Expression of MDM2 during mammary tumorigenesis.
The MDM2 oncoprotein encodes a 90 kDa nuclear phosphoprotein capable of abrogating the growth suppressive functions of p53 and pRb tumor suppressor proteins by direct interaction. Alternative splicing of MDM2 protein coding sequences has been documented during tumor progression in human ovarian and bladder carcinomas. The aim of this study was to determine whether alternative splicing of MDM2 occurs during breast tumorigenesis in mice and humans and whether protein coding sequences were affected. Specimens representing normal and malignant breast tissues from the murine D2 mammary tumor model system and human breast carcinomas were examined. Three distinct mdm2 mRNA transcripts of 3.3, 1.6 and 1.5 kb were detected in normal and malignant murine mammary tissues by Northern blot analysis using a full-length mdm2 cDNA probe. Additional Northern blot analysis using a probe derived from exon 12 of murine mdm2 demonstrated that the 1.5 and 1.6 kb transcripts lack sequences encoding the C-terminus of the protein. No evidence of internal deletions of protein coding sequences of mdm2 was detected in any of the normal mammary tissues or D2 murine mammary tumors examined by reverse transcription PCR (RT-PCR). Three distinct MDM2 transcripts of 6.7, 4.7 and 1.9 kb were detected in malignant human breast tissue by Northern blot analysis using a cDNA probe specific for the complete open reading frame of human MDM2. However, a cDNA probe specific for the last exon of human MDM2 hybridized only to the 6.7 and 4.7 kb transcripts, demonstrating that the 1.9 kb transcript lacked protein coding sequences contained in exon 12. Similarly, no internal deletions were detected in a panel of malignant human breast tissues using RT-PCR and analogous primers within human MDM2. Therefore, breast tumors differ from other solid tumors reported previously in that no internal deletions of MDM2 protein coding sequences were observed. However, the data document the presence of multiple MDM2 mRNA transcripts in both normal and malignant breast tissues. A subset of MDM2 transcripts were shown to lack the last exon which contains sequences coding for the RING and zinc fingers and domains which are targets for caspase-3 mediated proteolytic degradation and are required to target p53 for proteosomal degradation.


Risk prediction for new-onset atrial fibrillation using the Minnesota code electrocardiography classification system.
Few risk models are available to predict future onset of atrial fibrillation (AF) in workers. We aimed to develop risk prediction models for new-onset AF, using annual health checkup (HC) data with electrocardiogram findings. We retrospectively included 56,288 factory or office workers (mean age = 51.5 years, 33.0% women) who underwent a HC at a medical center and fulfilled the following criteria; age ≥ 40 years, no history of AF, and greater than 1 annual follow-up HC in 2013-2016. Using Cox models with the Akaike information criterion, we developed and compared prediction models for new-onset AF with and without the Minnesota code information. We externally validated the discrimination accuracy of the models in a general Japanese population cohort, the Hisayama cohort. During the median 3.0-year follow-up, 209 (0.37%) workers developed AF. Age, sex, waist circumference, blood pressure, LDL cholesterol, and γ-GTP were associated with new-onset of AF. Using the Minnesota code information, the AUC significantly improved from 0.82 to 0.84 in the derivation cohort and numerically improved from 0.78 to 0.79 in the validation cohort, and from 0.77 to 0.79 in the Hisayama cohort. The NRI and IDI significantly improved in all and male subjects in both the derivation and validation cohorts, and in female subjects in both the validation and the Hisayama cohorts. We developed useful risk model with Minnesota code information for predicting new-onset AF from large worker population validated in the original and external cohorts, although study interpretation is limited by small improvement of AUC.


A cost evaluation of the Georgia Stroke and Heart Attack Prevention Program.
Hypertension is a leading cause of stroke, coronary artery disease, heart attack, and heart and kidney failure in the United States, all of which contribute to the rising costs of health care. The Georgia Stroke and Heart Attack Prevention Program is an education and direct service program for low-income patients with hypertension. This project evaluated the cost-effectiveness of the program compared with the following two alternative scenarios: no treatment for high blood pressure and the typical hypertension treatment received in the private sector nationwide (usual care). We estimated the preventive treatment costs and number of adverse health events averted (hemorrhagic and ischemic stroke, heart disease, and kidney failure) associated with the Georgia Stroke and Heart Attack Prevention Program in two Georgia health districts. We used program cost and service usage data obtained from the Georgia Department of Human Resources and probabilities and costs of expected adverse events published in peer-reviewed sources. We compared program costs and number of expected adverse health events averted with those expected from 1) no preventive care and 2) usual care for high blood pressure. The Georgia Stroke and Heart Attack Prevention Program was less costly and resulted in better health outcomes than either no preventive care or usual care. Compared with no preventive care in the two districts, the program was estimated to result in 54% fewer expected adverse events; compared with usual care, the program was estimated to result in 46% fewer expected adverse events. Combining the costs of preventive treatment with the costs of expected adverse events, the Georgia Stroke and Heart Attack Prevention Program cost an average of 486 dollars per patient annually, compared with average annual costs of 534 dollars for no care and 624 dollars for usual care. Maintaining a publicly financed stroke and heart attack prevention program is more cost-effective and results in greater health benefits than other plausible scenarios. Because the benefits of this program accrue to both the state and federal governments through reduced Medicaid and indigent care expenditures, both the state and federal governments have a financial incentive to support the program.


Assessing spatial associations between thermal stress and mortality in Hong Kong: a small-area ecological study.
Physiological equivalent temperature (PET) is a widely used index to assess thermal comfort of the human body. Evidence on how thermal stress-related health effects vary with small geographical areas is limited. The objectives of this study are (i) to explore whether there were significant patterns of geographical clustering of thermal stress as measured by PET and mortality and (ii) to assess the association between PET and mortality in small geographical areas. A small area ecological cross-sectional study was conducted at tertiary planning units (TPUs) level. Age-standardized mortality rates (ASMR) and monthly deaths at TPUs level for 2006 were calculated for cause-specific diseases. A PET map with 100 m × 100 m resolution for the same period was derived from Hong Kong Urban Climatic Analysis Map data and the annual and monthly averages of PET for each TPU were computed. Global Moran's I and local indicator of spatial association (LISA) analyses were performed. A generalized linear mixed model was used to model monthly deaths against PET adjusted for socio-economic deprivation. We found positive spatial autocorrelation between PET and ASMR. There were spatial correlations between PET and ASMR, particularly in the north of Hong Kong Island, most parts of Kowloon, and across New Territories. A 1°C change in PET was associated with an excess risk (%) of 2.99 (95% CI: 0.50-5.48) for all natural causes, 4.75 (1.14-8.36) for cardiovascular, 7.39 (4.64-10.10) for respiratory diseases in the cool season, and 4.31 (0.12 to 8.50) for cardiovascular diseases in the warm season. Variations between TPUs in PET had an important influence on cause-specific mortality, especially in the cool season. PET may have an impact on the health of socio-economically deprived population groups. Our results suggest that targeting policy interventions at high-risk areas may be a feasible option for reducing PET-related mortality.


Economic Impact of Alvimopan Considering Varying Definitions of Postoperative Ileus.
Prolonged postoperative ileus (POI) is the predominant cause of extended hospitalization after bowel resection surgery. Alvimopan accelerates gastrointestinal recovery, potentially reducing health care costs. We examined the value of alvimopan in reducing prolonged POI and length of stay for patients undergoing abdominal surgery using different definitions of POI. We developed a decision analytic model to examine costs and outcomes associated with postoperative treatment with either an accelerated care pathway (ACP) only or alvimopan+ACP. To represent an overall perspective for alvimopan, data from four phase 3 bowel resection trials and one phase 4 radical cystectomy trial were used to populate the model with 3 different definitions of POI. The period analyzed included start of surgery to 7 days post discharge. Costs were obtained from standard US costing sources and are reported in 2015 US dollars. Due to variations in published definitions of POI, alternative definitions based on adverse event reports, NG tube insertion, and time to food toleration were examined. The combined clinical trial data included 1,003 ACP and 1,013 alvimopan+ACP patients. When POI was reported as an adverse event, the incidence of POI was significantly lower with alvimopan+ACP (n = 70 [7%]) vs ACP alone (n = 148 [15%]; p < 0.0001). Time to discharge order written was shorter for patients with POI who were treated with alvimopan+ACP than with ACP (202 ± 115 hours vs 266 ± 138 hours; p < 0.0001). As a result, costs were $731 lower with alvimopan+ACP ($17,835) vs ACP ($18,566). Alternative definitions of POI produced similar results. The addition of alvimopan to existing treatment pathways for patients undergoing abdominal surgery can reduce overall hospital costs.


Higher Plasma Viremia in the Febrile Phase Is Associated With Adverse Dengue Outcomes Irrespective of Infecting Serotype or Host Immune Status: An Analysis of 5642 Vietnamese Cases.
One of the generally accepted constructs of dengue pathogenesis is that clinical disease severity is at least partially dependent upon plasma viremia, yet data on plasma viremia in primary versus secondary infections and in relation to clinically relevant endpoints remain limited and contradictory. Using a large database comprising detailed clinical and laboratory characterization of Vietnamese participants enrolled in a series of research studies executed over a 15-year period, we explored relationships between plasma viremia measured by reverse transcription-polymerase chain reaction and 3 clinically relevant endpoints-severe dengue, plasma leakage, and hospitalization-in the dengue-confirmed cases. All 4 dengue serotypes and both primary and secondary infections were well represented. In our logistic regression models we allowed for a nonlinear effect of viremia and for associations between viremia and outcome to differ by age, serotype, host immune status, and illness day at study enrollment. Among 5642 dengue-confirmed cases we identified 259 (4.6%) severe dengue cases, 701 (12.4%) patients with plasma leakage, and 1441 of 4008 (40.0%) patients recruited in outpatient settings who were subsequently hospitalized. From the early febrile phase onwards, higher viremia increased the risk of developing all 3 endpoints, but effect sizes were modest (ORs ranging from 1.12-1.27 per 1-log increase) compared with the effects of a secondary immune response (ORs, 1.67-7.76). The associations were consistent across age, serotype, and immune status groups, and in the various sensitivity and subgroup analyses we undertook. Higher plasma viremia is associated with increased dengue severity, regardless of serotype or immune status.


