Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies.
This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins on-chip with the dialysis method and allowing in situ single-crystal or serial crystallography experiments at room temperature. The protocol details the fabrication process of the microchips, the manipulation of the on-chip crystallization experiments and the treatment of the in situ collected X-ray diffraction data for the structural elucidation of the protein sample. The main feature of this microfabrication procedure lies on the integration of a commercially available, semipermeable regenerated cellulose dialysis membrane in between two layers of the chip. The molecular weight cut-off of the embedded membrane varies depending on the molecular weight of the macromolecule and the precipitants. The device exploits the advantages of microfluidic technology, such as the use of minute volumes of samples (<1 µL) and fine tuning over transport phenomena. The chip coupled them with the dialysis method, providing precise and reversible control over the crystallization process and can be used for investigating phase diagrams of proteins at the microliter scale. The device is patterned using a photocurable thiolene-based resin with soft imprint lithography on an optically transparent polymeric substrate. Moreover, the background scattering of the materials composing the microchips and generating background noise was evaluated rendering the chip compatible for in situ X-ray diffraction experiments. Once protein crystals are grown on-chip up to an adequate size and population uniformity, the microchips can be directly mounted in front of the X-ray beam with the aid of a 3D printed holder. This approach addresses the challenges rising from the use of cryoprotectants and manual harvesting in conventional protein crystallography experiments through an easy and inexpensive manner. Complete X-ray diffraction data sets from multiple, isomorphous lysozyme crystals grown on-chip were collected at room temperature for structure determination.


Comparison of bilevel positive airway pressure and average volume-assured pressure support mode in terms of patient compliance and treatment success in hypercapnic patients. A cross-sectional study.
It is necessary for an effective NIV application to provide proper modality selection, sufficient minute ventilation (MV), also the amount of leakage on the circuit must be minimized and patient-ventilator adaptation must be achieved. 30 patients with acute respiratory failure as a result of either internal or postoperative reasons were included in the study. Patient comfort was analyzed with a scale ranging from 0 to 2. Firstly the patient was used for two hours in BIPAP modality, after then the AVAPS modality (Period Av) was applied by setting the required rates the same mask. During BIPAP and AVAPS, arterial blood gases analysis, comfort scale and hemodynamic parameters were recorded in the 30th minute, 1st hour and 2nd hour. According to the assessment of arterial blood gases, the pH changes of both periods were statistically significant compared to their baseline values (p=0.001). Treatment compliance of the patients was significantly better at AVAPS modality at all times (p = 0.015, p = 0.008, p = 0.008, respectively). According to the results obtained from this study, the AVAPS modality has positive effects on pH and gas variation and patient comfort; therefore, it can be confidently used in clinical practice. Average Volume Assured Pressure Support, Bilevel Continuous Positive Airway Pressure, Intensive Care Units, Noninvasive Ventilation, Patients Compliance.


Loss of Mitochondrial Pyruvate Carrier 2 in the Liver Leads to Defects in Gluconeogenesis and Compensation via Pyruvate-Alanine Cycling.
Pyruvate transport across the inner mitochondrial membrane is believed to be a prerequisite for gluconeogenesis in hepatocytes, which is important for the maintenance of normoglycemia during prolonged food deprivation but also contributes to hyperglycemia in diabetes. To determine the requirement for mitochondrial pyruvate import in gluconeogenesis, mice with liver-specific deletion of mitochondrial pyruvate carrier 2 (LS-Mpc2(-/-)) were generated. Loss of MPC2 impaired, but did not completely abolish, hepatocyte conversion of labeled pyruvate to TCA cycle intermediates and glucose. Unbiased metabolomic analyses of livers from fasted LS-Mpc2(-/-) mice suggested that alterations in amino acid metabolism, including pyruvate-alanine cycling, might compensate for the loss of MPC2. Indeed, inhibition of pyruvate-alanine transamination further reduced mitochondrial pyruvate metabolism and glucose production by LS-Mpc2(-/-) hepatocytes. These data demonstrate an important role for MPC2 in controlling hepatic gluconeogenesis and illuminate a compensatory mechanism for circumventing a block in mitochondrial pyruvate import.


Different Positron Emission Tomography Tau Tracers Bind to Multiple Binding Sites on the Tau Fibril: Insight from Computational Modeling.
Using the recently reported cryo-EM structure for the tau fibril [ Fitzpatrick et al. (2017) Nature 547, 185-190 ], which is a potential target concerning Alzheimer's disease, we present the first molecular modeling studies on its interaction with various positron emission tomography (PET) tracers. Experimentally, based on the binding assay studies, at least three different high-affinity binding sites have been reported for tracers in the tau fibril. Herein, through integrated modeling using molecular docking, molecular dynamics, and binding free energy calculations, we provide insight into the binding patterns of various tracers to the tau fibril. We suggest that there are four different high-affinity binding sites available for many of the studied tracers showing varying binding affinity to different binding sites. Thus, PBB3 binds most strongly to site 4, and interestingly, this site is not a preferable site for any other tracers. For THK5351, our data show that it strongly binds to sites 3 and 1, the former one being more preferable. We also find that MK6240 and T807 bind to site 1 specifically. The modeling data also give some insight into whether a tracer bound to a specific site can be replaced by others or not. For example, the displacement of T807 by PBB3 as reported experimentally can also be explained and attributed to the larger binding affinity of the latter compound in all binding sites. The binding free energy results explain very well the small binding affinity of THK523 compared to all the aryl quinoline moieties containing THK tracers. The ability of certain tau tracers, like FDDNP and THK523, to bind to amyloid fibrils has also been investigated. Furthermore, such off-target interaction of tau tracers with amyloid beta fibrils has been validated using a quantum mechanical fragmentation approach.


The 'methylamine oxidase' system of an obligate methylotroph.
The terminal respiratory oxidase was solubilized from membranes of organism 4025, an obligate methylotroph. The partially purified oxidase is probably a cytochrome co. It does not oxidize amicyanin, but it oxidizes 'azurin' and cytochromes cH and cL. By using a complete 'methylamine oxidase' system reconstituted from pure methylamine dehydrogenase, purified oxidase and soluble blue copper proteins and cytochromes, it was confirmed that amicyanin is essential for methylamine oxidation; it could not be replaced by 'azurin' or cytochrome cH or cL. It was shown that the usual mediator between amicyanin and the oxidase is cytochrome cH, with 'azurin' able to replace it during growth at the high copper concentrations required for optimum growth of this unusual methylotroph.


Differences in body composition between infants of South Asian and European ancestry: the London Mother and Baby Study.
South Asian children and adults have a more adipose body composition compared with those of European ancestry. This is thought to be related to their increased risk of metabolic disorders. However, little is known about how early in life such differences are manifest. To determine whether there are differences in fat mass (FM) and fat-free mass (FFM) between UK-born South Asians and White Europeans in infancy. Design A cross-sectional study of 30 South Asian and 30 White European infants aged 6-12 weeks. Mothers were recruited from clinics in London, and infants' FM and FFM were determined using air-displacement plethysmography (PeaPod(®)). In early infancy South Asians had less FFM than White Europeans [0.34 kg less, 95% confidence interval (CI): 0.15, 0.52], with a considerably weaker indication of them also having more FM (0.02 kg more, 95% CI: -0.14, 0.18). These differences persisted when the overall smaller body size of South Asian infants was taken into account. For a given total infant weight, the balance of body composition of South Asians was shifted by 0.16 kg (95% CI: 0.06, 0.25) from FFM to FM. The ethnic differences in the amount of FFM were almost completely accounted for by ethnic differences in the rate of growth in utero and length of gestation. The characteristic differences in body composition observed between adult South Asians and White Europeans are apparent in early infancy. Of particular note is that this is the first study to demonstrate that South Asians compared with White Europeans have reduced FFM in infancy. The early manifestation of this phenotype suggests that it is either genetic and/or determined through exposure to maternal physiology, rather than a consequence of behaviours or diet in childhood or at older ages.


Travel medicine and mHealth technology: a study using smartphones to collect health data during travel.
mHealth methodology such as smartphone applications offers new opportunities to capture the full range of health risks during travel in real time. Our study aims to widen the scope of travel health research in tropical and subtropical destinations by using a smartphone application to collect detailed information on health behaviours, clinical symptoms, accidents and environmental factors during travel. We enrolled travel clinic clients in Zurich and Basel ≥18 years of age travelling to Thailand for <5 weeks. Sociodemographic, clinical and risk behaviour information was collected pre-travel. Participants were equipped with a smartphone and an application that (1) actively administers a daily self-report questionnaire on the health risks, behaviours and symptoms the traveller encountered, and (2) passively collects information on the traveller's location and environmental conditions by transformation of raw GPS data. A prospective cohort of 101 travellers planning travel to Thailand between January and June 2015 was recruited. Of the 101 enrolled travellers, 75 (74.3%) answered at least one questionnaire during travel, 10 (9.9%) had technical difficulties and 16 (15.8%) dropped out. Those who completed questionnaires were a median of 27.0 years old (range 18-57). Travellers filled out a median of 12.0 questionnaires during their trip (range 1-30), corresponding to a median completion rate of 85.0% days of travel. The typical example of a healthy female traveller shows that many and diverse health issues arise during a trip that clusters on certain days. The rich data on behaviour and local environment may be used to explain the occurrence and clustering of health issues. Use of a smartphone app to collect health information is technically feasible and acceptable amongst a traveller population, minimizes recall bias and greatly increases the quality and quantity of data collected during travel. mHealth technology shows great potential for innovation in travel medicine.


