Mutations in the 5' end of the human immunodeficiency virus type 1 polypurine tract affect RNase H cleavage specificity and virus titer.
The RNase H activity of retroviral reverse transcriptases (RTs) degrades viral genomic RNA after it has been copied into DNA, removes the tRNA used to initiate negative-strand DNA synthesis, and generates and removes the polypurine tract (PPT) primer used to initiate positive-strand DNA synthesis. The cleavages that remove the tRNA and that generate and remove the PPT primer must be specific to generate linear viral DNAs with ends that are appropriate for integration into the host cell genome. The crystal structure of human immunodeficiency virus type 1 (HIV-1) RT in a complex with an RNA/DNA duplex derived from the PPT revealed that the 5' end of the PPT deviates from traditional Watson-Crick base pairing. This unusual structure may play a role in the proper recognition of the PPT by HIV-1 RT. We made substitution mutations in the 5' end of the PPT and determined their effects on virus titer. The results indicated that single and double mutations in the 5' end of the PPT had modest effects on virus replication in a single-cycle assay. More complex mutations had stronger effects on virus titer. Analysis of the two-long-terminal-repeat circle junctions derived from infecting cells with the mutant viruses indicated that the mutations affected RNase H activity, resulting in the retention of PPT sequences on viral DNA. The mutants tested preferentially retained specific segments of the PPT, suggesting an effect on cleavage specificity. These results suggest that structural features of the PPT are important for its recognition and cleavage in vivo.


Identification of putative calorie restriction mimetics using mammalian gene expression profiles.
Obesity is a risk factor for cardiovascular diseases, diabetes and cancer. In theory, the obesity problem could be solved by the adherence to a calorie-restricted diet, but that is not generally achieved in practice. An alternative is a pharmacological approach, using compounds that trigger the same metabolic changes associated with calorie restriction. Here, I expand in the pharmacological direction by identifying compounds that induce liver gene signature profiles that mimic those induced by calorie restriction. Using gene expression profiles from mice and rat, I identify corticosteroids, PPAR agonists and some antibacterial/antifungal as candidate compounds mimicking the response to calorie restriction in the liver gene signatures.


Facilitators and Barriers to HIV Screening: A Qualitative Meta-Synthesis.
Accomplishments in biomedical research and technology, combined with innovative community and clinically based interventions, have expanded HIV testing globally. However, HIV screening and receipt of results remains a challenge in some areas. To optimize the benefits of HIV screening, it is imperative that there is a better understanding of the barriers to and motivators of testing for HIV infection. This study is a meta-synthesis of the qualitative literature on HIV screening and receipt of results; 128 unique publications had implications for HIV screening and receipt of results. A socioecological perspective provided an appropriate approach for synthesizing the literature. Three levels of influence emerged: individual attributes, interpersonal attributes, and broader patterns of influence. Findings were reviewed and found to have implications for continued engagement in the HIV treatment cascade. Recommendations to enhance HIV screening and to ensure receipt of results are proposed and discussed.


Noninvasive cineangiography by magnetic resonance global coherent free precession.
Cardiovascular disease is primarily diagnosed using invasive X-ray cineangiography. Here we introduce a new concept in magnetic resonance imaging (MRI) that, for the first time, produces similar images noninvasively and without a contrast agent. Protons in moving blood are 'tagged' every few milliseconds as they travel through an arbitrary region in space. Simultaneous with ongoing tagging of new blood, previously tagged blood is maintained in a state of global coherent free precession (GCFP), which allows acquisition of consecutive movie frames as the heart pushes blood through the vascular bed. Body tissue surrounding the moving blood is never excited and therefore remains invisible. In 18 subjects, pulsating blood could be seen flowing through three-dimensional (3D) space for distances of up to 16 cm outside the stationary excitation region. These data underscore that our approach noninvasively characterizes both anatomy and blood flow in a manner directly analogous to invasive procedures.


Does genotype and equol-production status affect response to isoflavones? Data from a pan-European study on the effects of isoflavones on cardiovascular risk markers in post-menopausal women.
The increase in CVD incidence following the menopause is associated with oestrogen loss. Dietary isoflavones are thought to be cardioprotective via their oestrogenic and oestrogen receptor-independent effects, but evidence to support this role is scarce. Individual variation in response to diet may be considerable and can obscure potential beneficial effects in a sample population; in particular, the response to isoflavone treatment may vary according to genotype and equol-production status. The effects of isoflavone supplementation (50 mg/d) on a range of established and novel biomarkers of CVD, including markers of lipid and glucose metabolism and inflammatory biomarkers, have been investigated in a placebo-controlled 2 x 8-week randomised cross-over study in 117 healthy post-menopausal women. Responsiveness to isoflavone supplementation according to (1) single nucleotide polymorphisms in a range of key CVD genes, including oestrogen receptor (ER) alpha and beta and (2) equol-production status has been examined. Isoflavones supplementation was found to have no effect on markers of lipids and glucose metabolism. Isoflavones improve C-reactive protein concentrations but do not affect other plasma inflammatory markers. There are no differences in response to isoflavones according to equol-production status. However, differences in HDL-cholesterol and vascular cell adhesion molecule 1 response to isoflavones v. placebo are evident with specific ERbeta genotypes. In conclusion, isoflavones have beneficial effects on C-reactive protein, but not other cardiovascular risk markers. However, specific ERbeta gene polymorphic subgroups may benefit from isoflavone supplementation.


Human and climate drivers of global biomass burning variability.
Biomass burning is one of the most critical factors impacting vegetation and atmospheric trends, with important societal implications, particularly when extreme weather conditions occur. Trends and factors of burned area (BA) have been analysed at regional and global scales, but little effort has been dedicated to study the interannual variability. This paper aimed to better understand factors explaining this variation, under the assumption that the more human control of fires the more frequently they occur, as burnings will be less dependent of weather cycles. Interannual variability of BA was estimated from the coefficient of variation of the annual BA (BA_CV) estimated from satellite data at 250 m, covering the period from 2001 to 2018. These data and the explanatory variables were resampled at 0.25-degree resolution for global analysis. Relations between this variable and explanatory factors, including human and climate drivers, were estimated using Random Forest (RF) and generalized additive models (GAM). BA_CV was negatively related to BA_Mean, implying that areas with higher average BA have lower variability as well. Interannual BA variability decreased when maximum temperature (TMAX) and actual and potential evapotranspiration (AET, PET) increased, cropland and livestock density increased and the human development index (HDI) values decreased. GAM models indicated interesting links with AET, PET and precipitation, with negative relation with BA_CV for the lower ranges and positive for the higher ones, the former indicating fuel limitations of fire activity, and the latter climate constrains. For the global RF model, TMAX, AET and HDI were the main drivers of interannual variability. As originally hypothesised, BA_CV was more dependent on human factors (HDI) in those areas with medium to large BA occurrence, particularly in tropical Africa and Central Asia, while climatic factors were more important in boreal regions, but also in the tropical regions of Australia and South America.


Occupational level of the father and alcohol consumption during adolescence; patterns and predictors.
This paper describes and attempts to explain the association between occupational level of the father and high alcohol consumption among a cohort of New Zealand adolescents from age 11 to 21. Data were obtained from the longitudinal Dunedin multidisciplinary health and development study. At each measurement wave, those who then belonged to the quartile that reported the highest usual amount of alcohol consumed on a typical drinking occasion were categorised as high alcohol consumers. Potential predictors of high alcohol consumption included environmental factors, individual factors, and educational achievement measured at age 9, 11, or 13. Longitudinal logistic GEE analyses described and explained the relation between father's occupation and adolescent alcohol consumption. Dunedin, New Zealand. About 1000 children were followed up from birth in 1972 until adulthood. A significant association between fathers' occupation and adolescent alcohol consumption emerged at age 15. Overall adolescents from the lowest occupational group had almost twice the odds of being a large consumer than the highest occupational group. The association between father's occupation and high alcohol consumption during adolescence was explained by the higher prevalence of familial alcohol problems and friends approving of alcohol consumption, lower intelligence scores, and lower parental attachment among adolescents from lower occupational groups. Socioeconomic background affects adolescent alcohol consumption substantially. This probably contributes to cumulation of disadvantage. Prevention programmes should focus on adolescents from lower socioeconomic groups and make healthier choices the easier choices by means of environmental change.


A Pilot Study to Determine the Consistency of Simultaneous Sleep Actigraphy Measurements Comparing All Four Limbs of Patients with Parkinson Disease.
Wrist actigraphy is a form of objective sleep measurement that has gained a central role in sleep research and clinical settings. Guidelines for actigraphy recommend placing the monitor on the non-dominant wrist, however, this potentially will be the most involved limb for someone with Parkinson disease, and so alternative placement would be preferred. To-date, there is little published about sleep actigraphy use in Parkinson disease (PD). This study examines the degree of sleep actigraphy score variation in persons with PD when monitors are placed simultaneously on all four limbs. In this study, four participants wore a sleep actigraph on each limb for seven nights. Data from the four actigraphs was compared within each participant to determine the degree of consistency. We found that all of the participants' sleep efficiency and total sleep time scores were higher in the lower limb than upper limb. There was no notable difference in sleep variables between the dominant arm and non-dominant arm. We concluded that simultaneous actigraphy measurement did not notably vary between dominant and non-dominant arms. However, a discrepancy was seen between upper limbs and lower limbs actigraph scores. Further study is warranted to develop guidelines for sleep actigraphy use in this population.


Attitudes in Patients with Autosomal Dominant Polycystic Kidney Disease Toward Prenatal Diagnosis and Preimplantation Genetic Diagnosis.
No recommendations currently exist regarding implementation of both prenatal diagnosis and preimplantation genetic diagnosis (PGD) for autosomal dominant polycystic kidney disease (ADPKD). This study evaluated attitudes in ADPKD patients with either chronic kidney disease (CKD) stages I-IV or end-stage renal failure (ESRF) toward prenatal diagnosis and PGD. Ninety-six ADPKD patients were recruited from an outpatient clinic, wards, and dialysis units. Thirty-eight patients had ESRF and 58 had CKD stages I-IV. Participants were given an information sheet on prenatal diagnosis and PGD and subsequently completed a questionnaire. The median age of participants was 51.5 years. Seventeen percent of ADPKD patients with CKD and 18% of ADPKD patients with ESRF would consider prenatal diagnosis and termination of pregnancy for ADPKD. Fifty percent with CKD would have opted for PGD (or might consider it in the future) were it available and funded by the UK National Health Service, compared to 63% in the ESRF group (p = 0.33). Sixty-nine percent in the CKD group and 68% in the ESRF group believed that PGD should be offered to other patients. There was a spectrum of attitudes among this cohort. A proportion of patients believe that PGD should be made available to prospective parents with this disease. The discrepancy between the low proportion (17% CKD, 18% ESRF) who would consider prenatal diagnosis and termination of pregnancy and the higher number who hypothetically express an intention or wish to access PGD (50% CKD and 63% ESRF) indicates far greater acceptability for diagnostic methods that occur before embryo implantation. It is not known how the development of methods to identify patients whose renal function is likely to decline rapidly and treatments altering the natural history of ADPKD will affect these attitudes.