Mercury levels in sediments and mangrove oysters, Crassostrea rizophorae, from the north coast of Villa Clara, Cuba.
Total mercury levels were quantified in sediments and oyster tissues (Crassostrea rizophorae) from the Sagua la Grande River estuary and offshore mangrove keys 19 km downstream of a chlor-alkali plant (CAP) in Villa Clara, Cuba. Relatively elevated total mercury levels were found in sediments from the estuary itself, ranging from 0.507 to 1.81 μg g(-1) dry weight. However, levels were lower in sediments from the keys farther from the estuary. Oyster mercury levels were always acceptable for human consumption, although levels significantly correlated in sediments and oysters across sampling sites (p < 0.05), which suggests that mercury from the CAP is impacting coastal water quality conditions.


Enhancement of NK Cell Cytotoxicity Induced by Long-Term Living in Negatively Charged-Particle Dominant Indoor Air-Conditions.
Investigation of house conditions that promote health revealed that negatively charged-particle dominant indoor air-conditions (NCPDIAC) induced immune stimulation. Negatively charged air-conditions were established using a fine charcoal powder on walls and ceilings and utilizing forced negatively charged particles (approximate diameter: 20 nm) dominant in indoor air-conditions created by applying an electric voltage (72 V) between the backside of the walls and the ground. We reported previously that these conditions induced a slight and significant increase of interleukin-2 during a 2.5-h stay and an increase of NK cell cytotoxicity when examining human subjects after a two-week night stay under these conditions. In the present study, seven healthy volunteers had a device installed to create NCPDIAC in the living or sleeping rooms of their own homes. Every three months the volunteers then turned the NCPDIAC device on or off. A total of 16 ON and 13 OFF trials were conducted and their biological effects were analyzed. NK activity increased during ON trials and decreased during OFF trials, although no other adverse effects were found. In addition, there were slight increases of epidermal growth factor (EGF) during ON trials. Furthermore, a comparison of the cytokine status between ON and OFF trials showed that basic immune status was stimulated slightly during ON trials under NCPIADC. Our overall findings indicate that the NCPDIAC device caused activation of NK activity and stimulated immune status, particularly only on NK activity, and therefore could be set in the home or office buildings.


R-Ras is a global regulator of vascular regeneration that suppresses intimal hyperplasia and tumor angiogenesis.
R-Ras is a small GTPase of the Ras family that regulates cell survival and integrin activity. Despite a number of in vitro studies, the in vivo function of R-Ras remains unclear. Here, we used R-Ras-null mice to explore the in vivo function of this small GTPase. Our results show a role for R-Ras as a regulator of vascular differentiation that primarily affects the remodeling of blood vessels. We show that R-Ras-null mice, although otherwise phenotypically normal, mount excessive vascular responses. We found that in vivo R-Ras expression is largely confined to fully differentiated smooth muscle cells, including those of blood vessels, and to endothelial cells. Challenging the R-Ras-null mice with arterial injury or tumor implantation showed exaggerated neointimal thickening in response to the injury and increased angiogenesis in the tumors. In wild-type mice, R-Ras expression was greatly reduced in hyperplastic neointimal smooth muscle cells and in angiogenic endothelial cells. Forced expression of activated R-Ras suppressed mitogenic and invasive activities of growth factor-stimulated vascular cells. These results establish an unexpected role for R-Ras in blood vessel homeostasis and suggest that R-Ras signaling may offer a target for therapeutic intervention in vascular diseases.


Chronic inflammatory demyelinating polyneuropathy associated with metastatic malignant melanoma of unknown primary origin.
Paraneoplastic neurological syndromes (PNS) are rare manifestations of malignant disease. It is currently believed that most, if not all, PNS are autoimmune in nature. Proteins expressed on the surface of tumor cells trigger an immune response that cross-reacts with similar proteins in the nervous system, resulting in damage. Moreover, recent studies propose that the appearance of autoimmune phenomena in patients with malignant melanoma implies a strong antitumoral activity and constitutes a marker of improvement in overall survival. We describe a most unusual case of chronic inflammatory demyelinating polyneuropathy (CIDP) occurring in a patient with metastatic malignant melanoma of unknown primary origin who received treatment with interferon alpha-2b. We also review the relevant literature about this infrequent association, discuss on its pathogenesis and underline its importance as surrogate marker, useful to verify the antineoplastic treatment efficacy.


Effects of surface nano-topography on human osteoblast filopodia.
Anchorage-dependent cells growing over a substratum require stable adhesion areas on the surface for the next cellular activities. The adhesion is achieved by some contact points called focal adhesions. Because focal adhesions were distributed randomly, a trial to control the positions of focal adhesion with a specific order may cause interesting effects like as cytoskeleton rearrangement, which may induce and transfer new signals to the nucleus. Here, we cultured human osteoblasts over two sorts of nanopatterned surfaces with different pattern densities fabricated by using laser interference lithography and the nanoimprinting technique. Of the two nanopatterns, cells over the nanopattern with low pattern density showed relatively higher adaptation to the topography with guided filopodia protrusion. However, cells over the dense nanopattern showed difficulty in finding suitable paths for migration, as judged from the activities of filopodium formation and the presence of a shovel-like feature at the tip of each filopodium.


Trauma team activation varies across Dutch emergency departments: a national survey.
Tiered trauma team response may contribute to efficient in-hospital trauma triage by reducing the amount of resources required and by improving health outcomes. This study evaluates current practice of trauma team activation (TTA) in Dutch emergency departments (EDs). A survey was conducted among managers of all 102 EDs in the Netherlands, using a semi-structured online questionnaire. Seventy-two questionnaires were analysed. Most EDs use a one-team system (68 %). EDs with a tiered-response receive more multi trauma patients (p < 0.01) and have more trauma team alerts per year (p < 0.05) than one-team EDs. The number of trauma team members varies from three to 16 professionals. The ED nurse usually receives the pre-notification (97 %), whereas the decision to activate a team is made by an ED nurse (46 %), ED physician (30 %), by multiple professionals (20 %) or other (4 %). Information in the pre-notification mostly used for trauma team activation are Airway-Breathing-Circulation (87 %), Glasgow Coma Score (90 %), and Revised Trauma Score (85 %) or Paediatric Trauma Score (86 %). However, this information is only available for 75 % of the patients or less. Only 56 % of the respondents were satisfied with their current in-hospital trauma triage system. Trauma team activation varies across Dutch EDs and there is room for improvement in the trauma triage system used, size of the teams and the professionals involved. More direct communication and more uniform criteria could be used to efficiently and safely activate a specific trauma team. Therefore, the implementation of a revised national consensus guideline is recommended.


Classification of upper limb center-out reaching tasks by means of EEG-based continuous decoding techniques.
One of the current challenges in brain-machine interfacing is to characterize and decode upper limb kinematics from brain signals, e.g. to control a prosthetic device. Recent research work states that it is possible to do so based on low frequency EEG components. However, the validity of these results is still a matter of discussion. In this paper, we assess the feasibility of decoding upper limb kinematics from EEG signals in center-out reaching tasks during passive and active movements. The decoding of arm movement was performed using a multidimensional linear regression. Passive movements were analyzed using the same methodology to study the influence of proprioceptive sensory feedback in the decoding. Finally, we evaluated the possible advantages of classifying reaching targets, instead of continuous trajectories. The results showed that arm movement decoding was significantly above chance levels. The results also indicated that EEG slow cortical potentials carry significant information to decode active center-out movements. The classification of reached targets allowed obtaining the same conclusions with a very high accuracy. Additionally, the low decoding performance obtained from passive movements suggests that discriminant modulations of low-frequency neural activity are mainly related to the execution of movement while proprioceptive feedback is not sufficient to decode upper limb kinematics. This paper contributes to the assessment of feasibility of using linear regression methods to decode upper limb kinematics from EEG signals. From our findings, it can be concluded that low frequency bands concentrate most of the information extracted from upper limb kinematics decoding and that decoding performance of active movements is above chance levels and mainly related to the activation of cortical motor areas. We also show that the classification of reached targets from decoding approaches may be a more suitable real-time methodology than a direct decoding of hand position.


Zero-mode waveguides for single-molecule analysis at high concentrations.
Optical approaches for observing the dynamics of single molecules have required pico- to nanomolar concentrations of fluorophore in order to isolate individual molecules. However, many biologically relevant processes occur at micromolar ligand concentrations, necessitating a reduction in the conventional observation volume by three orders of magnitude. We show that arrays of zero-mode waveguides consisting of subwavelength holes in a metal film provide a simple and highly parallel means for studying single-molecule dynamics at micromolar concentrations with microsecond temporal resolution. We present observations of DNA polymerase activity as an example of the effectiveness of zero-mode waveguides for performing single-molecule experiments at high concentrations.


Measuring economic outcomes of alcohol treatment using the Economic Form 90.
This article assesses the ability of the economic outcome measures in the Economic Form 90 to detect differences across levels of alcohol dependence as measured by the Alcohol Dependence Scale. We used baseline data from the Combining Medications and Behavioral Interventions (COMBINE) Study, a large, multisite clinical trial, to assess the extent to which the economic items on the Economic Form 90 instrument can detect differences across levels of alcohol dependence. After adjusting for differences in demographic characteristics, the Economic Form 90 can detect significant differences across a range of dependence severity levels for the economic outcomes of inpatient medical care, emergency-department medical care, behavioral health care, being on parole or probation, and missed workdays, conditional on being employed. We did not detect significant differences across dependence severity for employment status, outpatient medical care, other criminal justice involvement, or motor vehicle accidents. The Economic Form 90 can identify differences in many economic outcomes associated with differing levels of alcohol dependence. This suggests that the Economic Form 90 may be useful in assessing changes in economic outcomes that result from changes in alcohol dependence.