GTXOP: a game theoretic approach for QoS provisioning using transmission opportunity tuning.
In unsupervised contention-based networks such as EDCA mode of IEEE 802.11(e)(s), upon winning the channel, each node gets a transmission opportunity (TXOP) in which the node can transmit multiple frames consequently without releasing the channel. Adjusting TXOP can lead to better bandwidth utilization and QoS provisioning. To improve WLAN throughput performance, EDCA packet bursting can be used in 802.11e, meaning that once a station has gained an EDCA-TXOP, it can be allowed to transmit more than one frame without re-contending for the channel. Following the access to the channel, the station can send multiple frames as long as the total access time does not exceed the TXOP Limit. This mechanism can reduce the network overhead and increase the channel utilization instead. However, packet bursting may cause unfairness in addition to increasing jitter, delay and loss. To the best of the authors' knowledge, although TXOP tuning has been investigated through different methods, it has not been considered within a game theory framework. In this study, based on the analytical models of EDCA, a game theoretic approach called GTXOP is proposed to determine TXOP dynamically (i.e. according to the dynamisms of WLAN networks and the number of nodes in the network). Using GTXOP, each node can choose its TXOP autonomously, such that in addition to QoS improvement, the overall network performance is also improved.


Huntingtin mediates dendritic transport of β-actin mRNA in rat neurons.
Transport of mRNAs to diverse neuronal locations via RNA granules serves an important function in regulating protein synthesis within restricted sub-cellular domains. We recently detected the Huntington's disease protein huntingtin (Htt) in dendritic RNA granules; however, the functional significance of this localization is not known. Here we report that Htt and the huntingtin-associated protein 1 (HAP1) are co-localized with the microtubule motor proteins, the KIF5A kinesin and dynein, during dendritic transport of β-actin mRNA. Live cell imaging demonstrated that β-actin mRNA is associated with Htt, HAP1, and dynein intermediate chain in cultured neurons. Reduction in the levels of Htt, HAP1, KIF5A, and dynein heavy chain by lentiviral-based shRNAs resulted in a reduction in the transport of β-actin mRNA. These findings support a role for Htt in participating in the mRNA transport machinery that also contains HAP1, KIF5A, and dynein.


Mechanisms of MAPK activation by bradykinin in vascular smooth muscle cells.
Vascular smooth muscle cell (VSMC) proliferation is a prominent feature of the atherosclerotic process occurring after endothelial injury. A vascular wall kallikrein-kinin system has been described. The contribution of this system to vascular disease is undefined. In the present study we characterized the signal transduction pathway leading to mitogen-activated protein kinase (MAPK) activation in response to bradykinin (BK) in VSMC. Addition of 10(-10)-10(-7) M BK to VSMC resulted in a rapid and concentration-dependent increase in tyrosine phosphorylation of several 144- to 40-kDa proteins. This effect of BK was abolished by the B(2)-kinin receptor antagonist HOE-140, but not by the B(1)-kinin receptor antagonist des-Arg(9)-Leu(8)-BK. Immunoprecipitation with anti-phosphotyrosine antibodies followed by immunoblot revealed that 10(-9) M BK induced tyrosine phosphorylation of focal adhesion kinase (p125(FAK)). BK (10(-8) M) promoted the association of p60(src) with the adapter protein growth factor receptor binding protein-2 and also induced a significant increase in MAPK activity. Pertussis and cholera toxins did not inhibit BK-induced MAPK tyrosine phosphorylation. Protein kinase C downregulation by phorbol 12-myristate 13-acetate and/or inhibitors to protein kinase C, p60(src) kinase, and MAPK kinase inhibited BK-induced MAPK tyrosine phosphorylation. These findings provide evidence that activation of the B(2)-kinin receptor in VSMC leads to generation of multiple second messengers that converge to activate MAPK. The activation of this crucial kinase by BK provides a strong rationale to investigate the mitogenic actions of BK on VSMC proliferation in disease states of vascular injury.


I Can See It in Your Eyes: Gaze as an Implicit Cue of Uncanniness and Task Performance in Repeated Interactions With Robots.
Over the past years, extensive research has been dedicated to developing robust platforms and data-driven dialog models to support long-term human-robot interactions. However, little is known about how people's perception of robots and engagement with them develop over time and how these can be accurately assessed through implicit and continuous measurement techniques. In this paper, we explore this by involving participants in three interaction sessions with multiple days of zero exposure in between. Each session consists of a joint task with a robot as well as two short social chats with it before and after the task. We measure participants' gaze patterns with a wearable eye-tracker and gauge their perception of the robot and engagement with it and the joint task using questionnaires. Results disclose that aversion of gaze in a social chat is an indicator of a robot's uncanniness and that the more people gaze at the robot in a joint task, the worse they perform. In contrast with most HRI literature, our results show that gaze toward an object of shared attention, rather than gaze toward a robotic partner, is the most meaningful predictor of engagement in a joint task. Furthermore, the analyses of gaze patterns in repeated interactions disclose that people's mutual gaze in a social chat develops congruently with their perceptions of the robot over time. These are key findings for the HRI community as they entail that gaze behavior can be used as an implicit measure of people's perception of robots in a social chat and of their engagement and task performance in a joint task.


The plant ER: a dynamic organelle composed of a large number of discrete functional domains.
The endoplasmic reticulum (ER) of plants is comprised of a three-dimensional network of continuous tubules and sheets that underlies the plasma membrane, courses through the cytoplasm, and links up with the nuclear envelope. Aside from discussing the dynamic properties of this versatile and adaptable organelle, the review highlights the structure and the functional properties of 16 types of morphologically defined ER membrane domains. Owing to their labile or transient nature, several of these domains can only be visualized reliably through the use of ultrarapid freezing techniques. The ER domains discussed are: the lamin receptor domain; the nuclear pores; the nuclear envelope-ER gates, the microtubule nucleation domains; the protein and oil body-forming domains; the vacuole-forming ER; the actin-binding, the plasma membrane-anchoring and the vacuole and mitochondrion-attachment domains; the lipid recycling ER cisternae and the plasmodesmata. Preliminary evidence suggests that this list will have to be expanded in the near future. Understanding the assembly, the functional roles, and the developmental regulation of these domains has implications both for understanding cell structure and function, and for exploiting plants for agricultural and biotechnological purposes.


13C tracer experiments and metabolite balancing for metabolic flux analysis: comparing two approaches
Conventional metabolic flux analysis uses the information gained from determination of measurable fluxes and a steady-state assumption for intracellular metabolites to calculate the metabolic fluxes in a given metabolic network. The determination of intracellular fluxes depends heavily on the correctness of the assumed stoichiometry including the presence of all reactions with a noticeable impact on the model metabolite balances. Determination of fluxes in complex metabolic networks often requires the inclusion of NADH and NADPH balances, which are subject to controversial debate. Transhydrogenation reactions that transfer reduction equivalents from NADH to NADPH or vice versa can usually not be included in the stoichiometric model, because they result in singularities in the stoichiometric matrix. However, it is the NADPH balance that, to a large extent, determines the calculated flux through the pentose phosphate pathway. Hence, wrong assumptions on the presence or activity of transhydrogenation reactions will result in wrong estimations of the intracellular flux distribution. Using 13C tracer experiments and NMR analysis, flux analysis can be performed on the basis of only well established stoichiometric equations and measurements of the labeling state of intracellular metabolites. Neither NADH/NADPH balancing nor assumptions on energy yields need to be included to determine the intracellular fluxes. Because metabolite balancing methods and the use of 13C labeling measurements are two different approaches to the determination of intracellular fluxes, both methods can be used to verify each other or to discuss the origin and significance of deviations in the results. Flux analysis based entirely on metabolite balancing and flux analysis, including labeling information, have been performed independently for a wild-type strain of Aspergillus oryzae producing alpha-amylase. Two different nitrogen sources, NH4+ and NO3-, have been used to investigate the influence of the NADPH requirements on the intracellular flux distribution. The two different approaches to the calculation of fluxes are compared and deviations in the results are discussed. Copyright 1998 John Wiley & Sons, Inc.


Preventable deaths in childhood, according to actions of the Unified Health System, Brazil.
To analyze the mortality trend of children under five years of age living in Brazil and regions, using the "Brazilian List of Preventable Causes of Death." Ecological time-series study of mortality rate due to preventable and non-preventable causes, with corrections for ill-defined causes and underreporting of deaths from 2000 to 2013. In Brazil, preventable death rates (5.1% per year) had a higher decrease compared with non-preventable ones (2.5% per year). Preventable causes associated with proper care during pregnancy had the highest concentration of deaths in 2013 (12,267) and the second lowest average percentage reduction in the year (2.1%) and for the period (24.4%). The South and Southeast regions had the lowest mortality rates in childhood. However, the Northeast region had the highest decrease in reducible child mortality (6.1% per year) and the Midwest, the lowest (3.5% per year). The decrease in childhood mortality rates was expected in the last decade, suggesting the progress in the response of health systems, in addition to improvements in health conditions and social determinants. Special attention should be given to pregnancy-related causes, i.e., expand the quality of prenatal care, in particular, due to fetal and newborn deaths resulted from maternal conditions, which increased significantly in the period (8,3% per year).


