clinical software on the delivery of high quality consultations. A study comparing two computerized medical record systems in a nurse run heart clinic in a general practice setting.
A rating scale was developed to assess the contribution made by computer software towards the delivery of a quality consultation, with the purpose of informing the development of the next generation of systems. Two software programmes were compared, using this scale to test their ability to enable or inhibit the delivery of an ideal consultation with a patient with heart disease. The context was a general practice based, nurse run clinic for the secondary prevention of heart disease. One of the programmes was customized for this purpose; the other was a standard general practice programme. Consultations were video-recorded, and then assessed by an expert panel using the new assessment tool. Both software programmes were oriented towards the implementation of the evidence, rather than facilitating patient-centred practice. The rating scale showed, not surprisingly, significantly greater support from the customized software in the consultation in five out of eight areas. However, the scale's reliability measured by Cronbach's Alpha, was sub-optimal. With further refinement, this rating scale may become a useful tool that will inform software developers of the effectiveness of their programmes in the consultation, and suggest where they need development.


6-Methyluracil derivatives as acetylcholinesterase inhibitors for treatment of Alzheimer's disease.
Alzheimer's disease (AD) is the major age-related progressive neurodegenerative disorder. The brain of AD patients suffers from loss of cholinergic neurons and decreased number of synapses [1]. AD is caused by an imbalance between Aβ production and clearance, resulting in increased amount of Aβ in various forms [2]. Reduction of Aβ production and increasing clearance of Aβ pathogenic forms are key targets in the development of potential therapeutic agents for AD treatment. Unfortunately, only nosotropic approaches for treatment of AD are currently effective in humans. These approaches mainly focus on the inhibition of brain acetyl-cholinesterase (AChE) to increase lifetime of cerebral acetylcholine [3]. It is important to emphasize that AChE itself promotes the formation of Aβ fibrils in vitro and Aβ plaques in the cerebral cortex of transgenic mouse models of AD [4]. This property of AChE results from interaction between Aβ and the peripheral anionic site of the enzyme (PAS) [5]. Dual binding site inhibitors of both catalytic active site (CAS) and PAS can simultaneously improve cognition and slow down the rate of Aβ-induced neural degeneration. Unfortunately, the assortment of AChE PAS ligands is still extremely limited. To study putative advantages of AChE non-charged PAS inhibitors based on 6-methyluracil derivatives for the treatment of Alzheimer's disease. In vitro studies. Concentration of drug producing 50% of AChE/BuChE activity inhibition (IC50) was measured using the method of Ellman et al. [6]. Toxicological experiments were performed using IP injection of the different compounds in mice. LD50, dose (in mg/kg) causing lethal effects in 50% of animals was taken as a criterion of toxicity [7]. The ability of compound to block in vitro AChE-induced Aβ1-40 aggregation was studied using a thioflavin T (ThT) fluorescent probe [8].In vivo biological assays. For in vivo blood-brain barrier permeation assay brains were removed 30 min after IP injection of LD50 dose of tested compound injection. The inhibitory potency was measured using the method of Ellman.Scopolamine and transgenic models of AD were used to evaluate the influence of compound 35 on spatial memory performance.Water solution of scopolamine was injected to mice (ip) 20 minutes before starting memory test during 14 days [9]. Mice were assigned to 7 groups, including 4 groups receiving injection (ip) of compound in different dosages, donepezil-treated mice (donepezil is conventionally used to treat Alzheimer's disease), positive and negative control groups. Double transgenic (APP/PS1) mice expressing a chimeric mouse/human amyloid precursor protein and a mutant of human presenilin-1 [10] were assigned to 4 groups, including transgenic animals injected (ip) with compound 35 or donepezil solution, positive (transgenes injected with water) and negative (wild-type mice) controls.To evaluate spatial memory performance, mice were trained on a reward alternation task using a conventional T-maze [11]. The criterion for a mouse having learned the rewarded alternation task was 3 consecutive days of at least 5 correct responses out of the 6 free trials.For β-amyloid peptide load was evaluated quantitatively as a number and summary area of Thioflavine S fluorescent spots in cerebral cortex and hippocampal images using Image J program. Statistical analyses were performed using the Mann-Whitney test. We evaluated the acute toxicity of the most active compounds. The most potent AChE inhibitor compound 35 (IC50 (AChE) = 5 ± 0.5 nM) exhibited the lowest LD50 values (51 mg/kg) and inhibited brain AChE by more than 71 ± 1%. Compound 35 at 10 nM, exhibited a significant (35 ± 9%) inhibitory activity toward human AChE-induced Aβ aggregation.Scopolamine injection induced significant decrease in correct choice percentage in T-maze, as well as decrease in percentage of mice reaching criterion for learning the task by day 14. This memory deficit was relieved to some extent either by compound 35 (5 mg/kg) or donepezil (reference compound) treatment (0.75 mg/kg). Interestingly, higher doses of compound 35 (10 and 15 mg/kg) produced less therapeutic effect on spatial memory deficit.Group of APP/PS1 mice showed 3 times lower percentage of reaching behavioral criterion and lower percentage of correct choice in T-maze alternation task comparing to WT mice, whereas compound 35 (5 mg/kg) or Donepezil treatment effectively improved these parameters in APP/PS1 mice.Compound 35 treatment (5 mg/kg) during 14 days significantly reduced percentage of summary area and number of β-amyloid peptide (βAP) deposits visualized in sections of cerebral cortex, dentate gyrus, and hippocampal CA3 area in APP/PS1 mice. The most prominent reduction of βAP load by compound 35 treatment was found in CA3 area and cerebral cortex. Meanwhile, Donepezil treatment (1 mg/kg) during 14 days significantly reduced βAP load in cerebral cortex but not in dentate gyrus and CA3 area. Experiments showed that the most potent AChE inhibitor compound 35 (6-methyluracil derivative) permeated the blood-brain barrier, improved working memory in the APP/PS1 transgenic mice and significantly reduced the number and area of Aβ plaques in the brain. Thus, compound 35 is a promising candidate as a bi-functional inhibitor of AChE for treatment of AD.