Microbial community composition shapes enzyme patterns in topsoil and subsoil horizons along a latitudinal transect in Western Siberia.
Soil horizons below 30 cm depth contain about 60% of the organic carbon stored in soils. Although insight into the physical and chemical stabilization of soil organic matter (SOM) and into microbial community composition in these horizons is being gained, information on microbial functions of subsoil microbial communities and on associated microbially-mediated processes remains sparse. To identify possible controls on enzyme patterns, we correlated enzyme patterns with biotic and abiotic soil parameters, as well as with microbial community composition, estimated using phospholipid fatty acid profiles. Enzyme patterns (i.e. distance-matrixes calculated from these enzyme activities) were calculated from the activities of six extracellular enzymes (cellobiohydrolase, leucine-amino-peptidase, N-acetylglucosaminidase, chitotriosidase, phosphatase and phenoloxidase), which had been measured in soil samples from organic topsoil horizons, mineral topsoil horizons, and mineral subsoil horizons from seven ecosystems along a 1500 km latitudinal transect in Western Siberia. We found that hydrolytic enzyme activities decreased rapidly with depth, whereas oxidative enzyme activities in mineral horizons were as high as, or higher than in organic topsoil horizons. Enzyme patterns varied more strongly between ecosystems in mineral subsoil horizons than in organic topsoils. The enzyme patterns in topsoil horizons were correlated with SOM content (i.e., C and N content) and microbial community composition. In contrast, the enzyme patterns in mineral subsoil horizons were related to water content, soil pH and microbial community composition. The lack of correlation between enzyme patterns and SOM quantity in the mineral subsoils suggests that SOM chemistry, spatial separation or physical stabilization of SOM rather than SOM content might determine substrate availability for enzymatic breakdown. The correlation of microbial community composition and enzyme patterns in all horizons, suggests that microbial community composition shapes enzyme patterns and might act as a modifier for the usual dependency of decomposition rates on SOM content or C/N ratios.


Communication over the network of binary switches regulates the activation of A2A adenosine receptor.
Dynamics and functions of G-protein coupled receptors (GPCRs) are accurately regulated by the type of ligands that bind to the orthosteric or allosteric binding sites. To glean the structural and dynamical origin of ligand-dependent modulation of GPCR activity, we performed total ~ 5 μsec molecular dynamics simulations of A2A adenosine receptor (A2AAR) in its apo, antagonist-bound, and agonist-bound forms in an explicit water and membrane environment, and examined the corresponding dynamics and correlation between the 10 key structural motifs that serve as the allosteric hotspots in intramolecular signaling network. We dubbed these 10 structural motifs "binary switches" as they display molecular interactions that switch between two distinct states. By projecting the receptor dynamics on these binary switches that yield 2(10) microstates, we show that (i) the receptors in apo, antagonist-bound, and agonist-bound states explore vastly different conformational space; (ii) among the three receptor states the apo state explores the broadest range of microstates; (iii) in the presence of the agonist, the active conformation is maintained through coherent couplings among the binary switches; and (iv) to be most specific, our analysis shows that W246, located deep inside the binding cleft, can serve as both an agonist sensor and actuator of ensuing intramolecular signaling for the receptor activation. Finally, our analysis of multiple trajectories generated by inserting an agonist to the apo state underscores that the transition of the receptor from inactive to active form requires the disruption of ionic-lock in the DRY motif.


Removal or Revision of Vaginal Mesh Used for the Treatment of Stress Urinary Incontinence.
Synthetic mesh slings are the most common surgical treatment for female stress urinary incontinence (SUI). However, the US Food and Drug Administration has released warnings that question the safety of vaginal mesh. To measure the incidence of mesh removal or revision after SUI procedures and to determine whether significant surgeon and patient risk factors exist. Population-based retrospective cohort study that included all adult women undergoing an incident procedure for SUI with synthetic mesh in Ontario, Canada, from April 1, 2002, through December 31, 2012 (N = 59 887). The end of potential follow-up was March 31, 2013. Data were analyzed from November 1, 2014, through February 28, 2015. Yearly volume of mesh-based procedures for SUI performed by the treating surgeons and their surgical specialty. The primary outcome was a composite of surgical procedures related to removal or revision of mesh slings (owing to erosion, fistula, pain, or retention). We hypothesized a priori that surgeon volume would be inversely correlated with complications. Among the identified 59 887 women who underwent a mesh-based procedure for SUI, the median age was 52 (interquartile range [IQR], 45-63) years. High-volume surgeons (≥75th percentile of yearly mesh-based procedures) were less likely to perform a simultaneous hysterectomy (performed in 11.5% vs 16.5% of patients; standardized difference, 0.14), were more likely to work in an academic center (28.9% vs 16.3% of patients; standardized difference, 0.30), and saw the patient less frequently in the year before the procedure (median, 2 [IQR, 1-3] vs 3 [IQR, 2-4] visits; standardized difference, 0.26). Complications were treated in 1307 women (2.2%), and the 10-year cumulative incidence rate was 3.29 (95% CI, 3.05-3.53). In our multivariable survival model, patients of high-volume surgeons had a significantly lower risk (95% CI) for experiencing our composite outcome (hazard ratio [HR], 0.73 [0.65-0.83]; absolute risk reduction, 0.63% [0.36%-0.92%]; P < .01). Gynecologists were not significantly associated with more complications compared with urologists (HR, 0.94 [95% CI, 0.83-1.08]; P = .38). Among our secondary exposures of interest, multiple mesh-based SUI procedures increased the risk for complications (HR, 4.73 [95% CI, 3.62-6.17]; P < .01). However, traditional high-risk patient features did not increase the risk (HR, 0.58 [95% CI, 0.08-4.13]; P = .59). Ten years after SUI mesh surgery, 1 of every 30 women may require a second procedure for mesh removal or revision. Patients of lower-volume surgeons have a 37% increased likelihood of having a complication. These findings support the recommendations of the US Food and Drug Administration related to the use of vaginal mesh for treatment of SUI.


Editing at the crossroad of innate and adaptive immunity.
Genetic information can be altered through the enzymatic modification of nucleotide sequences. This process, known as editing, was originally identified in the mitochondrial RNA of trypanosomes and later found to condition events as diverse as neurotransmission and lipid metabolism in mammals. Recent evidence reveals that editing enzymes may fulfill one of their most essential roles in the defense against infectious agents: first, as the mediators of antibody diversification, a step crucial for building adaptive immunity, and second, as potent intracellular poisons for the replication of viruses. Exciting questions are raised, which take us to the depth of the intimate relations between vertebrates and the microbial underworld.


A scoping review protocol on social participation of indigenous elders, intergenerational solidarity and their influence on individual and community wellness.
Indigenous elders have traditionally played an important role in maintaining social cohesion within their communities. Today, part of this role has been taken over by government social and healthcare services, but they are having limited success in addressing social challenges. Increasing elders' social participation and intergenerational solidarity might foster community development and benefit young people, families, communities and the elders themselves. However, knowledge of the contribution of elders' social participation and intergenerational solidarity to wellness is scattered and needs to be synthesised. This protocol presents a scoping review on the social participation of indigenous elders, intergenerational solidarity and their influence on individual and community wellness. This scoping review protocol is based on an innovative methodological framework designed to gather information from the scientific and grey literature and from indigenous sources. It was developed by an interdisciplinary team including indigenous scholars/researchers, knowledge users and key informants. In addition to searching information databases in fields such as public health and indigenous studies, an advisory committee will ensure that information is gathered from grey literature and indigenous sources. The protocol was approved by the Ethics Review Board of the Université du Québec en Abitibi-Témiscamingue and the First Nations of Quebec and Labrador Health and Social Services Commission. The comprehensive synthesis of the scientific and grey literature and indigenous sources proposed in this protocol will not only raise awareness within indigenous communities and among healthcare professionals and community organisations, but will also enable decision-makers to better meet the needs of indigenous people. The innovative methodological framework proposed in this scoping review protocol will yield richer information on the contribution of elders to community wellness. This work is an essential preliminary step towards developing research involving indigenous communities, drawing on the social participation of elders and intergenerational solidarity.


Relations among loneliness, social anxiety, and problematic Internet use.
The model of problematic Internet use advanced and tested in the current study proposes that individuals' psychosocial well-being, along with their beliefs about interpersonal communication (both face-to-face and online) are important cognitive predictors of negative outcomes arising from Internet use. The study examined the extent to which social anxiety explains results previously attributed to loneliness as a predictor of preference for online social interaction and problematic Internet use. The results support the hypothesis that the relationship between loneliness and preference for online social interaction is spurious, and that social anxiety is the confounding variable.


Rhnull: a rare blood group phenotype.
Rhnull phenotype is a rare blood group characterized by the lack of expression of all Rh antigens (D, C, c, E and e) on the red cells. The phenotype is further classified into the regulator and amorph type based on underlying genetic defect. The clinical significance of its recognition is that such patients suffer from Rhnull syndrome associated with osmotically fragile red cells called stomatocytes with subsequent chronic haemolytic anaemia of varying degree. Another importance is that such subjects readily form alloantibodies on exposure to Rh antigens. We report herein rare Rhaull phenotype in a young pregnant female which was detected as a part of routine antenatal work-up for red cell antibody screening and identification.


Redox-Active Phenanthrenequinone Triangles in Aqueous Rechargeable Zinc Batteries.
Aqueous rechargeable zinc batteries (ZBs) have received considerable attention recently for large-scale energy storage systems in terms of rate performance, cost, and safety. Nevertheless, these ZBs still remain a subject for investigation, as researchers search for cathode materials enabling high performance. Among the various candidate cathode materials for ZBs, quinone compounds stand out as candidates because of their high specific capacity, sustainability, and low cost. Quinone-based cathodes, however, suffer from the critical limitation of undergoing dissolution during battery cycling, leading to a deterioration in battery life. To address this problem, we have introduced a redox-active triangular phenanthrenequinone-based macrocycle (PQ-Δ) with a rigid geometry and layered superstructure. Notably, we have confirmed that Zn2+ ions, together with H2O molecules, can be inserted into the PQ-Δ organic cathode, and, as a consequence, the interfacial resistance between the cathode and electrolytes is decreased effectively. Density functional theory calculations have revealed that the low interfacial resistance can be attributed mainly to decreasing the desolvation energy penalty as a result of the insertion of hydrated Zn2+ ions in the PQ-Δ cathode. The combined effects of the insertion of hydrated Zn2+ ions and the robust triangular structure of PQ-Δ serve to achieve a large reversible capacity of 210 mAh g-1 at a high current density of 150 mA g-1, along with an excellent cycle-life, that is, 99.9% retention after 500 cycles. These findings suggest that the utilization of electron-active organic macrocycles, combined with the low interfacial resistance associated with the solvation of divalent carrier ions, is essential for the overall performance of divalent battery systems.