Imaging markers of disability in aquaporin-4 immunoglobulin G seropositive neuromyelitis optica: a graph theory study.
Neuromyelitis optica spectrum disorders lack imaging biomarkers associated with disease course and supporting prognosis. This complex and heterogeneous set of disorders affects many regions of the central nervous system, including the spinal cord and visual pathway. Here, we use graph theory-based multimodal network analysis to investigate hypothesis-free mixed networks and associations between clinical disease with neuroimaging markers in 40 aquaporin-4-immunoglobulin G antibody seropositive patients (age = 48.16 ± 14.3 years, female:male = 36:4) and 31 healthy controls (age = 45.92 ± 13.3 years, female:male = 24:7). Magnetic resonance imaging measures included total brain and deep grey matter volumes, cortical thickness and spinal cord atrophy. Optical coherence tomography measures of the retina and clinical measures comprised of clinical attack types and expanded disability status scale were also utilized. For multimodal network analysis, all measures were introduced as nodes and tested for directed connectivity from clinical attack types and disease duration to systematic imaging and clinical disability measures. Analysis of variance, with group interactions, gave weights and significance for each nodal association (hyperedges). Connectivity matrices from 80% and 95% F-distribution networks were analyzed and revealed the number of combined attack types and disease duration as the most connected nodes, directly affecting changes in several regions of the central nervous system. Subsequent multivariable regression models, including interaction effects with clinical parameters, identified associations between decreased nucleus accumbens (β = -0.85, P = 0.021) and caudate nucleus (β = -0.61, P = 0.011) volumes with higher combined attack type count and longer disease duration, respectively. We also confirmed previously reported associations between spinal cord atrophy with increased number of clinical myelitis attacks. Age was the most important factor associated with normalized brain volume, pallidum volume, cortical thickness and the expanded disability status scale score. The identified imaging biomarker candidates warrant further investigation in larger-scale studies. Graph theory-based multimodal networks allow for connectivity and interaction analysis, where this method may be applied in other complex heterogeneous disease investigations with different outcome measures.


Factors Associated With Intentions to Engage in Vaginal Intercourse Among Sexually Abstinent Missouri High School Freshmen.
We examine personal characteristics, alcohol consumption, normative beliefs, household factors, and extracurricular engagement associated with intentions to have intercourse before marriage among abstinent students. Data were analyzed from 245 freshmen enrolled in a school-based abstinence-only-until-marriage program. Two binary logistic regression analyses identified factors associated with intentions to engage in intercourse before marriage and within the next year. Approximately 21% and 14% of participants reported intentions to have intercourse. Respondents participated in 2.2 (standard deviation [SD] = 1.2) extracurricular activities. Freshmen who were male, perceived their friends to approve of premarital sex, and consumed alcohol were more likely to report intentions to have intercourse. For every additional extracurricular activity in which freshmen participated, they were less likely to report intentions to have intercourse within the next year (odds ratio [OR] = 0.56). Factors such as extracurricular activities provide youth with opportunities to build supportive relationships, connect with peers and role models, and positively engage in schools and communities. Extracurricular activities typically already exist, have funding, and are generally well-supported. School-based strategies can increase teenagers' autonomy by providing a variety of activities to participate in and reduce unsupervised time. This, in turn, has the potential to decrease sexual risk taking behaviors.


Nothing Iffy about HIF in the Hypothalamus.
Two crucial biological processes are (1) the sensing and coordination of responses to low oxygen levels and (2) the control of food intake and energy expenditure. The hypoxia-inducible factor (HIF) family of proteins is known to regulate responses to low oxygen, whereas neuropeptides derived from proopiomelanocortin (POMC) are implicated in the control of food intake and energy expenditure. It is now becoming apparent that these two apparently disparate processes may be linked, with the exciting discovery that HIF proteins can act in the brain to regulate food intake and energy expenditure as reported in the current issue of PLoS Biology. This primer discusses the traditional role of HIF proteins in terms of responding to oxygen levels in the periphery and also their new role in coordinating responses to nutrients in the brain through regulation of POMC.


AAN position statement: The COVID-19 pandemic and the ethical duties of the neurologist.
Patients, clinicians, and hospitals have undergone monumental changes during the coronavirus disease 2019 (COVID-19) pandemic. This crisis has forced us to consider the obligations that we neurologists have to our individual patients as well as the greater community. By returning to our fundamental understanding of these duties, we can ensure that we are providing the most ethically appropriate contingency and crisis care possible. We recommend specific adaptations to both the inpatient and outpatient settings, as well as changes to medical and trainee education. Furthermore, we explore the daunting but potentially necessary implementation of scare resource allocation protocols. As the pandemic evolves, we will need to adapt continuously to these rapidly changing circumstances and consider both national and regional standards and variation.


Inactivating the beta 2-microglobulin locus in mouse embryonic stem cells by homologous recombination.
We have inactivated, by gene targeting, the endogenous beta 2-microglobulin gene in a mouse embryonic stem cell line. A cloned fragment of the beta 2-microglobulin gene with the coding sequence disrupted by the insertion of the neomycin-resistance gene was used to transfect the embryonic stem cells. G418-resistant colonies were selected and then screened using the polymerase chain reaction to identify those in which the incoming DNA had integrated into the embryonic stem cell genome by homologous recombination. Of a total of 234 G418-resistant colonies screened, 2 correctly targeted colonies were identified. Chimeric mice carrying the inactivated beta 2-microglobulin gene have been obtained from both of these targeted embryonic cell lines. Breeding of offspring from such animals will allow investigation of the effects of homozygous loss of beta 2-microglobulin.


Developmental Trajectories of Subjective Social Status.
Subjective social status (SSS), a person's sense of their (or for youth, abstract their family's) position in the socioeconomic hierarchy, is strongly related to health in adults but not health in adolescence. Understanding this developmental discrepancy requires first understanding the developmental trajectory of SSS. The objective of this study was to identify the number and shape of SSS trajectories as adolescents transition to adulthood and explore if trajectory membership affects health. Using data from 7436 assessments from the Princeton School District Study, a decade long cohort study of non-Hispanic black and white youth, latent class growth models with 3 to 7 SSS trajectories were developed. Model fit, trajectory structure, and shape were used to guide optimal model selection. Using this optimal model, the associations of trajectory membership with BMI and depressive symptoms in young adulthood were explored. The 5-class model was optimal. In this model, trajectories were persistent high (7.8%),mid–high (32.2%), middle (43.4%), low–lower (7.4%), and high–low (9.1%). Non-Hispanic black race/ethnicity, lower household income, and low parent education were associated with membership in this high–low trajectory. High–low trajectory membership was associated with higher BMI and depressive symptoms in non-Hispanic white subjects but was not associated with depressive symptoms. It was associated with lower BMI only after adjustment for BMI in adolescence in non-Hispanic black subjects. SSS is relatively stable in adolescence and the transition to adulthood, and it generally reflects objective markers of social advantage. However, socially disadvantaged youth with high SSS in early adolescence may be at increased health risk.


Recruitment, attrition and intervention completion in clinical trials of psychosocial interventions involving people with early and emerging psychosis: a systematic review protocol.
Psychosocial interventions for people experiencing early and emerging psychosis have demonstrated efficacy in reducing symptom severity and supporting recovery; however, much remains unknown about optimising treatment and future research trials are required. Gaining a better understanding of feasibility in trials of psychosocial interventions involving this population would inform the design and planning of future research and support the development of high-quality evidence. The aim of this systematic review is to evaluate the recruitment rate, study attrition rates and intervention completion of psychosocial intervention randomised controlled trial studies involving people with early and emerging psychosis. The systematic review will be reported in adherence with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis Protocols 2015 guideline. The Cochrane Library, PubMed, Medline, PsycINFO, Web of Science and CINAHL databases will be searched from inception to September 2021 to identify potentially relevant studies. The title and abstracts of returned records will be assessed for eligibility against the inclusion/exclusion criteria by two reviewers, independently, and records which appear eligible will be included. The full texts of included records will then be assessed using the same procedure. Qualitative and quantitative synthesis will be undertaken. Proportion meta-analyses will be used to calculate the recruitment rate, study attrition rate and intervention completion rate, while subgroup analyses will explore differences among subgroups of study and intervention characteristics. This study will collate and analyse anonymised data from published research and therefore, ethical approval is not necessary. Study results will be disseminated via publication in academic journals.


A single-dose placebo- and moxifloxacin-controlled study of the effects of temsirolimus on cardiac repolarization in healthy adults.
Temsirolimus, a selective inhibitor of mammalian target of rapamycin, is an approved treatment for patients with advanced renal cell carcinoma (RCC). This study assessed the effect of intravenous (i.v.) temsirolimus 25 mg, the recommended dose for patients with RCC, on the corrected QT (QTc) interval. This 3-period crossover study enrolled healthy subjects. In periods 1 and 2, subjects received i.v. placebo either alone or with open-label oral moxifloxacin. In period 3, subjects received a single dose of temsirolimus 25 mg. The primary statistical objective was to estimate the effect of temsirolimus compared with placebo on change from time-matched baseline QTc at the end of infusion (0.5 h). Assay sensitivity was evaluated by the effect of moxifloxacin on change from time-matched baseline QTc compared with placebo. In total, 58 subjects were enrolled. Temsirolimus had no effect on QTc interval in the primary analysis. At 11 of 12 secondary time points, the upper bound for the temsirolimus QTc 90% confidence intervals for the time-matched change from baseline difference from placebo was <10 ms, with no evidence of QTc trends or relationship to concentrations of temsirolimus or its major metabolite, sirolimus. Moxifloxacin, the positive control, produced a significant increase in the QTc interval compared with placebo 0.5-4 h post-dose (P < 0.0001). No subject had a QTc interval exceeding 450 ms or a change from baseline of >30 ms. Therapeutic exposure to temsirolimus is not associated with clinically significant changes in QTc intervals in healthy adults.


