Derivation and validation of a clinical prediction score for isolation of inpatients with suspected pulmonary tuberculosis.
The use of a clinical prediction score to improve the practice of instituting airborne-transmission precautions in patients with suspected tuberculosis holds promise for increasing appropriate isolation and decreasing unnecessary isolation. The objective of this study was to derive and validate a clinical prediction score for patients with suspected tuberculosis. We used a case-control study design to evaluate differences between patients with a diagnosis of tuberculosis and those placed under airborne precautions who had negative culture results. We developed risk scores based on a multivariable analysis of independently significant factors associated with tuberculosis. Subsequently, we evaluated the sensitivity and specificity of the score in a separate (validation) cohort of patients. Within our population, we found 4 clinical factors associated with tuberculosis: chronic symptoms (odds ratio [OR], 10.2 [95% confidence interval {CI}, 2.95-35.4]), upper lobe disease on chest radiograph (OR, 5.27 [95% CI, 1.6-17.23]), foreign-born status (OR, 7.01 [95% CI, 2.1-23.8]), and immunocompromised state other than human immunodeficiency virus infection (OR, 8.14 [95% CI, 2.08-31.8]). Shortness of breath (OR, 0.13 [95% CI, 0.04-0.45]) was found to be associated with non-tuberculosis diagnoses and considered a negative predictor in the model. Using a cut-off point to maximize sensitivity, we applied the prediction rule to the validation cohort, resulting in a sensitivity of 97% and a specificity of 42%. The tuberculosis prediction rule derived from our patient population could improve utilization of airborne precautions. Clinical prediction rules continue to show their utility for improvement in isolation practices in different demographic areas.


Development of a new Sonovue™ contrast-enhanced ultrasound approach reveals temporal and age-related features of muscle microvascular responses to feeding.
Compromised limb blood flow in aging may contribute to the development of sarcopenia, frailty, and the metabolic syndrome. We developed a novel contrast-enhanced ultrasound technique using Sonovue™ to characterize muscle microvasculature responses to an oral feeding stimulus (15 g essential amino acids) in young (∼20 years) and older (∼70 years) men. Intensity-time replenishment curves were made via an ultrasound probe "fixed" over the quadriceps, with intermittent high mechanical index destruction of microbubbles within muscle vasculature. This permitted real-time measures of microvascular blood volume (MBV), microvascular flow velocity (MFV) and their product, microvascular blood flow (MBF). Leg blood flow (LBF) was measured by Doppler and insulin by enzyme-linked immunosorbent assay. Steady-state contrast concentrations needed for comparison between different physiological states were achieved <150 sec from commencing Sonovue™ infusion, and MFV and MBV measurements were completed <120 sec thereafter. Interindividual coefficients of variation in MBV and MFV were 35-40%, (N = 36). Younger men (N = 6) exhibited biphasic vascular responses to feeding with early increases in MBV (+36%, P < 0.008 45 min post feed) reflecting capillary recruitment, and late increases in MFV (+77%, P < 0.008) and MBF (+130%, P < 0.007 195 min post feed) reflecting more proximal vessel dilatation. Early MBV responses were synchronized with peak insulin but not increased LBF, while later changes in MFV and MBF occurred with insulin at post absorptive values but alongside increased LBF. All circulatory responses were absent in old men (N = 7). Thus, impaired postprandial circulation could impact age-related declines in muscle glucose disposal, protein anabolism, and muscle mass.


Structural analysis of herpes simplex virus by optical super-resolution imaging.
Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.


Multivitamins, individual vitamin and mineral supplements, and risk of diabetes among older U.S. adults.
Understanding the relationship between multivitamin use and diabetes risk is important given the wide use of multivitamin supplements among U.S. adults. We prospectively examined supplemental use of multivitamins and individual vitamins and minerals assessed in 1995-1996 in relation to self-reported diabetes diagnosed after 2000 among 232,007 participants in the National Institutes of Health-American Association of Retired Persons Diet and Health Study. Multivitamin use was assessed by a food-frequency questionnaire at baseline. Odds ratios (ORs) and 95% CIs were calculated by logistic regression models, adjusted for potential confounders. In total, 14,130 cases of diabetes diagnosed after 2000 were included in the analysis. Frequent use of any multivitamins was not associated with risk of diabetes after adjustment for potential confounders and uses of individual supplements. Compared with nonusers of any multivitamins, the multivariate ORs among users were 1.07 (95% CI 0.94-1.21) for taking vitamins less than once per week, 0.97 (0.88-1.06) for one to three times per week, 0.92 (0.84-1.00) for four to six times per week, and 1.02 (0.98-1.06) for seven or more times per week (P for trend = 0.64). Significantly lower risk of diabetes was associated with the use of vitamin C or calcium supplements. The multivariate ORs comparing daily users with nonusers were 0.91 (0.86-0.97) for vitamin C supplements and 0.85 (0.80-0.90) for calcium supplements. Use of vitamin E or other individual vitamin and mineral supplements were not associated with diabetes risk. In this large cohort of U.S. older adults, multivitamin use was not associated with diabetes risk. The findings of lower diabetes risk among frequent users of vitamin C or calcium supplements warrant further evaluations.


[Hospital readmissions: A reliable quality indicator?].
The percentage of readmissions within 30 days after discharge is an official quality indicator for Dutch hospitals in 2016. In this commentary the authors argue why readmissions cannot be regarded as a reliable way of assessing quality of healthcare in a hospital. To date, policy makers have been struggling with its precise definition and the indicator has not been properly formulated yet. It does not distinguish between planned and unplanned readmissions and does not take into account the 'preventability'. Therefore the authors believe that the indicator in its current form might falsely interpret the quality of care of a hospital and it is questionable to use readmissions as a quality indicator.


Four-dimensional MR microscopy of the mouse heart using radial acquisition and liposomal gadolinium contrast agent.
Magnetic resonance microscopy (MRM) has become an important tool for small animal cardiac imaging. In relation to competing technologies (microCT and ultrasound), MR is limited by spatial resolution, temporal resolution, and acquisition time. All three of these limitations have been addressed by developing a four-dimensional (4D) (3D plus time) radial acquisition (RA) sequence. The signal-to-noise ratio (SNR) has been optimized by minimizing the echo time (TE) (300 us). The temporal resolution and throughput have been improved by center-out trajectories resulting in repetition time (TR) <2.5 ms. The contrast has been enhanced through the use of a liposomal blood pool agent that reduces the T(1) of the blood to <400 ms. We have developed protocols for three specific applications: 1) high-throughput with spatial resolution of 87 x 87 x 352 um(3) (voxel volume = 2.7 nL) and acquisition time of 16 min; 2) high-temporal resolution with spatial resolution of 87 x 87 x 352 um(3) (voxel volume = 2.7 nL) and temporal resolution at 4.8 ms and acquisition time of 32 minutes; and 3) high-resolution isotropic imaging at 87 x 87 x 87 um(3) (voxel volume = 0.68 nL) and acquisition time of 31 min. The 4D image arrays allow direct measure of cardiac functional parameters dependent on chamber volumes, e.g., ejection fraction (EF), end diastolic volume (EDV), and end systolic volume (ESV).


Mice expressing T4826I-RYR1 are viable but exhibit sex- and genotype-dependent susceptibility to malignant hyperthermia and muscle damage.
Mutation T4825I in the type 1 ryanodine receptor (RYR1(T4825I/+)) confers human malignant hyperthermia susceptibility (MHS). We report a knock-in mouse line that expresses the isogenetic mutation T4826I. Heterozygous RYR1(T4826I/+) (Het) or homozygous RYR1(T4826I/T4826I) (Hom) mice are fully viable under typical rearing conditions but exhibit genotype- and sex-dependent susceptibility to environmental conditions that trigger MH. Hom mice maintain higher core temperatures than WT in the home cage, have chronically elevated myoplasmic[Ca(2+)](rest), and present muscle damage in soleus with a strong sex bias. Mice subjected to heat stress in an enclosed 37°C chamber fail to trigger MH regardless of genotype, whereas heat stress at 41°C invariably triggers fulminant MH in Hom, but not Het, mice within 20 min. WT and Het female mice fail to maintain euthermic body temperature when placed atop a bed whose surface is 37°C during halothane anesthesia (1.75%) and have no hyperthermic response, whereas 100% Hom mice of either sex and 17% of the Het males develop fulminant MH. WT mice placed on a 41°C bed maintain body temperature while being administered halothane, and 40% of the Het females and 100% of the Het males develop fulminant MH within 40 min. Myopathic alterations in soleus were apparent by 12 mo, including abnormally distributed and enlarged mitochondria, deeply infolded sarcolemma, and frequent Z-line streaming regions, which were more severe in males. These data demonstrate that an MHS mutation within the S4-S5 cytoplasmic linker of RYR1 confers genotype- and sex-dependent susceptibility to pharmacological and environmental stressors that trigger fulminant MH and promote myopathy.


[Caseload of poisoning among children treated by the Rio Grande do Sul State Toxicology Information Center (CIT/RS), Brazil].
This was an analytical, descriptive, cross-sectional study to establish the profile of poisonings among children 0 to 4 years of age reported to the Rio Grande do Sul State Toxicology Information Center (CIT/RS) in Porto Alegre, Brazil, 2003. A total of 1,311 cases were selected and data were extracted on children who had suffered individual accidental poisoning inside the home. From a systematic random sample of 286 patients, 21 interviews were held with the children's parents or guardians, using a structured instrument. The typical profile of poisoning featured: < or=1 year age bracket; male gender; oral exposure; poisonous agent located on living room or bedroom floor; furniture height < 30cm; time of poisoning from 18:01 to 22:00; time between poisoning and search for emergency medical care less than 30 minutes; analgesics as poisonous agent; medical assessment of purported poisoning as non-toxic, evolving to cure. The data indicate relevant characteristics as to the time of poisoning and place where poisonous agents were located. Definition of a profile helps promote and expand childhood poisoning prevention campaigns by public health agencies.