[Chagas' disease in a urban population of the health district of Rio Verde, Mato Grosso do Sul State, Brazil].
In order to evaluate some aspects of the Chagas' disease on urban population of the 12 municipalities of the sanitarian district of Rio Verde, Mato Grosso do Sul state, on 1998 we carried out a serological survey investigating anti-T. cruzi IgG by indirect immunofluorescence on blood samples collected on filter paper of 14,709 resident persons, confirmed by indirect hemagglutination and ELISA in serum. The parasitaemia was evaluated by indirect xenodiagnosis on 134 chagasic patients and the morbidity evaluated by anamnesis, physical exam and electrocardiogram in 191 pairs of chagasic/non-chagasic patients. The seropositivity was 1.83% (0.93% in autochtonous, 5.01% in alochtonous), xenodiagnosis was positive in 17.2% of the individuals (12.3% in autochtonous, 20.8% in alochtonous) and 24.6% of the seropositive patients presented chronic chagasic cardiopathy (19.1% in autochtonous, 27.8 in alochtonous). The analysis of the dates showed that alochtonous population is the main responsible for presence of the infection and morbidity of Chagas disease in the studied area.


'I wash until it feels right' the phenomenology of stopping criteria in obsessive-compulsive washing.
Recent elaborations of cognitive behavioral theory in OCD suggest that difficulties in deciding when to stop a compulsive action may be related to the use of counter-productive termination criteria by obsessional patients [Salkovskis, P. M. (1999). Understanding and treating obsessive-compulsive disorder. Behaviour Research and Therapy, 37, s29-s52]. Such criteria are characterized by their subjective nature, i.e. a primarily internal reference point (e.g. 'just right' feelings), and are conceptualized as the "top level" of a general strategy involving elevated evidence requirements. Thirty-eight obsessional washers, 41 obsessionals with other problems and 43 healthy controls were interviewed about and rated two situations varying in the degree of urgency to wash; they also washed their hands in a behavioral test. Washers reported using subjective criteria more frequently and rated them as more important for the termination of the washes than the other groups in questionnaire, interview and laboratory data. Both obsessional groups considered more criteria before stopping than the healthy controls, suggesting that using multiple criteria is a general strategy. The data are consistent with the predictions of the elaborated cognitive-behavioral model of OCD. They indicate that the use of subjective criteria and elevated evidence requirements is affected by the perceived significance of the situation in a similar way for obsessional and non-obsessional individuals.


The lethal giant larvae tumour suppressor mutation requires dMyc oncoprotein to promote clonal malignancy.
Neoplastic overgrowth depends on the cooperation of several mutations ultimately leading to major rearrangements in cellular behaviour. Precancerous cells are often removed by cell death from normal tissues in the early steps of the tumourigenic process, but the molecules responsible for such a fundamental safeguard process remain in part elusive. With the aim to investigate the molecular crosstalk occurring between precancerous and normal cells in vivo, we took advantage of the clonal analysis methods that are available in Drosophila for studying the phenotypes due to lethal giant larvae (lgl) neoplastic mutation induced in different backgrounds and tissues. We observed that lgl mutant cells growing in wild-type imaginal wing discs show poor viability and are eliminated by Jun N-terminal Kinase (JNK)-dependent cell death. Furthermore, they express very low levels of dMyc oncoprotein compared with those found in the surrounding normal tissue. Evidence that this is a cause of lgl mutant cells elimination was obtained by increasing dMyc levels in lgl mutant clones: their overgrowth potential was indeed re-established, with mutant cells overwhelming the neighbouring tissue and forming tumourous masses displaying several cancer hallmarks. Moreover, when lgl mutant clones were induced in backgrounds of slow-dividing cells, they upregulated dMyc, lost apical-basal cell polarity and were able to overgrow. Those phenotypes were abolished by reducing dMyc levels in the mutant clones, thereby confirming its key role in lgl-induced tumourigenesis. Furthermore, we show that the eiger-dependent Intrinsic Tumour Suppressor pathway plays only a minor role in eliminating lgl mutant cells in the wing pouch; lgl-/- clonal death in this region is instead driven mainly by dMyc-induced Cell Competition. Our results provide the first evidence that dMyc oncoprotein is required in lgl tumour suppressor mutant tissue to promote invasive overgrowth in larval and adult epithelial tissues. Moreover, we show that dMyc abundance inside versus outside the mutant clones plays a key role in driving neoplastic overgrowth.


Emerging aspects of gut sulfur amino acid metabolism.
This review discusses the recent evidence indicating that sulfur amino acid metabolism in gastrointestinal tissues may be linked to human health and gut disease. Studies indicate that the gastrointestinal tract metabolizes 20% of dietary methionine and that its main metabolic fate is transmethylation to homocysteine and transsulfuration to cysteine. The gastrointestinal tract accounts for approximately 25% of whole-body transmethylation and transsulfuration and is a site of net homocysteine release. The production of homocysteine within the intestinal mucosa may contribute to the inflammatory response and endothelial cell dysfunction in patients with inflammatory bowel disease. Studies also show that the availability of S-adenosylmethionine as a precursor for methylation reactions and polyamines plays a key role in epigenetic DNA methylation, gene expression and colon carcinogenesis. Cysteine derived from the diet and methionine transsulfuration is a functional constituent of antioxidant systems and impacts several elements of redox status that regulate epithelial intracellular signaling, proliferation and survival. Further studies are warranted to establish how local production of homocysteine, S-adenosylmethionine and antioxidants contributes to the development of gastrointestinal diseases and whether dietary intervention with folate and cysteine is an efficacious approach to prevention and treatment.


Critical illness-induced bone loss is related to deficient autophagy and histone hypomethylation.
Survivors of critical illness are at increased risk of fractures. This may be due to increased osteoclast formation during critical illness, leading to trabecular bone loss. Such bone loss has also been observed in Paget's disease, and has been related to deficient autophagy. Deficient autophagy has also been documented in vital organs and skeletal muscle of critically ill patients. The objective of this study was to investigate whether deficient autophagy can be linked to critical illness-induced bone loss. Osteoclasts grown in vitro and their precursor cells isolated from peripheral blood of critically ill patients and from matched healthy volunteers were analysed for the expression of autophagy genes (SQSTM1, Atg3 and Atg7), and proteins (p62, Atg-5, and microtubule-associated protein light chain 3-II (LC3-II)) and for autophagy and epigenetic signalling factors via PCR arrays and were treated with the autophagy inducer rapamycin. The effect of rapamycin was also investigated at the tissue level in an in vivo rabbit model of critical illness. Many more osteoclasts formed in vitro from the blood precursor cells isolated from critically ill patients, which accumulated p62, and displayed reduced expression of Atg5, Atg7, and LC3-II compared to healthy controls, suggesting deficient autophagy, whilst addition of rapamycin reduced osteoclast formation. PCR arrays revealed a down-regulation of histone methyltransferases coupled with an up-regulation of negative regulators of autophagy. Critically ill rabbits displayed a reduction in trabecular and cortical bone, which was rescued with rapamycin. Deficient autophagy in osteoclasts and their blood precursor cells at least partially explained aberrant osteoclast formation during critical illness and was linked to global histone hypomethylation. Treatment with the autophagy activator Rapamycin reduced patient osteoclast formation in vitro and reduced the amount of bone loss in critically ill rabbits in vivo. These findings may help to develop novel therapeutic targets to prevent critical illness-induced bone loss.


The role of copper(II) in the aggregation of human amylin.
Amylin is a 37-residue peptide hormone produced by the islet β-cells of pancreas and the formation of amylin aggregates is strongly associated with β-cell degeneration in type 2 diabetes, as demonstrated by more than 95% of patients exhibiting amylin amyloid upon autopsy. It is widely recognized that metal ions such as copper(II) have been implicated in the aggregation process of amyloidogenic peptides such as Aβ and α-synuclein and there is evidence that amylin self-assembly is also largely affected by copper(II). For this reason, in this work, the role of copper(II) in the aggregation of amylin has been investigated by several different experimental approaches. Mass spectrometric investigations show that copper(II) induces significant changes in the amylin structure, which decrease the protein fibrillogenesis as observed by ThT measurements. Accordingly, solid-state NMR experiments together with computational analysis carried out on a model amylin fragment confirmed the non-fibrillogenic nature of the copper(II) induced aggregated structure. Finally, the presence of copper(II) is also shown to have a major influence on amylin proneness to be degraded by proteases and cytotoxicity studies on different cell cultures are reported.


Nordihydroguaiaretic acid increases endothelial nitric oxide synthase expression via the transcription factor AP-1.
It has been previously reported that the antioxidant compound nordihydroguaiaretic acid (NDGA) increases endothelial nitric oxide synthase (eNOS) expression in cultured bovine aortic endothelial cells. However, the exact mechanism for this effect was unresolved. Thus, the purpose of this study was to further elucidate the effect of NDGA on eNOS protein expression and enzymatic activity in fetal pulmonary arterial endothelial cells (FPAECs), and to identify the transcription factors involved in this regulation. Following overnight exposure to 0-32 microM NDGA, we observed a 2- to 2.5-fold increase in eNOS protein expression in FPAECs, with a similar increase observed in eNOS activity. For eNOS gene promoter analysis, we initially used two promoter-reporter constructs: a 1.6 kb promoter fragment and an 840 bp construct, both of which include an AP-1-specific binding site. NDGA exposure induced a significant increase in eNOS promoter activity in both constructs. However, the NDGA-mediated increase was abolished when we used either a truncated promoter construct lacking the AP-1 element or a construct in which the AP-1 binding site was mutated. AP-1 binding efficiency was also determined by electrophoretic mobility shift assay, where we observed an increase in AP-1 binding in FPAECs treated with NDGA while the binding of AP-1 was found to be decreased when a mutated AP-1 consensus sequence was used. Further, supershift analyses indicated that the AP-1 complex consisted of c-Jun and FosB. We therefore conclude that NDGA antioxidant activity regulates eNOS expression via AP-1 and that antioxidant therapy could potentially be used to increase eNOS expression in diseases, such as persistent pulmonary hypertension of the newborn, where eNOS expression and activity are known to be reduced.