Aggregated platelets enhance adherence of Candida yeasts to endothelium.
The adherence of Candida albicans yeasts to human umbilical vein endothelium to subendothelial extracellular matrix (ECM) was investigated. Yeasts added to confluent endothelium in citrated platelet-poor plasma adhered on the average of 1 colony forming unit (cfu) per culture well. When platelets were added as platelet-rich plasma, a significant increase of yeast adherence was not seen. However, when endothelium was contracted by treatment with 2 mM EDTA, resulting in exposure of ECM, yeast adherence was increased to 10 cfu/well. When platelets were added with these yeasts, the number of adhering yeasts was further increased to 23 cfu/well (P less than .01). This represented an increase in adherence of yeasts of 230%. When the endothelial cells were completely removed and ECM exposed, platelets were found to likewise augment yeast adherence. Platelets, when added to the ECM, formed aggregates to which the yeasts firmly adhered. Likewise, when platelets were aggregated by adenosine diphosphate and mixed with yeasts, yeasts were shown to bind avidly to aggregated platelets, whereas yeasts did not adhere to unactivated, discoid platelets. Thus, exposed subendothelial ECM induces the aggregation of platelets and yeasts bind avidly to these platelet aggregates.


Smart technology vs. face-to-face physical activity interventions in older adults: a systematic review protocol.
The objective of this review is to determine the effect of physical activity interventions delivered via smart technology compared with face-to-face interventions for improving physical activity and physical function in older adults. Physical activity is a modifiable risk factor for multiple noncommunicable diseases and reduces the risk of premature mortality. Despite this, one in four adults does not meet recommended levels of physical activity. This pattern of inactivity increases with age. Smart technology, such as wearables, tablets, or laptops, is one solution for improving physical activity. Research has shown that different smart technology solutions can increase physical activity in older adults. While individual studies support smart technology to increase physical activity, there are no systematic reviews comparing the effects of smart technology with traditional face-to-face physical activity interventions. We will include randomized controlled trials of physical activity interventions delivered via smart technology (eg, wearables, tablets, computers) compared with face-to-face (ie, in person) interventions for community-dwelling older adults aged 60 years or older. We will search four databases (AMED, CINAHL, Embase, MEDLINE) from inception for relevant studies. All abstracts and full texts will be screened independently and in duplicate. Risk of bias, data extraction, and quality assessment will be completed in the same manner. If possible, a meta-analysis will be performed of the primary outcomes of physical activity, physical function, and adherence rate. Subgroup analyses will be conducted by type of physical activity, and type of smart technology, where possible. PROSPERO CRD42020135232.


Synthesis gas: a raw material for industrial chemicals.
Rapid increases in the price of imported crude oil have accelerated a shift in the raw material base for chemical feedstocks from natural gas to crude oil to coal. Widespread use of ethylene as a feedstock has depended on the availability at attractive prices of natural gas and petroleum. As the resource base shifts from natural gas and petroleum to coal, ethylene is being replaced by synthesis gas (a mixture of hydrogen and carbon monoxide of varying composition), which can be manufactured directly from any of these carbonaceous sources. This trend is expected to accelerate in the 1980's. Organics likely to be produced from synthesis gas include ethanol, ethylene glycol, and vinyl acetate.


Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus.
Bassoon is a core scaffold protein of the presynaptic active zone. In brain synapses, the C-terminus of Bassoon is oriented toward the plasma membrane and its N-terminus is oriented toward synaptic vesicles. At the Golgi-apparatus, Bassoon is thought to assemble active zone precursor structures, but whether it is arranged in an orderly fashion is unknown. Understanding the topology of this large scaffold protein is important for models of active zone biogenesis. Using stimulated emission depletion nanoscopy in cultured hippocampal neurons, we found that an N-terminal intramolecular tag of recombinant Bassoon, but not C-terminal tag, colocalized with markers of the trans-Golgi network (TGN). The N-terminus of Bassoon was located between 48 and 69 nm away from TGN38, while its C-terminus was located between 100 and 115 nm away from TGN38. Sequences within the first 95 amino acids of Bassoon were required for this arrangement. Our results indicate that, at the Golgi-apparatus, Bassoon is oriented with its N-terminus toward and its C-terminus away from the trans Golgi network membrane. Moreover, they suggest that Bassoon is an extended molecule at the trans Golgi network with the distance between amino acids 97 and 3,938, estimated to be between 46 and 52 nm. Our data are consistent with a model, in which the N-terminus of Bassoon binds to the membranes of the trans-Golgi network, while the C-terminus associates with active zone components, thus reflecting the topographic arrangement characteristic of synapses also at the Golgi-apparatus.


Pseudomonas aeruginosa infections: factors relating to mortality with emphasis on resistance pattern and antimicrobial treatment.
A retrospective case-control study was conducted to investigate the risk factors for death among intensive care unit patients with Pseudomonas aeruginosa infection. Out of 131 patients investigated, 67 (51.1%) died within 30 days of being diagnosed with this infection. The mean duration of hospital stay before this diagnosis was 28.5 +/- 26.5 days. No association was found between bacterial resistance and death in this study (multiresistant p= 0.26; panresistant p= 0.42), but the adequacy of the initial treatment was inversely proportional to the degree of resistance. There was a tendency towards greater mortality among patients who received combination therapy (empirical p= 0.09; definitive p= 0.08), despite the greater frequency of appropriate treatment in these patients and the similar degree of severity in the two groups. This finding may be explained by pharmacodynamic parameters that were not studied or by the extensive use of aminoglycosides in the combination therapy, which play a controversial role in combination therapy due to their potential for renal toxicity. The multivariate analysis in our study demonstrated that age [odds ratio (OR) 1.04], septic shock (OR 15.4) and hypoalbuminemia (OR 0.32) were independent risk factors for death.


Overexpression of Bcl-2 protects from ultraviolet B-induced apoptosis but promotes hair follicle regression and chemotherapy-induced alopecia.
Hair follicle (HF) growth and regression is an exquisitely regulated process of cell proliferation followed by massive cell death and is accompanied by cyclical expression of the apoptosis regulatory gene pair, Bcl-2 and Bax. To further investigate the role of Bcl-2 expression in the control of hair growth and keratinocyte apoptosis, we have used transgenic mice that overexpress human Bcl-2 in basal epidermis and in the outer root sheath under the control of the human keratin-14 promoter (K14/Bcl-2). When irradiated with ultraviolet B (UVB) light, K14/Bcl-2 mice developed about 5-10-fold fewer sunburn cells (ie, apoptotic keratinocytes) in the basal layer of the epidermis, compared to wild-type mice, whereas cultures of primary keratinocytes from transgenic mice were completely resistant to UVB-induced histone formation, at doses that readily induced histone release from wild-type cells. K14/Bcl-2 mice show no alteration of neonatal hair follicle morphogenesis or of the onset of the first wave of HF regression (catagen). However, compared to wild-type controls, K14/Bcl-2 mice subsequently displayed a significant acceleration of spontaneous catagen progression. During chemotherapy-induced alopecia, follicular dystrophy was promoted in K14/Bcl-2 mice. Thus, although K14-driven overexpression of Bcl-2 protected murine epidermal keratinocytes from UVB-induced apoptosis, it surprisingly promoted catagen- and chemotherapy-associated keratinocyte apoptosis.


Transcriptional control signals of a herpes simplex virus type 1 late (gamma 2) gene lie within bases -34 to +124 relative to the 5' terminus of the mRNA.
The cis-acting DNA sequences required for regulated expression of a herpes simplex virus type 1 (HSV-1) late (gamma 2) gene were studied by using viruses containing specific deletions in the 5' transcribed noncoding and upstream regions of the HSV-1 glycoprotein C (gC) gene, a model gamma 2 gene. Nine mutant viruses which had variable 5' and 3' deletions within bases -569 to +124 relative to the 5' terminus of the gC mRNA were isolated. The mutants were isolated by a simple in situ hybridization screening procedure not requiring any prior selective pressure for or against expression of the gC gene. Analysis of RNA extracted from cells infected with individual mutants showed that the DNA sequences required for regulated expression of this gamma 2 gene lay within bases -34 to +124. This 158-base-pair fragment was sufficient to confer accurate and quantitative expression of gC mRNA and to maintain the stringent requirement on viral DNA replication for expression of this gene. Moreover, it was found that sequences located between -34 and +14 contained signals essential for expression of gC. To determine whether the -34 to +124 sequences would function as a gamma 2 promoter when moved to another region of the HSV-1 genome, the 158-base-pair fragment was substituted for the normal thymidine kinase promoter-regulatory sequences in the thymidine-kinase gene locus. Transcription of this chimeric gene was regulated as a gamma 2 gene in that its expression in infected cells was dependent on viral DNA synthesis. The only recognizable consensus sequence upstream of the transcription initiation site for this gene was the TATAAA sequence at -30.


[Manganese DPDP as a contrast medium for MR tomography of focal liver lesions. Tolerance and image quality in 20 patients].
Twenty patients with focal liver lesions (18 metastases, 1 hepatocellular carcinoma, 1 cholangiocarcinoma) were given manganese DPDP as part of a multicentric phase II study of paramagnetic hepatobiliary MR contrast media. 5 mumol/kg manganese DPDP were injected into 10 patients in a concentration of 50 mumol/ml or 10 mumol/ml (3 ml/min). Blood pressure, pulse rate, ECG, respiratory rate, body temperature, blood and serum parameters and the patients' subjective feelings were recorded. MRI was performed with 1.5 T using T1- and T2-weighted sequences. 6 patients reported 8 side effects (flushing, feeling of warmth, metallic taste); 7 of these were produced by the 50 mumol concentration. Two hours after injection there was a significant reduction in alkaline phosphatase which was no longer present after 24 hours. On T1-weighted images manganese DPDP resulted in marked improvement in the contrast difference between the lesions and the liver parenchyma which resulted in a marked increase in the signal to noise ratio. Comparing the two concentrations, better results were obtained by the lower concentration. Extrahepatic uptake was found in the gallbladder, duodenum, pancreas, kidneys, gastric mucosa and myocardium. Manganese DPDP in a concentration of 10 mumol/ml and a dose of 5 mumol/kg is a well tolerated contrast medium which improves the demonstration of focal liver lesions in view of its distribution and uptake. The mechanisms for the transitory side effects require further studies.