The cotranslational maturation program for the type II membrane glycoprotein influenza neuraminidase.
The earliest steps in nascent protein maturation greatly affect its overall efficiency. Constraints placed on maturing proteins at these early stages limit available conformations and help to direct the native maturation process. For type II membrane proteins, these cotranslational constraints include N- and C-terminal membrane tethering, chaperone binding, and disulfide bond formation. The cotranslational maturation process for the type II membrane glycoprotein influenza neuraminidase (NA) was investigated to provide a deeper understanding of these initial endoplasmic reticulum events. The type II orientation provides experimental advantages to monitor the first maturation steps. Calnexin was shown to cotranslationally interact with NA prior to calreticulin. These interactions were required for the efficient maturation of NA as it prematurely formed intramolecular disulfides and aggregated when calnexin and calreticulin interactions were abolished. Lectin chaperone binding slowed the NA maturation process, increasing its fidelity. Carbohydrates were required for NA maturation in a regio-specific manner. A subset of NA formed intermolecular disulfides and oligomerized cotranslationally. This fraction increased in the absence of calnexin and calreticulin binding. NA dimerization also occurred for an NA mutant lacking the critical large loop disulfide bond, indicating that dimerization did not require proper NA oxidation. The strict evaluation of proper maturation carried out by the quality control machinery was instilled at the tetramerization step. This study illustrates the type II membrane protein maturation process and shows how important cotranslational events contribute to the proper cellular maturation of glycoproteins.


Cross-priming of CD8+ T cells stimulated by virus-induced type I interferon.
CD8+ T cell responses can be generated against antigens that are not expressed directly within antigen-presenting cells (APCs), through a process known as cross-priming. To initiate cross-priming, APCs must both capture extracellular antigen and receive specific activation signals. We have investigated the nature of APC activation signals associated with virus infection that stimulate cross-priming. We show that infection with lymphocytic choriomeningitis virus induces cross-priming by a mechanism dependent on type I interferon (IFN-alpha/beta). Activation of cross-priming by IFN-alpha/beta was independent of CD4+ T cell help or interaction of CD40 and CD40 ligand, and involved direct stimulation of dendritic cells. These data identify expression of IFN-alpha/beta as a mechanism for the induction of cross-priming during virus infections.


Impaired immune tolerance to Porphyromonas gingivalis lipopolysaccharide promotes neutrophil migration and decreased apoptosis.
Periodontitis, a chronic inflammatory disease of the tissues supporting the teeth, is characterized by an exaggerated host immune and inflammatory response to periopathogenic bacteria. Toll-like receptor activation, cytokine network induction, and accumulation of neutrophils at the site of inflammation are important in the host defense against infection. At the same time, induction of immune tolerance and the clearance of neutrophils from the site of infection are essential in the control of the immune response, resolution of inflammation, and prevention of tissue destruction. Using a human monocytic cell line, we demonstrate that Porphyromonas gingivalis lipopolysaccharide (LPS), which is a major etiological factor in periodontal disease, induces only partial immune tolerance, with continued high production of interleukin-8 (IL-8) but diminished secretion of tumor necrosis factor alpha (TNF-α) after repeated challenge. This cytokine response has functional consequences for other immune cells involved in the response to infection. Primary human neutrophils incubated with P. gingivalis LPS-treated naïve monocyte supernatant displayed a high migration index and increased apoptosis. In contrast, neutrophils treated with P. gingivalis LPS-tolerized monocyte supernatant showed a high migration index but significantly decreased apoptosis. Overall, these findings suggest that induction of an imbalanced immune tolerance in monocytes by P. gingivalis LPS, which favors continued secretion of IL-8 but decreased TNF-α production, may be associated with enhanced migration of neutrophils to the site of infection but also with decreased apoptosis and may play a role in the chronic inflammatory state seen in periodontal disease.


Prognosis of chronic granulomatous disease.
The records of 28 patients with chronic granulomatous disease born over a 32 year period were reviewed. The characteristics of the group, and the frequency with which various clinical and laboratory features had been recorded, was assessed. Nine patients were known to have died, in most cases of progressive suppurative infection. Actuarial analysis showed 50% survival through the third decade of life. The long term survival of patients developing symptoms after the end of the first year of life was significantly better than that of patients whose illness started in infancy. Our data confirm that the severity of chronic granulomatous disease is not uniform, and that the prognosis for long term survival is better than that suggested in earlier reports. Early onset may be a poor prognostic sign and invasive aspergillosis is a life threatening complication. In the absence of curative treatment, trials to assess the effectiveness of interferon gamma are necessary and early antenatal diagnosis should be offered to as many affected families as possible.


Risk factors associated with human cystic echinococcosis in Florida, Uruguay: results of a mass screening study using ultrasound and serology.
Sonographic evidence of asymptomatic Echinococcus granulosus lesions in the liver was found in 156 of 9,515 persons in the Department of Florida, Uruguay. The sensitivity of ELISA and latex agglutination serology compared with ultrasound was 47.6% and 28.1%, respectively, and specificity was > 85%. There was a significant positive association between positive sonography and a personal history of previous but treated Echinococcus infection while those that were seropositive but ultrasound-negative were significantly more likely to have a personal history of infection or a history of infection in their family. Prevalence of infection increased significantly with age. There was no correlation between echinococcosis and dog ownership or home slaughter of sheep but offal disposal was important, with an increased prevalence of infection of 3.2%, 2.8%, and 3.1%, respectively, in persons feeding offal to dogs or burying or burning it compared with a prevalence of 0.8-1.5% in those using other methods of disposal. Almost half the population, when questioned, seemed to have sound knowledge about E. granulosus and described correct treatment of E. granulosus in dogs but this did not affect prevalence. There was a significant positive association between infection and the presence of a fenced fruit/vegetable garden and use of rural waters, particularly the cachimba (a small dam) and the aljibe (a cistern or tank) that collect rainwater from the ground surface and roofs, respectively.


Transfection mechanisms of polyplexes, lipoplexes, and stealth liposomes in α₅β₁ integrin bearing DLD-1 colorectal cancer cells.
Receptor targeted, PEGylated transfection agents can improve stability and delivery specificity of current cationic lipid and polymer based nonviral gene delivery vehicles, but their mode of transfection is poorly understood. We therefore investigated the transfection mechanisms of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP)/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) lipoplexes, branched polyethylenimine (bPEI) polyplexes, and bPEI encapsulated in either PEGylated (stealth) nontargeted liposomes or PR_b peptide (targeted to α5β1 integrin) functionalized stealth liposomes in DLD-1 colorectal cancer cells in vitro with gene expression assays, flow cytometry and confocal microscopy. DOTAP/DOPE and PR_b functionalized stealth liposomes mediated higher gene expression compared to nontargeted stealth liposomes and bPEI. However DOTAP/DOPE was internalized slowly leading to lower levels of DNA uptake. In contrast, despite high internalization of bPEI polyplexes, gene expression levels were low as DNA was unable to escape from the endosomes. Nontargeted stealth liposomes also mediated low gene expression due to low amounts of DNA internalized and slow internalization kinetics. PR_b functionalized stealth liposomes struck an optimal balance among these transfection agents with efficient transfection arising from fast integrin mediated internalization kinetics, high amounts of DNA uptake, and endosomal escape. We found α5β1 integrin to be a valuable target for gene delivery and that the caveolar endocytic pathway may offer an advantage to receptor targeted PEGylated transfection agents in DLD-1 cells.


COVID-19 Impacts at a Small Mid-Atlantic Liberal-Arts College with Implications for STEM Education.
During the COVID-19 pandemic, with very little preparation and within a brief span of 48 hours, the Wesley College STEM faculty and students triaged into a remote-only form of instruction. Wesley College STEM student COVID-19 impact surveys showed underlying gaps in economic equity, increased family responsibilities, struggles to stay motivated, social isolation, and higher levels of psychological stress. Yet, the crisis demonstrated new ways in which technology can be harnessed and allowed STEM students to reconsider how jobs and skills should be aligned. A STEM faculty COVID-19 check-in survey and interview responses revealed a quick realization that faculty could not rely solely on Wesley's Jenzebar learning management system (MyWesley). To engage their students and to create a supportive learning environment, STEM faculty sought new strategies and approaches for a diverse set of STEM learners. For synchronous e-teaching, the faculty used the Microsoft-Teams and the Zoom video conferencing platforms. Faculty only adopted MyWesley to execute dedicated asynchronous tasks (laboratory assignments, reports, exams). The STEM students were overwhelmingly positive about STEM faculty availability during the crisis. Still, both faculty and students indicated a much stronger preference for the face-to-face delivery of their course content via a traditional classroom setting.


An Atomistic Look into Bio-inspired Nanoparticles and their Molecular Interactions with Cells.
Nanoparticles (NPs) have sizes that approach those of pathogens and they can interact with the membranes of eukaryotic cells in an analogous fashion. Typically, NPs are taken up by the cell via the plasma membrane by receptor-mediated processes and subsequently interact with various endomembranes. Unlike pathogens, however, NPs lack the remarkable specificity gained during the evolutionary process and their design and optimization remains an expensive and time-consuming undertaking, especially considering the limited information available on their molecular interactions with cells. In this context, molecular dynamics (MD) simulations have emered as a promising strategy to investigate the mechanistic details of the interaction of NPs with mammalian or viral membranes. In particular, MD simulations have been extensively used to study the uptake process of NPs into the cell, focusing on membrane vesiculation, endocytic routes, or passive permeation processes. While such work is certainly relevant for understanding NP-cell interactions, it remains very difficult to determine the correspondence between generic models and the actual NP. Here, we review how chemically-specific MD simulations can provide rational guidelines towards further bio-inspired NP optimization.