Bullying involvement, psychological distress, and short sleep duration among adolescents.
Previous research has found links between involvement in bullying and sleep duration in adolescents. However, little is known about the factors that might mediate these relationships. This study examined the associations between cyberbullying victimization and school bullying involvement (bully, victim, bully-victim) with short sleep duration in a large sample of middle and high school students and tested whether psychological distress mediates these relationships. Data on 5061 students (49% females; mean age = 15.1 years) from the 2015 Ontario Student Drug Use and Health Survey were used. Participants self-reported their sleep duration and their levels of bullying involvement over the past year. Psychological distress was assessed using the Kessler 6 (K6) scale. Covariates in multiple linear regression analyses included age, sex, racial background, socioeconomic status, and substance use. Being a victim of cyberbullying (β = - 1.179; 95% CI - 0.238; - 0.120) or school bullying (β = - 0.119; 95% CI - 0.190; - 0.049) was associated with short sleep duration. Mediation analyses suggested that psychological distress fully mediates the relationships between being cyberbullied, a school bullying victim or bully-victim with short sleep duration. There was a complementary mediation by psychological distress on the relationship between being a bully at school and short sleep duration. These results suggest that higher levels of bullying involvement place adolescents at risk of developing higher psychological distress which, in turn, can lead to short sleep duration. Longitudinal research is necessary to confirm the mediating role of psychological distress on the relationship between bullying involvement and short sleep duration among adolescents.


A phase II, multicenter trial of weekly topotecan in patients with recurrent platinum-sensitive epithelial cancers of the ovary and peritoneum.
The purpose of this study was to evaluate the response rate and toxicity of weekly topotecan in patients with recurrent platinum-sensitive epithelial cancers of the ovary and peritoneum. Thirty-nine platinum-sensitive recurrent ovarian cancer patients received topotecan (4 mg/m(2)) intravenously day 1, day 8, day 15, every 28 days. Colony-stimulating factors were excluded from the study. Clinical response was assessed by clinical, serologic, and radiographic measures at the conclusion of cycle four. Patients received 136 cycles of topotecan (median = 3; range 1-6) and were evaluated for response and toxicity. Median number of prior regimens was one. Grade 3/4 neutropenia developed in 3 (7.7%) patients. Grade 3 thrombocytopenia was seen in one (2.6%) patient, with no incidence of grade 4 thrombocytopenia. There was no evidence of grade 3 anemia, but one patient (2.6%) was associated with grade 4 anemia. There was no grade 3 or 4 neuropathy. We encountered 18 dose reductions following less than or equal to grade 2 myelosuppression, necessitating the removal of eight (20.5%) patients prior to cycle four. Twenty-one (53.8%) patients were removed from the study due to disease progression. Following the completion of cycle four, four (10.3%) patients demonstrated stable disease and four (10.3%) patients exhibited a partial response. There were no complete responses. Median disease-free survival was 12 weeks. Weekly topotecan (4 mg/m(2)) demonstrated modest activity and was moderately well tolerated. However, the significant number of dose reductions and high incidence of patients who demonstrated disease progression suggests additional modifications with this specific regimen are necessary.