Pregnancy-Related Complications in Women with Recurrent Pregnancy Loss: A Prospective Cohort Study.
The aim of this prospective cohort study was to determine whether women with recurrent pregnancy loss (RPL) have an increased risk of pregnancy complications compared to normal pregnant women. A total of 1092 singleton pregnancies were followed, 431 in women with RPL and 661 in normal healthy women. The prevalence of the following complications was observed: threatened miscarriage, miscarriage, cervical insufficiency, chromosomal/genetic abnormalities, fetal anomalies, oligohydramnios, polyhydramnios, fetal growth restriction, intrauterine fetal death, gestational diabetes mellitus (GDM), preeclampsia, placenta previa, abruptio placentae, pregnancy-related liver disorders, and preterm premature rupture of the membranes. The odds ratio and 95% CI for each pregnancy complication considered were determined by comparing women with RPL and normal healthy women. Women with RPL had an overall rate of pregnancy complications higher than normal women (OR = 4.37; 95% CI: 3.353-5.714; p < 0.0001). Their risk was increased for nearly all the conditions considered. They also had an increased risk of multiple concomitant pregnancy complications (OR = 4.64; 95% CI: 3.10-6.94, p < 0.0001). Considering only women with RPL, women with ≥3 losses had a higher risk of pregnancy complications than women with two losses (OR = 1.269; 95% CI: 1.112-2.386, p < 0.02). No differences were found in the overall risk of pregnancy complications according to the type, explained or unexplained, of RPL. Women with secondary RPL had an increased risk of GDM than women with primary RPL. Pregnancy in women with RPL should be considered at high risk.


Moyamoya syndrome associated with neurofibromatosis Type 1: perioperative and long-term outcome after surgical revascularization.
Children with neurofibromatosis Type 1 (NF1) can present with progressive arteriopathy of the branches of the internal carotid artery consistent with moyamoya syndrome. Clinical symptoms, radiographic evidence of ischemia, and the potential for disease progression may necessitate surgical revascularization to minimize the risk of stroke and progressive neurological deficits. This study aims to evaluate the presentation and surgical outcomes of these patients by reviewing clinical, radiographic, and angiographic data. A retrospective review was conducted of clinical and radiographic records of all children with NF1 who were diagnosed with moyamoya syndrome and underwent surgical revascularization between January 1988 and April 2012 at Boston Children's Hospital. During this period, 39 patients (27 female and 12 male, ages 0.2-19.3 years) had both NF1 and moyamoya syndrome, of whom 32 underwent surgical revascularization with pial synangiosis. Of the 32 patients treated by surgical revascularization, 21 (66%) manifested ischemic symptoms and 18 (56%) had radiographic evidence of prior stroke at the time of moyamoya diagnosis. In total, 25 of 32 patients developed neurological symptoms prior to surgical intervention. Only 1 patient presented with hemorrhage. The average age at first surgery was 8.1 years (range 0.5-15.6 years). Perioperative complications in the first 7 days included stroke (n = 2), transient ischemic attack (n = 1), and infection (n = 1). Twenty-two patients had more than 6 months of follow-up, with an average clinical and radiographic postoperative follow-up period of 80.2 months (range 9.4-257.1 months). Of those patients with long-term follow-up, 21 (95%) of 22 demonstrated stable or improved neurological status despite radiographic evidence of moyamoya progression in 48% of patients. Children with NF1-associated moyamoya syndrome are often diagnosed prior to development of fixed neurological deficits as a consequence of imaging studies obtained for other manifestations of NF1. The clinical, radiographic, and angiographic features in this population are comparable to primary moyamoya disease, with the exception of patients treated with cranial irradiation, who may be at greater risk for both stroke as well as perioperative complications. Despite radiographic evidence of progressive stenosis in 48% of patients, nearly all demonstrated stable or improved neurological status after surgical revascularization. Surgical revascularization for children with NF1 appears safe and is protective against further ischemic and neurological damage, with a 27-fold reduction in stroke rate.


PUF60: a prominent new target of the autoimmune response in dermatomyositis and Sjögren's syndrome.
Autoantibodies are used clinically to phenotype and subset patients with autoimmune rheumatic diseases. We detected a novel 60 kDa autoantibody specificity by immunoblotting using a dermatomyositis (DM) patient's serum. Our objective was to identify the targeted autoantigen and to evaluate disease specificity and clinical significance of this new autoantibody. A new 60 kDa specificity was detected by immunoblotting HeLa cell lysates. The targeted autoantigen was identified as poly(U)-binding-splicing factor 60 kDa (PUF60) using (i) a human protein array and (ii) two-dimensional gel electrophoresis and liquid chromatography tandem mass spectrometry peptide sequencing. Anti-PUF60 antibodies were assayed by ELISA using sera from patients with primary Sjögren's syndrome (SS; n=84), systemic lupus erythematosus (SLE; n=71), DM (n=267), polymyositis (n=45), inclusion body myositis (n=45) and healthy controls (n=38). PUF60 was identified as a new autoantigen. Anti-PUF60 antibodies were present in 25/84 (30%) patients with SS, 6/71 (8.5%) patients with SLE and 2/38 (5.0%) control subjects (SS vs controls, p=0.002; SLE vs controls, p=0.711). Anti-PUF60 antibodies were present in 48/267 (18.0%) patients with DM versus 4/45 (8.9%) and 5/45 (11.1%) patients with inclusion body myositis and polymyositis, respectively. The antibody was significantly associated with anti-Ro52 antibodies, rheumatoid factor and hyperglobulinemia in the patients with primary SS. In patients with DM, the antibody was associated with anti-transcription intermediary factor 1 gamma seropositivity and Caucasian race. PUF60 represents a novel autoantigen in patients with SS and DM. PUF60 antibodies are associated with distinct clinical features and different immune responses in different diseases.


Presentation and clinical course of patients ultimately succumbing to carcinoma of the prostate.
Ninety-six patients who expired due to carcinoma of the prostate were identified within the primary treatment area of Brooke Army Medical Center between the years of 1981 and 1988. Although 71 patients were noted to have abnormal prostate examinations at the time of diagnosis, 11 had been noted to have a palpably normal prostate within three years of tumor diagnosis. Twenty-four patients had a palpably normal prostate at the time of diagnosis. Fifteen patients had been previously noted to have an abnormal prostate but biopsy in 11 of these had been benign. These data suggest that digital rectal examination of the prostate will not detect a number of patients who will ultimately die of carcinoma of the prostate.


Corticosterone induces discrete epigenetic signatures in the dorsal and ventral hippocampus that depend upon sex and genotype: focus on methylated Nr3c1 gene.
The genomic effects of circulating glucocorticoids are particularly relevant in cortico-limbic structures, which express a high concentration of steroid hormone receptors. To date, no studies have investigated genomic differences in hippocampal subregions, namely the dorsal (dHPC) and ventral (vHPC) hippocampus, in preclinical models treated with exogenous glucocorticoids. Chronic oral corticosterone (CORT) in mouse is a pharmacological approach that disrupts the activity of the hypothalamic-pituitary-adrenal axis, increases affective behavior, and induces genomic changes after stress in the HPC of wildtype (WT) mice and mice heterozygous for the gene coding for brain-derived neurotrophic factor Val66Met (hMet), a variant associated with genetic susceptibility to stress. Using RNA-sequencing, we investigated the genomic signatures of oral CORT in the dHPC and vHPC of WT and hMet male and female mice, and examined sex and genotype differences in response to oral CORT. Males under CORT showed lower glycemia and increased anxiety- and depression-like behavior compared to females that showed instead opposite affective behavior in response to CORT. Rank-rank-hypergeometric overlap (RRHO) was used to identify genes from a continuous gradient of significancy that were concordant across groups. RRHO showed that CORT-induced differentially expressed genes (DEGs) in WT mice and hMet mice converged in the dHPC of males and females, while in the vHPC, DEGs converged in males and diverged in females. The vHPC showed a higher number of DEGs compared to the dHPC and exhibited sex differences related to glucocorticoid receptor (GR)-binding genes and epigenetic modifiers. Methyl-DNA-immunoprecipitation in the vHPC revealed differential methylation of the exons 1C and 1F of the GR gene (Nr3c1) in hMet females. Together, we report behavioral and endocrinological sex differences in response to CORT, as well as epigenetic signatures that i) differ in the dHPC and vHPC,ii) are distinct in males and females, and iii) implicate differential methylation of Nr3c1 selectively in hMet females.


Midlife vascular risk factors and midlife cognitive status in relation to prevalence of mild cognitive impairment and dementia in later life: The Atherosclerosis Risk in Communities Study.
The interplay between midlife vascular risk factors and midlife cognitive function with later life mild cognitive impairment (MCI) and dementia (DEM) is not well understood. In the Atherosclerosis Risk in Communities Study, cardiovascular risk factors and cognition were assessed in midlife, ages 45-64 years. In 2011-2013, 20-25 years later, all consenting Atherosclerosis Risk in Communities participants underwent a cognitive and neurological evaluation and were given adjudicated diagnoses of cognitively normal, MCI, or DEM. In 5995 participants with complete covariate data, midlife diabetes, hypertension, obesity, and hypercholesterolemia were associated with late-life MCI and DEM. Low midlife cognition function was also associated with greater likelihood of late-life MCI or DEM. Both midlife vascular risk factors and midlife cognitive function remained associated with later life MCI or DEM when both were in the model. Later life MCI and DEM were independently associated with midlife vascular risk factors and midlife cognition.