Genetic Polymorphisms of rs3077 and rs9277535 in HLA-DP associated with Systemic lupus erythematosus in a Chinese population.
Although the SLE risk gene loci of HLA-DR and HLA-DQ within the major histocompatibility complex (MHC) region has been gradually revealed by recent Genome-Wide Association studies (GWAS), the association of HLA-DP polymorphisms with SLE was minimally reported. Considering that the variants in rs3077 and rs9277535 in the HLA-DP region could influence the immune response by affecting antigen presentation of HLA class II molecules to CD4+ T cells, the present study aimed to explore the role of HLA-DP polymorphisms in SLE. In total, samples from 335 SLE patients and 635 healthy controls were collected and genotyped by a polymerase chain reaction-high resolution melting (PCR-HRM) assay. A significant positive correlation was observed between the SNP rs3077, rs9277535 of HLA-DP and SLE susceptibility (rs3077, OR = 0.74, 95%CI = 0.60-0.91, P = 0.004; rs9277535, OR = 0.72, 95%CI = 0.59-0.88, P = 0.001). Rs3077 polymorphism was corelated to IL-17, INF-γ and cutaneous vasculitis (P = 0.037, P = 0.020 and P = 0.006, respectively). Additionally, rs3077 AA genotype carriers showed lower concentration of inflammatory cytokines and lower cutaneous vasculitis incidence than did the other two genotype. No significant association was observed between rs9277535 and cytokines or any clinical features. In conclusion, HLA-DP polymorphisms (rs3077 and rs9277535) were associated with SLE susceptibility and the levels of some inflammatory cytokines in SLE patients.


Vitamin k2, a naturally occurring menaquinone, exerts therapeutic effects on both hormone-dependent and hormone-independent prostate cancer cells.
In recent years, several studies have shown that vitamin k2 (VK2) has anticancer activity in a variety of cancer cells. The antitumor effects of VK2 in prostate cancer are currently not known. In the present study, we sought to characterize the anticancer potential of VK2 in both androgen-dependent and -independent prostate cancer cells. Our investigations show that VK2 is able to suppress viability of androgen-dependent and androgen-independent prostate cancer cells via caspase-3 and -8 dependent apoptosis. We also show that VK2 treatment reduces androgen receptor expression and PSA secretion in androgen-dependent prostate cancer cells. Our results also implicate VK2 as a potential anti-inflammatory agent, as several inflammatory genes are downregulated in prostate cancer cells following treatment with VK2. Additionally, AKT and NF-kB levels in prostate cancer cells are reduced significantly when treated with VK2. These findings correlated with the results of the Boyden chamber and angiogenesis assay, as VK2 treatment reduced cell migration and angiogenesis potential of prostate cancer cells. Finally, in a nude mice model, VK2 administration resulted in significant inhibition of both androgen-dependent and androgen-independent tumor growth. Overall, our results suggest that VK2 may be a potential therapeutic agent in the treatment of prostate cancer.


Massive turnover of functional sequence in human and other mammalian genomes.
Despite the availability of dozens of animal genome sequences, two key questions remain unanswered: First, what fraction of any species' genome confers biological function, and second, are apparent differences in organismal complexity reflected in an objective measure of genomic complexity? Here, we address both questions by applying, across the mammalian phylogeny, an evolutionary model that estimates the amount of functional DNA that is shared between two species' genomes. Our main findings are, first, that as the divergence between mammalian species increases, the predicted amount of pairwise shared functional sequence drops off dramatically. We show by simulations that this is not an artifact of the method, but rather indicates that functional (and mostly noncoding) sequence is turning over at a very high rate. We estimate that between 200 and 300 Mb (∼6.5%-10%) of the human genome is under functional constraint, which includes five to eight times as many constrained noncoding bases than bases that code for protein. In contrast, in D. melanogaster we estimate only 56-66 Mb to be constrained, implying a ratio of noncoding to coding constrained bases of about 2. This suggests that, rather than genome size or protein-coding gene complement, it is the number of functional bases that might best mirror our naïve preconceptions of organismal complexity.


Long non-coding RNA-derived peptides are immunogenic and drive a potent anti-tumour response.
Protein arginine methyltransferase (PRMT) 5 is over-expressed in a variety of cancers and the master transcription regulator E2F1 is an important methylation target. We have explored the role of PRMT5 and E2F1 in regulating the non-coding genome and report here a striking effect on long non-coding (lnc) RNA gene expression. Moreover, many MHC class I protein-associated peptides were derived from small open reading frames in the lncRNA genes. Pharmacological inhibition of PRMT5 or adjusting E2F1 levels qualitatively altered the repertoire of lncRNA-derived peptide antigens displayed by tumour cells. When presented to the immune system as either ex vivo-loaded dendritic cells or expressed from a viral vector, lncRNA-derived peptides drove a potent antigen-specific CD8 T lymphocyte response, which translated into a significant delay in tumour growth. Thus, lncRNA genes encode immunogenic peptides that can be deployed as a cancer vaccine.


Genome position and gene amplification.
Amplifications, regions of focal high-level copy number change, lead to overexpression of oncogenes or drug resistance genes in tumors. Their presence is often associated with poor prognosis; however, the use of amplification as a mechanism for overexpression of a particular gene in tumors varies. To investigate the influence of genome position on propensity to amplify, we integrated a mutant form of the gene encoding dihydrofolate reductase into different positions in the human genome, challenged cells with methotrexate and then studied the genomic alterations arising in drug resistant cells. We observed site-specific differences in methotrexate sensitivity, amplicon organization and amplification frequency. One site was uniquely associated with a significantly enhanced propensity to amplify and recurrent amplicon boundaries, possibly implicating a rare folate-sensitive fragile site in initiating amplification. Hierarchical clustering of gene expression patterns and subsequent gene enrichment analysis revealed two clusters differing significantly in expression of MYC target genes independent of integration site. These studies suggest that genome context together with the particular challenges to genome stability experienced during the progression to cancer contribute to the propensity to amplify a specific oncogene or drug resistance gene, whereas the overall functional response to drug (or other) challenge may be independent of the genomic location of an oncogene.


Analysis of hematopoietic cell specific protein, M-DOCK.
Full length cDNA of M-DOCK was isolated at Kazusa DNA Institute and its sequence has been deposited as KIAA0209. The cDNA sequence of M-DOCK contains a 5490 bp open reading frame that encodes a 1830-aa protein with a calculated molecular mass of 212 kDa. The amino acid sequence of M-DOCK has 62.3% identity with DOCK180, excluding the carboxyl terminal variable regions. DOCK180, which was originally identified as a major protein bound to Crk oncogene product, is an archetype of a family of proteins including Ced-5 of C. elegans and Mbc of D. melanogaster. DOCK180 is localized at focal adhesions and regulates cell morphology through the activation of Rac. Expression of DOCK180 is ubiquitous except in hematopoietic cells. Here, the author reports on the characterization of M-DOCK protein, which is closely related to DOCK180 but expressed only in the hematopoietic cells. We immunized rabbits with bacterially-expressed GST-M-DOCK protein, and obtained an anti-serum specific to M-DOCK. Tissue distribution of M-DOCK was examined by Northern blotting and Western blotting. M-DOCK was expressed abundantly in peripheral blood leukocytes. M-DOCK expression was detected also in the cell lines derived from T-cells, B-cells, and monocytes, but never in the adherent cells. Most of the lymphocytes and macrophages in human organs expressed M-DOCK diffusely in the cytoplasm, which was analyzed by immunohistochemistry using an anti-M-DOCK antibody. We previously reported that membrane-targeted form of DOCK180 induced spreading of NIH 3T3 cells. By contrast, overexpression of M-DOCK rounded up flat NRK cells. This suggests that M-DOCK may regulate cell detachment of adherent cells. These results demonstrate that M-DOCK is a new member of DOCK180-family proteins and regulates cell shape in suspension cells.


Imaging mass spectrometry technology and application on ganglioside study; visualization of age-dependent accumulation of C20-ganglioside molecular species in the mouse hippocampus.
Gangliosides are particularly abundant in the central nervous system (CNS) and thought to play important roles in memory formation, neuritogenesis, synaptic transmission, and other neural functions. Although several molecular species of gangliosides have been characterized and their individual functions elucidated, their differential distribution in the CNS are not well understood. In particular, whether the different molecular species show different distribution patterns in the brain remains unclear. We report the distinct and characteristic distributions of ganglioside molecular species, as revealed by imaging mass spectrometry (IMS). This technique can discriminate the molecular species, raised from both oligosaccharide and ceramide structure by determining the difference of the mass-to-charge ratio, and structural analysis by tandem mass spectrometry. Gangliosides in the CNS are characterized by the structure of the long-chain base (LCB) in the ceramide moiety. The LCB of the main ganglioside species has either 18 or 20 carbons (i.e., C18- or C20-sphingosine); we found that these 2 types of gangliosides are differentially distributed in the mouse brain. While the C18-species was widely distributed throughout the frontal brain, the C20-species selectively localized along the entorhinal-hippocampus projections, especially in the molecular layer (ML) of the dentate gyrus (DG). We revealed development- and aging-related accumulation of the C-20 species in the ML-DG. Thus it is possible to consider that this brain-region specific regulation of LCB chain length is particularly important for the distinct function in cells of CNS.


Enantioselective capillary electrochromatography: recent developments and new trends.
Since its development in the early 1970s, CEC has been studied quite extensively, but unfortunately its use is still mostly located at an academic level. Reasons for this are the limited availability of commercially available stationary phases (SPs) and columns, along with some practical limitations, such as column fragility, lack of column robustness and reproducibility. Nevertheless, CEC maintains a place among the separation techniques, probably because of its unique feature to combine two separation principles. Also in the field of chiral separations, CEC is often used as a separation technique and already showed its potential for this kind of analyses. This overview will focus on the recent applications, i.e. between 2006 and 2010, in enantioselective analysis by means of CEC. For the selected applications, the used SPs (chiral selectors) and their potential for future method development or screening purposes will be evaluated and critically discussed.