Pressure and perverse flights to familiarity.
Under pressure, people often prefer what is familiar, which can seem safer than the unfamiliar. We show that such favoring of familiarity can lead to choices precisely contrary to the source of felt pressure, thus exacerbating, rather than mitigating, its negative consequences. In Experiment 1, time pressure increased participants' frequency of choosing to complete a longer but incidentally familiar task option (as opposed to a shorter but unfamiliar alternative), resulting in increased felt stress during task completion. In Experiment 2, pressure to reach a performance benchmark in a chosen puzzle increased participants' frequency of choosing an incidentally familiar puzzle that both augured and delivered objectively worse performance (i.e., fewer points obtained). Participants favored this familiar puzzle even though familiarity was established through unpleasant prior experience. This "devil you know" preference under pressure contrasted with disfavoring of the negatively familiar option in a pressure-free situation. These results demonstrate that pressure-induced flights to familiarity can sometimes aggravate rather than ameliorate pressure, and can occur even when available evidence points to the suboptimality of familiar options.


Probiotic Treatment Decreases the Number of CD14-Expressing Cells in Porcine Milk Which Correlates with Several Intestinal Immune Parameters in the Piglets.
Modulating the mucosal immune system of neonates by probiotic treatment of their mothers is a promising approach which can only be investigated through the use of animal models. Here, we used sows and their piglets to investigate the impact of a bacterial treatment on the sow's milk and on the neonate piglet intestinal immune system. In previous experiments, feed supplementation of sows with the probiotic Enterococcus faecium NCIMB 10415 during pregnancy and lactation had been shown to affect intestinal microbiota and cytokine expression of the offspring during the suckling and weaning periods. We therefore investigated the composition of the milk from treated sows in comparison to samples from a control group. In treated sows, the amount of lactose increased, and the somatic cell numbers were reduced. In all milk samples, the percentage of cells expressing membranous CD14 (mCD14) was greater than the fractions of immune cells, indicating expression of mCD14 on mammary epithelial cells. However, in the milk of E. faecium-treated sows, mCD14(+) cells were reduced. Furthermore, the number of CD14(+) milk cells was positively correlated with the percentages of B cells and activated T cells in the ileal MLN of the piglets. This study provides evidence for the expression of mCD14 by the porcine mammary epithelium, and suggests an immunological effect of mCD14(+) milk cells on the piglets' intestinal immune system. Our study further suggests that mCD14(+) mammary epithelial cell populations can be modulated by probiotic feed supplementation of the sow.


Cannabidiol Prevents the Expression of the Locomotor Sensitization and the Metabolic Changes in the Nucleus Accumbens and Prefrontal Cortex Elicited by the Combined Administration of Cocaine and Caffeine in Rats.
In the last years, clinical and preclinical researchers have increased their interest in non-psychotomimetic cannabinoids, like cannabidiol (CBD), as a strategy for treating psychostimulant use disorders. However, there are discrepancies in the pharmacological effects and brain targets of CBD. We evaluated if CBD was able to prevent the locomotor sensitization elicited by cocaine and caffeine co-administration. The effect of CBD on putative alterations in the metabolic activity of the medial prefrontal cortex (mPFC) and nucleus accumbens (NAc), and its respective subregions (cingulated, prelimbic, and infralimbic cortices, and NAc core and shell) associated to the behavioral response, was also investigated. Rats were intraperitoneally and repeatedly treated with CBD (20 mg/kg) or its vehicle, followed by the combination of cocaine and caffeine (Coc+Caf; 5 mg/kg and 2.5 mg/kg, respectively) or saline for 3 days. After 5 days of withdrawal, all animals were challenged with Coc+Caf (day 9). Locomotor activity was automatically recorded and analyzed by a video-tracking software. The metabolic activity was determined by measuring cytochrome oxidase-I (CO-I) staining. Locomotion was significantly and similarly increased both in Veh-Coc+Caf- and CBD-Coc+Caf-treated animals during the pretreatment period (3 days); however, on day 9, the expression of the sensitization was blunted in CBD-treated animals. A hypoactive metabolic response and a hyperactive metabolic response in mPFC and NAc subregions respectively were observed after the behavioral sensitization. CBD prevented almost all these changes. Our findings substantially contribute to the understanding of the functional changes associated with cocaine- and caffeine-induced sensitization and the effect of CBD on this process.


Improving accuracy in estimation of artery-wall displacement by referring to center frequency of RF echo.
Noninvasive measurement of mechanical properties, such as elasticity, of the arterial wall, is useful for diagnosis of atherosclerosis. The elasticity of the arterial wall can be estimated by combining measurement of displacement of the arterial wall with that of blood pressure. In general, the displacement of the arterial wall is estimated from the phase shift of radio frequency (RF) echoes between two consecutive frames using a correlation estimator with quadrature demodulated complex signals. Recently, digitized data of broadband RF echoes are available in modern diagnostic equipment. The Fourier transform can be used to estimate the phase of the RF echo at each frequency within the RF frequency bandwidth. Therefore, the phase shifts between RF echoes of two consecutive frames can be estimated at multiple frequencies. In this estimation, due to object displacement, the RF echo is time shifted in comparison with that of the previous frame. However, the position of the time window for the Fourier transform is not changed between two consecutive frames. This change in relative position between the RF echo and the time window has a strong influence on the estimation of the artery-wall displacement, resulting in error. To suppress this error, the phase shift should be estimated at the actual RF center frequency. In this paper, this error suppression was investigated through simulation experiments and in vivo experiments on the human carotid artery.


A divalent FHA/BRCT-binding mechanism couples the MRE11-RAD50-NBS1 complex to damaged chromatin.
The MRE11-RAD50-NBS1 (MRN) complex accumulates at sites of DNA double-strand breaks in large chromatin domains flanking the lesion site. The mechanism of MRN accumulation involves direct binding of the Nijmegen breakage syndrome 1 (NBS1) subunit to phosphorylated mediator of the DNA damage checkpoint 1 (MDC1), a large nuclear adaptor protein that interacts directly with phosphorylated H2AX. NBS1 contains an FHA domain and two BRCT domains at its amino terminus. Here, we show that both of these domains participate in the interaction with phosphorylated MDC1. Point mutations in key amino acid residues of either the FHA or the BRCT domains compromise the interaction with MDC1 and lead to defects in MRN accumulation at sites of DNA damage. Surprisingly, only mutation in the FHA domain, but not in the BRCT domains, yields a G2/M checkpoint defect, indicating that MDC1-dependent chromatin accumulation of the MRN complex at sites of DNA breaks is not required for G2/M checkpoint activation.


Biochemical parameters for the diagnosis of mitochondrial respiratory chain deficiency in humans, and their lack of age-related changes.
It is now widely acknowledged that a large number of human diseases originate from respiratory-chain dysfunctions. Because the molecular bases of these diseases are still poorly known, a biochemical approach has to be used in the screening procedures for the diagnoses of these conditions. Assessment of respiratory-chain function in human samples faces several problems: (i) the small size of available samples, (ii) the determination of discriminating parameters, and (iii) the interfering factors, such as age and physical activity. The present study focuses on isolated mitochondria prepared from a minute amount (100-200 mg) of skeletal-muscle biopsies from 201 patients between 0 and 65 years. Whereas 42 patients presented an isolated complex (C)I, CII, CIII or CIV deficiency, no respiratory-chain dysfunction or indirect evidence for a mitochondrial disorder could be attested in 159 of these patients. In this reference group, there was little correlation between enzyme activities and age, whatever the age class considered, 0-3 or 0-65 years of age. However, a confident handling of data points was largely hampered by the marked scattering of enzyme activities measured in the reference population. Activity ratios between the various respiratory-chain complexes presenting a much reduced scattering may be considered as diagnostic tools. As to the effect of age, no correlation with any of the enzyme-activity ratios could be shown. Use of age-matched controls for the diagnosis of respiratory-chain disorders may therefore be avoided, enzyme-activity ratios being highly discriminating and age-independent parameters.


Food Safety Knowledge, Attitudes, and Behaviors of Native American Families with Young Children: A Mixed Methods Study.
Children are at increased risk for foodborne illness due to underdeveloped immune system. Limited research has been reported on food safety knowledge of Native American families with children 10 years of age and younger. This study was conducted to determine the food safety knowledge, attitudes, and behaviors of the main food preparer in these families by collecting quantitative and qualitative data simultaneously in a mixed method approach. A food safety knowledge survey created using FightBAC!™ concepts was administered prior to focus groups discussions held in Native American communities using a script based upon the Health Belief Model. Quantitative data were analyzed using SPSS. Qualitative data were coded by three reviewers independently and then compared jointly for themes. Over three fourths of participants (n = 102) were female with an average age of 38.3 years. Over one half of participants were unemployed (54 %), lived on reservations (54 %), and 86 % had a high school degree or higher level of education. The following four themes emerged from the eight focus groups (n = 66): food can make one sick, I am not in control when others handle food, I know how to safely prepare foods for my family, and I do not have time or best equipment (for food safety). Mixed method analysis revealed that participants were aware of the severity and susceptibility for foodborne illness but were confident in preparing foods safely for their family. A food safety education program for Native American food preparers with young children is needed to prevent foodborne illness (FBI) in this population and promote safe food handling practice.


Asymmetric synthesis of (-)-martinellic acid.
A high-yielding total asymmetric synthesis of (-)-martinellic acid is reported. The conjugate addition of lithium (R)-N-allyl-N-(α-methyl-4-methoxybenzyl)amide to tert-butyl (E)-3-[2'-(N,N-diallylamino)-5'-bromophenyl]propenoate and alkylation of the resultant β-amino ester have been used as the key steps to install the C(9b) and C(3a) stereogenic centers, respectively, and a highly diastereoselective Wittig reaction/intramolecular Michael addition was then used to create the C(4) stereogenic center within this tricyclic molecular architecture.