Structure of FIV capsid C-terminal domain demonstrates lentiviral evasion of genetic fragility by coevolved substitutions.
Viruses use a strategy of high mutational rates to adapt to environmental and therapeutic pressures, circumventing the deleterious effects of random single-point mutations by coevolved compensatory mutations, which restore protein fold, function or interactions damaged by initial ones. This mechanism has been identified as contributing to drug resistance in the HIV-1 Gag polyprotein and especially its capsid proteolytic product, which forms the viral capsid core and plays multifaceted roles in the viral life cycle. Here, we determined the X-ray crystal structure of C-terminal domain of the feline immunodeficiency virus (FIV) capsid and through interspecies analysis elucidate the structural basis of co-evolutionarily and spatially correlated substitutions in capsid sequences, which when otherwise uncoupled and individually substituted into HIV-1 capsid impair virion assembly and infectivity. The ability to circumvent the deleterious effects of single amino acid substitutions by cooperative secondary substitutions allows mutational flexibility that may afford viruses an important survival advantage. The potential of such interspecies structural analysis for preempting viral resistance by identifying such alternative but functionally equivalent patterns is discussed.


Variation in Antiosteoporotic Drug Prescribing and Spending Across Spain. A Population-Based Ecological Cross-Sectional Study.
Introduction: Evidence has shown that utilization of antiosteoporotic medications does not correspond with risk, and studies on other therapies have shown that adequacy of pharmaceutical prescribing might vary between regions. Nevertheless, very few studies have addressed the variability in osteoporotic drug consumption. We aimed to describe variations in pharmaceutical utilization and spending on osteoporotic drugs between Health Areas (HA) in Spain. Methods: Population-based cross-sectional ecological study of expenditure and utilization of the five therapeutic groups marketed for osteoporosis treatment in Spain in 2009. Small area variation analysis (SAVA) methods were used. The units of analysis were the 168 HA of 13 Spanish regions, including 7.2 million women aged 50 years and older. The main outcomes were the defined daily dose (DDD) per 1000 inhabitants and day (DDD/1000/Day) dispensed according to the pharmaceutical claims reimbursed, and the expenditure on antiosteoporotics at retail price per woman ≥50 years old and per year. Results: The average osteoporosis drug consumption was 116.8 DDD/1000W/Day, ranging from 78.5 to 158.7 DDD/1000W/Day between the HAs in the 5th and 95th percentiles. Seventy-five percent of the antiosteoporotics consumed was bisphosphonates, followed by raloxifene, strontium ranelate, calcitonins, and parathyroid hormones including teriparatide. Regarding variability by therapeutic groups, biphosphonates showed the lowest variation, while calcitonins and parathyroid hormones showed the highest variation. The annual expenditure on antiosteoporotics was €426.5 million, translating into an expenditure of €59.2 for each woman ≥50 years old and varying between €38.1 and €83.3 between HAs in the 5th and 95th percentiles. Biphosphonates, despite accounting for 79% of utilization, only represented 63% of total expenditure, while parathyroid hormones with only 1.6% of utilization accounted for 15% of the pharmaceutical spending. Conclusion: This study highlights a marked geographical variation in the prescription of antiosteoporotics, being more pronounced in the case of costly drugs such as parathyroid hormones. The differences in rates of prescribing explained almost all of the variance in drug spending, suggesting that the difference in prescription volume between territories, and not the price of the drugs, is the main source of variation in this setting. Data on geographical variation of prescription can help guide policy proposals for targeting areas with inadequate antiosteoporotic drug use.


Marginal masticatory mucosa dimensional changes in immediate post-extractive implants: a 2 year prospective cohort study.
The present two-year prospective cohort study was undertaken to evaluate marginal masticatory mucosa dimensional changes around immediate post-extractive implants positioned transgingivally with a non-submerged healing screw. Twenty-one immediate post-extractive implants from 21 patients were enrolled, peri-implant gap was filled with bovine bone mineral, and soft tissue was allowed to heal around a non-submerged healing screw. Post-extractive socket dimension was recorded. Intraoperative (T0) vertical distances: bone margin level (BML) from the bone margin to the implant platform and mucosal margin height (MMH) from marginal mucosa to implant platform were taken; MMH measurement was repeated 4 months later (T4). Horizontal mucosal level (HML): from customized stent to marginal mucosa at 0, 4, 12, and 24 months postoperatively (T0, T4, T12, T24) and vertical mucosal level (VML): from the stent to marginal mucosa at T4, T12, T24 were registered. One implant failed at 3 weeks; in the remaining 20 cases the MMH, coronally positioned with respect to the BML ≅2 mm at T0, showed a statistically significant vertical contraction of the mucosa at T4. Other vertical mucosal measurements (VML) did not show further changes over time. HML measures showed a, statistically significant, shrinkage of the mucosa on the transverse plane between T0/T12 and T0/T24 and between T4/T12 and T4/T24. Immediate post-extractive implant inserted transgingivally with a non-submerged healing screw and internal peri-implant gap filled with bovine bone mineral may favor an early and stable peri-implant soft tissue healing over 2 years.


Wild birds respond to flockmate loss by increasing their social network associations to others.
Understanding the consequences of losing individuals from wild populations is a current and pressing issue, yet how such loss influences the social behaviour of the remaining animals is largely unexplored. Through combining the automated tracking of winter flocks of over 500 wild great tits (Parus major) with removal experiments, we assessed how individuals' social network positions responded to the loss of their social associates. We found that the extent of flockmate loss that individuals experienced correlated positively with subsequent increases in the number of their social associations, the average strength of their bonds and their overall connectedness within the social network (defined as summed edge weights). Increased social connectivity was not driven by general disturbance or changes in foraging behaviour, but by modifications to fine-scale social network connections in response to losing their associates. Therefore, the reduction in social connectedness expected by individual loss may be mitigated by increases in social associations between remaining individuals. Given that these findings demonstrate rapid adjustment of social network associations in response to the loss of previous social ties, future research should examine the generality of the compensatory adjustment of social relations in ways that maintain the structure of social organization.


Determination of the catalytic mechanism for mitochondrial malate dehydrogenase.
The kinetics of malate dehydrogenase (MDH) catalyzed oxidation/reduction of L-malate/oxaloacetate is pH-dependent due to the proton generated/taken up during the reaction. Previous kinetic studies on the mitochondrial MDH did not yield a consensus kinetic model that explains both substrate and pH dependency of the initial velocity. In this study, we propose, to our knowledge, a new kinetic mechanism to explain kinetic data acquired over a range of pH and substrate concentrations. Progress curves in the forward and reverse reaction directions were obtained under a variety of reactant concentrations to identify associated kinetic parameters. Experiments were conducted at physiologically relevant ionic strength of 0.17 M, pH ranging between 6.5 and 9.0, and at 25 °C. The developed model was built on the prior observation of proton uptake upon binding of NADH to MDH, and that the MDH-catalyzed oxidation of NADH may follow an ordered bi-bi mechanism with NADH/NAD binding to the enzyme first, followed by the binding of oxaloacetate/L-malate. This basic mechanism was expanded to account for additional ionic states to explain the pH dependency of the kinetic behavior, resulting in what we believe to be the first kinetic model explaining both substrate and pH dependency of the reaction velocity.


A safe and sensitive enterovirus A71 infection model based on human SCARB2 knock-in mice.
Enterovirus A71 infection has become a severe threat for global public health. Vaccines for controlling and preventing Enterovirus A71 epidemics are highly demanded, however, vaccine evaluation has been hindered by the lack of suitable Enterovirus A71 infection animal models. Here we established an hSCARB2 knockin mouse model for real-time monitoring of enterovirus A71 infection in vivo. This model was sensitive to the infection of both replication-competent virus rEV71(FY)-EGFP and single round pseudotype virus pEV71(FY)-Luc. The intensity of bioluminescence correlated well with viral loads in infected tissues (R=0.86, P<0.01). Pathological changes recapitulated human infectious and clinical features of enterovirus A71, including both general characteristics of "hand foot and mouth" and the severe symptoms in the CNS. A formalin-inactivated enterovirus A71 vaccine can elicit antibodies in R26-hSCARB2 mice, which play effective roles in protecting knockin mice against enterovirus A71 infection as indicated by bioluminescence. Therefore, this work provides a safe, sensitive and visualizing model for exploring mechanisms of enterovirus A71 infection and examining human enterovirus A71 vaccines and antiviral therapies.


Who Really Wins? Efficacy of a Croatian Youth Gambling Prevention Program.
This paper reports on the development and pilot evaluation of a Croatian school-based youth gambling prevention program "Who really wins?". The program is aimed at minimizing risk and enhancing protective factors related to youth gambling. A short-term evaluation of the program was conducted with a sample of 190 first and second year high-school students (67.6% boys, aged 14-17 years; average age 15.61). An experimental design with two groups (Training vs. No Training) and two measurement sessions (pre-test and post-test sessions) was used to evaluate change in problem gambling awareness, cognitive distortions, knowledge of the nature of random events as well as in social skills. Results showed significant changes in the post-test sessions, which can be attributed to changes in the Training group. We observed a decrease in risk factors, namely better knowledge about gambling and less gambling related cognitive distortions. Immediate effects on protective factors such as problem solving skills, refusal skills, and general self-efficacy were not observed. Findings also show program effects to be the same for both boys and girls, students from different types of schools, for those with different learning aptitudes, as well as for those at different risk levels with regard to their gambling, which speaks in favour of the program's universality. The program had no iatrogenic effects on behaviour change and shows promise as an effective tool for youth gambling prevention. Future research and a long-term evaluation are needed to determine whether the observed changes are also linked to behavioural change.