Effects of gonadotrophin-releasing hormone agonists on apoptosis of granulosa cells.
Granulosa cells are known to contribute to maturation of oocytes, and most of the growth factors exert their action via granulosa cells. It has been established that granulosa cell death during follicular atresia and luteolysis results from apoptosis. However, the precise mechanistic pathways of granulosa cell apoptosis have not yet been defined. In this study, we determined the proportions of apoptosis in granulosa cells treated with two kinds of gonadotrophin-releasing hormone agonists (GnRHa): buserelin and leuprorelin depot. The incidences of DNA fragmentation of human granulosa cells treated with buserelin and leuprorelin were 54.33% and 39.02%, respectively. The proportions of apoptotic bodies were 6.04% and 4.29%, respectively. There was a significant difference in the proportions of DNA fragmentation between the two kinds of GnRHa-treated granulosa cells. The apoptosis pathway and associated protein expression in granulosa cells treated with GnRHa were also determined. The Bax molecule, a pro-apoptosis protein, was expressed in granulosa cells undergoing apoptosis. In contrast, Bcl-2, an anti-apoptosis protein, could not be detected in the same group of granulosa cells. The distribution of cytochrome c determined via immunostaining showed a diffuse pattern, which most likely indicated that cytochrome c was translocated from mitochondria into the cytoplasm. Western blotting showed the expressions of caspase-9 and caspase-3 in patients' granulosa cells. The GnRHa effects on granulosa cells indicated a higher incidence of DNA fragmentation and apoptotic bodies in the buserelin-treated than in the leuprorelin depot-treated group. The granulosa cells go through the mitochondria-dependent apoptosis pathway; the indicated pro-apoptosis protein Bax was expressed and induced cytochrome c release from mitochondria, which then activated caspase-9 and caspase-3 until cell death occurred.


Computing gap free Pareto front approximations with stochastic search algorithms.
Recently, a convergence proof of stochastic search algorithms toward finite size Pareto set approximations of continuous multi-objective optimization problems has been given. The focus was on obtaining a finite approximation that captures the entire solution set in some suitable sense, which was defined by the concept of epsilon-dominance. Though bounds on the quality of the limit approximation-which are entirely determined by the archiving strategy and the value of epsilon-have been obtained, the strategies do not guarantee to obtain a gap free approximation of the Pareto front. That is, such approximations A can reveal gaps in the sense that points f in the Pareto front can exist such that the distance of f to any image point F(a), a epsilon A, is "large." Since such gap free approximations are desirable in certain applications, and the related archiving strategies can be advantageous when memetic strategies are included in the search process, we are aiming in this work for such methods. We present two novel strategies that accomplish this task in the probabilistic sense and under mild assumptions on the stochastic search algorithm. In addition to the convergence proofs, we give some numerical results to visualize the behavior of the different archiving strategies. Finally, we demonstrate the potential for a possible hybridization of a given stochastic search algorithm with a particular local search strategy-multi-objective continuation methods-by showing that the concept of epsilon-dominance can be integrated into this approach in a suitable way.


Drug-induced amplification of nanoparticle targeting to tumors.
Nanomedicines have the potential to significantly impact cancer therapy by improving drug efficacy and decreasing off-target effects, yet our ability to efficiently home nanoparticles to disease sites remains limited. One frequently overlooked constraint of current active targeting schemes is the relative dearth of targetable antigens within tumors, which restricts the amount of cargo that can be delivered in a tumor-specific manner. To address this limitation, we exploit tumor-specific responses to drugs to construct a cooperative targeting system where a small molecule therapeutic modulates the disease microenvironment to amplify nanoparticle recruitment in vivo. We first administer a vascular disrupting agent, ombrabulin, which selectively affects tumors and leads to locally elevated presentation of the stress-related protein, p32. This increase in p32 levels provides more binding sites for circulating p32-targeted nanoparticles, enhancing their delivery of diagnostic or therapeutic cargos to tumors. We show that this cooperative targeting system recruits over five times higher doses of nanoparticles to tumors and decreases tumor burden when compared with non-cooperative controls. These results suggest that using nanomedicine in conjunction with drugs that enhance the presentation of target antigens in the tumor environment may be an effective strategy for improving the diagnosis and treatment of cancer.


Phenobarbital administration every eight hours: improvement of seizure management in idiopathic epileptic dogs with decreased phenobarbital elimination half-life.
Estimated prevalence of canine idiopathic epilepsy is 0.6 per cent in the first-opinion canine population in the UK. Phenobarbital monotherapy has been reported to reduce/eradicate seizure activity in 60-93 per cent of idiopathic epileptic dogs (IEDs). The objective of this study was to evaluate safety and efficacy of the administration of phenobarbital orally every eight hours in IEDs with phenobarbital elimination half-life less than 20 hours. Medical records of 10 IEDs in which steady state trough serum phenobarbital levels were within the reference range and phenobarbital elimination half-life had become less than 20 hours following prolonged administration every 12 hours were reviewed. Side effects and seizure frequency when phenobarbital was administered every 12 hours or 8 hours were compared. In all dogs the side effects of the antiepileptic medication treatment improved. When phenobarbital was administered every eight hours, 9/10 dogs experienced improvement in seizure frequency and 8/10 dogs maintained seizure freedom for a period three times longer than the longest interictal interval period previously recorded. Reduction in the severity and number of clusters of seizures was recorded in one of the remaining two dogs. The administration of phenobarbital orally every eight hours in IEDs with decreased phenobarbital elimination half-life appears safe and can improve seizure management. The results of this study were presented in abstract form (poster) for the 28th symposium of the European Society of Veterinary Neurology - European College of Veterinary Neurology (ESVN), September 18-19, 2015, Amsterdam, Netherlands.


Lamarck, evolution, and the inheritance of acquired characters.
Scientists are not always remembered for the ideas they cherished most. In the case of the French biologist Jean-Baptiste Lamarck, his name since the end of the nineteenth century has been tightly linked to the idea of the inheritance of acquired characters. This was indeed an idea that he endorsed, but he did not claim it as his own nor did he give it much thought. He took pride instead in advancing the ideas that (1) nature produced successively all the different forms of life on earth, and (2) environmentally induced behavioral changes lead the way in species change. This article surveys Lamarck's ideas about organic change, identifies several ironies with respect to how his name is commonly remembered, and suggests that some historical justice might be done by using the adjective "Lamarckian" to denote something more (or other) than a belief in the inheritance of acquired characters.


Fluorescence correlation spectroscopy in a reverse Kretchmann surface plasmon assisted microscope.
Fluorescence Correlation Spectroscopy (FCS) demands a high rate of photon detection per molecule, low background, and large fluctuations of fluorescence associated with translational motion. The new approach presented here, Surface Plasmon Assisted Microscope (SPAM), meets these requirements by drastically limiting the observation volume. In this method, the observational layer is made so thin that fluctuations are mostly due to the axial motion of molecules. This is conveniently realized by placing a sample on a thin metal film and illuminating it with a laser beam through an aqueous medium. The excited fluorophores close to the surface couple (via near-field interactions) to surface plasmons in the metal. Propagated surface plasmons decouple on opposite side of the metal film as a far-field radiation and emit in directional manner. Fluorescence is collected with a high Numerical Aperture objective. A confocal aperture inserted in its conjugate image plane reduces lateral dimensions of the detection volume to a diffraction limit. The thickness of the detection layer is reduced further by metal quenching of excited fluorophores at a close proximity (about 30 nm) to the surface. We used a suspension of fluorescent microspheres to show that FCS-SPAM is an efficient method to measure molecular diffusion.


Circadian influences on myocardial infarction.
Components of circadian rhythm maintenance, or "clock genes," are endogenous entrainable oscillations of about 24 h that regulate biological processes and are found in the suprachaismatic nucleus (SCN) and many peripheral tissues, including the heart. They are influenced by external cues, or Zeitgebers, such as light and heat, and can influence such diverse phenomena as cytokine expression immune cells, metabolic activity of cardiac myocytes, and vasodilator regulation by vascular endothelial cells. While it is known that the central master clock in the SCN synchronizes peripheral physiologic rhythms, the mechanisms by which the information is transmitted are complex and may include hormonal, metabolic, and neuronal inputs. Whether circadian patterns are causally related to the observed periodicity of events, or whether they are simply epi-phenomena is not well established, but a few studies suggest that the circadian effects likely are real in their impact on myocardial infarct incidence. Cycle disturbances may be harbingers of predisposition and subsequent response to acute and chronic cardiac injury, and identifying the complex interactions of circadian rhythms and myocardial infarction may provide insights into possible preventative and therapeutic strategies for susceptible populations.


Immunohistochemical Analysis of the Harbor Porpoise Using Antipodoplanin Antibody PMab-237.
Podoplanin (PDPN)/T1 alpha is known as a specific marker of lymphatic endothelial cells and type I alveolar cells. Sensitive and specific monoclonal antibodies (mAbs) for PDPN are needed for immunohistochemical analyses. Recently, we developed an anticetacean PDPN mAb, PMab-237. Herein, immunohistochemical analyses showed that PMab-237 strongly detected pulmonary type I alveolar cells, renal podocytes, and lymphatic endothelial cells of the harbor porpoise. These findings suggest that PMab-237 may be useful for immunohistochemical analyses for cetacean tissues.


Disseminated histoplasmosis: a comparative study between patients with acquired immunodeficiency syndrome and non-human immunodeficiency virus-infected individuals.
We studied 52 patients with disseminated histoplasmosis, 30 with the acquired immunodeficiency syndrome (AIDS) (cohort 1) and 22 not co-infected with the human immunodeficiency virus (cohort 2). Demographic, clinical, laboratory, mycologic findings, as well as antifungal therapy and highly active antiretroviral (HAART), were analyzed. Skin lesions were significantly higher in cohort 1 than in cohort 2 (P = 0.001). Anemia, leukopenia, and an elevated erythrocyte sedimentation rate were also more pronounced in cohort 1 than in cohort 2 (P < 0.001). Histoplasma capsulatum was isolated more often in cohort 1 than in cohort 2 (P < 0.05) patients, but antibodies to H. capsulatum were detected more frequently in cohort 2 than in cohort 1 (P < 0.05). Itraconazole treatment was less effective in cohort 1 than in cohort 2 (P = 0.012). In cohort 1 patients, HAART improved response to antifungals when compared with individuals not given HAART (P = 0.003), who exhibited higher mortality rates (P = 0.025). Cohort 1 patients who were given dual antifungal and anti-retroviral therapies responded as well as the non-HIV patients in cohort 2, who were treated only with itraconazole. These results indicate the need to promote restoration of the immune system in patients with AIDS and histoplasmosis.