Effect of dietary supplements containing antioxidants on attenuation of muscle damage in exercising sled dogs.
To determine whether dietary antioxidants would attenuate exercise-induced increases in plasma creatine kinase (CK) activity in sled dogs. 41 trained adult sled dogs. Dogs, randomly assigned to 2 groups, received the same base diet throughout the study. After 8 weeks on that diet, 1 group (21 dogs) received a daily supplement containing vitamins E (457 U) and C (706 mg) and beta-carotene (5.1 mg), and a control group (20 dogs) received a supplement containing minimal amounts of antioxidants. After 3 weeks, both groups performed identical endurance exercise on each of 3 days. Blood samples were collected before and 3 weeks after addition of supplements and after each day of exercise. Plasma was analyzed for vitamins E and C, retinol, uric acid, triglyceride, and cholesterol concentrations, total antioxidant status (TAS), and CK activity. Feeding supplements containing antioxidants caused a significant increase in vitamin E concentration but did not change retinol or vitamin C concentrations orTAS. Exercise caused significantly higher CK activity, but did not cause a significant difference in CK activity between groups. Exercise was associated with significantly lower vitamin E, retinol, and cholesterol concentrations and TAS but significantly higher vitamin C, triglyceride, and uric acid concentrations in both groups. Use of supplements containing the doses of antioxidants used here failed to attenuate exercise-induced increases in CK activity. Muscle damage in sled dogs, as measured by plasma CK activity, may be caused by a mechanism other than oxidant stress.


Rapid activity prediction of HIV-1 integrase inhibitors: harnessing docking energetic components for empirical scoring by chemometric and artificial neural network approaches.
Improving performance of scoring functions for drug docking simulations is a challenging task in the modern discovery pipeline. Among various ways to enhance the efficiency of scoring function, tuning of energetic component approach is an attractive option that provides better predictions. Herein we present the first development of rapid and simple tuning models for predicting and scoring inhibitory activity of investigated ligands docked into catalytic core domain structures of HIV-1 integrase (IN) enzyme. We developed the models using all energetic terms obtained from flexible ligand-rigid receptor dockings by AutoDock4, followed by a data analysis using either partial least squares (PLS) or self-organizing maps (SOMs). The models were established using 66 and 64 ligands of mercaptobenzenesulfonamides for the PLS-based and the SOMs-based inhibitory activity predictions, respectively. The models were then evaluated for their predictability quality using closely related test compounds, as well as five different unrelated inhibitor test sets. Weighting constants for each energy term were also optimized, thus customizing the scoring function for this specific target protein. Root-mean-square error (RMSE) values between the predicted and the experimental inhibitory activities were determined to be <1 (i.e. within a magnitude of a single log scale of actual IC50 values). Hence, we propose that, as a pre-functional assay screening step, AutoDock4 docking in combination with these subsequent rapid weighted energy tuning methods via PLS and SOMs analyses is a viable approach to predict the potential inhibitory activity and to discriminate among small drug-like molecules to target a specific protein of interest.


Bacterial community structure and functional contributions to emergence of health or necrotizing enterocolitis in preterm infants.
Preterm infants represent a unique patient population that is born functionally immature and must accomplish development under the influence of a hospital environment. Neonatal necrotizing enterocolitis (NEC) is an inflammatory intestinal disorder affecting preterm infants. The purpose of this study was to evaluate the progression of intestinal microbiota community development between preterm infants who remained healthy compared to preterm infants who developed NEC. Weekly fecal samples from ten preterm infants, five with NEC and five matched healthy controls were obtained. Bacterial DNA from individual fecal samples was subjected to sequencing of 16S rRNA-based inventories using the 454 GS-FLX platform. Fecal samples from control infants demonstrated a temporal pattern in their microbiota, which converged toward that of a healthy full term breast-fed infant. Microbiota development in NEC patients diverged from controls beginning three weeks prior to diagnosis. Shotgun metagenomic sequencing was performed to identify functional differences in the respective microbiota of fecal samples from a set of twins in which one twin developed NEC and one did not. The majority of the differentially abundant genes in the NEC patient were associated with carbohydrate metabolism and mapped to members of the family Enterobacteriaceae. This may indicate an adaptation of the community to an altered profile of substrate availability for specific members as a first step towards the development of NEC. We propose that the microbial communities as a whole may metabolize milk differently, resulting in differential substrate availability for specific microbial groups. Additional differentially represented gene sets of interest were related to antibiotic resistance and vitamin biosynthesis. Our results suggest that there is a temporal component to microbiome development in healthy preterm infants. Thus, bacteriotherapy for the treatment or prevention of NEC must consider this temporal component of the microbial community in addition to its taxonomic composition and functional content.


Arabidopsis SEIPIN Proteins Modulate Triacylglycerol Accumulation and Influence Lipid Droplet Proliferation.
The lipodystrophy protein SEIPIN is important for lipid droplet (LD) biogenesis in human and yeast cells. In contrast with the single SEIPIN genes in humans and yeast, there are three SEIPIN homologs in Arabidopsis thaliana, designated SEIPIN1, SEIPIN2, and SEIPIN3. Essentially nothing is known about the functions of SEIPIN homologs in plants. Here, a yeast (Saccharomyces cerevisiae) SEIPIN deletion mutant strain and a plant (Nicotiana benthamiana) transient expression system were used to test the ability of Arabidopsis SEIPINs to influence LD morphology. In both species, expression of SEIPIN1 promoted accumulation of large-sized lipid droplets, while expression of SEIPIN2 and especially SEIPIN3 promoted small LDs. Arabidopsis SEIPINs increased triacylglycerol levels and altered composition. In tobacco, endoplasmic reticulum (ER)-localized SEIPINs reorganized the normal, reticulated ER structure into discrete ER domains that colocalized with LDs. N-terminal deletions and swapping experiments of SEIPIN1 and 3 revealed that this region of SEIPIN determines LD size. Ectopic overexpression of SEIPIN1 in Arabidopsis resulted in increased numbers of large LDs in leaves, as well as in seeds, and increased seed oil content by up to 10% over wild-type seeds. By contrast, RNAi suppression of SEIPIN1 resulted in smaller seeds and, as a consequence, a reduction in the amount of oil per seed compared with the wild type. Overall, our results indicate that Arabidopsis SEIPINs are part of a conserved LD biogenesis machinery in eukaryotes and that in plants these proteins may have evolved specialized roles in the storage of neutral lipids by differentially modulating the number and sizes of lipid droplets.


Hold the antioxidants and improve plasma lipids?
Intrahepatic proteolysis is a major determinant of secretion of ApoB-containing lipoproteins into plasma. Stimulation of post-ER presecretory proteolysis (PERPP) of ApoB by n-3 polyunsaturated fatty acids has been found to result in reduced secretion of VLDL particles by hepatocytes. A new study has shown that this stimulation is promoted by pro-oxidant conditions that result in increased hepatic lipid hydroperoxide content. Conversely, PERPP is suppressed by antioxidants and by saturated fatty acids, which are not susceptible to lipid peroxidation. Hence reduction of oxidative stress may have the unexpected side effect of increasing plasma lipid levels.


Quercetin Isolated from Toona sinensis Leaves Attenuates Hyperglycemia and Protects Hepatocytes in High-Carbohydrate/High-Fat Diet and Alloxan Induced Experimental Diabetic Mice.
The development of diabetes mellitus is related to oxidant stress induced by a high carbohydrate/high-fat diet (HFD). Quercetin, as a major bioactive component in Toona sinensis leaves (QTL), is a natural antioxidant. However, the exact mechanism by which QTL ameliorate diabetes mellitus is still unknown. In this study, we investigated the hypoglycemic effects and hepatocytes protection of QTL on HFD and alloxan induced diabetic mice. Intragastric administration of QTL significantly reduced body weight gain, serum glucose, insulin, total cholesterol, triglyceride, low density lipoprotein-cholesterol, alanine aminotransferase, and aspartate aminotransferase serum levels compared to those of diabetic mice. Furthermore, it significantly attenuated oxidative stress, as determined by lipid peroxidation, nitric oxide content, and inducible nitric oxide synthase activity and as a result attenuated liver injury. QTL also significantly suppressed the diabetes-induced activation of the p65/NF-κB and ERK1/2/MAPK pathways, as well as caspase-9 and caspase-3 levels in liver tissues of diabetic mice. Finally, micrograph analysis of liver samples showed decreased cellular organelle injury in hepatocytes of QTL treated mice. Taken together, QTL can be viewed as a promising dietary agent that can be used to reduce the risk of diabetes mellitus and its secondary complications by ameliorating oxidative stress in the liver.


Identification of PARP-1, Histone H1 and SIRT-1 as New Regulators of Breast Cancer-Related Aromatase Promoter I.3/II.
Paracrine interactions between malignant estrogen receptor positive (ER+) breast cancer cells and breast adipose fibroblasts (BAFs) stimulate estrogen biosynthesis by aromatase in BAFs. In breast cancer, mainly the cAMP-responsive promoter I.3/II-region mediates excessive aromatase expression. A rare single nucleotide variant (SNV) in this promoter region, which caused 70% reduction in promoter activity, was utilized for the identification of novel regulators of aromatase expression. To this end, normal and mutant promoter activities were measured in luciferase reporter gene assays. DNA-binding proteins were captured by DNA-affinity and identified by mass spectrometry. The DNA binding of proteins was analyzed using electrophoretic mobility shift assays, immunoprecipitation-based in vitro binding assays and by chromatin immunoprecipitation in BAFs in vivo. Protein expression and parylation were analyzed by western blotting. Aromatase activities and RNA-expression were measured in BAFs. Functional consequences of poly (ADP-ribose) polymerase-1 (PARP-1) knock-out, rescue or overexpression, respectively, were analyzed in murine embryonic fibroblasts (MEFs) and the 3T3-L1 cell model. In summary, PARP-1 and histone H1 (H1) were identified as critical regulators of aromatase expression. PARP-1-binding to the SNV-region was crucial for aromatase promoter activation. PARP-1 parylated H1 and competed with H1 for DNA-binding, thereby inhibiting its gene silencing action. In MEFs (PARP-1 knock-out and wild-type) and BAFs, PARP-1-mediated induction of the aromatase promoter showed bi-phasic dose responses in overexpression and inhibitor experiments, respectively. The HDAC-inhibitors butyrate, panobinostat and selisistat enhanced promoter I.3/II-mediated gene expression dependent on PARP-1-activity. Forskolin stimulation of BAFs increased promoter I.3/II-occupancy by PARP-1, whereas SIRT-1 competed with PARP-1 for DNA binding but independently activated the promoter I.3/II. Consistently, the inhibition of both PARP-1 and SIRT-1 increased the NAD+/NADH-ratio in BAFs. This suggests that cellular NAD+/NADH ratios control the complex interactions of PARP-1, H1 and SIRT-1 and regulate the interplay of parylation and acetylation/de-acetylation events with low NAD+/NADH ratios (reverse Warburg effect), promoting PARP-1 activation and estrogen synthesis in BAFs. Therefore, PARP-1 inhibitors could be useful in the treatment of estrogen-dependent breast cancers.