Heat-related respiratory hospital admissions in Europe in a changing climate: a health impact assessment.
Respiratory diseases are ranked second in Europe in terms of mortality, prevalence and costs. Studies have shown that extreme heat has a large impact on mortality and morbidity, with a large relative increase for respiratory diseases. Expected increases in mean temperature and the number of extreme heat events over the coming decades due to climate change raise questions about the possible health impacts. We assess the number of heat-related respiratory hospital admissions in a future with a different climate. A Europe-wide health impact assessment. An assessment for each of the EU27 countries. Heat-related hospital admissions under a changing climate are projected using multicity epidemiological exposure-response relationships applied to gridded population data and country-specific baseline respiratory hospital admission rates. Times-series of temperatures are simulated with a regional climate model based on four global climate models, under two greenhouse gas emission scenarios. Between a reference period (1981-2010) and a future period (2021-2050), the total number of respiratory hospital admissions attributed to heat is projected to be larger in southern Europe, with three times more heat attributed respiratory hospital admissions in the future period. The smallest change was estimated in Eastern Europe with about a twofold increase. For all of Europe, the number of heat-related respiratory hospital admissions is projected to be 26 000 annually in the future period compared with 11 000 in the reference period. The results suggest that the projected effects of climate change on temperature and the number of extreme heat events could substantially influence respiratory morbidity across Europe.


Measuring attitudes towards smoking in the Community Intervention Trial for Smoking Cessation (COMMIT).
We present the development of indices using baseline data from the Evaluation Survey for the Community Intervention Trial for Smoking Cessation (COMMIT). The indices are designed to measure two primary attitude constructs that relate to smoking behavior: beliefs about smoking as a public health problem (SPHP); and norms and values concerning smoking (NVS). Two general approaches to index construction, the rational method and the factor analytic method, were used. Item analysis suggested good internal consistency for both indices (alpha > 0.75). Seven subconstructs emerged from the factor analysis accounting for 55.0% of the total variance. The SPHP and NVS items uniquely identify with four factors and three factors, respectively, confirming the validity of the two indices. Confirmatory factor analyses of a different data set provided further validation. Validity was also assessed by an examination of the relationships between index scores and smoking status. Smokers reported significantly higher scores than non-smokers on the two measures indicating, as anticipated, that smokers have more favorable attitudes towards smoking than non-smokers. These findings suggest that the two a priori constructs of SPHP and NVS are empirically distinguishable components of attitudes towards smoking, and that the indices developed here are reliable and valid measures of those constructs.


Activation of protein kinase C alters the intracellular distribution and mobility of cardiac Na+ channels.
Na(+) current derived from expression of the cardiac isoform SCN5A is reduced by receptor-mediated or direct activation of protein kinase C (PKC). Previous work has suggested a possible role for loss of Na(+) channels at the plasma membrane in this effect, but the results are controversial. In this study, we tested the hypothesis that PKC activation acutely modulates the intracellular distribution of SCN5A channels and that this effect can be visualized in living cells. In human embryonic kidney cells that stably expressed SCN5A with green fluorescent protein (GFP) fused to the channel COOH-terminus (SCN5A-GFP), Na(+) currents were suppressed by an exposure to PKC activation. Using confocal microscopy, colocalization of SCN5A-GFP channels with the plasma membrane under control and stimulated conditions was quantified. A separate population of SCN5A channels containing an extracellular epitope was immunolabeled to permit temporally stable labeling of the plasma membrane. Our results demonstrated that Na(+) channels were preferentially trafficked away from the plasma membrane by PKC activation, with a major contribution by Ca(2+)-sensitive or conventional PKC isoforms, whereas stimulation of protein kinase A (PKA) had the opposite effect. Removal of the conserved PKC site Ser(1503) or exposure to the NADPH oxidase inhibitor apocynin eliminated the PKC-mediated effect to alter channel trafficking, indicating that both channel phosphorylation and ROS were required. Experiments using fluorescence recovery after photobleaching demonstrated that both PKC and PKA also modified channel mobility in a manner consistent with the dynamics of channel distribution. These results demonstrate that the activation of protein kinases can acutely regulate the intracellular distribution and molecular mobility of cardiac Na(+) channels in living cells.