Reported use of intravesical therapy for non-muscle-invasive bladder cancer (NMIBC): results from the Bladder Cancer Advocacy Network (BCAN) survey.
Study Type - Therapy (patterns of practice) Level of Evidence 2b. What's known on the subject? and What does the study add? Claims-based analyses suggest unexplained and potentially problematic variation in treatment intensity adherence to guidelines-recommended care in NMIBC. Previous physician surveys prior to the contemporary Clinical Practice Guidelines (CPGs) reported associations between variation in NMIBC care and practice type, years in practice, and other physician-related factors. In the largest physician survey addressing the management of NMIBC, and the first to examine these questions after the promulgation of contemporary CPGs, US urologists report grade-specific utilization consistent with CPG recommendations, at rates higher than suggested by recent claims-based analyses. As with prior studies, practice type and years in practice were significantly associated with variation in practices. Further research is needed to reconcile these findings with administrative claims data. To determine self-reported practices of use of intravesical chemo- and immunotherapy for patients with non-muscle-invasive bladder cancer (NMIBC) • To evaluate the extent to which respondent characteristics were associated with any observed variation. Guidelines recommend intravesical therapy (IVT) in the management of NMIBC, but recent claims-based analyses suggest exceedingly low rates of use of some of these therapies. An electronic survey was developed by members of the Bladder Cancer Advocacy Network (BCAN) to elicit self-reported use of management strategies for NMIBC. • The survey was circulated to urologists via the American Urological Association, Society for Urologic Oncology and Large Urology Group Practice Association distribution lists. • In all, 512 respondents completed the survey. In all, 63% reported routine perioperative mitomycin-c (MMC) after transurethral resection of bladder tumour (80% academic vs 54% private practice, P < 0.001). • Whereas 5% of respondents reported routine induction therapy with all new low-grade (LG) diagnoses, 99% reported routinely doing so in new high-grade (HG) cases; most commonly with single-agent bacille Calmette-Guérin (BCG) (94% vs 9% BCG/interferon and 5% MMC). • Reported induction therapy was higher in the setting of high-volume (77%) or frequently recurrent (44%) LG disease. • In all, 89% reported routinely using maintenance therapy for HG vs 29% for LG disease. • Routine biopsy after BCG, even with normal cystoscopy, was endorsed by 28% (39% academic vs 22% private practice, P < 0.001). Urologists report grade-specific use of IVT for NMIBC, at rates higher than suggested in some claims-based analyses. • Further study is needed to corroborate these self-reported patterns of care with lower rates of use suggested by claims-based analyses.


Osteogenic differentiation of human mesenchymal stem cells on α5 integrin binding peptide hydrogels is dependent on substrate elasticity.
The extracellular matrix plays a crucial role in controlling human mesenchymal stem cell (hMSC) biology including differentiation, and α5β1 integrin signaling plays an important role during osteogenic differentiation of hMSCs. Here, peptide-functionalized hydrogels were used to examine the role of α5β1 integrin signaling in inducing osteogenic differentiation in hMSCs. Further, the role of substrate elasticity was also studied. A thiolene chemistry was used to functionalize poly(ethylene glycol) hydrogels with a pendant peptide moieity, c(RRETAWA), as previous studies have shown that RRETAWA containing peptides bind to the α5β1 integrin with very high specificity. Notably, hMSC attachment to c(RRETAWA)-functionalized hydrogels was found to occur primarily through α5 integrins, as the number of attached cells was significantly reduced to ~20% upon blocking the α5 integrin during culture. To investigate the interplay between stiffness and c(RRETAWA) concentration, hydrogels were formulated with Young's moduli of ~2 kPa (soft) and ~25 kPa (stiff) and c(RRETAWA) concentrations of 0.1 mM and 1 mM. Stiff substrates led to ~3.5 fold higher hMSC attachment and ~3 fold higher cell area in comparison to soft substrates. hMSCs formed robust and larger focal adhesions on stiff substrates at 1 mM c(RRETAWA) compared to soft substrates. Alkaline phosphatase (ALP) activity in hMSCs cultured on stiff gels at 0.1 mM and 1 mM c(RRETAWA) was increased 2.5 and 3.5 fold, respectively after 14 days in growth media. hMSCs did not show an increase in ALP activity when cultured on soft gels. Further, gene expression of osteogenic related genes, core binding factor-1, osteopontin and Collagen-1a at day 14 in hMSCs cultured on stiff gels at 1 mM c(RRETAWA) were increased 10, 7 and 4 fold, respectively, while on soft gels, gene expression was at basal levels. Collectively, these results demonstrate that the combination of high substrate stiffness and α5β1 integrin signaling stimulated by c(RRETAWA) is sufficient to induce osteogenic differentiation of hMSCs without requiring the addition of soluble factors.


Exposure of human lung cancer cells to 8-chloro-adenosine induces G2/M arrest and mitotic catastrophe.
8-Chloro-adenosine (8-Cl-Ado) is a potent chemotherapeutic agent whose cytotoxicity in a variety of tumor cell lines has been widely investigated. However, the molecular mechanisms are uncertain. In this study, we found that exposure of human lung cancer cell lines A549 (p53-wt) and H1299 (p53-depleted) to 8-Cl-Ado induced cell arrest in the G2/M phase, which was accompanied by accumulation of binucleated and polymorphonucleated cells resulting from aberrant mitosis and failed cytokinesis. Western blotting showed the loss of phosphorylated forms of Cdc2 and Cdc25C that allowed progression into mitosis. Furthermore, the increase in Ser10-phosphorylated histone H3-positive cells revealed by fluorescence-activated cell sorting suggested that the agent-targeted cells were able to exit the G2 phase and enter the M phase. Immunocytochemistry showed that microtubule and microfilament arrays were changed in exposed cells, indicating that the dynamic instability of microtubules and microfilaments was lost, which may correlate with mitotic dividing failure. Aberrant mitosis resulted in mitotic catastrophe followed by varying degrees of apoptosis, depending on the cell lines. Thus, 8-Cl-Ado appears to exert its cytotoxicity toward cells in culture by inducing mitotic catastrophe.


HMM-ModE--improved classification using profile hidden Markov models by optimising the discrimination threshold and modifying emission probabilities with negative training sequences.
Profile Hidden Markov Models (HMM) are statistical representations of protein families derived from patterns of sequence conservation in multiple alignments and have been used in identifying remote homologues with considerable success. These conservation patterns arise from fold specific signals, shared across multiple families, and function specific signals unique to the families. The availability of sequences pre-classified according to their function permits the use of negative training sequences to improve the specificity of the HMM, both by optimizing the threshold cutoff and by modifying emission probabilities to minimize the influence of fold-specific signals. A protocol to generate family specific HMMs is described that first constructs a profile HMM from an alignment of the family's sequences and then uses this model to identify sequences belonging to other classes that score above the default threshold (false positives). Ten-fold cross validation is used to optimise the discrimination threshold score for the model. The advent of fast multiple alignment methods enables the use of the profile alignments to align the true and false positive sequences, and the resulting alignments are used to modify the emission probabilities in the original model. The protocol, called HMM-ModE, was validated on a set of sequences belonging to six sub-families of the AGC family of kinases. These sequences have an average sequence similarity of 63% among the group though each sub-group has a different substrate specificity. The optimisation of discrimination threshold, by using negative sequences scored against the model improves specificity in test cases from an average of 21% to 98%. Further discrimination by the HMM after modifying model probabilities using negative training sequences is provided in a few cases, the average specificity rising to 99%. Similar improvements were obtained with a sample of G-Protein coupled receptors sub-classified with respect to their substrate specificity, though the average sequence identity across the sub-families is just 20.6%. The protocol is applied in a high-throughput classification exercise on protein kinases. The protocol has the potential to maximise the contributions of discriminating residues to classify proteins based on their molecular function, using pre-classified positive and negative sequence training data. The high specificity of the method, and increasing availability of pre-classified sequence data holds the potential for its application in sequence annotation.


MicroRNA-124 inhibits proliferation and metastasis of esophageal cancer via negatively regulating NRP1.
MicroRNAs are a kind of endogenous, non-coding RNAs, which exert a significant role in pathological processes. Previous studies have reported that microRNA-124 is a tumor suppressor. The specific effect of microRNA-124 on esophageal cancer, however, has not been fully elucidated. This study aims to explore the role of microRNA-124 in esophageal cancer and its underlying mechanism. MicroRNA-124 expressions in 75 esophageal cancer tissues, paracancerous tissues, and esophageal cancer cell lines were detected by qRT-PCR (quantitative Real-Time Polymerase Chain Reaction). The relationship between microRNA-124 expression, clinical progression, pathological indicators, and prognosis of patients with esophageal cancer was analyzed. For in vitro experiments, we performed CCK-8 (cell counting kit-8), colony formation and transwell assay to detect cell proliferation, migration, and invasion abilities after microRNA-124 overexpression in TE-1 and EC-109 cells, respectively. Western blot was utilized to explore the regulatory role of microRNA-124 in esophageal cancer cells. MicroRNA-124 was downregulated in esophageal cancer tissues than that of paracancerous tissues. Patients with esophageal cancer who had lower expression level of microRNA-124 presented higher tumor stage and metastasis incidence, as well as lower survival rate. In vitro studies demonstrated a decreased cell proliferation and migration abilities after microRNA-124 overexpression. Western blot results showed upregulated PI3K and AKT, and downregulated PTEN in esophageal cancer cells after overexpression of microRNA-124. Furthermore, microRNA-124 was confirmed to negatively regulate NRP1, so as to participate in the development of esophageal cancer. MicroRNA-124 is downregulated in esophageal cancer tissues, which is remarkably correlated to the development, pathological grade, and poor prognosis of esophageal cancer. Overexpressed microRNA-124 is capable of inhibiting the malignant progression of esophageal cancer via negatively regulating NRP1.