Effect of ageing on peak expiratory flow variability and nocturnal exacerbations in bronchial asthma.
The aim of this study was to evaluate the role of ageing on variability of airflow obstruction and on the specific pattern of nocturnal exacerbations ("morning dipping" of peak expiratory flow (PEF)) in asthma. Two groups of stable asthmatics (Group A: 23 patients, aged 14-47 yrs; Group B: 20 patients, aged 53-74 yrs), that were similar for duration of disease, degree of obstruction and response to bronchodilators, were studied. PEF was monitored four times daily for 2 weeks, and amplitude of variation and "morning dip" were calculated. Both PEF amplitude and "morning dip" were greater in the older patients (p<0.005). Amplitude was negatively correlated with baseline forced expiratory volume in one second (FEV1) in both groups (p<0.01); multiple correlation with age, duration of disease, baseline FEV1 and degree of reversibility was significant in the older group only. Morning dip was inversely correlated to baseline FEV1 (p<0.01) and reversibility (p<0.05). Nocturnal symptoms were reported by all of the five Group A "dippers", but by only 6 of the 13 Group B "dippers" (p<0.04). Ageing contributes to increased variability of airway calibre in asthma. Aged patients are potentially at risk because functional evidence of nocturnal asthma may frequently be accompanied by a poor subjective awareness. Therefore, a more extensive practice of PEF monitoring is recommended in aged asthmatics.


Consumption of Cuban Policosanol Improves Blood Pressure and Lipid Profile via Enhancement of HDL Functionality in Healthy Women Subjects: Randomized, Double-Blinded, and Placebo-Controlled Study.
Policosanol has been reported to improve blood pressure, lipid profile, and HDL functionality via inhibition of cholesteryl ester transfer protein (CETP) both in vitro and in vivo in zebrafish and human models. However, there are limited reports and randomized, double-blinded trials on policosanol that could advocate the blood pressure-lowering effect in prehypertensive participants. Therefore, we performed in vitro, in vivo, and ex vivo experiments to provide more substantial and concrete data on the blood pressure-lowering effect of policosanol. Consumption of policosanol for 8 weeks enhanced plasma antioxidant activity. In the policosanol group, plasma total cholesterol (TC) and triglyceride (TG) levels were reduced up to 20% and 14%, respectively, and HDL-C level was elevated up to 1.3-fold compared to that at week 0. TG/HDL-C and cholesteryl ester transfer protein (CETP) activities were reduced up to 36% and 20%, respectively. Uptake of oxidized LDL in macrophages was reduced as oxidized species levels were reduced, and HDL2-associated paraoxonase activities were enhanced by 60% compared to those at week 0. Encapsulation of policosanol into reconstituted HDL (PCO-rHDL) enhanced cholesterol efflux activity and insulin secretion capacity. In conclusion, consumption of policosanol for 8 weeks in healthy female subjects resulted in lowered blood pressure and CETP activity via elevation of HDL/apoA-I contents and enhancement of HDL functionalities, including cholesterol efflux and insulin secretion. These functional enhancements of HDL can contribute to the prevention of aging-related diseases, hypertension, and stroke.


Bile reflux measurement and its contribution to the severity of reflux esophagitis.
Gastroesophageal reflux disease (GERD) may occur with acid, bile or in a mixed form. Endoscopic injury and mucosal metaplasia are a known sequelae to pathological GERD. The aim of the study was to determine the contribution of acid and duodenogastroesophageal reflux to endoscopic severity in patients with GERD and Barrett's esophagus (BE). Ninety-one patients complaining of reflux symptoms were studied with upper gastrointestinal endoscopy and graded to nonerosive reflux disease (NERD), erosive reflux disease (ERD) and BE. Esophageal manometry and simultaneous ambulatory 24-h esophageal pH and bilirubin monitoring (Bilitec 2000) were performed in all patients. Seventy-one patients (78.0%) had ERD (Savary-Miller (grade I-III), 11 patients (12.1%) had NERD and 9 patients (9.9%) had BE, which were suspected endoscopically and diagnosed by histological esophageal biopsy. Combined 24-h esophageal bilirubin and pH monitoring revealed the following: 39 patients (42.9%) had mixed acid and bile reflux, 16 (17.6%) had pathological acid reflux alone, 18 (19.8%) had bile reflux alone and 18 patients (19.8%) showed no evidences of abnormal reflux. The percentage of the total time of the bilirubin absorbance > 0.14 in 71 patients with ERD was (8.18+/-11.28%) and in 9 patients with BE was (15.48+/-30.48%), which was significantly greater than that in 11 patients with NERD (4.48+/-8.99%), P<0.05 and P=0.01 respectively. All the BE patients had abnormal esophageal bile reflux (bile alone (3 patients)); and mixed bile and acid (6 patients)); 44 of 71 patients (61.97%) with ERD had abnormal esophageal bile reflux (alone (13 patients) and mixed bile and acid (31 patients)); meanwhile, 15 of them (21.2%) had abnormal acid exposure alone. Despite 11 patients having NERD, four patients (36.4%) had abnormal esophageal bile reflux and two of them had mixed reflux of bile with acid. We believe that the Bilitec method reliably identifies the presence of bilirubin and quantitatively detects the duodenogastroesophageal reflux of bile. Mixed reflux (acid and bile) is the chief pattern of reflux in our GERD patients. Bile reflux either alone or along with acid reflux contributes to the severity of erosive and nonerosive reflux diseases as well as in BE.


Artificial Olfactory Neuron for an In-Sensor Neuromorphic Nose.
A neuromorphic module of an electronic nose (E-nose) is demonstrated by hybridizing a chemoresistive gas sensor made of a semiconductor metal oxide (SMO) and a single transistor neuron (1T-neuron) made of a metal-oxide-semiconductor field-effect transistor (MOSFET). By mimicking a biological olfactory neuron, it simultaneously detects a gas and encoded spike signals for in-sensor neuromorphic functioning. It identifies an odor source by analyzing the complicated mixed signals using a spiking neural network (SNN). The proposed E-nose does not require conversion circuits, which are essential for processing the sensory signals between the sensor array and processors in the conventional bulky E-nose. In addition, they do not have to include a central processing unit (CPU) and memory, which are required for von Neumann computing. The spike transmission of the biological olfactory system, which is known to be the main factor for reducing power consumption, is realized with the SNN for power savings compared to the conventional E-nose with a deep neural network (DNN). Therefore, the proposed neuromorphic E-nose is promising for application to Internet of Things (IoT), which demands a highly scalable and energy-efficient system. As a practical example, it is employed as an electronic sommelier by classifying different types of wines.


A hybrid cellular automaton model of solid tumor growth and bioreductive drug transport.
Bioreductive drugs are a class of hypoxia selective drugs that are designed to eradicate the hypoxic fraction of solid tumors. Their activity depends upon a number of biological and pharmacological factors and we used a mathematical modeling approach to explore the dynamics of tumor growth, infusion, and penetration of the bioreductive drug Tirapazamine (TPZ). An in-silico model is implemented to calculate the tumor mass considering oxygen and glucose as key microenvironmental parameters. The next stage of the model integrated extra cellular matrix (ECM), cell-cell adhesion, and cell movement parameters as growth constraints. The tumor microenvironments strongly influenced tumor morphology and growth rates. Once the growth model was established, a hybrid model was developed to study drug dynamics inside the hypoxic regions of tumors. The model used 10, 50 and 100 \mu {\rm M} as TPZ initial concentrations and determined TPZ pharmacokinetic (PK) (transport) and pharmacodynamics (cytotoxicity) properties inside hypoxic regions of solid tumor. The model results showed that diminished drug transport is a reason for TPZ failure and recommend the optimization of the drug transport properties in the emerging TPZ generations. The modeling approach used in this study is novel and can be a step to explore the behavioral dynamics of TPZ.


Changing atmospheric acidity as a modulator of nutrient deposition and ocean biogeochemistry.
Anthropogenic emissions to the atmosphere have increased the flux of nutrients, especially nitrogen, to the ocean, but they have also altered the acidity of aerosol, cloud water, and precipitation over much of the marine atmosphere. For nitrogen, acidity-driven changes in chemical speciation result in altered partitioning between the gas and particulate phases that subsequently affect long-range transport. Other important nutrients, notably iron and phosphorus, are affected, because their soluble fractions increase upon exposure to acidic environments during atmospheric transport. These changes affect the magnitude, distribution, and deposition mode of individual nutrients supplied to the ocean, the extent to which nutrient deposition interacts with the sea surface microlayer during its passage into bulk seawater, and the relative abundances of soluble nutrients in atmospheric deposition. Atmospheric acidity change therefore affects ecosystem composition, in addition to overall marine productivity, and these effects will continue to evolve with changing anthropogenic emissions in the future.


A Synthetic Virus-Like Particle Streptococcal Vaccine Candidate Using B-Cell Epitopes from the Proline-Rich Region of Pneumococcal Surface Protein A.
Alternatives to the well-established capsular polysaccharide-based vaccines against Streptococcus pneumoniae that circumvent limitations arising from limited serotype coverage and the emergence of resistance due to capsule switching (serotype replacement) are being widely pursued. Much attention is now focused on the development of recombinant subunit vaccines based on highly conserved pneumococcal surface proteins and virulence factors. A further step might involve focusing the host humoral immune response onto protective protein epitopes using as immunogens structurally optimized epitope mimetics. One approach to deliver such epitope mimetics to the immune system is through the use of synthetic virus-like particles (SVLPs). SVLPs are made from synthetic coiled-coil lipopeptides that are designed to spontaneously self-assemble into 20-30 nm diameter nanoparticles in aqueous buffer. Multivalent display of epitope mimetics on the surface of SVLPs generates highly immunogenic nanoparticles that elicit strong epitope-specific humoral immune responses without the need for external adjuvants. Here, we set out to demonstrate that this approach can yield vaccine candidates able to elicit a protective immune response, using epitopes derived from the proline-rich region of pneumococcal surface protein A (PspA). These streptococcal SVLP-based vaccine candidates are shown to elicit strong humoral immune responses in mice. Following active immunization and challenge with lethal doses of streptococcus, SVLP-based immunogens are able to elicit significant protection in mice. Furthermore, a mimetic-specific monoclonal antibody is shown to mediate partial protection upon passive immunization. The results show that SVLPs combined with synthetic epitope mimetics may have potential for the development of an effective vaccine against Streptococcus pneumoniae.