Electrical cortical stimulation of the human prefrontal cortex evokes complex visual hallucinations.
Complex visual hallucinations are a well-known feature of electrical stimulation or epileptic discharge in the temporal lobe. It has been proposed that these visual hallucinations result from an electrical interference with the ventral visual processing stream in the lateral temporal lobe and the memory system in medial temporal structures, which explains their frequent visual and mnestic features. Even though recent studies have demonstrated visual and memory functions in the prefrontal cortex, up to now epileptic discharge or electrical stimulation of prefrontal structures has only rarely been reported to induce visual phenomena. We report on two patients undergoing invasive presurgical epilepsy evaluation in which electrical cortical stimulation of the left prefrontal cortex repeatedly induced complex visual hallucinations. Interestingly, the induced visual responses differed with respect to their spatial organization: whereas those evoked on the inferior frontal gyrus were perceived in the whole visual field, complex visual responses on the middle frontal gyrus were restricted to the contralateral hemispace. Based on the spatial organization of the visual experiences in our patients, animal work, and neuroimaging data it might be suggested that specific subregions of the human prefrontal cortex might contain separate visual and mnemonic processing mechanisms.


Networks of energetic and metabolic interactions define dynamics in microbial communities.
Microorganisms form diverse communities that have a profound impact on the environment and human health. Recent technological advances have enabled elucidation of community diversity at high resolution. Investigation of microbial communities has revealed that they often contain multiple members with complementing and seemingly redundant metabolic capabilities. An understanding of the communal impacts of redundant metabolic capabilities is currently lacking; specifically, it is not known whether metabolic redundancy will foster competition or motivate cooperation. By investigating methanogenic populations, we identified the multidimensional interspecies interactions that define composition and dynamics within syntrophic communities that play a key role in the global carbon cycle. Species-specific genomes were extracted from metagenomic data using differential coverage binning. We used metabolic modeling leveraging metatranscriptomic information to reveal and quantify a complex intertwined system of syntrophic relationships. Our results show that amino acid auxotrophies create additional interdependencies that define community composition and control carbon and energy flux through the system while simultaneously contributing to overall community robustness. Strategic use of antimicrobials further reinforces this intricate interspecies network. Collectively, our study reveals the multidimensional interactions in syntrophic communities that promote high species richness and bolster community stability during environmental perturbations.


Lineages evolved under stronger sexual selection show superior ability to invade conspecific competitor populations.
Despite limitations on offspring production, almost all multicellular species use sex to reproduce. Sex gives rise to sexual selection, a widespread force operating through competition and choice within reproduction, however, it remains unclear whether sexual selection is beneficial for total lineage fitness, or if it acts as a constraint. Sexual selection could be a positive force because of selection on improved individual condition and purging of mutation load, summing into lineages with superior fitness. On the other hand, sexual selection could negate potential net fitness through the actions of sexual conflict, or because of tensions between investment in sexually selected and naturally selected traits. Here, we explore these ideas using a multigenerational invasion challenge to measure consequences of sexual selection for the overall net fitness of a lineage. After applying experimental evolution under strong versus weak regimes of sexual selection for 77 generations with the flour beetle Tribolium castaneum, we measured the overall ability of introductions from either regime to invade into conspecific competitor populations across eight generations. Results showed that populations from stronger sexual selection backgrounds had superior net fitness, invading more rapidly and completely than counterparts from weak sexual selection backgrounds. Despite comprising only 10% of each population at the start of the invasion experiment, colonizations from strong sexual selection histories eventually achieved near-total introgression, almost completely eliminating the original competitor genotype. Population genetic simulations using the design and parameters of our experiment indicate that this invasion superiority could be explained if strong sexual selection had improved both juvenile and adult fitness, in both sexes. Using a combination of empirical and modeling approaches, our findings therefore reveal positive and wide-reaching impacts of sexual selection for net population fitness when facing the broad challenge of invading competitor populations across multiple generations.