Where Two Are Fighting, the Third Wins: Stronger Selection Facilitates Greater Polymorphism in Traits Conferring Competition-Dispersal Tradeoffs.
A major conundrum in evolution is that, despite natural selection, polymorphism is still omnipresent in nature: Numerous species exhibit multiple morphs, namely several abundant values of an important trait. Polymorphism is particularly prevalent in asymmetric traits, which are beneficial to their carrier in disruptive competitive interference but at the same time bear disadvantages in other aspects, such as greater mortality or lower fecundity. Here we focus on asymmetric traits in which a better competitor disperses fewer offspring in the absence of competition. We report a general pattern in which polymorphic populations emerge when disruptive selection increases: The stronger the selection, the greater the number of morphs that evolve. This pattern is general and is insensitive to the form of the fitness function. The pattern is somewhat counterintuitive since directional selection is excepted to sharpen the trait distribution and thereby reduce its diversity (but note that similar patterns were suggested in studies that demonstrated increased biodiversity as local selection increases in ecological communities). We explain the underlying mechanism in which stronger selection drives the population towards more competitive values of the trait, which in turn reduces the population density, thereby enabling lesser competitors to stably persist with reduced need to directly compete. Thus, we believe that the pattern is more general and may apply to asymmetric traits more broadly. This robust pattern suggests a comparative, unified explanation to a variety of polymorphic traits in nature.


Regulation of recombination at yeast nuclear pores controls repair and triplet repeat stability.
Secondary structure-forming DNA sequences such as CAG repeats interfere with replication and repair, provoking fork stalling, chromosome fragility, and recombination. In budding yeast, we found that expanded CAG repeats are more likely than unexpanded repeats to localize to the nuclear periphery. This positioning is transient, occurs in late S phase, requires replication, and is associated with decreased subnuclear mobility of the locus. In contrast to persistent double-stranded breaks, expanded CAG repeats at the nuclear envelope associate with pores but not with the inner nuclear membrane protein Mps3. Relocation requires Nup84 and the Slx5/8 SUMO-dependent ubiquitin ligase but not Rad51, Mec1, or Tel1. Importantly, the presence of the Nup84 pore subcomplex and Slx5/8 suppresses CAG repeat fragility and instability. Repeat instability in nup84, slx5, or slx8 mutant cells arises through aberrant homologous recombination and is distinct from instability arising from the loss of ligase 4-dependent end-joining. Genetic and physical analysis of Rad52 sumoylation and binding at the CAG tract suggests that Slx5/8 targets sumoylated Rad52 for degradation at the pore to facilitate recovery from acute replication stress by promoting replication fork restart. We thereby confirmed that the relocation of damage to nuclear pores plays an important role in a naturally occurring repair process.


Modification of spontaneous emission in Bragg onion resonators.
We formulated an analytical model and analyzed the modification of spontaneous emission in Bragg onion resonators. We consider both the case of a single light emitter and a uniformly distributed ensemble of light emitters within the resonator. We obtain an expression for the average radiation rate of the light emitters ensemble and discuss the modification of the average radiation rate as a function of cavity parameters such as the core radius, the number of Bragg cladding layers, the index contrast of the Bragg cladding, and the refractive index of surrounding medium. We also consider the possibility of non-exponential decay of the light emitter ensemble due to the strong dependence of spontaneous emission on the location and polarization of individual light emitter. We conclude that Bragg onion resonators can both enhance and inhibit spontaneous emission by several orders of magnitude. This property can have significant impact in the field of cavity quantum electrodynamics (QED).


ATP release guides neutrophil chemotaxis via P2Y2 and A3 receptors.
Cells must amplify external signals to orient and migrate in chemotactic gradient fields. We find that human neutrophils release adenosine triphosphate (ATP) from the leading edge of the cell surface to amplify chemotactic signals and direct cell orientation by feedback through P2Y2 nucleotide receptors. Neutrophils rapidly hydrolyze released ATP to adenosine that then acts via A3-type adenosine receptors, which are recruited to the leading edge, to promote cell migration. Thus, ATP release and autocrine feedback through P2Y2 and A3 receptors provide signal amplification, controlling gradient sensing and migration of neutrophils.


[Hopes for future practice of junior orthopedic surgeons in France: a sociological study].
Medical demographics have become a major public health issue in France since the publication of the Berlan report in 2002 which announced an upcoming shortage in the next 20 years. But demographic projections cannot be limited to an accounting of training and retirement figures. Other factors such as demographic distribution, changing societal aspirations (free time), modes and sites of occupational activities, the influence of feminization, and initial training and its effects on professional practice must be taken into consideration. The purpose of this work was to study these parameters in orthopedic and traumatology surgery. This was a sociological study designed to ascertain the aspirations of junior orthopedic surgeons. Forty-seven interns in orthopedic surgery in the Paris area were invited to complete a questionnaire in March 2003. Forty-two responded. The choice of orthopedics was an intellectual choice. The attractiveness of the specialty was particularly important during the final years of medical school. Fewer junior surgeons would rather practice in a university hospital setting, more preferring a mixed setting with a private (fees) and a public (salary) component. The desire for "free time" was particularly important. Fewer juniors wanted to practice in general hospitals. For orthopedic surgeons, private practice and payment by fees are the cornerstones of medical practice.


An open-label pilot study of cannabis-based extracts for bladder dysfunction in advanced multiple sclerosis.
The majority of patients with multiple sclerosis (MS) develop troublesome lower urinary tract symptoms (LUTS). Anecdotal reports suggest that cannabis may alleviate LUTS, and cannabinoid receptors in the bladder and nervous system are potential pharmacological targets. In an open trial we evaluated the safety, tolerability, dose range, and efficacy of two whole-plant extracts of Cannabis sativa in patients with advanced MS and refractory LUTS. Patients took extracts containing delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD; 2.5 mg of each per spray) for eight weeks followed by THC-only (2.5 mg THC per spray) for a further eight weeks, and then into a long-term extension. Assessments included urinary frequency and volume charts, incontinence pad weights, cystometry and visual analogue scales for secondary troublesome symptoms. Twenty-one patients were recruited and data from 15 were evaluated. Urinary urgency, the number and volume of incontinence episodes, frequency and nocturia all decreased significantly following treatment (P <0.05, Wilcoxon's signed rank test). However, daily total voided, catheterized and urinary incontinence pad weights also decreased significantly on both extracts. Patient self-assessment of pain, spasticity and quality of sleep improved significantly (P <0.05, Wilcoxon's signed rank test) with pain improvement continuing up to median of 35 weeks. There were few troublesome side effects, suggesting that cannabis-based medicinal extracts are a safe and effective treatment for urinary and other problems in patients with advanced MS.


Ex vivo culture of primary human fallopian tube epithelial cells.
Epithelial ovarian cancer is a leading cause of female cancer mortality in the United States. In contrast to other women-specific cancers, like breast and uterine carcinomas, where death rates have fallen in recent years, ovarian cancer cure rates have remained relatively unchanged over the past two decades (1). This is largely due to the lack of appropriate screening tools for detection of early stage disease where surgery and chemotherapy are most effective (2, 3). As a result, most patients present with advanced stage disease and diffuse abdominal involvement. This is further complicated by the fact that ovarian cancer is a heterogeneous disease with multiple histologic subtypes (4, 5). Serous ovarian carcinoma (SOC) is the most common and aggressive subtype and the form most often associated with mutations in the BRCA genes. Current experimental models in this field involve the use of cancer cell lines and mouse models to better understand the initiating genetic events and pathogenesis of disease (6, 7). Recently, the fallopian tube has emerged as a novel site for the origin of SOC, with the fallopian tube (FT) secretory epithelial cell (FTSEC) as the proposed cell of origin (8, 9). There are currently no cell lines or culture systems available to study the FT epithelium or the FTSEC. Here we describe a novel ex vivo culture system where primary human FT epithelial cells are cultured in a manner that preserves their architecture, polarity, immunophenotype, and response to physiologic and genotoxic stressors. This ex vivo model provides a useful tool for the study of SOC, allowing a better understanding of how tumors can arise from this tissue, and the mechanisms involved in tumor initiation and progression.


Costimulatory molecules in the developing human gastrointestinal tract: a pathway for fetal allergen priming.
Antigen-specific responses can be detected in umbilical cord blood mononuclear cells. The fetal immune system must therefore attain a level of maturity compatible with the initiation of such responses as well as be exposed to antigen. We sought to assess the expression of costimulatory molecules in fetal gut and the presence of cytokines in amniotic fluid at this time as a preliminary analysis of the suitability of the fetal gut as a site of antigen priming during intrauterine life. Human fetal gut was analyzed for cells expressing costimulatory molecules through use of immunohistochemistry. Amniotic fluid was studied by ELISA, for cytokines regulating the nature of the response, and as a source of the common dietary antigen ovalbumin. MHC class II--positive cells were abundant over the period examined (11-24 weeks of gestation), other surface antigens showing spatial and temporal variation in expression. From 11 to 14 weeks of gestation, CD68-positive and CD40-positive cells, like MHC class II--positive cells, were present throughout the lamina propria; few CD3-positive cells (T cells) were observed. With the emergence of lymphoid aggregates (14-16 weeks), CD83-positive cells (dendritic cells) and CD20-positive cells (B cells) could be detected in fetal gut; however, expression was restricted to the lymphoid aggregates. In contrast, MHC class II, CD40, and CD68 continued to be expressed in the lamina propria. CD28-positive cells were also evident from 14 weeks of gestation, occurring throughout the lamina propria and lymphoid aggregates; this corresponded to the increasing numbers of CD3-positive cells. The occasional CD86-positive, CD40L-positive, or CTLA4-positive cell could be seen in or around lymphoid aggregates after 14 weeks of gestation. Lymphoid follicles forming after 16 weeks of gestation contained MHC class II--positive, CD83-positive, CD20-positive, CD40-positive, CD86-positive, CD3-positive, CD28-positive, CD40L-positive, and CTLA4-positive cells. MHC class II--positive, CD40-positive, CD68-positive, CD3-positive, and CD28-positive cells continued to be present in the lamina propria at this time. At all times studied, CD14 was not expressed in the lamina propria or lymphoid follicles. Prostaglandin E(2), TGF beta(1), and IL-10 dominated the amniotic fluid cytokine milieu, and ovalbumin was also detectable in amniotic fluid from 3 of 26 women who had detectable circulating levels. Of the costimulatory molecules studied, CD40 was the most abundant. However, both of the ligand families studied (CD40-CD40L and CD86-CD28/CD152) could provide the costimulatory signals required for the initiation of antigenspecific reactivity in the gastrointestinal tract of the human fetus as early as 16 weeks of gestation. The cytokine milieu would favor the development of T(H)2-type reactivity to antigens, such as ovalbumin, that are present at this time.