Seeded Amplification of Chronic Wasting Disease Prions in Nasal Brushings and Recto-anal Mucosa-Associated Lymphoid Tissues from Elk by Real-Time Quaking-Induced Conversion.
Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since been detected across North America and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of its spread; this is especially true in cases of relocation/reintroduction or prevalence studies of large or protected herds, where depopulation may be contraindicated. This study evaluated the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay of recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brushings collected antemortem. These findings were compared to results of immunohistochemistry (IHC) analysis of ante- and postmortem samples. RAMALT samples were collected from populations of farmed and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni;n= 323), and nasal brush samples were collected from a subpopulation of these animals (n= 205). We hypothesized that the sensitivity of RT-QuIC would be comparable to that of IHC analysis of RAMALT and would correspond to that of IHC analysis of postmortem tissues. We found RAMALT sensitivity (77.3%) to be highly correlative between RT-QuIC and IHC analysis. Sensitivity was lower when testing nasal brushings (34%), though both RAMALT and nasal brush test sensitivities were dependent on both thePRNPgenotype and disease progression determined by the obex score. These data suggest that RT-QuIC, like IHC analysis, is a relatively sensitive assay for detection of CWD prions in RAMALT biopsy specimens and, with further investigation, has potential for large-scale and rapid automated testing of antemortem samples for CWD.


Dynamic study of the blood viscosity of insulin-dependent diabetics by means of an artificial pancreas.
The blood viscosity of 15 insulin-dependent, poorly controlled, diabetic subjects was determined by using a microviscosimeter at low shear rates, with cylindrical cuvettes of the Couette type. It was found that the blood viscosity of these diabetics was more elevated than that of control patients (p less than 0.001). In ten diabetics, the return to a strict metabolic control over a period of more than 24 h by means of an artificial pancreas resulted in a significant systematic lowering of blood viscosity. These results suggest that the metabolic control of diabetes influences blood viscosity, and they underline the importance of an artificial pancreas for the dynamic study of the factors affecting blood viscosity during the course of diabetes.


Role of IFN-gamma in an experimental murine model of West Nile virus-induced seizures.
Seizures are a major complication of viral encephalitis. However, the mechanisms of seizure-associated neuronal dysfunction remain poorly understood. We report that intranasal inoculation with West Nile virus (WNV) (Sarafend) causes limbic seizures in C57BL/6 mice, but not in interferon (IFN)-gamma-deficient (IFN-gamma-/-) mice. Both strains showed similar levels of virus in the brain, as well as similar concentrations of the cytokines, tumor necrosis factor and interleukin-6, both of which can alter neuronal excitability. Experiments in chimeric IFN-gamma-/- mice reconstituted with IFN-gamma-producing leukocytes showed that IFN-gamma is not required during central nervous system infection for limbic seizure development, suggesting a role for IFN-gamma in the developing brain. This was supported responses to pentylenetetrazole, kainic acid (KA), and N-methyl-d-aspartate (NMDA). Both strains of mice exhibited similar behavior after pentylenetetrazole challenge. However, while NMDA and KA treatment resulted in characteristic seizures in C57BL/6 mice, these responses were diminished (NMDA treatment) or absent (KA treatment) in IFN-gamma-/- mice. Furthermore, NMDA-receptor blockade with MK-801 in WNV-infected C57BL/6 mice abrogated seizures and prolonged survival. Our data show that IFN-gamma plays an important role in the development of the excitatory seizure pathways in the brain and that these cascades become pathogenic in encephalitic WNV infection.


[Nanosilver - harmful effects of biological activity].
Nanosilver, also identified as colloidal silver, has been known and used for ages to combat diseases or prolong food freshness. It usually occurs in the form of a suspension consisting of particles of size < 100 nm. Due to its specific properties, silver nanoparticles are used in many technologies to produce medical devices, textiles, conductive materials or photovoltaic cells. The growing popularity of nanosilver applications increases the number of people occupationally exposed to this substance. Potential exposure routes for silver nanoparticles are through dermal, oral and inhalation pathways. Silver nanoparticles may be absorbed through the lungs, intestine, and through the skin into circulation and thus may reach such organs as the liver, kidney, spleen, brain, heart and testes. Nanosilver may cause mild eyes and skin irritations. It can also act as a mild skin allergen. Inhalation of silver nanoparticles mainly affects the lungs and liver. It has been demonstrated that silver nanoparticles may be genotoxic to mammalian cells. There are some alarming reports on the adverse effects of silver nanoparticles on reproduction of experimental animals. Exposure to silver nanoparticles may exert a neurotoxic effect and affect cognitive functions, causing the impairment of short-term and working memory. Maximum admissible concentration (MAC) for the inhalable fraction of silver of 0.05 mg/m3 is currently binding in Poland. In light of toxicological studies of silver nanoparticles it seems reasonable to update the hygiene standards for silver with nanoparticles as a separate fraction.


Proteomic analysis of GTP cyclohydrolase 1 multiprotein complexes in cultured normal adult human astrocytes under both basal and cytokine-activated conditions.
GTP cyclohydrolase 1 (GCH1) is the rate-limiting enzyme of a metabolic pathway synthesizing tetrahydrobiopterin (BH(4)), the cofactor dimerizing and activating inducible nitric oxide synthase (NOS-2). GCH1 protein expression and enzyme activity are minimal in cultured, phenotypically stable, untreated normal adult human astrocytes (NAHA), but are strongly induced, together with NOS-2, by a mixture of three proinflammatory cytokines (IL-1beta, TNF-alpha, and IFN-gamma--the CM-trio) released by microglia under brain-damaging conditions. The resulting hyper-production of NO severely harms neurons. In this study, using MALDI-TOF/MS, PMF, Western immunoblotting (WB), and antibody microarrays we identified several proteins coimmunoprecipitating with GCH1. Under basal conditions, GCH1 was associated with various adaptor/regulator molecules involved in G-protein-coupled receptors signalling, protein serine/threonine phosphatase 2Cbeta (PP2Cbeta), and serine-threonine kinases like Ca(2+) calmodulin kinases (CaMKs), casein kinases (CKs), cAMP-dependent kinases (PKAs), and mitogen-activated protein kinases (MAPKs). Exposure to the three cytokines' mixture (CM-trio) significantly changed, within the 48-72 h required for the induction and activation of GCH1, the levels and identities of some of the 0 h-associated proteins: after 72 h CK-IIalpha tended to dissociate from, whereas MAPK12 and JNK3 were strongly associated with fully active GCH1. These findings provide a first enticing glimpse into the intricate mechanisms regulating GCH1 activation by proinflammatory cytokines in NAHA, and may have therapeutic implications.


Experiences of progestin-only pill users in the United States and attitudes toward over-the-counter access.
Removing the prescription requirement and making oral contraceptive pills available over the counter (OTC) could increase contraceptive access in the United States. Despite current efforts to make a progestin-only pill (POP) available OTC, there are no qualitative data exploring the experiences of POP users and their perspectives on making POPs available OTC. We conducted six online, asynchronous focus group discussions with 36 POP users between July and November 2020. We stratified focus group discussion placement based on three reasons for POP use: breastfeeding, having a contraindication to estrogen (for a reason other than breastfeeding), and for any other reason. We thematically analyzed these data using inductive and deductive coding. The majority of participants described their overall POP experience as positive, mostly because they experienced minor or no side effects and thought the pill was effective. Participants overwhelmingly supported OTC availability of POPs and expressed interest in purchasing an OTC POP. Over one-third of participants had very limited knowledge or held inaccurate assumptions about POPs before they started using this method and many stressed the need to disseminate accurate information and resources about POPs to the public. Most focus group discussion participants were satisfied with POPs and supported OTC access. While misconceptions and concerns about POPs should be addressed, an OTC POP has the potential to be a safe, effective, and convenient contraceptive option in the United States.


Pancreaticoduodenectomy for periampullary adenocarcinoma.
This study evaluates the indications for and effects of pancreaticoduodenectomy (102 patients) or total pancreatectomy (15 patients) with extensive lymph node dissection performed upon 117 patients for treatment of periampullary adenocarcinoma. Presenting symptoms and postoperative morbidity and mortality rates were recorded. Cumulative survival rates were evaluated in relation to origin, size, and staging of tumor. Postoperative follow-up of clinical symptoms was done after one year. The postoperative mortality rate after Whipple's operation was 8 percent (eight patients). The median survival period was 1.1 year and the overall five year survival rate was 15 percent (confidence limits, 5 to 25 percent). The five year survival rate for patients without tumor extension beyond the pancreas was 25 percent (confidence limits, 5 to 50 percent), and in patients with adenocarcinoma of the ampulla of Vater, 34 percent (confidence limits, 3 to 65 percent). The median survival rate in patients with adenocarcinoma of the ampulla of Vater was 3.3 years, which was significantly longer than in the other patients. Fifty-nine patients with distant spread could be divided into 14 patients with para-aortic lymph node metastases who had a significantly shorter survival period than 45 patients without para-aortic lymph node metastases (p = 0.004). Most patients surviving more than one year were doing well, although 60 percent needed exocrine pancreatic substitution therapy. Resection of periampullary carcinoma provides a better palliation and survival rate than nonoperative biliary drainage or bypass operation. An improved preoperative verification of para-aortic metastases could restrict resection to patients with a prognostic five year survival rate of more than 25 percent and a postoperative mortality rate of less than 5 percent.