The Hospital Zambrano Hellion venous thromboembolism rapid response team (PREVENTION-team): Improving pulmonary embolism and deep venous thrombosis patient care.
Fast-track worldwide reperfusion programs improve outcomes in ST-elevation myocardial infarction and stroke. Similar programs called Program Evaluation and Review Technique (PERT) focus on submassive and massive pulmonary embolism (PE) excluding deep venous thrombosis (DVT). PREVENTION-team (Hospital Zambrano Hellion Venous Thromboembolism [VTE] Rapid Response). Fast-track stratification, diagnostics, and treatment (60-90 min) to improve proximal DVT and submassive and massive PE patients care. Increase diagnosis rate of low-risk PE and distal DVT; exploration of cause; long-term anticoagulation; identify high-risk profile for chronic complications; community-based support groups and patient education to extend the concept of the thrombosis-free hospital to thrombosis-free home. The team includes cardiologists, vascular medicine, angiologist, echocardiographer, cardiovascular imaging, and interventional cardiologists. The team will be accessible 24 h a day, 7 days a week, 365 days a year, and base on previous national experience. The cardiology fellow on call will be responsible for activation and evaluation. We will design several tools to accelerate these processes. Risk stratification and therapeutic approach will be based on clinical presentation, echocardiogram, and biomarkers findings. According to PERT stratification based on resources and medical specialties, Hospital Zambrano Hellion has level 1 PERT. PREVENTION-team links physicians with different expertise, provide fast, efficient, and time-saving treatment, potentially saving lives and reducing bleeding and -chronic complications in VTE patients. Finally, establishing a network in our hospital and health system to improve VTE patients care. To the best of our knowledge, this is the first rapid response team focused on VTE in Mexico.


Dynamic shifts of limited working memory resources in human vision.
Our ability to remember what we have seen is very limited. Most current views characterize this limit as a fixed number of items-only four objects-that can be held in visual working memory. We show that visual memory capacity is not fixed by the number of objects, but rather is a limited resource that is shared out dynamically between all items in the visual scene. This resource can be shifted flexibly between objects, with allocation biased by selective attention and toward targets of upcoming eye movements. The proportion of resources allocated to each item determines the precision with which it is remembered, a relation that we show is governed by a simple power law, allowing quantitative estimates of resource distribution in a scene.


Eltrombopag for the treatment of thrombocytopenia in patients with malignant and non-malignant hematologic disorders.
Eltrombopag (EP) is an orally bioavailable, non-peptide, thrombopoietin receptor (TPO-R) agonist developed to stimulate platelet production. EP is a small hydrazone molecule which interacts with the transmembrane domain of TPO-R and promotes megakaryopoiesis, and a subsequent increase in platelet number. To date, multiple large clinical trials have demonstrated the ability of EP to reduce the burden of thrombocytopenia and its associated side effects in patients with chronic immune thrombocytopenia purpura and patients with hepatitis-C related thrombocytopenia. Given these promising results and the morbidity associated with thrombocytopenia in cancer patients, there is significant interest in investigating the role of EP for thrombocytopenia secondary to myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). In this review, the authors address the potential utility of EP for patients with AML and MDS with thrombocytopenia. The review provides an overview of the rationale for the development of EP in AML and MDS, and the mechanism(s) of action of EP. The authors focus on preclinical data describing the effectiveness of EP as both a platelet-stimulating, and an anti-leukemia agent and describe the use of EP in clinical trials. EP has the potential to be an effective supportive care agent, improving platelet counts and decreasing thrombocytopenia-related morbidity, in patients with AML and MDS. Large, randomized clinical trials are needed to assess the efficacy of EP in reducing the duration and severity of thrombocytopenia, as well assess the clinical utility of EP as an anti-leukemia agent.


Sealing the broken barrier in IBD: intestinal permeability, epithelial cells and junctions.
Inflammatory bowel diseases (IBD) are considered barrier diseases. After misleading initial results, the pathogenic importance of a disturbed mucosa is now widely accepted, largely because a certain percentage of first-degree relatives of patients with IBD do have permeability alterations, as assessed by oral markers. In the presence of a normal appearing gut mucosa, functional alterations of the highly dynamic inter-enterocyte tight junctions have to be considered to be responsible for the observed alterations. Indeed, various alterations of the transmembrane and intracytoplasmic proteins have been reported in IBD. An important therapeutic goal is to maintain disease remission by preservation of the correct organization of these complexes. Of the potential therapeutic approaches, the various anti-TNF agents are the best-studied agents, but other treatments may tighten the gut through as yet unknown mechanisms.


Physical association and coordinate function of the H3 K4 methyltransferase MLL1 and the H4 K16 acetyltransferase MOF.
A stable complex containing MLL1 and MOF has been immunoaffinity purified from a human cell line that stably expresses an epitope-tagged WDR5 subunit. Stable interactions between MLL1 and MOF were confirmed by reciprocal immunoprecipitation, cosedimentation, and cotransfection analyses, and interaction sites were mapped to MLL1 C-terminal and MOF zinc finger domains. The purified complex has a robust MLL1-mediated histone methyltransferase activity that can effect mono-, di-, and trimethylation of H3 K4 and a MOF-mediated histone acetyltransferase activity that is specific for H4 K16. Importantly, both activities are required for optimal transcription activation on a chromatin template in vitro and on an endogenous MLL1 target gene, Hox a9, in vivo. These results indicate an activator-based mechanism for joint MLL1 and MOF recruitment and targeted methylation and acetylation and provide a molecular explanation for the closely correlated distribution of H3 K4 methylation and H4 K16 acetylation on active genes.