Measurement of blood protease kinetic parameters with self-assembled monolayer ligand binding assays and label-free MALDI-TOF MS.
We report novel ligand binding assay (LBA) surface modalities that permit plasma protease catalytic efficiency (kcat/km) determination by MALDI-TOF MS without the use of liquid chromatography or internal standards such as chemical or metalized labels. Two model LBAs were constructed on planar self-assembled monolayers (SAMs) and used to evaluate the clinically relevant metalloprotease ADAMTS-13 kinetics in plasma. The SAM chemistries were designed to improve biosampling efficiency by minimization of nonspecific adsorption of abundant proteins present at ~100,000× the concentration of the endogenous enzyme. In the first protocol, in-solution digestion of the ADAMTS-13 substrate (vWFh) was performed with immunoaffinity enrichment of the reaction substrate and product to SAM arrays. The second configuration examined protease kcat/km via a surface digestion modality where different substrates were covalently immobilized to the SAM at controlled surface density for optimized protease screens. The results show the MALDI-TOF MS LBA platforms provide limits of quantitation to ~1% protease activity (~60 pM enzyme concentration) in <1 h analysis time, a ~16× improvement over other MS-based LBA formats. Implementation of a vacuum-sublimed MALDI matrix provided good MALDI-TOF MS intra- and interday repeatability, ~1.2 and ~6.6% RSD, respectively. Platform reliability permitted kcat/km determination without internal standards with observed values ~10× improved versus conventional fluorophoric assays. Application of the assays to 12 clinical plasma samples demonstrated proof-of-concept for clinical applications. Overall, this work demonstrates that rationally designed surface chemistries for MALDI-TOF MS may serve as an alternative, label-free methodology with potential for a wide range of biotechnology applications related to targeted enzyme molecular diagnostics.


Sodium fluorescein is a probe substrate for hepatic drug transport mediated by OATP1B1 and OATP1B3.
The aim of this study was to characterize the in vitro hepatic uptake kinetics of sodium fluorescein (NaFluo) and identify the transporters involved. NaFluo exhibited saturable uptake kinetics in suspended rat and human hepatocytes as reflected by K(m) values of 22.5 and 14.1 µM, and V(max) values of 98.3 and 5.8 pmol/(million cells • min), respectively. Coincubation with known inhibitors (e.g. rifampicin) of organic anion transporting polypeptide (OATP/Oatp; SLCO gene family) significantly decreased NaFluo uptake in hepatocytes. In contrast, neither inhibitors/substrates of the organic cation transporter or organic anion transporter family nor depletion of extracellular sodium resulted in significant inhibition of NaFluo uptake. To explore the contribution of individual uptake transporters, NaFluo uptake was determined in Chinese hamster ovary cells transfected with OATP1B1, OATP1B3, and OATP2B1. Transporter-mediated uptake of NaFluo was observed in OATP1B1- and OATP1B3-transfected cells (K(m) = 4.2 and 10.9 µM; V(max) = 30.9 and 135 [pmol/(mg protein • min)], respectively). NaFluo can be used as a probe substrate to study Oatp/OATP1B-mediated drug interactions in fluorescence-based in vitro transport assays of rat and human liver. Labeling of drugs or bile salts with a fluorescein moiety can be expected to result in fluorescent conjugates with substantially altered hepatic uptake characteristics as compared with the unconjugated compounds.


Assessing the performance of the photo-acoustic infrared gas monitor for measuring CO(2), N(2)O, and CH(4) fluxes in two major cereal rotations.
Rapid, precise, and globally comparable methods for monitoring greenhouse gas (GHG) fluxes are required for accurate GHG inventories from different cropping systems and management practices. Manual gas sampling followed by gas chromatography (GC) is widely used for measuring GHG fluxes in agricultural fields, but is laborious and time-consuming. The photo-acoustic infrared gas monitoring system (PAS) with on-line gas sampling is an attractive option, although it has not been evaluated for measuring GHG fluxes in cereals in general and rice in particular. We compared N2 O, CO2 , and CH4 fluxes measured by GC and PAS from agricultural fields under the rice-wheat and maize-wheat systems during the wheat (winter), and maize/rice (monsoon) seasons in Haryana, India. All the PAS readings were corrected for baseline drifts over time and PAS-CH4 (PCH4 ) readings in flooded rice were corrected for water vapor interferences. The PCH4 readings in ambient air increased by 2.3 ppm for every 1000 mg cm(-3) increase in water vapor. The daily CO2 , N2 O, and CH4 fluxes measured by GC and PAS from the same chamber were not different in 93-98% of all the measurements made but the PAS exhibited greater precision for estimates of CO2 and N2 O fluxes in wheat and maize, and lower precision for CH4 flux in rice, than GC. The seasonal GC- and PAS-N2 O (PN2 O) fluxes in wheat and maize were not different but the PAS-CO2 (PCO2 ) flux in wheat was 14-39% higher than that of GC. In flooded rice, the seasonal PCH4 and PN2 O fluxes across N levels were higher than those of GC-CH4 and GC-N2 O fluxes by about 2- and 4fold, respectively. The PAS (i) proved to be a suitable alternative to GC for N2 O and CO2 flux measurements in wheat, and (ii) showed potential for obtaining accurate measurements of CH4 fluxes in flooded rice after making correction for changes in humidity.


Ambient air pollution and low birthweight: a European cohort study (ESCAPE).
Ambient air pollution has been associated with restricted fetal growth, which is linked with adverse respiratory health in childhood. We assessed the effect of maternal exposure to low concentrations of ambient air pollution on birthweight. We pooled data from 14 population-based mother-child cohort studies in 12 European countries. Overall, the study population included 74 178 women who had singleton deliveries between Feb 11, 1994, and June 2, 2011, and for whom information about infant birthweight, gestational age, and sex was available. The primary outcome of interest was low birthweight at term (weight <2500 g at birth after 37 weeks of gestation). Mean concentrations of particulate matter with an aerodynamic diameter of less than 2·5 μm (PM2·5), less than 10 μm (PM10), and between 2·5 μm and 10 μm during pregnancy were estimated at maternal home addresses with temporally adjusted land-use regression models, as was PM2·5 absorbance and concentrations of nitrogen dioxide (NO2) and nitrogen oxides. We also investigated traffic density on the nearest road and total traffic load. We calculated pooled effect estimates with random-effects models. A 5 μg/m(3) increase in concentration of PM2·5 during pregnancy was associated with an increased risk of low birthweight at term (adjusted odds ratio [OR] 1·18, 95% CI 1·06-1·33). An increased risk was also recorded for pregnancy concentrations lower than the present European Union annual PM2·5 limit of 25 μg/m(3) (OR for 5 μg/m(3) increase in participants exposed to concentrations of less than 20 μg/m(3) 1·41, 95% CI 1·20-1·65). PM10 (OR for 10 μg/m(3) increase 1·16, 95% CI 1·00-1·35), NO2 (OR for 10 μg/m(3) increase 1·09, 1·00-1·19), and traffic density on nearest street (OR for increase of 5000 vehicles per day 1·06, 1·01-1·11) were also associated with increased risk of low birthweight at term. The population attributable risk estimated for a reduction in PM2·5 concentration to 10 μg/m(3) during pregnancy corresponded to a decrease of 22% (95% CI 8-33%) in cases of low birthweight at term. Exposure to ambient air pollutants and traffic during pregnancy is associated with restricted fetal growth. A substantial proportion of cases of low birthweight at term could be prevented in Europe if urban air pollution was reduced. The European Union.


Human CLASP2 specifically regulates microtubule catastrophe and rescue.
Cytoplasmic linker-associated proteins (CLASPs) are microtubule-associated proteins essential for microtubule regulation in many cellular processes. However, the molecular mechanisms underlying CLASP activity are not understood. Here, we use purified protein components and total internal reflection fluorescence microscopy to investigate the effects of human CLASP2 on microtubule dynamics in vitro. We demonstrate that CLASP2 suppresses microtubule catastrophe and promotes rescue without affecting the rates of microtubule growth or shrinkage. Strikingly, when CLASP2 is combined with EB1, a known binding partner, the effects on microtubule dynamics are strongly enhanced. We show that synergy between CLASP2 and EB1 is dependent on a direct interaction, since a truncated EB1 protein that lacks the CLASP2-binding domain does not enhance CLASP2 activity. Further, we find that EB1 targets CLASP2 to microtubules and increases the dwell time of CLASP2 at microtubule tips. Although the temporally averaged microtubule growth rates are unaffected by CLASP2, we find that microtubules grown with CLASP2 display greater variability in growth rates. Our results provide insight into the regulation of microtubule dynamics by CLASP proteins and highlight the importance of the functional interplay between regulatory proteins at dynamic microtubule ends.


Hyperantithrombotic, noncytoprotective Glu149Ala-activated protein C mutant.
Activated protein C (APC) reduces mortality in severe sepsis patients. APC exerts anticoagulant activities via inactivation of factors Va and VIIIa and cytoprotective activities via endothelial protein C receptor and protease-activated receptor-1. APC mutants with selectively altered and opposite activity profiles, that is, greatly reduced anticoagulant activity or greatly reduced cytoprotective activities, are compared here. Glu149Ala-APC exhibited enhanced in vitro anticoagulant and in vivo antithrombotic activity, but greatly diminished in vitro cytoprotective effects and in vivo reduction of endotoxin-induced murine mortality. Thus, residue Glu149 and the C-terminal region of APC's light chain are identified as functionally important for expression of multiple APC activities. In contrast to Glu149Ala-APC, 5A-APC (Lys191-193Ala + Arg229/230Ala) with protease domain mutations lacked in vivo antithrombotic activity, although it was potent in reducing endotoxin-induced mortality, as previously shown. These data imply that APC molecular species with potent antithrombotic activity, but without robust cytoprotective activity, are not sufficient to reduce mortality in endotoxemia, emphasizing the need for APC's cytoprotective actions, but not anticoagulant actions, to reduce endotoxin-induced mortality. Protein engineering can provide APC mutants that permit definitive mechanism of action studies for APC's multiple activities, and may also provide safer and more effective second-generation APC mutants with reduced bleeding risk.