Differential mechanisms of action of the trace amines octopamine, synephrine and tyramine on the porcine coronary and mesenteric artery.
Trace amines such as p-tyramine, p-octopamine and p-synephrine are found in low concentrations in animals and plants. Consumption of pre-workout supplements containing these plant-derived amines has been associated with cardiovascular side effects. The aim of this study was to determine the mechanisms of action of these trace amines on porcine isolated coronary and mesenteric arteries. Noradrenaline caused contraction of mesenteric arteries and relaxation of coronary arteries. In both tissues, all three trace amines induced contractions with similar potencies and responses were unaffected by the β-adrenoceptor antagonist propranolol (1 µM), the nitric oxide synthase inhibitor L-NNA (100 µM), or the TAAR-1 antagonist, EPPTB (100 nM). However, the contractile responses of mesenteric arteries, but not coronary arteries, were significantly reduced by depletion of endogenous noradrenaline. Mesenteric responses to all three amines were abolished in the presence of prazosin (1 µM) whereas residual contractile responses remained in the coronary artery which were inhibited by a high concentration (100 µM) of EPPTB. The results suggest complex responses of the coronary artery to the trace amines, with activity at α1-adrenoceptors and potentially TAARs other than TAAR-1. In contrast the actions of the amines on the mesenteric artery appeared to involve indirect sympathomimetic actions and direct actions on α1-adrenoceptors.


Predictability and hierarchy in Drosophila behavior.
Even the simplest of animals exhibit behavioral sequences with complex temporal dynamics. Prominent among the proposed organizing principles for these dynamics has been the idea of a hierarchy, wherein the movements an animal makes can be understood as a set of nested subclusters. Although this type of organization holds potential advantages in terms of motion control and neural circuitry, measurements demonstrating this for an animal's entire behavioral repertoire have been limited in scope and temporal complexity. Here, we use a recently developed unsupervised technique to discover and track the occurrence of all stereotyped behaviors performed by fruit flies moving in a shallow arena. Calculating the optimally predictive representation of the fly's future behaviors, we show that fly behavior exhibits multiple time scales and is organized into a hierarchical structure that is indicative of its underlying behavioral programs and its changing internal states.


A fully automated image analysis method to quantify lung fibrosis in the bleomycin-induced rat model.
Intratracheal administration of bleomycin induces fibrosis in the lung, which is mainly assessed by histopathological grading that is subjective. Current literature highlights the need of reproducible and quantitative pulmonary fibrosis analysis. If some quantitative studies looked at fibrosis parameters separately, none of them quantitatively assessed both aspects: lung tissue remodeling and collagenization. To ensure reliable quantification, support vector machine learning was used on digitalized images to design a fully automated method that analyzes two important aspects of lung fibrosis: (i) areas having substantial tissue remodeling with appearance of dense fibrotic masses and (ii) collagen deposition. Fibrotic masses were identified on low magnification images and collagen detection was performed at high magnification. To insure a fully automated application the tissue classifier was trained on several independent studies that were performed over a period of four years. The detection method generates two different values that can be used to quantify lung fibrosis development: (i) percent area of fibrotic masses and (ii) percent of alveolar collagen. These two parameters were validated using independent studies from bleomycin- and saline-treated animals. A significant change of these lung fibrosis quantification parameters- increased amount of fibrotic masses and increased collagen deposition- were observed upon intratracheal administration of bleomycin and subsequent significant beneficial treatments effects were observed with BIBF-1120 and pirfenidone.


Mutual information for testing gene-environment interaction.
Despite current enthusiasm for investigation of gene-gene interactions and gene-environment interactions, the essential issue of how to define and detect gene-environment interactions remains unresolved. In this report, we define gene-environment interactions as a stochastic dependence in the context of the effects of the genetic and environmental risk factors on the cause of phenotypic variation among individuals. We use mutual information that is widely used in communication and complex system analysis to measure gene-environment interactions. We investigate how gene-environment interactions generate the large difference in the information measure of gene-environment interactions between the general population and a diseased population, which motives us to develop mutual information-based statistics for testing gene-environment interactions. We validated the null distribution and calculated the type 1 error rates for the mutual information-based statistics to test gene-environment interactions using extensive simulation studies. We found that the new test statistics were more powerful than the traditional logistic regression under several disease models. Finally, in order to further evaluate the performance of our new method, we applied the mutual information-based statistics to three real examples. Our results showed that P-values for the mutual information-based statistics were much smaller than that obtained by other approaches including logistic regression models.


Pre-administration of luteoline attenuates neonatal sevoflurane-induced neurotoxicity in mice.
Sevoflurane is a widely used inhaled anesthetic, which triggers neuroapoptosis and oxidative damage in the developing central nervous system and cognitive dysfunction later in life. However, no effective therapeutic strategy for sevoflurane-induced deleterious effects is well developed. The purpose of the present study was to explore whether luteoline could attenuate neonatal sevoflurane exposure-triggered neurotoxicity. In this study, six-day-old C57BL/6 mice were pretreated with luteoline (30, 60 mg/kg) intraperitoneally for 30 min before exposed to 3% sevoflurane 6 h consecutively. We first examined the effects of luteoline on hippocampal neuron apoptosis, inflammation and oxidative stress 18 h post anesthesia. The spatial learning and memory performance was measured using Morris water maze test from postnatal day 31 to 38. The results showed that luteoline ameliorated neuronal apoptosis as evidenced by decrease of apoptotic cells, downregulation of the cleavage levels of caspase-3 and PRAP, and inactivation of caspase-3. Moreover, luteoline significantly decreased protein expressions of inflammatory cytokines (IL-1β, IL-18 and TNF-α), inhibited NF-кB/NLRP3 pathway (NF-кB, NLRP3, ASC and caspase-1) and suppressed NF-кB activity. Our analyses indicated that luteoline had a significant effect on decreasing the contents of ROS and MDA, elevating the activity of SOD, and ultimately improving spatial learning and memory deficits of mice. In summary, our findings confirm that the attenuation of luteoline on sevoflurane-induced spatial learning and memory impairment later is associated with inhibition of hippocampal neuron apoptosis, inflammation and oxidative stress early. Luteoline might be a potential therapeutic for sevoflurane anesthesia-induced neurobehavioral dysfunction.


The blaNDM-1-Carrying IncA/C2 Plasmid Underlies Structural Alterations and Cointegrate Formation In Vivo.
In 2012, a carbapenemase-producing Salmonella enterica serovar Corvallis isolate carrying a blaNDM-1 multiresistance IncA/C2 plasmid, apart from IncHI2 and ColE-like plasmids, was detected in a wild bird in Germany. In a recent broiler chicken infection study, we observed transfer of this blaNDM-1-carrying IncA/C2 plasmid to other Enterobacteriaceae Here, we focused on the stability of this plasmid and gained insight into the type and frequency of its structural alterations after an in vivo passage in a broiler chicken infection study.


Prevalence and risk factors for anxiety and depression disorders in workers with work-related musculoskeletal strain or sprain in British Columbia, Canada: a comparison of men and women using administrative health data.
To examine the prevalence and risk factors for medically treated anxiety and depression disorders among men and women with musculoskeletal strain or sprain work injury in British Columbia, Canada. A retrospective population-based cohort of accepted workers' compensation lost-time claims from 2000 to 2013 was constructed using linked administrative health data. Anxiety and depression disorders were identified using diagnoses from physician, hospital and pharmaceutical records. The 1-year period prevalence was estimated for the year before and the year after injury. Sociodemographic, clinical and work-related risk factors for prevalent and new onset anxiety and depression disorders were examined using multinomial regression. 13.2% of men and 29.8% of women had medically treated anxiety, depression or both in the year before injury. Only a slight increase (~2%) in the prevalence of these disorders was observed in the year after injury. Somatic and mental comorbidities were both strong risk factors for pre-existing and new onset anxiety and depression for both men and women, but these relationships were stronger for men. Anxiety and depression disorders including those from prior to injury are common in workers with musculoskeletal strain or sprain and are associated with a complicated clinical profile. Gender-sensitive and sex-sensitive mental healthcare is an important consideration for work disability management.


Sedation with intravenous midazolam in the pediatric intensive care unit.
Physical and emotional distress can have important effects on patients in the pediatric intensive care unit (ICU). Intravenous (IV) infusion of benzodiazepines is an important adjunct to assisted ventilation and other potentially distressing ICU procedures. Combined with intermittent or continuous infusion of opioids, the benzodiazepines provide smooth control of anxiety, pain, and agitation. Intravenous midazolam (Versed Roche Laboratories) is distinguished from diazepam (Valium, Roche Products) by its water solubility, short elimination half-life, and generally short duration of action. These pharmacological properties, which are also shared, in part, with the more slowly eliminated drug lorazepam (Ativan, Wyeth-Ayerst), facilitate titration of the rate of infusion against patient response and permit regulation of the depth of sedation. The major adverse effects of long-term benzodiazepine infusion are withdrawal symptoms and, occasionally, delayed awakening. The dosage needed to initiate and maintain sedation must be adjusted to body weight, degree of sedation desired, and concomitant medications, as well as to underlying health and cardiovascular status. Benzodiazepines, such as midazolam and lorazepam, represent important choices among drugs used for sedation in the pediatric ICU.