Positron emission tomography imaging of CD105 expression with 89Zr-Df-TRC105.
High tumor microvessel density correlates with a poor prognosis in multiple solid tumor types. The clinical gold standard for assessing microvessel density is CD105 immunohistochemistry on paraffin-embedded tumor specimens. The goal of this study was to develop an (89)Zr-based PET tracer for noninvasive imaging of CD105 expression. TRC105, a chimeric anti-CD105 monoclonal antibody, was conjugated to p-isothiocyanatobenzyl-desferrioxamine (Df-Bz-NCS) and labeled with (89)Zr. FACS analysis and microscopy studies were performed to compare the CD105 binding affinity of TRC105 and Df-TRC105. PET imaging, biodistribution, blocking, and ex-vivo histology studies were performed on 4T1 murine breast tumor-bearing mice to evaluate the pharmacokinetics and tumor-targeting of (89)Zr-Df-TRC105. Another chimeric antibody, cetuximab, was used as an isotype-matched control. FACS analysis of HUVECs revealed no difference in CD105 binding affinity between TRC105 and Df-TRC105, which was further validated by fluorescence microscopy. (89)Zr labeling was achieved with high yield and specific activity. Serial PET imaging revealed that the 4T1 tumor uptake of (89)Zr-Df-TRC105 was 6.1 ± 1.2, 14.3 ± 1.2, 12.4 ± 1.5, 7.1 ± 0.9, and 5.2 ± 0.3 %ID/g at 5, 24, 48, 72, and 96 h after injection, respectively (n = 4), higher than all organs starting from 24 h after injection, which provided excellent tumor contrast. Biodistribution data as measured by gamma counting were consistent with the PET findings. Blocking experiments, control studies with (89)Zr-Df-cetuximab, and ex-vivo histology all confirmed the in vivo target specificity of (89)Zr-Df-TRC105. We report here the first successful PET imaging of CD105 expression with (89)Zr as the radiolabel. Rapid, persistent, CD105-specific uptake of (89)Zr-Df-TRC105 in the 4T1 tumor was observed.


Genetic variation in the human insulin gene.
Four recombinant lambda phages containing nucleotide sequences complementary to a cloned human preproinsulin DNA probe have been isolated from human DNA. Restriction analyses in conjunction with Southern hybridizations reveal two types of gene sequences. One isolate of each type was subjected to complete nucleotide sequence determination. The sequences contain the entire preproinsulin messenger RNA region, two intervening sequence. 260 nucleotides upstream from the messenger RNA capping site, and 35 nucleotides beyond the polyadenylate attachment site. Our results strongly suggest that these two gene types are allelic variants of a single insulin gene.


Cisplatin plus oral etoposide (EoP) combination is more effective than paclitaxel in patients with advanced breast cancer pretreated with anthracyclines: a randomised phase III trial of Turkish Oncology Group.
Our objective was to determine whether oral etoposide and cisplatin combination (EoP) is superior to paclitaxel in the treatment of advanced breast cancer (ABC) patients pretreated with anthracyclines. From December 1997 to August 2003, 201 patients were randomised, 100 to EoP and 101 to paclitaxel arms. Four patients in each arm were ineligible. The doses of etoposide and cisplatin were 50 mg p.o. twice a day for 7 days and 70 mg m(-2) intravenously (i.v.) on day 1, respectively, and it was 175 mg m(-2) on day 1 for paclitaxel. Both treatments were repeated every 3 weeks. A median of four cycles of study treatment was given in both arms. The response rate obtained in the EoP arm was significantly higher (36.3 vs 22.2%; P=0.038). Median response duration was longer for the EoP arm (7 vs 4 months) (P=0.132). Also, time to progression was significantly in favour of the EoP arm (5.5 vs 3.9 months; P=0.003). Median overall survival was again significantly longer in the EoP arm (14 vs 9.5 months; P=0.039). Toxicity profile of both groups was similar. Two patients in each arm were lost due to febrile neutropenia. The observed activity and acceptable toxicity of EoP endorses the employment of this combination in the treatment of ABC following anthracyclines.