Modeling DNA affinity landscape through two-round support vector regression with weighted degree kernels.
A quantitative understanding of interactions between transcription factors (TFs) and their DNA binding sites is key to the rational design of gene regulatory networks. Recent advances in high-throughput technologies have enabled high-resolution measurements of protein-DNA binding affinity. Importantly, such experiments revealed the complex nature of TF-DNA interactions, whereby the effects of nucleotide changes on the binding affinity were observed to be context dependent. A systematic method to give high-quality estimates of such complex affinity landscapes is, thus, essential to the control of gene expression and the advance of synthetic biology. Here, we propose a two-round prediction method that is based on support vector regression (SVR) with weighted degree (WD) kernels. In the first round, a WD kernel with shifts and mismatches is used with SVR to detect the importance of subsequences with different lengths at different positions. The subsequences identified as important in the first round are then fed into a second WD kernel to fit the experimentally measured affinities. To our knowledge, this is the first attempt to increase the accuracy of the affinity prediction by applying two rounds of string kernels and by identifying a small number of crucial k-mers. The proposed method was tested by predicting the binding affinity landscape of Gcn4p in Saccharomyces cerevisiae using datasets from HiTS-FLIP. Our method explicitly identified important subsequences and showed significant performance improvements when compared with other state-of-the-art methods. Based on the identified important subsequences, we discovered two surprisingly stable 10-mers and one sensitive 10-mer which were not reported before. Further test on four other TFs in S. cerevisiae demonstrated the generality of our method. We proposed in this paper a two-round method to quantitatively model the DNA binding affinity landscape. Since the ability to modify genetic parts to fine-tune gene expression rates is crucial to the design of biological systems, such a tool may play an important role in the success of synthetic biology going forward.


Painful Granulomatous Hashimoto Thyroiditis Uncovered Systemic Histoplasmosis.
Background. Granulomatous thyroiditis is usually viral and very rarely due to fungal infection. Fungal thyroiditis affects the severely immunocompromised and only rarely immunocompetent individuals. Case report. A 32-year-old man with known Hashimoto thyroiditis was consulted for a 1-week history of neck pain, dysphagia, cough, asthenia, loss of appetite, and night sweats. Imaging showed an enlarged right thyroid lobe. Laboratory studies confirmed hypothyroidism. He underwent a thyroid lobectomy. Microscopic examination showed granulomas in a background of Hashimoto thyroiditis. Silver stain revealed numerous yeasts consistent with Histoplasma. Subsequent serology was also positive. Further evaluations were negative for acquired or constitutional immunodeficiencies. He was started on oral itraconazole with resolution of all constitutional symptoms. Conclusions. We report a second case of concurrent Hashimoto and Histoplasma thyroiditis in an immunocompetent patient. Systemic manifestations of fungal infection are confounded by systemic symptoms of thyroid dysfunction. Fungal thyroiditis is a manifestation of systemic fungal infection requiring prolonged systemic therapy to prevent complications or fatal outcomes.


Microbial diversity in anaerobic sediments at Rio Tinto, a naturally acidic environment with a high heavy metal content.
The Tinto River is an extreme environment located at the core of the Iberian Pyritic Belt (IPB). It is an unusual ecosystem due to its size (100 km long), constant acidic pH (mean pH, 2.3), and high concentration of heavy metals, iron, and sulfate in its waters, characteristics that make the Tinto River Basin comparable to acidic mine drainage (AMD) systems. In this paper we present an extensive survey of the Tinto River sediment microbiota using two culture-independent approaches: denaturing gradient gel electrophoresis and cloning of 16S rRNA genes. The taxonomic affiliation of the Bacteria showed a high degree of biodiversity, falling into 5 different phyla: Proteobacteria, Firmicutes, Bacteroidetes, Acidobacteria, and Actinobacteria; meanwhile, all the Archaea were affiliated with the order Thermoplasmatales. Microorganisms involved in the iron (Acidithiobacillus ferrooxidans, Sulfobacillus spp., Ferroplasma spp., etc.), sulfur (Desulfurella spp., Desulfosporosinus spp., Thermodesulfobium spp., etc.), and carbon (Acidiphilium spp., Bacillus spp., Clostridium spp., Acidobacterium spp., etc.) cycles were identified, and their distribution was correlated with physicochemical parameters of the sediments. Ferric iron was the main electron acceptor for the oxidation of organic matter in the most acid and oxidizing layers, so acidophilic facultative Fe(III)-reducing bacteria appeared widely in the clone libraries. With increasing pH, the solubility of iron decreases and sulfate-reducing bacteria become dominant, with the ecological role of methanogens being insignificant. Considering the identified microorganisms-which, according to the rarefaction curves and Good's coverage values, cover almost all of the diversity-and their corresponding metabolism, we suggest a model of the iron, sulfur, and organic matter cycles in AMD-related sediments.


Transport of fluorescein in the ocular posterior segment in retinitis pigmentosa.
The function of the blood-retinal barrier was assessed by vitreous fluorophotometry in 12 patients with various genetic types of retinitis pigmentosa and in 11 normal subjects. The measurements were corrected to minimize the effect of artifacts. We evaluated the inward penetration of fluorescein sodium across the blood-retinal barrier and determined the outward permeability coefficient for fluorescein by interpreting the data with a pharmacokinetic computer model. Patients with retinitis pigmentosa had increased inward permeability and decreased outward permeability to fluorescein in comparison with values of normal subjects.


[Interaction between chemical substances contaminating a gaseous medium of hermetically sealed compartments].
We have analyzed pertinent literature data and hypothesized that chemicals in an enclosed environment may interact (products of polymer outgassing, volatile metabolites of man). We have demonstrated that photochemical processes, thermochemical degradation and oxidation of chemicals in an enclosed atmosphere may lead to the formation of new highly toxic substances. The chemical composition of an enclosed atmosphere may be influenced by the technological processes of air purification. We have carried out experiments in a small enclosure to study ammonia and carbon dioxide interaction. When the concentration of ammonia is about 100 mg/m3, that of carbon dioxide is 2.5 vol. % and relative humidity of the atmosphere is 40-50%, the formation reaction of ammonia carbonate compounds proceeds very slowly, producing no significant changes in the environmental toxicological characteristics. It is recommended to continue the study of the interaction of chemicals at low concentrations in an enclosed environment.


Inhibition of MiR-122 Decreases Cerebral Ischemia-reperfusion Injury by Upregulating DJ-1-Phosphatase and Tensin Homologue Deleted on Chromosome 10 (PTEN)/Phosphonosinol-3 Kinase (PI3K)/AKT.
BACKGROUND Ischemia-reperfusion injury is caused by a blood reperfusion injury in ischemic brain tissue, and usually occurs in the treatment stage of ischemic disease, which can aggravate brain tissue injury. MiR-122 is closely related to ischemia-reperfusion injury in the myocardium, kidney, and liver; however, the role in cerebral ischemia-reperfusion injury has not been established. MATERIAL AND METHODS In this study, cerebral ischemia-reperfusion injury was established in a rat model, and the control group was a sham-operated group. After ischemia-reperfusion injury for 6, 12, and 24 hours, brain tissue specimens were collected and the expression of miR-122 and DJ-1 were determined using quantitative real-time polymerase chain reaction. Flow cytometry was used to determine the reactive oxygen species (ROS) content. The modified Neurological Severity Score (mNSS) scale was used to evaluate the sensory and motor function defects of the rats. The malondialdehyde (MDA), superoxide dismutase (SOD), and enzyme activity were determined. The rats in the cerebral ischemia-reperfusion injury model were divided into 2 groups (antagomir-NC group and antagomir miR-122 group). Brain neuron RN-c cells were divided into the following 4 groups: antagomir-NC, antagomir miR-122, pIRES2-blank, and pIRES2-DJ-1. Seventy-two hours after transfection, ischemia-reperfusion treatment was carried out and conventional cultured RN-c cells were used as the control group. Flow cytometry was used to detect apoptosis and western blot was used to detect the expression of DJ-1, PTEN, AKT, and p-AKT. RESULTS The expression of miR-122 increased significantly in the process of ischemia-reperfusion damage after cerebral infarction, while the expression of DJ-1 decreased significantly. Downregulation of miR-122 significantly increased the expression of DJ-1, enhanced the activity of the PTEN/PI3K/AKT pathway, reduced cell apoptosis, and alleviated cerebral ischemia-reperfusion injury. CONCLUSIONS Inhibition of miR-122 can decrease cerebral ischemia-reperfusion injury by upregulating DJ-1-PTEN/PI3K/AKT pathway.


Streamlined regulation and gene loss as adaptive mechanisms in Prochlorococcus for optimized nitrogen utilization in oligotrophic environments.
Prochlorococcus is one of the dominant cyanobacteria and a key primary producer in oligotrophic intertropical oceans. Here we present an overview of the pathways of nitrogen assimilation in Prochlorococcus, which have been significantly modified in these microorganisms for adaptation to the natural limitations of their habitats, leading to the appearance of different ecotypes lacking key enzymes, such as nitrate reductase, nitrite reductase, or urease, and to the simplification of the metabolic regulation systems. The only nitrogen source utilizable by all studied isolates is ammonia, which is incorporated into glutamate by glutamine synthetase. However, this enzyme shows unusual regulatory features, although its structural and kinetic features are unchanged. Similarly, urease activities remain fairly constant under different conditions. The signal transduction protein P(II) is apparently not phosphorylated in Prochlorococcus, despite its conserved amino acid sequence. The genes amt1 and ntcA (coding for an ammonium transporter and a global nitrogen regulator, respectively) show noncorrelated expression in Prochlorococcus under nitrogen stress; furthermore, high rates of organic nitrogen uptake have been observed. All of these unusual features could provide a physiological basis for the predominance of Prochlorococcus over Synechococcus in oligotrophic oceans.