Rectal and Bladder Temperatures vs Forehead Core Temperatures Measured With SpotOn Monitoring System.
Methods and frequency of temperature monitoring in intensive care unit patients vary widely. The recently available SpotOn system uses zero-heat-flux technology and offers a noninvasive method for continuous monitoring of core temperature of critical care patients at risk for alterations in body temperature. To evaluate agreement between and precision of a zero-heat-flux thermometry system (SpotOn) and continuous rectal and urinary bladder thermometry during fever and defervescence in adult patients in intensive care units. Prospective comparison of SpotOn vs rectal and urinary bladder thermometry in eligible patients enrolled in a randomized clinical trial on the effect of acetaminophen on core body temperature and hemodynamic status. A total of 748 paired temperature measurements from 38 patients who had both SpotOn monitoring and either continuous rectal or continuous bladder thermometry were analyzed. Temperatures during the study were from 36.6°C to 39.9°C. The mean difference for SpotOn compared with bladder thermometry was -0.07°C (SD, 0.24°C; 95% limits of agreement, ± 0.47°C [-0.54°C, 0.40°C]). The mean difference for SpotOn compared with rectal thermometry was -0.24°C (SD, 0.29°C; 95% limits of agreement, ± 0.57°C [-0.81°C, 0.33°C]). Most differences in temperature between methods were within ± 0.5°C in both groups (96% bladder and 85% rectal). The SpotOn thermometry system has excellent agreement and good precision and is a potential alternative for noninvasive continuous monitoring of core temperature in critical care patients, especially when alternative methods are contraindicated or not available.


Surface properties of spider silk particles in solution.
Recombinant spider silk proteins, such as eADF4(C16), can be used for various applications. Colloidal particles of eADF4(C16) show potential as drug delivery systems. Tuning the colloidal properties of suspensions of eADF4(C16) particles represents a major prerequisite for their use in pharmaceutical formulations. In this study we determined the surface properties concerning inter-particle interactions by means of electrophoretic mobility and direct force measurements. The surface charge of eADF4(C16) spider silk particles was determined as a function of ionic strength and pH, respectively. The resulting electrophoretic mobility can be described using the O'Brien and White theory and is directly related to the amino acid sequence of the protein. We determined the extension of a fuzzy protein layer protruding into the solution by direct force measurements using a colloidal probe technique. This soft layer leads to deviations in the electrophoretic mobility and is responsible for additional repulsive forces at small separation distances. These steric forces lead to a stabilization of the particle suspension at high ionic strength.


Effect of aging on animal response to chronic fluoride exposure.
This study was conducted to test the hypothesis that physiological changes which occur during aging increase the biological impact of fluoride and reduce the threshold of safe fluoride exposure. Four groups of rats were fed a low-fluoride diet (< 1.2 ppm) ad libitum and received 0, 5, 15, or 50 ppm fluoride in their drinking water. Animals were killed after three, six, 12, or 18 months of treatment. Blood and urine were monitored for biochemical markers of tissue function, and plasma, urine, feces, and representative tissues were analyzed for fluoride. In addition, bone marrow cells from animals killed after 18 months of treatment were examined for frequency of sister chromatid exchange (SCE), a marker of genetic damage. Study results indicated that, within treatment groups, fluoride intake, excretion, and retention did not change significantly between three and 18 months. Fluoride concentration in soft tissues did not change with treatment duration in the fluoride-treated animals. Mineralized tissue fluoride concentration and the total fluoride in the carcasses increased continually as the animals aged. In spite of significant, dose-related differences in tissue fluoride levels which occurred in all age groups in this study, there were no indications that increased fluoride in the tissues caused any adverse physiological or genotoxic effects. None of the monitored clinical "wellness" markers of tissue integrity and function was altered by fluoride in a clinically significant manner. Therefore, there was no evidence from this study that aging reduces the threshold of safe chronic fluoride exposure.


Decreased persistence to cholinesterase inhibitor therapy with concomitant use of drugs that can impair cognition.
To assess persistence with cholinesterase inhibitor therapy 6 months after the start of treatment, and to determine whether the likelihood of persistence is associated with the coprescription of drugs that can impair cognition. Retrospective cohort study. Community (home residence) or long-term care facility. A total of 1183 patients enrolled in the Rhode Island Medicaid program, aged 45 years or older, who were dispensed a cholinesterase inhibitor from January 1, 2000-June 30, 2002. Patients were considered persistent with treatment if they filled at least five prescriptions for a 1-month supply of the same cholinesterase inhibitor, without an extended gap in days between refills. We compared rates of persistence among patients receiving and those not receiving drugs that can impair cognition. Covariates assessed were patient age, sex, race, and care setting. Approximately one in four patients discontinued cholinesterase inhibitor therapy within 6 months. Patients aged 85 years or older were more persistent than younger patients (77% vs 71%, p<0.05). Caucasian patients were more likely to be persistent than non-Caucasian patients (74% vs 52%, p<0.001). Patients living in the community were less likely to persist than those residing in long-term care facilities (58% vs 76%, p<0.001). After adjusting for race and care setting, patients who were prescribed drugs that can impair cognition were more likely not to have persisted with cholinesterase inhibitor therapy at 6 months than those who did not receive such drugs (odds ratio 1.56, 95% confidence interval 1.13-2.16). A substantial percentage of patients who began receiving cholinesterase inhibitor therapy had discontinued the therapy within 6 months. Many patients also received prescriptions for agents that can impair cognition. Our findings indicated a modest but statistically significant increase in likelihood of treatment discontinuation among patients who also received prescriptions for drugs that can impair cognition. Iatrogenic causes of dementia are important to recognize and address so that therapies for enhancing cognition can be fully effective.


Differential serotonin2 receptor recovery in mature and senescent rat brain after irreversible receptor modification: effect of chronic reserpine treatment.
We report here the differential time course of recovery of [3H]ketanserin-labeled serotonin2 (5-HT2) receptors in frontal cortex of mature (4 months old) and senescent (28 months old) male Fischer 344 rats. We provide evidence that the irreversible modification of 5-HT2 serotonin receptors by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline occurs at the ligand binding site and that this treatment does not appear to affect the interaction of these receptors with their guanine nucleotide regulatory protein. Senescent rats exhibited significantly reduced (-20%) maximum receptor density levels of 5-HT2 receptors compared with their mature counterparts. The time course of recovery of [3H] ketanserin binding to 5-HT2 receptors after irreversible receptor modification by a single peripheral injection of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline was significantly slower in senescent rats. The slower recovery was a function of decreases in both the receptor production rate and the degradation rate constant in senescent rats compared with mature rats. Interestingly, chronic reserpine treatment resulted in a significant decrease in control maximum receptor density values in both mature (-17%) and senescent (-18%) rats. Although the receptor production rate and receptor degradation rate constants in both mature and senescent reserpinized rats were slightly decreased, there was no significant change in the overall rates of receptor recovery when compared with their respective age-matched nonreserpinized counterparts. These data indicate that both the steady state levels and "turnover" of 5-HT2 receptors are decreased in senescence and that these receptors can be down-regulated by chronic reserpine treatment.


Viruses: making friends with old foes.
The study of viruses has traditionally focused on their roles as infectious agents and as tools for understanding cell biology. Viruses are now finding a new expanded role as nanoplatforms with applications in materials science and medicine. Viruses form highly symmetrical monodisperse architectures and are ideal templates for engineering multifunctionality, including multivalent display of surface ligands and encapsulation of inorganic and organic materials. These developments assure that viruses will find applications as versatile nanoscale materials.


Optimal magnetic resonance imaging of the spine.
Magnetic resonance (MR) imaging is generally considered to be the best imaging modality for the spine because of its excellent tissue contrast and multiplanar imaging capability; however, good technique is vital in order to avoid nondiagnostic or even misleading images. The possibility of imaging in multiple planes and using many different pulse sequences means that judgment is required in order to gain maximum diagnostic information within a reasonable scanning time. Spinal MR imaging technique for small animals is reviewed with emphasis on technical aspects including patient positioning, selection of pulse sequences, and image planes.


Molecular dosimetry of ethylene oxide: formation and persistence of N-(2-hydroxyethyl)valine in hemoglobin following repeated exposures of rats and mice.
The formation of N-(2-hydroxyethyl)valine (HEVal) in hemoglobin was investigated in male F344 rats (10/group) and B6C3F1 mice (20/group) exposed to 0, 3, 10, 33, 100, or 300 (rats only) ppm ethylene oxide (ETO) by inhalation for 6 h/day for 4 weeks (5 days/week) or exposed to 100 (mice) or 300 ppm (rats) ETO for 1 or 3 days, or 1, 2, or 4 weeks (5 days/week). The persistence of HEVal was studied in animals killed up to 10 days after cessation of the 4-week time-course studies. HEVal was determined by a modified Edman degradation and quantitation of the resulting pentafluorophenylthiohydantoin derivative, using gas chromatography-mass spectrometry. The resulting experimental data were compared to simulations derived with a model for the formation and removal of hemoglobin adducts (T.R. Fennell, S.C.J. Sumner, and V.E. Walker, Cancer Epidemiol., Biomarkers & Prev., 1: 213-219, 1992). Repeated exposures of rats and mice for 4 weeks to 300 and 100 ppm ETO, respectively, led to an accumulation of HEVal that was 14 (rats) and 15 (mice) times greater than that found after 1 day of exposure [28 +/- 2 (SE) and 9.4 +/- 0.4 (SE) pmol HEVal/mg globin in rats and mice, respectively]. After cessation of exposures, HEVal was lost faster than predicted by the normal erythrocyte life span alone. An initial phase of rapid decline in HEVal concentrations was consistent with the removal of older, more heavily alkylated populations of RBCs, accompanied by a burst of erythropoiesis. The dose-response curves for HEVal were linear between 3 and 33 ppm ETO, with 3.5 +/- 0.2 and 3.4 +/- 0.3 pmol adduct/mg globin formed in rats and mice, respectively, after 4 weeks of exposure to 3 ppm ETO. Above 33 ppm ETO, the slope of the dose-response curves increased. Comparison of the dose response for HEVal in rats exposed to ETO for 4 weeks to the dose-response for N tau-(2-hydroxyethyl)histidine in rats exposed to the same concentrations of ETO for 2 years (S. Osterman-Golkar et al., Teratog. Carcinog. Mutagen., 3: 395-405, 1983) suggested that exposures to ETO can reduce the life span of erythrocytes in a concentration- and time-dependent manner. Correlation of the experimental data and simulations for the formation and removal of HEVal demonstrated that perturbations in erythropoiesis and RBC life span complicate the estimation of exposures to ETO when estimates are based upon hemoglobin adduct measurements in heavily exposed individuals.(ABSTRACT TRUNCATED AT 400 WORDS)