An electron-microscopical analysis of capture and initial stages of penetration of nematodes by Arthrobotrys oligospora.
A detailed analysis was made of the capture and subsequent penetration of nematodes by the nematophagous fungus Arthrobotrys oligospora using different electron-microscopical techniques. Capture of nematodes by this fungus occurred on complex hyphal structures (traps) and was effectuated by an adhesive coating, present on these trap cells. The adhesive layer was largely fibrillar in nature and was absent on cells of normal hyphae. Following capture, penetration hyphae were formed at those sites where the trap cell wall was anchored to the nematode cuticle by the adhesive. New walls of these hyphae were formed underneath the original trap cell walls, which were partly hydrolysed to allow growth and development of the penetration tubes through the adhesive coating towards the cuticle. Our observations indicated that the cuticle of the nematode was subsequently penetrated by the penetration tubes by mechanical means. After penetration a large infection bulb was formed from which trophic hyphae arose. Cytochemical experiments indicated that the sites of penetration of the cuticle were intensely stained for acid phosphatase activity. At later stages of infection activity of this enzyme was present throughout the nematode contents; the enzyme was most probably secreted by complex membranous structures associated with the cytoplasmic membrane of the infection bulb and the trophic hyphae.


Natural inorganic nanoparticles--formation, fate, and toxicity in the environment.
The synthesis, stability, and toxicity of engineered metal nanoparticles (ENPs) have been extensively studied during the past two decades. In contrast, research on the formation, fate, and ecological effects of naturally-occurring nanoparticles (NNPs) has become a focus of attention only recently. The natural existence of metal nanoparticles and their oxides/sulfides in waters, wastewaters, ore deposits, mining regions, and hydrothermal vents, as exemplified by the formation of nanoparticles containing silver and gold (AgNPs and AuNPs), Fe, Mn, pyrite (FeS2), Ag2S, CuS, CdS, and ZnS, is dictated largely by environmental conditions (temperature, pH, oxic/anoxic, light, and concentration and characteristics of natural organic matter (NOM)). Examples include the formation of nanoparticles containing pyrite, Cu and Zn-containing pyrite, and iron in hydrothermal vent black smoker emissions. Metal sulfide nanoparticles can be formed directly from their precursor ions or indirectly by sulfide ion-assisted transformation of the corresponding metal oxides under anaerobic conditions. This tutorial focuses on the formation mechanisms, fate, and toxicity of natural metal nanoparticles. Natural waters containing Ag(I) and Au(III) ions in the presence of NOM generate AgNPs and AuNPs under thermal, non-thermal, and photochemical conditions. These processes are significantly accelerated by existing redox species of iron (Fe(II)/Fe(III)). NOM, metal-NOM complexes, and reactive oxygen species (ROS) such as O2˙(-), ˙OH, and H2O2 are largely responsible for the natural occurrence of nanoparticles. AgNPs and AuNPs emanating from Ag(I)/Au(III)-NOM reactions are stable for several months, thus indicating their potential to be transported over long distances from their point of origin. However, endogenous cations present in natural waters can destabilize the nanoparticles, with divalent cations (e.g., Ca(2+), Mg(2+)) being more influential than their monovalent equivalents (e.g., Na(+), K(+)). The toxicity of NNPs may differ from that of ENPs because of differences in the coatings on the nanoparticle surfaces. An example of this phenomenon is presented and is briefly discussed.


Distribution and abundance of parasites of the rhodesgrass mealybug, Antonina graminis: reassessment of a classic example of biological control in the Southeastern United States.
Control of the rhodesgrass mealybug, Antonina graminis Maskell (Hemiptera: Pseudococcidae), by the encyrtid wasp Neodusmetia sangwani is considered a textbook example of classical biological control. However, recent evidence suggests that A. graminis is abundant in the southeastern United States and no recent surveys have been conducted to determine the status of N. sangwani or other A. graminis parasites. A survey was conducted and it was found that N. sangwani was uncommon overall, occurring at only 20 percent of survey sites. In addition, N. sangwani exhibited a patchy geographic distribution. Possible causes for these results are that N. sangwani has not dispersed widely since its introduction, or that the imported fire ant, Solenopsis invicta, is interfering with biological control. These results suggest that a reevaluation of the efficacy of biological control may be necessary. The survey also found two other encyrtid wasps utilizing A. graminis as a host. One, Acerophagus sp., is apparently native and was nearly as frequent as N. sangwani, while the other, Pseudectroma sp., is apparently introduced and relatively rare.


Opposing actions of rosiglitazone and resveratrol on mineralization in human vascular smooth muscle cells.
Arteriosclerotic vascular disease is a major cardiac health problem in westernized countries and the primary cause of mortality in diabetic patients. Recent data have raised serious safety concerns with the antidiabetic rosiglitazone, a thiazolidinedione with peroxisome proliferator-activated receptor γ (PPAR-γ) agonistic activity, in regard to cardiovascular risks. A common feature of atherosclerosis is vascular mineralization. The latter is formed by vascular smooth muscle cells (VSMC) through complex processes that are similar to mineralization in bone. The aim of the current study was to investigate the effect of rosiglitazone on mineralization in cultured human VSMCs. We found that rosiglitazone stimulated mineralization by, at least in part, induction of caspase-dependent apoptosis. Furthermore, rosiglitazone-induced oxidative stress was correlated with stimulated osteoblast-like differentiation of VSMCs. Treatment of rosiglitazone-supplemented VSMC cultures with the caloric restriction mimetic and antioxidant resveratrol diminished rosiglitazone-induced oxidative stress, osteoblast-like differentiation and mineralization. In conclusion, this study reveals novel insights into the relationship of rosiglitazone and cardiovascular events by providing a model that links rosiglitazone-induced osteoblast-like differentiation, oxidative stress and apoptosis with mineralization in VSMCs. In addition, we position resveratrol in this model acting to reduce rosiglitazone-induced oxidative stress, osteoblast-like VSMC differentiation and mineralization.


Insights on the Functions and Ecophysiological Relevance of the Diverse Carbonic Anhydrases in Microalgae.
Carbonic anhydrases (CAs) exist in all kingdoms of life. They are metalloenzymes, often containing zinc, that catalyze the interconversion of bicarbonate and carbon dioxide-a ubiquitous reaction involved in a variety of cellular processes. So far, eight classes of apparently evolutionary unrelated CAs that are present in a large diversity of living organisms have been described. In this review, we focus on the diversity of CAs and their roles in photosynthetic microalgae. We describe their essential role in carbon dioxide-concentrating mechanisms and photosynthesis, their regulation, as well as their less studied roles in non-photosynthetic processes. We also discuss the presence in some microalgae, especially diatoms, of cambialistic CAs (i.e., CAs that can replace Zn by Co, Cd, or Fe) and, more recently, a CA that uses Mn as a metal cofactor, with potential ecological relevance in aquatic environments where trace metal concentrations are low. There has been a recent explosion of knowledge about this well-known enzyme with exciting future opportunities to answer outstanding questions using a range of different approaches.


Improving Pediatric Preparedness in Critical Access Hospital Emergency Departments: Impact of a Longitudinal In Situ Simulation Program.
Critical access hospitals (CAH) see few pediatric patients. Many of these hospitals do not have access to physicians with pediatric training. We sought to evaluate the impact of an in situ pediatric simulation program in the CAH emergency department setting on care team performance during resuscitation scenarios. Five CAHs conducted 6 high-fidelity pediatric simulations over a 12-month period. Team performance was evaluated using a validated 35-item checklist representing commonly expected resuscitation team interventions. Checklists were scored by assigning zero point for "yes" and 1 point for "no". A lower final score meant more items on the list had been completed. The Kruskal-Wallis rank test was used to assess for differences in average scores among institutions. A linear mixed effects model with a random institution intercept was used to examine trends in average scores over time. P < 0.05 was considered significant. The Kruskal-Wallis rank test showed no difference in average scores among institutions. (P = 0.90). Checklist scores showed a significant downward trend over time, with a scenario-to-scenario decrease of 0.022 (P < 0.01). One hundred percent of providers surveyed in the last month stated they would benefit from ongoing scenarios. Regularly scheduled pediatric simulations in the CAH emergency department setting improved team performance over time on expected resuscitation tasks. The program was accepted by providers. Implementation of simulation-based training programs can help address concerns regarding pediatric preparedness in the CAH setting. A future project will look at the impact of the program on patient care and safety.


Association of environment and policy characteristics on children's moderate-to-vigorous physical activity and time spent sedentary in afterschool programs.
Afterschool programs are an important setting in which to promote children's physical activity. This study examines the association of environmental and policy characteristics on the moderate-to-vigorous physical activity and sedentary behavior of children attending afterschool programs. A total of 1302 children attending 20 afterschool programs across South Carolina wore accelerometers (ActiGraph GT3X+) for up to 4non-consecutive days. Policy-level characteristics were evaluated using the Healthy Afterschool Program Index-Physical Activity scale. Physical activity space was measured using a measuring wheel (indoor, ft(2)) and Geographical Information Systems software (outdoor, acres). The structure (free-play or organized) of activity opportunities was evaluated via direct observation. Time spent in moderate-to-vigorous physical activity and sedentary, both indoors and outdoors, was estimated using accelerometry. For every 5000 ft(2) of utilized indoor activity space an additional 2.4 and 3.3 min/day of sedentary behavior was observed among boys and girls, respectively. A higher ratio of free-play to organized play was associated with higher indoor sedentary behavior among boys and girls (3.9 min/day and 10.0 min/day, respectively). For every 1 acre of outdoor activity space used, an additional 2.7 min/day of moderate-to-vigorous physical activity was observed for boys. A higher free-play to organized play ratio was associated with higher outdoor moderate-to-vigorous physical activity for boys and girls (4.4 and 3.4 min/day increase, respectively). Policy characteristics were unrelated to moderate-to-vigorous physical activity levels and time spent sedentary. Findings indicate that policies and size of activity space had limited influence on moderate-to-vigorous physical activity and sedentary behavior, suggesting that a programmatic structure may be a more effective option to improve moderate-to-vigorous physical activity levels of children attending afterschool programs.