The clinical utility of screening of biochemical parameters in elite athletes: analysis of 100 cases.
To determine the clinical utility of screening for biochemical parameters in elite athletes. A prospective sequential case series. The Department of Sports Medicine at the Australian Institute of Sport. 100 elite athletes from 11 sports (56 male and 44 female athletes, mean age 19 years, range 16-27), undergoing routine medical screening. Initial and follow-up assessment of the following biochemical parameters in association with clinical assessment; serum iron, ferritin, transferrin, percent transferrin saturation, sodium, potassium, chloride, calcium, magnesium, phosphate, urate, urea and creatinine, total protein, albumin, creatine kinase (CK), lactate dehydrogenase, aspartate aminotransaminase (AST), alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transpeptidase, total bilirubin, cholesterol and triglycerides (non-fasting), and random glucose. 18 athletes showed no abnormalities on biochemical screening. 194 abnormal results were found in 82 athletes. 115 abnormalities were noted in 46 male and 79 in 36 female athletes. In 43 individual tests, the results did not return to normal on repeat testing. The most common abnormalities were increases in AST (27%), phosphate (13%), CK (13%), urea (12%) and bilirubin (12%). Three cases of hypercholesterolaemia and one case of haemochromatosis were identified, and one athlete, who was asymptomatic, was diagnosed with Epstein-Barr virus infection, which was suspected because of an abnormal liver function test. The other abnormalities found appeared not to be of clinical significance. Most abnormalities found on routine biochemical screening in elite athletes are of no clinical significance, therefore such testing should, if used only for clinical purposes, be abandoned. When athletes are tested for iron status it would be prudent to include assessment of serum cholesterol in those with a family history of hyperlipidaemia.


Loss of CDKN2B promotes p53-dependent smooth muscle cell apoptosis and aneurysm formation.
Genomewide association studies have implicated allelic variation at 9p21.3 in multiple forms of vascular disease, including atherosclerotic coronary heart disease and abdominal aortic aneurysm. As for other genes at 9p21.3, human expression quantitative trait locus studies have associated expression of the tumor suppressor gene CDKN2B with the risk haplotype, but its potential role in vascular pathobiology remains unclear. Here we used vascular injury models and found that Cdkn2b knockout mice displayed the expected increase in proliferation after injury, but developed reduced neointimal lesions and larger aortic aneurysms. In situ and in vitro studies suggested that these effects were attributable to increased smooth muscle cell apoptosis. Adoptive bone marrow transplant studies confirmed that the observed effects of Cdkn2b were mediated through intrinsic vascular cells and were not dependent on bone marrow-derived inflammatory cells. Mechanistic studies suggested that the observed increase in apoptosis was attributable to a reduction in MDM2 and an increase in p53 signaling, possibly due in part to compensation by other genes at the 9p21.3 locus. Dual inhibition of both Cdkn2b and p53 led to a reversal of the vascular phenotype in each model. These results suggest that reduced CDKN2B expression and increased smooth muscle cell apoptosis may be one mechanism underlying the 9p21.3 association with aneurysmal disease.


Design and methods of the Ludwig-McGill longitudinal study of the natural history of human papillomavirus infection and cervical neoplasia in Brazil. Ludwig-McGill Study Group.
This article reports on a large longitudinal study, begun in 1993, of the natural history of human papillomavirus (HPV) infection and cervical neoplasia in a population of low-income women in São Paulo, Brazil, a city with one of the highest risks worldwide for cervical cancer. Known as the Ludwig-McGill cohort study, the epidemiological investigation focuses on persistent infection with oncogenic HPV types as the precursor event leading to cervical neoplasia. The objectives of this study are to: 1) study the epidemiology of persistent cervical HPV infection in asymptomatic women, 2) investigate whether persistent HPV infection increases risk of low-grade and high-grade cervical lesions, 3) search for determinants of persistent HPV infection, 4) search for molecular variants of HPV that may be associated with an increased risk of lesions, 5) investigate whether viral burden is correlated with persistent infections and with lesion risk, 6) study the antibody response to HPV as a predictor of persistence and lesion progression, and 7) examine the role of HLA typing and codon 72 p53 gene polymorphism in mediating HPV persistence and lesion severity. The study accrued 2,528 female subjects through March 1997. Subjects were followed up every 4 months in the first year, with twice-yearly return visits to take place in subsequent years. Participants undergo a questionnaire-based interview, have a cervical specimen taken for Pap cytology and HPV testing, and have a blood sample drawn for HPV antibody testing. A cervicography is performed once in the first year and every two years thereafter. In this article we describe the design and methods of the study, provide baseline cohort characteristics, and present a preliminary assessment of the prognostic value of baseline HPV status.


Development and characterization of a novel hybrid tissue engineering-based scaffold for spinal cord injury repair.
Spinal cord injury (SCI) represents a significant health and social problem, and therefore it is vital to develop novel strategies that can specifically target it. In this context, the objective of the present work was to develop a new range of three-dimensional (3D) tubular structures aimed at inducing the regeneration within SCI sites. Up to six different 3D tubular structures were initially developed by rapid prototyping: 3D bioplotting-based on a biodegradable blend of starch. These structures were then further complemented by injecting Gellan Gum, a polysaccharide-based hydrogel, in the central area of structures. The mechanical properties of these structures were assessed using dynamic mechanical analysis, under both dry and wet conditions, and their morphologies/porosities were analyzed using micro-computed tomography and scanning electron microscopy. Biological evaluation was carried out to determine their cytotoxicity, using both minimum essential medium (MEM) extraction and MTS tests, as well as by encapsulation of an oligodendrocyte-like cell (M03-13 cell line) within the hydrogel phase. The histomorphometric analysis showed a fully interconnected network of pores with porosity ranging from 70% to 85%. Scaffolds presented compressive modulus ranging from 17.4 to 62.0 MPa and 4.42 to 27.4 MPa under dry and wet conditions, respectively. Cytotoxicity assays revealed that the hybrid starch/poly-epsilon-caprolactone/Gellan Gum scaffolds were noncytotoxic, as they did not cause major alterations on cell morphology, proliferation, and metabolic viability. Moreover, preliminary cell encapsulation assays showed that the hybrid scaffolds could support the in vitro culture of oligodendrocyte-like cells. Finally, preliminary in vivo studies conducted in a hemisection rat SCI model revealed that the above-referred structures were well integrated within the injury and did not trigger chronic inflammatory processes. The results herein presented indicate that these 3D systems might be of use in future SCI regeneration approaches.


[Dental decay and tooth loss at the high school level in Mexican students].
To determine the prevalence of dental decay and tooth loss, and to consider the treatment needs for students at the Mexican National Autonomous University (UNAM). A cross sectional study was designed including 77 191 students corresponding to the years of entrance to high school 2003, 2004 and 2005. Oral health data were obtained from the DMF-T index included in the Automatized Medical Exam (AME), a student self-answer instrument administered by the General Medical Services Department, UNAM. The analysis of the AME was univaried with the purpose of identifying the distribution and frequency of the variables. The prevalence of dental decay and tooth loss was 48.0% and 34.2%, respectively. The DMF-T index was 5. The treatment needs for dental decay and tooth loss for a student were considered as at least one lost tooth. Approximately half of the students from UNAM high schools need at least restorations and dental prosthesis. This suggests that oral health programs have not yet reached the expected health indexes according to international regulations, as well as the need to develop new research projects directed toward identifying disease severity and the factors that are detrimental to oral health.


A tale of two bridges: effect of the bloodless bridge on renal function and blood pressure in neonates managed with venoarterial extracorporeal membrane oxygenation.
To investigate if a change in bridge design of the extracorporeal membrane oxygenation (ECMO) circuit had an impact on renal function and blood pressure in neonates requiring venoarterial ECMO support. : Retrospective chart review. A tertiary care neonatal intensive care unit and ECMO center. The medical records of neonates admitted to the neonatal intensive care unit and treated with venoarterial ECMO were reviewed. Data were collected on 50 consecutive neonates treated previous to (prebridge group) and following (postbridge group) transition to a new bridge design on the ECMO circuit. None. Gestational age, gender, racial distribution, and use of hypertensive therapy were similar between the two groups. Daily blood urea nitrogen, serum creatinine, urine output, fluid balance, and average and maximum systolic and mean arterial blood pressures were recorded for the first 3 days on bypass. The postbridge group had lower maximum mean arterial blood pressure and systolic blood pressure on day 2 of ECMO and lower average mean arterial blood pressure and systolic blood pressure on days 2 and 3 of ECMO. These differences remained significant after controlling for covariates in a multiple regression model. A higher percentage of patients were hypertensive (mean arterial blood pressure >60) in the prebridge group compared with the postbridge group. There were no differences in blood urea nitrogen, serum creatinine, fluid balance, and urine output between the two groups. Patients managed on venoarterial ECMO after the transition to the "bloodless" bridge had less hypertension compared with those managed before the bridge change. This may reflect improved maintenance of renal perfusion associated with transition to an ECMO bridge design that does not require intermittent circulation with associated arterial-venous shunting.


Variability in expression of antigens responsible for serotype specificity in Candida albicans.
The monoclonal antibody (mAb) B9E, which reacts with a cell wall surface determinant of Candida albicans serotype A, and a polyclonal monospecific antiserum against the antigen 6 (IF6) were used to investigate the expression of the antigens responsible for the serotype specificity in C. albicans under different growth conditions. By indirect immunofluorescence, both antibodies reacted with the cell wall surface of serotype A yeast cells and germ tubes grown in vitro but no reactivity was observed with serotype B yeast cells. In some cases, only a weak reactivity restricted to a zone close to the parent yeast cell was observed in serotype B germ tubes stained with mAb B9E. Both antibodies reacted strongly with yeast cells and germ tubes present in kidney abscesses from rabbits infected with both serotypes, but only serotype A yeast cells and germ tubes present in smears from patients with vulvovaginal candidiasis reacted with B9E and IF6 antibodies. The expression of antigens reactive with both antibodies was modulated by the pH of the environment in which the fungus was grown. Both antibodies showed a similar pattern of reactivity when studied with a spectrofluorometer. Serotype A yeast cells showed maximum reactivity when cells were grown on Sabouraud dextrose broth supplemented with yeast extract at pH 4.6. The lowest reactivity was observed in cells grown at pH 2.0. Conversely, the reactivity of serotype B yeast cells increased at alkaline pH values, the highest being in cells grown at pH values of 7.2 and 9.5.(ABSTRACT TRUNCATED AT 250 WORDS)


Depth-dependent changes in collagen organization in the human peripapillary sclera.
The collagen structure of the human peripapillary sclera plays a significant role in determining optic nerve head (ONH) biomechanics, and is therefore of interest in the study of glaucoma. The aim of the current work was to map the anisotropic collagen structure of the normal human peripapillary sclera as a function of tissue depth. Wide-angle x-ray scattering was used to quantify collagen fibril orientation at 0.5 mm intervals across six 150 μm-thick serial sections through the peripapillary sclera of eight normal European-derived human eyes. Two structural parameters were measured: 1) the relative number of fibrils preferentially aligned at a given angle within the tissue plane, 2) the degree of collagen alignment (anisotropy). The inner-most one-third of the peripapillary scleral stroma (nearest to the choroid) was characterised by collagen fibrils either randomly arranged or preferentially aligned radially with respect to the ONH. In contrast, the outer two-thirds of the tissue was dominated by a circumferential arrangement of collagen encircling the ONH. In all tissue regions the degree of collagen anisotropy peaked in the mid-stroma and progressively decreased towards the tissue surfaces, with the largest depth variations occurring in the inferior-nasal quadrant, and the smallest occurring in the superior-nasal quadrant. Significant, region-specific variations in collagen structure are present in the human peripapillary sclera as a function of depth. In normal eyes, the circumferential collagen fibril architecture is most prominent in the outer two-thirds of the stroma, possibly as a mechanical adaption to more effectively support the lamina cribrosa at the level of its insertion into the scleral canal wall.


Mass transport, atherogenesis, and risk.
If the local chemically-active interstitial concentration (chemical activity) of a particular atherogenic molecule (atherogen) is considered to be one of the fundamental driving forces in a system of reactions that produces an associated local atherogenic change, then the chemical activity may be used as a measure of the potential for lesion development or local "risk." A model of steady-state, combined, diffusive and convective transport of an assumed atherogen across a multilayered arterial wall was used to illustrate mechanisms by which transport processes interact with tissue barriers to determine the transmural (x axis) chemical activity distributions [a(x), mol cm-3]. The model was used to evaluate the effects of increased endothelial gap fractional area (increased endothelial permeability), elevated blood pressure, elevated serum concentration of the atherogen, internal elastica fenestration, and preexisting intimal thickening on a(x). It was found that sites along the arterial tree that are characterized by "mild" increases in endothelial gap fractional area and concomitant subjacent interstitial sieving of the atherogen developed high a(x), in some cases much higher than that (a0) of the blood, and therefore were at high risk even in the absence of other risk factors. Moreover, the risk at such sites was dramatically increased with hypertension and/or elevated serum atherogen concentration and/or preexisting intimal thickening. However, if either condition, i.e., opened endothelial intercellular junctions or subjacent interstitial sieving, was absent, these other risk factors were ineffective. Finally, the response of the model was shown to be consistent with a variety of well-known, as well as puzzling, observations suggesting that transport processes may play roles in determining both the architecture of the atherosclerotic lesion and its rate of development.


Physical and sexual violence and incident sexually transmitted infections.
To investigate whether women aged 13-35 who were victims of interpersonal violence were more likely than nonvictims to experience incident sexually transmitted infections (STIs). We examined 542 women aged 13-35 enrolled in Project PROTECT, a randomized clinical trial that compared two different methods of computer-based intervention to promote the use of dual methods of contraception. Participants completed a baseline questionnaire that included questions about their history of interpersonal violence and were followed for incident STIs over the 2-year study period. We compared the incidence of STIs in women with and without a history of interpersonal violence using bivariate analyses and multiple logistic regression. In the bivariate analyses, STI incidence was found to be significantly associated with African American race/ethnicity, a higher number of sexual partners in the past month, and a lower likelihood of avoidance of sexual partners who pressure to have sex without a condom. In both crude and adjusted regression analyses, time to STI incidence was faster among women who reported physical or sexual abuse in the year before study enrollment (HRR(adj) = 1.68, 95% CI 1.06, 2.65). Women with a recent history of abuse are at significantly increased risk of STI incidence than are nonvictims.


Predictors of hospitalization for heart failure and of all-cause mortality after atrioventricular nodal ablation and right ventricular pacing for atrial fibrillation.
Atrioventricular junction ablation (AVJA) is a highly effective treatment in patients with therapy refractory atrial fibrillation (AF) but renders the patient pacemaker dependent. We aimed to analyse the long-term incidence of hospitalization for heart failure (HF) and all-cause mortality in patients who underwent AVJA because of AF and to determine predictors for HF and mortality. We retrospectively enrolled 162 consecutive patients, mean age 67 ± 9 years, 48% women, who underwent AVJA because of symptomatic AF refractory to pharmacological treatment (n = 117) or unsuccessful repeated pulmonary vein isolation (n = 45). Hospitalization for HF occurred in 32 (20%) patients and 35 (22%) patients died, representing a cumulative incidence for hospitalization for HF and mortality over the first 2 years after AVJA of 9.1 and 5.2%, respectively. Hospitalization for HF occurred to the same extent in patients who failed pharmacological treatment as in patients with repeated pulmonary vein isolation (PVI), although the mortality was slightly higher in the former group. QRS prolongation ≥120 ms and left atrial diameter were independent predictors of hospitalization for HF, while hypertension and previous HF were independent predictors of death. The long-term hospitalization rate for HF and all-cause mortality was low, which implies that long-term ventricular pacing was not harmful in this patient population, including patients with unsuccessful repeated PVI.


Detection of clonal IGH gene rearrangements: summary of molecular oncology surveys of the College of American Pathologists.
The diagnosis of B-cell lymphoid malignancy can frequently be substantiated by detecting clonal immunoglobulin heavy chain (IGH) gene rearrangements, which is typically done by polymerase chain reaction (PCR) amplification and/or Southern blot analysis. To characterize current laboratory practice for the assessment of IGH rearrangements and to identify opportunities for improvement. The data from the Molecular Oncology Proficiency Survey distributed to participating laboratories by the Molecular Pathology Committee of the College of American Pathologists from 1998 through 2003 were analyzed. Thirty-nine proficiency survey specimens (29 positive and 10 negative for clonal IGH rearrangements) were distributed. For Southern blot analysis, 944 results were reported, with a successful response rate of 95%. For PCR detection, 2349 results were reported, with a successful response rate of 72%. A higher rate of successful responses by PCR was achieved using framework 3 primers in combination with other frameworks (82%) compared with framework 3 primers only (76%) and when fresh/frozen (72%) compared with paraffin-embedded (65%) tissues were analyzed. The performance of the participating laboratories was very good, by both Southern blot and PCR analysis. As expected, Southern blot analysis consistently detects a higher proportion of IGH rearrangements than PCR analysis. Further improvement and standardization of the IGH PCR assay is important if it is to replace Southern blot analysis as the standard method. Participation in this survey is a valuable tool for assessing laboratory performance and it directs our attention to areas where we may improve laboratory practice.


Analysis of multiple components involved in the interaction between Cryptococcus neoformans and Acanthamoeba castellanii.
Cryptococcus neoformans is an environmental fungus that can cause lethal meningoencephalitis in immunocompromised individuals. The mechanisms by which environmental microbes become pathogenic to mammals are still obscure, but different studies suggest that fungal virulence evolved from selection imposed by environmental predators. The soil-living Acanthamoeba castellanii is a well-known predator of C. neoformans. In this work, we evaluated the participation of C. neoformans virulence-associated structures in the interaction of fungal cells with A. castellanii. Fungal extracellular vesicles (EVs) and the polysaccharide glucuronoxylomannan (GXM) were internalized by A. castellanii with no impact on the viability of amoebal cells. EVs, but not free GXM, modulated antifungal properties of A. castellanii by inducing enhanced yeast survival. Phagocytosis of C. neoformans by amoebal cells and the pathogenic potential in a Galleria mellonella model were not affected by EVs, but previous interactions with A. castellanii rendered fungal cells more efficient in killing this invertebrate host. This observation was apparently associated with marked amoeba-induced changes in surface architecture and increased resistance to both oxygen- and nitrogen-derived molecular species. Our results indicate that multiple components with the potential to impact pathogenesis are involved in C. neoformans environmental interactions.


Incidence of posttraumatic stress disorder after traffic accidents in Germany.
Posttraumatic stress disorder (PTSD) is possibly an overlooked diagnosis of victims suffering from traffic accidents sustaining serious to severe injuries. This paper investigates the incidence of PTSD after traffic accidents in Germany. Data from an accident research unit were analyzed in regard to collision details, and preclinical and clinical data. Preclinical data included details on crash circumstances and estimated injury severity as well as data on victims' conditions (e.g. heart rate, blood pressure, consciousness, breath rate). Clinical data included initial assessment in the emergency department, radiographic diagnoses, and basic life parameters comparable to the preclinical data as well as follow-up data on the daily ward. Data were collected in the German-In-Depth Accident Research study, and included gender, type of accident (e.g. type of vehicle, road conditions, rural or urban area), mental disorder, and AIS (Abbreviated Injury Scale) head score. AIS represent a scoring system to measure the injury severity of traffic accident victims. A total 258 out of 32807 data sets were included in this analysis. Data on accident and victims was collected on scene by specialized teams following established algorithms. Besides higher AIS Head scores for male motorcyclists compared to all other subgroups, no significant correlation was found between the mean maximum AIS score and the occurrence of PTSD. Furthermore, there was no correlation between higher AIS head scores, gender, or involvement in road traffic accidents and PTSD. In our study the overall incidence of PTSD after road traffic accidents was very low (0.78% in a total of 32.807 collected data sets) when compared to other published studies. The reason for this very low incidence of PTSD in our patient sample could be seen in an underestimation of the psychophysiological impact of traffic accidents on patients. Patients suffering from direct experiences of traumatic events such as a traffic accident and presenting with signs of clinically significant distress or impairment in social interactions should be treated in a team approach including not only trauma surgeons and surgical skilled staff but also psychophysiological experienced physicians.


Project HOPE: online social network changes in an HIV prevention randomized controlled trial for African American and Latino men who have sex with men.
We examined whether and how an HIV prevention diffusion-based intervention spread throughout participants' online social networks and whether changes in social network ties were associated with increased HIV prevention and testing behaviors. We randomly assigned 112 primarily racial/ethnic minority men who have sex with men (MSM) to receive peer-delivered HIV (intervention) or general health (control) information over 12 weeks through closed Facebook groups. We recorded participants' public Facebook friend networks at baseline (September 2010) and follow-up (February 2011), and assessed whether changes in network growth were associated with changes in health engagement and HIV testing. Within-group ties increased in both conditions from baseline to follow-up. Among the intervention group, we found a significant positive relation between increased network ties and using social media to discuss sexual behaviors. We found a positive trending relationship between increased network ties and likelihood of HIV testing, follow-up for test results, and participation in online community discussions. No significant differences were seen within control groups. Among high-risk MSM, peer-led social media HIV prevention interventions can increase community cohesion. These changes appear to be associated with increased HIV prevention and testing behaviors.