Proteomic discovery of diverse immunity molecules in mammalian spermatozoa.
Ongoing proteomic analyses are providing a wealth of new data on the composition of the sperm proteome across a range of mammals and other taxa. Although molecular evolution and functional genomic analyses of the proteome have only begun recently, we now broadly understand the molecular composition of sperm. Systems level analysis has revealed a variety of molecular insights into sperm evolution and function, including a remarkable diversity of immunity-related proteins within the proteome. Using existing mammalian sperm proteomes as a starting point, we provide an overview of this important class of sperm proteins and what is known about their function in sperm maturation, sperm quality, sperm competition, and fertilization. The recent observation that many sperm immunity proteins are rapidly evolving, presumably under the influence of positive selection, suggests that they may be responding not only to selection associated with host immunity defense but also with pleiotropic functions in sperm. In addition to the documented role of sperm in the mediation of female immune response, we propose that the fundamental mechanisms involved in cell-cell recognition and binding in both immune processes and fertilization may underlie the multi-functionality of proteins in immunity and reproductive systems.


HIV and AIDS related knowledge, sources of information, and reported need for further education among dental students in Sudan--a cross sectional study.
Information on the HIV and AIDS-related knowledge among dental students provides a crucial foundation for efforts aimed at developing an appropriate dental curriculum on HIV and AIDS, and for attracting the attention of dental school educators towards the subject. Focusing on a census of dental students attending their 3rd, 4th and 5th study year at publicly - and privately funded dental faculties in Khartoum, this study aimed to assess the prevalence and socio-economic correlates of dental students' knowledge, sources of information and reported need for further education related to HIV and AIDS. At the time of the survey (March-May 2007), the total number of dental students registered was 782 of which 642 (response rate 82%, mean age 21.7 year, 72% girls) completed anonymous self-administered questionnaires in supervised class room settings. A total of 49% and 86% had correct sum scores with respect to knowledge of transmission through contamination and through shaking hands and eating, respectively. About half the dental students recognized a need for further education across HIV related issues, varying from 75% (basic HIV/AIDS related issues) to 84% (patient management). Only 38% of the students had correct sum scores regarding various occupational groups at risk for contacting HIV and AIDS. Multiple logistic regression analysis revealed that compared to privately funded dental school students, publicly funded dental school students were less likely to have correct knowledge about modes of HIV transmission (OR = 0.6) and occupational risk groups (OR = 0.6) and to have received information from lectures/health care workers (OR = 0.5). Students attending privately funded schools were more knowledgeable about various HIV related issues than students from publicly funded schools. About half of the students investigated had received HIV/AIDS information from various sources and reported need for further education. This suggests that students are not adequately prepared for treating patients with HIV infection and AIDS and that the dental school curriculum needs improvements. Differences between public and private dental schools should be considered when planning such improvements.


Controlling food, controlling relationships: exploring the meanings and dynamics of family food practices through the diary-interview approach.
Potential merits of a social practice perspective for examining the meanings and dynamics of family food include moving beyond individual behaviour, and exploring how practices emerge, develop and change. However, researchers have struggled to encourage reflection on mundane practices, and how to understand associated meanings. Drawing on a study of families in South Wales, this article reflects on the value of the diary-interview approach in addressing these methodological challenges, and aims to explore and understand the dynamics of control across family contexts. Contemporary practice theories distinguish between practices as 'performances' and practices as 'entities' and the diary-interview method facilitated an examination of these dimensions. Detailed accounts of daily 'performances' (through diaries), alongside reflection on underlying contexts and 'entities' (through interviews), illustrated the entanglement of control, practices and context. The article adds further complexity to the concept of practice 'bundles' which facilitated an understanding of how food was interrelated with other practices - across family contexts and across generations. Sociological approaches with a practices perspective at the core, offer potential for developing public health interventions by acknowledging: the relational meaning of food; the embeddedness of food within everyday practices; and the need to consider interventions across a range of policy areas.


Biochemical Verification of Tobacco Use and Abstinence: 2019 Update.
The changing prevalence and patterns of tobacco use, the advent of novel nicotine delivery devices, and the development of new biomarkers prompted an update of the 2002 Society for Research on Nicotine and Tobacco (SRNT) report on whether and how to apply biomarker verification for tobacco use and abstinence. The SRNT Treatment Research Network convened a group of investigators with expertise in tobacco biomarkers to update the recommendations of the 2002 SNRT Biochemical Verification Report. Biochemical verification of tobacco use and abstinence increases scientific rigor and is recommended in clinical trials of smoking cessation, when feasible. Sources, appropriate biospecimens, cutpoints, time of detection windows and analytic methods for carbon monoxide, cotinine (including over the counter tests), total nicotine equivalents, minor tobacco alkaloids, and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol are reviewed, as well as biochemical approaches to distinguishing cigarette smoking from use of electronic nicotine delivery devices (ENDS). Recommendations are provided for whether and how to use biochemical verification of tobacco use and abstinence. Guidelines are provided on which biomarkers to use, which biospecimens to use, optimal cutpoints, time windows to detection, and methodology for biochemical verifications. Use of combinations of biomarkers is recommended for assessment of ENDS use. Biochemical verification increases scientific rigor, but there are drawbacks that need to be assessed to determine whether the benefits of biochemical verification outweigh the costs, including the cost of the assays, the feasibility of sample collection, the ability to draw clear conclusions based on the duration of abstinence, and the variability of the assay within the study population. This paper provides updated recommendations from the 2002 SRNT report on whether and how to use biochemical markers in determining tobacco use and abstinence.


Analysis of core circadian feedback loop in suprachiasmatic nucleus of mCry1-luc transgenic reporter mouse.
The suprachiasmatic nucleus (SCN) coordinates circadian rhythms that adapt the individual to solar time. SCN pacemaking revolves around feedback loops in which expression of Period (Per) and Cryptochrome (Cry) genes is periodically suppressed by their protein products. Specifically, PER/CRY complexes act at E-box sequences in Per and Cry to inhibit their transactivation by CLOCK/BMAL1 heterodimers. To function effectively, these closed intracellular loops need to be synchronized between SCN cells and to the light/dark cycle. For Per expression, this is mediated by neuropeptidergic and glutamatergic extracellular cues acting via cAMP/calcium-responsive elements (CREs) in Per genes. Cry genes, however, carry no CREs, and how CRY-dependent SCN pacemaking is synchronized remains unclear. Furthermore, whereas reporter lines are available to explore Per circadian expression in real time, no Cry equivalent exists. We therefore created a mouse, B6.Cg-Tg(Cry1-luc)01Ld, carrying a transgene (mCry1-luc) consisting of mCry1 elements containing an E-box and E'-box driving firefly luciferase. mCry1-luc organotypic SCN slices exhibited stable circadian bioluminescence rhythms with appropriate phase, period, profile, and spatial organization. In SCN lacking vasoactive intestinal peptide or its receptor, mCry1 expression was damped and desynchronized between cells. Despite the absence of CREs, mCry1-luc expression was nevertheless (indirectly) sensitive to manipulation of cAMP-dependent signaling. In mPer1/2-null SCN, mCry1-luc bioluminescence was arrhythmic and no longer suppressed by elevation of cAMP. Finally, an SCN graft procedure showed that PER-independent as well as PER-dependent mechanisms could sustain circadian expression of mCry1. The mCry1-luc mouse therefore reports circadian mCry1 expression and its interactions with vasoactive intestinal peptide, cAMP, and PER at the heart of the SCN pacemaker.


A High Content Screen in Macrophages Identifies Small Molecule Modulators of STING-IRF3 and NFkB Signaling.
We screened a library of bioactive small molecules for activators and inhibitors of innate immune signaling through IRF3 and NFkB pathways with the goals of advancing pathway understanding and discovering probes for immunology research. We used high content screening to measure the translocation from the cytoplasm to nucleus of IRF3 and NFkB in primary human macrophages; these transcription factors play a critical role in the activation of STING and other pro-inflammatory pathways. Our pathway activator screen yielded a diverse set of hits that promoted nuclear translocation of IRF3 and/or NFkB, but the majority of these compounds did not cause activation of downstream pathways. Screening for antagonists of the STING pathway yielded multiple kinase inhibitors, some of which inhibit kinases not previously known to regulate the activity of this pathway. Structure-activity relationships (SARs) and subsequent chemical proteomics experiments suggested that MAPKAPK5 (PRAK) is a kinase that regulates IRF3 translocation in human macrophages. Our work establishes a high content screening approach for measuring pro-inflammatory pathways in human macrophages and identifies novel ways to inhibit such pathways; among the targets of the screen are several molecules that may merit further development as anti-inflammatory drugs.


Amelioration of intestinal dysmotility and stasis by octreotide early after small-bowel autotransplantation in dogs.
Intestinal dysmotility and stasis after intestinal transplantation are considered to promote bacterial overgrowth and translocation. Two prokinetic agents, KW5139 (13-leu-motilin) and the somatostatin analogue octreotide acetate, were studied to determine whether they can ameliorate intestinal dysmotility during the early postoperative period. Motility was recorded by multiple extraluminal strain-gauge transducers in 6 dogs on postoperative days 1, 3, 7, and 14. A barium meal study was performed with a separate group of 8 dogs on postoperative days 3 and 7. The agent KW5139 induced brief, weak contractions in the graft and had little effect on the dilated bowel; however, octreotide induced motor activity that propelled accumulated intestinal contents into the colon and reduced dilation of the transplanted bowel. Octreotide, but not KW5139, ameliorates intestinal dysmotility associated with bowel autotransplantation during the early postoperative period. Short-term administration of octreotide may be useful for the treatment of dysmotility following intestinal transplantation.