Endoscopic diagnosis of cervical esophageal heterotopic gastric mucosa with conventional and narrow-band images.
To compare the diagnostic yield of heterotopic gastric mucosa (HGM) in the cervical esophagus with conventional imaging (CI) and narrow-band imaging (NBI). A prospective study with a total of 760 patients receiving a CI examination (mean age 51.6 years; 47.8% male) and 760 patients undergoing NBI examination (mean age 51.2 years; 45.9% male). The size of HGM was classified as small (1-5 mm), medium (6-10 mm), or large (> 1 cm). A standardized questionnaire was used to obtain demographic characteristics, social habits, and symptoms likely to be related to cervical esophageal HGM, including throat symptoms (globus sensation, hoarseness, sore throat, and cough) and upper esophageal symptoms (dysphagia and odynophagia) at least 3 mo in duration. The clinicopathological classification of cervical esophageal HGM was performed using the proposal by von Rahden et al. Cervical esophageal HGM was found in 36 of 760 (4.7%) and 63 of 760 (8.3%) patients in the CI and NBI groups, respectively (P = 0.007). The NBI mode discovered significantly more small-sized HGM than CI (55% vs 17%; P < 0.0001). For the 99 patients with cervical esophageal HGM, biopsies were performed in 56 patients; 37 (66%) had fundic-type gastric mucosa, and 19 had antral-type mucosa. For the clinicopathological classification, 77 patients (78%) were classified as HGM I (asymptomatic carriers); 21 as HGM II (symptomatic without morphologic changes); and one as HGM III (symptomatic with morphologic change). No intraepithelial neoplasia or adenocarcinoma was found. NBI endoscopy detects more cervical esophageal HGM than CI does. Fundic-type gastric mucosa constitutes the most common histology. One-fifth of patients have throat or dysphagic symptoms.


Discrepancy between older clients' ability to read and comprehend and the reading level of written educational materials used by occupational therapists.
The match between the reading level of occupational therapy education materials and older clients' reading ability and comprehension was determined. The sociodemographic and literacy characteristics that influenced clients' reading ability and comprehension were investigated. The reading level of 110 written education materials (handouts, brochures, and information leaflets), distributed to older clients (65 years of age and older) by occupational therapists working in Queensland hospitals, was analyzed using the Flesch formula. The reading ability of 214 older persons (mean age 77 years, 63% female) was assessed using the Rapid Estimate of Adult Literacy in Medicine. Participants' comprehension of information of increasing reading difficulty was measured using the Cloze procedure. The written materials required a mean reading level between the ninth and tenth grades. Participants' mean reading ability was seventh to eighth grade. Therefore some materials may have been too difficult for participants to read and understand. Participants with a managerial or professional or clerical background (p= 0.001) and those who perceived they read well (p= 0.001) had a significantly higher reading ability. Older age was significantly related to poorer comprehension (p = 0.018), with participants 75 years of age and over having a mean comprehension score of 25.6 compared to 30.3 for those 65 to 74 years of age. Occupational therapists must analyze the reading level of the written education materials they develop for and use with clients by applying readability formulas. There should be a match between the reading level of written materials and clients' reading ability. Clients' reading ability may be assessed informally by discussing years of education and literacy habits or formally using reading assessments. Content and design characteristics should be considered when developing written education materials for clients.


Lumbosacral Epidural Primitive Neuroectodermal Tumor (PNET): Case Report and Literature Review.
The primitive neuroectodermal tumors (PNETs) of the spine are rare. They are usually intramedullary and reported in children. We herein report an epidural PNET of lumbosacral area presenting with the cauda equina syndrome in an adult. A 38-year-old woman presented to our emergency room with acute onset lower extremity weakness and urinary incontinence. Emergent magnetic resonance imaging revealed a mass lesion isointense on T1-weighted and heterogeneously hyperintense in T2-weighted images in the epidural lumbosacral area. The patient underwent emergent laminectomy of L1-L3 and total resection of the lesion. The patient"s neurological examination improved dramatically after the surgery and after 6-month of follow-up, she was neurologically intact. The histopathological and immunohistochemical evaluations revealed PNET. Epidural PNET of the spinal column, although rare, can present with an acute neurological deficit. Surgery remains the treatment of choice and immunohistochemistry is required for confirming the diagnosis.


Prospective Multicenter Comparison of Open and Robotic Radical Prostatectomy: The PROST-QA/RP2 Consortium.
Our goal was to evaluate the comparative effectiveness of robot-assisted laparoscopic prostatectomy (RALP) and open radical prostatectomy (ORP) in a multicenter study. We evaluated men with localized prostate cancer at 11 high-volume academic medical centers in the United States from the PROST-QA (2003-2006) and the PROST-QA/RP2 cohorts (2010-2013) with a pre-specified goal of comparing RALP (549) and ORP (545). We measured longitudinal patient-reported health-related quality of life (HRQOL) at pre-treatment and at 2, 6, 12, and 24 months, and pathological and perioperative outcomes/complications. Demographics, cancer characteristics, and margin status were similar between surgical approaches. ORP subjects were more likely to undergo lymphadenectomy (89% vs 47%; p <0.01) and nerve sparing (94% vs 89%; p <0.01). RALP vs ORP subjects experienced less mean intraoperative blood loss (192 vs 805 mL; p <0.01), shorter mean hospital stay (1.6 vs 2.1 days; p <0.01), and fewer blood transfusions (1% vs 4%; p <0.01), wound infections (2% vs 4%; p=0.02), other infections (1% vs 4%; p <0.01), deep venous thromboses (0.5% vs 2%; p=0.04), and bladder neck contractures requiring dilation (1.6% vs 8.3%; p <0.01). RALP subjects reported less pain (p=0.04), less activity interference (p <0.01) and higher incision satisfaction (p <0.01). Surgical approach (RALP vs ORP) was not a significant predictor of longitudinal HRQOL change in any HRQOL domain. In high-volume academic centers, RALP and ORP patients may expect similar long-term HRQOL outcomes. Overall, RALP patients have less pain, shorter hospital stays, and fewer post-surgical complications such as blood transfusions, infections, deep venous thromboses, and bladder neck contractures.


The Born in Guangzhou Cohort Study (BIGCS).
The Born in Guangzhou Cohort Study (BIGCS) is a large-scale prospective observational study investigating the role of social, biological and environmental influences on pregnancy and child health and development in an urban setting in southern China. Pregnant women who reside in Guangzhou and who attend Guangzhou Women and Children's Medical Center (GWCMC) for antenatal care in early pregnancy (<20 weeks' gestation) are eligible for inclusion. Study recruitment commenced in February 2012, with an overall participation rate of 76.3%. Study recruitment will continue until December 2018 to achieve the target sample size of 30,000 mother-child pairs. At 30 April 2016, a total of 75,422 questionnaires have been collected, while 14,696 live births have occurred with planned follow-up of cohort children until age 18 years. During the same period a total of 1,053,000 biological samples have been collected from participants, including maternal, paternal and infant blood, cord blood, placenta, umbilical cord, and maternal and infant stool samples. The dataset has been enhanced by record linkage to routine health and administrative records. We plan future record linkage to school enrolment and national examination records.


THE ARTHUS PHENOMENON : LOCAL ANAPHYLACTIC INFLAMMATION IN THE RABBIT BRAIN.
1. Sixteen out of seventeen rabbits actively sensitized to various antigens by repeated cerebral and intravenous injections showed upon intracerebral reinjection of the same antigen local anaphylactic inflammation of the brain at the site of inoculation. 2. Six out of twenty rabbits actively sensitized to either horse serum or egg albumen by extracerebral injections, showed, upon introduction of the homologous antigen into the cerebrum, local anaphylactic inflammation at the site of inoculation. 3. The pathological picture of the Arthus phenomenon in the brain of the rabbit resembled that seen in the skin, after allowing for differences in the fundamental structure of the tissues involved. 4. None of the control animals exhibited lesions comparable to those found in the experimental animals. A few controls showed slight hemorrhages due to mechanical injury of blood vessels. 5. Clinical symptoms of varying degrees of severity, often leading to the death of the rabbit were observed in the sensitized animals. These symptoms were referable to the site of injection.


Design, Synthesis, and Pharmacological Evaluation of 5,6-Disubstituted Pyridin-2(1H)-one Derivatives as Phosphodiesterase 10A (PDE10A) Antagonists.
We report the design and synthesis of novel 5,6-diarylated pyridin-2(1H)-one derivatives as pharmacophoric PDE10A inhibitors. This highly potent molecular scaffold was developed from an inactive diarylpyridine-2-amine derivative 3b by extensive and systematic analogue synthesis and SAR analysis. Further optimization of the scaffold resulted in identification of pyridin-2(1H)-one 18b as a lead compound with good potency (IC50 = 1.6 nM) and selectivity (>6000-fold) over other related PDEs but with a poor pharmacokinetic profile. Careful metabolite profiling of 18b revealed that poor systemic exposure in rats (Cmax = 44 ng/mL; AUC0-t = 359 ng · h/mL) at 10 mg/kg was due to the formation of O-glucuronide conjugate by phase 2 metabolism. The structure of the glucuronide metabolite was confirmed by retention time and LC-MS/MS fragmentation matching with the synthetic glucuronide 26. The problem of low exposure of 18b was effectively addressed by its conversion to an acetate prodrug 25b, which upon oral dosing resulted in an improved pharmacokinetic profile (Cmax = 359 ng.h/mL; AUC0-t = 2436 ng.h/mL) and a desirable brain to plasma ratio of 1.2. The prodrug 25b showed good efficacy in selected rodent models of psychosis.


DNA vaccines against human immunodeficiency virus type 1.
Development of a vaccine against human immunodeficiency virus type 1 (HIV-1) is the main hope for controlling the acquired immunodeficiency syndrome pandemic. An ideal HIV vaccine should induce neutralizing antibodies, CD4+ helper T cells, and CD8+ cytotoxic T cells. While the induction of broadly neutralizing antibodies remains a highly challenging goal, there are a number of technologies capable of inducing potent cell-mediated responses in animal models, which are now starting to be tested in humans. Naked DNA immunization is one of them. This review focuses on the stimulation of HIV-specific T cells and discusses in the context of the current 'state-of-art' of DNA vaccines, the areas where this technology might assist either alone or as a part of more complex vaccine formulations in the HIV vaccine development.