On the tool use behavior of the bonobo-chimpanzee last common ancestor, and the origins of hominine stone tool use.
The last common ancestor (LCA) shared by chimpanzees (Pan troglodytes) and bonobos (P. paniscus) was an Early Pleistocene African ape, which, based on the behavior of modern chimpanzees, may be assumed to be a tool-using animal. However, the character of tool use in the Pan lineage prior to the 20th century is largely unknown. Here, I use available data on wild bonobo tool use and emerging molecular estimates of demography during Pan evolution to hypothesise the plausible tool use behavior of the bonobo-chimpanzee LCA (or "Pancestor") at the start of the Pleistocene, over 2 million years ago. This method indicates that the common ancestor of living Pan apes likely used plant tools for probing, sponging, and display, but it did not use stone tools. Instead, stone tool use appears to have been independently invented by Western African chimpanzees (P. t. verus) during the Middle Pleistocene in the region of modern Liberia-Ivory Coast-Guinea, possibly as recently as 200,000-150,000 years ago. If this is the case, then the LCA of humans and chimpanzees likely also did not use stone tools, and this trait probably first emerged among hominins in Pliocene East Africa. This review also suggests that the consistently higher population sizes of Central African chimpanzees (P. t. troglodytes) over the past million years may have contributed to the increased complexity of wild tool use seen in this sub-species today.


Mechanisms of muscle injury after eccentric contraction.
Eccentric contractions of skeletal muscles produce injury and, ultimately, muscle strengthening. Current data suggests that the earliest events associated with injury are mechanical in nature and may be based primarily on the sarcomere strain experienced by the muscle. In this review, recent experimental data, primarily from rabbit dorsiflexor muscles, are used to provide general information regarding the factors that cause injury and means for preventing injury. Mechanical experiments reveal that excessive sarcomere strain is the primary cause of injury. We hypothesize that excessive strain permits extracellular or intracellular membrane disruption that may permit hydrolysis of structural proteins leading to the myofibrillar disruption that is commonly observed. Inflammation that occurs after injury actually further degrades the tissue, but prevention of the inflammation leads to a long-term loss in muscle function. Simple treatments such as increasing muscle oxidative capacity ("getting into shape") or cyclic stress-relaxation of tissue ("stretching out") have no measurable effect on the magnitude of muscle injury that occurs. Ultimately, an improved understanding of the damage mechanism may improve our ability to provide rehabilitative and strengthening prescriptions that have a rational scientific basis.


When in doubt follow your nose-a wayfinding strategy.
Route selection is governed by various strategies which often allow minimizing the required memory capacity. Previous research showed that navigators primarily remember information at route decision points and at route turns, rather than at intersections which required straight walking. However, when actually navigating the route or indicating directional decisions, navigators make fewer errors when they are required to walk straight. This tradeoff between location memory and route decisions accuracy was interpreted as a "when in doubt follow your nose" strategy which allows navigators to only memorize turns and walk straight by default, thus considerably reducing the number of intersections to memorize. These findings were based on newly learned routes. In the present study, we show that such an asymmetry in route memory also prevails for planning routes within highly familiar environments. Participants planned route sequences between locations in their city of residency by pressing arrow keys on a keyboard. They tended to ignore straight walking intersections, but they ignored turns much less so. However, for reported intersections participants were quicker at indicating straight walking than turning. Together with results described in the literature, these findings suggest that a "when in doubt follow your nose strategy" is applied also within highly familiar spaces and might originate from limited working memory capacity during planning a route.


Report of the workshop on biological dosimetry: increasing capacity for emergency response.
Recent events have brought increased attention to the possibility and dangers of a radiological terrorist threat and its potential implication on the national capacity for radiation accident preparedness. In such an event, there is a pressing need to rapidly identify severely irradiated individuals who require prompt medical attention from those who have not been exposed or have been subject to low doses. Initial dose assessment is a key component in rapid triage and treatment, however, the development of accurate methods for rapid dose assessment remains a challenge. In this report, the authors describe a recent workshop supported by the Chemical, Biological, Radiological-Nuclear and Explosives Research and Technology Initiative regarding the international effort to increase biological dosimetry capacity to effectively mount an emergency response in a mass casualty situation. Specifically, the focus of the workshop was on the current state of biological dosimetry capabilities and capacities in North America, recent developments towards increasing throughput for biological dosimetry and to identify opportunities for developing a North American Biological Dosimetry Network and forming partnerships and collaborations within Canada and the USA.


Complementing the Cancer-Immunity Cycle.
Reactivation of cytotoxic CD8+ T-cell responses has set a new direction for cancer immunotherapy. Neutralizing antibodies targeting immune checkpoint programmed cell death protein 1 (PD-1) or its ligand (PD-L1) have been particularly successful for tumor types with limited therapeutic options such as melanoma and lung cancer. However, reactivation of T cells is only one step toward tumor elimination, and a substantial fraction of patients fails to respond to these therapies. In this context, combination therapies targeting more than one of the steps of the cancer-immune cycle may provide significant benefits. To find the best combinations, it is of upmost importance to understand the interplay between cancer cells and all the components of the immune response. This review focuses on the elements of the complement system that come into play in the cancer-immunity cycle. The complement system, an essential part of innate immunity, has emerged as a major regulator of cancer immunity. Complement effectors such as C1q, anaphylatoxins C3a and C5a, and their receptors C3aR and C5aR1, have been associated with tolerogenic cell death and inhibition of antitumor T-cell responses through the recruitment and/or activation of immunosuppressive cell subpopulations such as myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs), or M2 tumor-associated macrophages (TAMs). Evidence is provided to support the idea that complement blocks many of the effector routes associated with the cancer-immunity cycle, providing the rationale for new therapeutic combinations aimed to enhance the antitumor efficacy of anti-PD-1/PD-L1 checkpoint inhibitors.


Support for the reproductive ground plan hypothesis of social evolution and major QTL for ovary traits of Africanized worker honey bees (Apis mellifera L.).
The reproductive ground plan hypothesis of social evolution suggests that reproductive controls of a solitary ancestor have been co-opted during social evolution, facilitating the division of labor among social insect workers. Despite substantial empirical support, the generality of this hypothesis is not universally accepted. Thus, we investigated the prediction of particular genes with pleiotropic effects on ovarian traits and social behavior in worker honey bees as a stringent test of the reproductive ground plan hypothesis. We complemented these tests with a comprehensive genome scan for additional quantitative trait loci (QTL) to gain a better understanding of the genetic architecture of the ovary size of honey bee workers, a morphological trait that is significant for understanding social insect caste evolution and general insect biology. Back-crossing hybrid European x Africanized honey bee queens to the Africanized parent colony generated two study populations with extraordinarily large worker ovaries. Despite the transgressive ovary phenotypes, several previously mapped QTL for social foraging behavior demonstrated ovary size effects, confirming the prediction of pleiotropic genetic effects on reproductive traits and social behavior. One major QTL for ovary size was detected in each backcross, along with several smaller effects and two QTL for ovary asymmetry. One of the main ovary size QTL coincided with a major QTL for ovary activation, explaining 3/4 of the phenotypic variance, although no simple positive correlation between ovary size and activation was observed. Our results provide strong support for the reproductive ground plan hypothesis of evolution in study populations that are independent of the genetic stocks that originally led to the formulation of this hypothesis. As predicted, worker ovary size is genetically linked to multiple correlated traits of the complex division of labor in worker honey bees, known as the pollen hoarding syndrome. The genetic architecture of worker ovary size presumably consists of a combination of trait-specific loci and general regulators that affect the whole behavioral syndrome and may even play a role in caste determination. Several promising candidate genes in the QTL intervals await further study to clarify their potential role in social insect evolution and the regulation of insect fertility in general.


UTRN on chromosome 6q24 is mutated in multiple tumors.
Though deletion of the long arm of chromosome 6 is one of the most common aberrations in tumors, its targeted gene(s) has not been convincingly identified. Using a functional screening approach, we found that UTRN (which encodes utrophin, a dystrophin-related protein) at 6q24, when expressed in an antisense orientation, induced cellular transformation, consistent with a tumor suppressor role. Northern blot analysis, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), and gene expression arrays all showed that UTRN expression was downregulated in primary tumors compared with matched normal tissues. Several UTRN neighbor genes were not affected in some tumors with UTRN downregulation, suggesting that UTRN was specifically targeted. RT-PCR, coupled with an in vitro transcription and translation assay, revealed inactivation mutations in 21/62 breast cancers, 4/20 neuroblastomas and 4/15 malignant melanomas. Most of the mutations were deletions involving one or more exons that led to the truncation of utrophin. Splicing errors were found in two cases, and nonsense mutation in one case. Overexpression of a wild-type UTRN in breast cancer cells inhibited tumor cell growth in vitro and reduced their tumor potential in nude mice. Our studies suggest that UTRN is a candidate tumor suppressor gene.


Does high-density lipoprotein protect vascular function in healthy pregnancy?
The maternal adaptation to pregnancy includes hyperlipidaemia, oxidative stress and chronic inflammation. In non-pregnant individuals, these processes are usually associated with poor vascular function. However, maternal vascular function is enhanced in pregnancy. It is not understood how this is achieved in the face of the adverse metabolic and inflammatory environment. Research into cardiovascular disease demonstrates that plasma HDL (high-density lipoprotein), by merit of its functionality rather than its plasma concentration, exerts protective effects on the vascular endothelium. HDL has vasodilatory, antioxidant, anti-thrombotic and anti-inflammatory effects, and can protect against endothelial cell damage. In pregnancy, the plasma HDL concentration starts to rise at 10 weeks of gestation, peaking at 20 weeks. The initial rise in plasma HDL occurs around the time of the establishment of the feto-placental circulation, a time when the trophoblast plugs in the maternal spiral arteries are released, generating oxidative stress. Thus there is the intriguing possibility that new HDL of improved function is synthesized around the time of the establishment of the feto-placental circulation. In obese pregnancy and, to a greater extent, in pre-eclampsia, plasma HDL levels are significantly decreased and maternal vascular function is reduced. Wire myography studies have shown an association between the plasma content of apolipoprotein AI, the major protein constituent of HDL, and blood vessel relaxation. These observations lead us to hypothesize that HDL concentration, and function, increases in pregnancy in order to protect the maternal vascular endothelium and that in pre-eclampsia this fails to occur.