Proliferation of Aedes aegypti in urban environments mediated by the availability of key aquatic habitats.
Aedes aegypti is the main vector of dengue, Zika, chikungunya, and yellow fever viruses. Controlling populations of vector mosquito species in urban environments is a major challenge and being able to determine what aquatic habitats should be prioritized for controlling Ae. aegypti populations is key to the development of more effective mosquito control strategies. Therefore, our objective was to leverage on the Miami-Dade County, Florida immature mosquito surveillance system based on requested by citizen complaints through 311 calls to determine what are the most important aquatic habitats in the proliferation of Ae. aegypti in Miami. We used a tobit model for Ae. aegypti larvae and pupae count data, type and count of aquatic habitats, and daily rainfall. Our results revealed that storm drains had 45% lower percentage of Ae. aegypti larvae over the total of larvae and pupae adjusted for daily rainfall when compared to tires, followed by bromeliads with 33% and garbage cans with 17%. These results are indicating that storm drains, bromeliads and garbage cans had significantly more pupae in relation to larvae when compared to tires, traditionally know as productive aquatic habitats for Ae. aegypti. Ultimately, the methodology and results from this study can be used by mosquito control agencies to identify habitats that should be prioritized in mosquito management and control actions, as well as to guide and improve policies and increase community awareness and engagement. Moreover, by targeting the most productive aquatic habitats this approach will allow the development of critical emergency outbreak responses by directing the control response efforts to the most productive aquatic habitats.


[The "Duisgurg model" of intensified control of nursing homes and homes for the aged by the Duisberg public health services in cooperation with the Duisburg social welfare offices].
This is a report on the joint cooperative control of nursing and old-age homes by the Public Health and Social Welfare Offices in Duisburg, Germany. A total of 40 of such homes were inspected and checked jointly by the physician and pharmacist of the local Public Health Office within a one-year period while collecting data on the intensity and quality of care in these homes. A total of 3883 inmates were covered. These data included, inter alia, information on the intensity of care, prevalence of urinary and fecal incontinence, feeding by means of percutaneous endoscopic gastrostomy, prevalence of bedridden patients and of cases of pressure sores. The data conveyed information on the overall situation (frequencies for all the Duisburg nursing and old-age homes) on the one hand, and for each individual home on the basis of its particular data on the other hand. It became evident that two-thirds of all the inmates of nursing homes were classified as belonging to nursing stages II and III. Urinary incontinence was present in 63 per cent of the cases, fecal incontinence in 43 per cent, whereas 32 per cent were severely confused. Of a total of 447 permanently bedridden patients (11.5 per cent of all the inmates) 131 had pressure sores or necroses of the heels (3.37 per cent of the total population). More than one-half of the pressure sores had been acquired by the patients during their hospital stay, i.e. before their referral to a home, whereas 56 inmates developed their pressure sores in the homes concerned.


Specific subdomains of Vav differentially affect T cell and NK cell activation.
The Vav protooncogene is a multidomain protein involved in the regulation of IL-2 gene transcription in T cells and the development of cell-mediated killing by cytotoxic lymphocytes. We have investigated the differential roles that specific protein subdomains within the Vav protooncogene have in the development of these two distinct cellular processes. Interestingly, a calponin homology (CH) domain mutant of Vav (CH-) fails to enhance NF-AT/AP-1-mediated gene transcription but is still able to regulate the development of cell-mediated killing. The inability of the CH- mutant to enhance NF-AT/AP-1-mediated transcription appears to be secondary to defective intracellular calcium, because 1) the CH- mutant has significantly reduced TCR-initiated calcium signaling, and 2) treatment with the calcium ionophore ionomycin or cotransfection with activated calcineurin restores NF-AT/AP-1-mediated gene transcription. The pleckstrin homology (PH) domain of Vav has also been implicated in regulating Vav activation. We found that deletion of the PH domain of Vav yields a protein that can neither enhance gene transcription from the NF-AT/AP-1 reporter nor enhance TCR- or FcR-mediated killing. In contrast, the PH deletion mutant of Vav is able to regulate the development of natural cytotoxicity, indicating a functional dichotomy for the PH domain in the regulation of these two distinct forms of killing. Lastly, mutation of three tyrosines (Y142, Y160, and Y174) within the acidic domain of Vav has revealed a potential negative regulatory site. Replacement of all three tyrosines with phenylalanine results in a hyperactive protein that increases NF-AT/AP-1-mediated gene transcription and enhances cell-mediated cytotoxicity. Taken together, these data highlight the differential roles that specific subdomains of Vav have in controlling distinct cellular functions. More broadly, the data suggest that separate lymphocyte functions can potentially be modulated by domain-specific targeting of Vav and other critical intracellular signaling molecules.


A comparison between follitropin α filled by mass and follitropin a filled by bioassay in the same egg donors.
This study assessed the ovarian stimulation characteristics of recombinant follitropin a filled by mass (rFSH-fbm) versus recombinant follitropin a filled by conventional bioassay (rFSH-bio) in the same egg donor patients. Eleven egg donors, who had two ovarian stimulation cycles for oocyte retrieval (total of 23 cycles), one with rFSH-bio (Gonal-f Multidose®) and the second one with rFSH-fbm (Gonal-f® RFF), were evaluated. The protocol of ovarian stimulation was exactly the same in both cycles, consisting of GnRH suppression (luteal phase) followed by exclusive stimulation with rFSH. Despite no differences in the number of days of rFSH treatment and in the total dosage of rFSH administered, the number of follicles >14 mm and the number of oocytes retrieved were significantly higher in the rFSH-fbm group (P = 0.01 and 0.04 respectively). The mean oestradiol peak values showed a trend in favour of rFSH-fbm (3123 versus 2405 pg/ml respectively). These results suggest that the consistency in dosing provided by follitropin a filled by mass as opposed to follitropin a filled by bioassay offers added value for the ovarian stimulation of oocyte donor patients.


Environmental Burkholderia cepacia complex isolates in human infections.
Members of the Burkholderia cepacia complex (Bcc), found in many environments, are associated with clinical infections. Examining diverse species and strains from different environments with multilocus sequence typing, we identified > 20% of 381 clinical isolates as indistinguishable from those in the environment. This finding links the natural environment with the emergence of many Bcc infections.


[Response of tree seedlings to altitudinal gradient and its seasonal variation in Ailao Mountain and Yulong Mountain, Yunnan Province, China.]
Tree seedlings under forests serve as an important part of forest ecosystem and play a role in the natural regeneration of forests. To understand the altitudinal distribution patterns of tree seedlings and the response to seasonal variation, we investigated the species composition and individual numbers of tree seedlings, and their seasonal dynamics in Ailao Mountain (subtropical region) and Yulong Mountain (subalpine region) in Yunnan Province. The results showed that the species richness of tree seedlings increased with altitude, peaked in the middle of the mountains, and followed a decline at higher altitudes, and the dominant species also changed significantly. Species richness of tree seedlings was significantly higher at the end of rainy season than that at the end of dry season, and the dominant species had obvious seasonal difference.


Managing postoperative pain in adult outpatients: a systematic review and meta-analysis comparing codeine with NSAIDs.
Analgesics that contain codeine are commonly prescribed for postoperative pain, but it is unclear how they compare with nonopioid alternatives. We sought to compare the effectiveness of codeine and nonsteroidal anti-inflammatory drugs (NSAIDs) for adults who underwent outpatient surgery. We conducted a systematic review and meta-analysis of randomized controlled trials comparing codeine and NSAIDs for postoperative pain in outpatient surgery. We searched MEDLINE and Embase from inception to October 2019 for eligible studies. Our primary outcome was the patient pain score, converted to a standard 10-point intensity scale. Our secondary outcomes were patient-reported global assessments and adverse effects. We used random-effects models and grading of recommendations assessment, development and evaluation (GRADE) to assess the quality of evidence. Forty studies, including 102 trial arms and 5116 patients, met inclusion criteria. The studies had low risk of bias and low-to-moderate heterogeneity. Compared with codeine, NSAIDs were associated with better pain scores at 6 hours (weighted mean difference [WMD] 0.93 points, 95% confidence interval [CI] 0.71 to 1.15) and at 12 hours (WMD 0.79, 95% CI 0.38 to 1.19). Stronger NSAID superiority at 6 hours was observed among trials where acetaminophen was coadministered at equivalent doses between groups (WMD 1.18, 95% CI 0.87 to 1.48). NSAIDs were associated with better global assessments at 6 hours (WMD -0.88, 95% CI -1.04 to -0.72) and at 24 hours (WMD -0.67, 95% CI -0.95 to -0.40), and were associated with fewer adverse effects, including bleeding events. We found that adult outpatients report better pain scores, better global assessments and fewer adverse effects when their postoperative pain is treated with NSAIDs than with codeine. Clinicians across all specialties can use this information to improve both pain management and opioid stewardship.


Emergent coordination underlying learning to reach to grasp with a brain-machine interface.
The development of coordinated reach-to-grasp movement has been well studied in infants and children. However, the role of motor cortex during this development is unclear because it is difficult to study in humans. We took the approach of using a brain-machine interface (BMI) paradigm in rhesus macaques with prior therapeutic amputations to examine the emergence of novel, coordinated reach to grasp. Previous research has shown that after amputation, the cortical area previously involved in the control of the lost limb undergoes reorganization, but prior BMI work has largely relied on finding neurons that already encode specific movement-related information. In this study, we taught macaques to cortically control a robotic arm and hand through operant conditioning, using neurons that were not explicitly reach or grasp related. Over the course of training, stereotypical patterns emerged and stabilized in the cross-covariance between the reaching and grasping velocity profiles, between pairs of neurons involved in controlling reach and grasp, and to a comparable, but lesser, extent between other stable neurons in the network. In fact, we found evidence of this structured coordination between pairs composed of all combinations of neurons decoding reach or grasp and other stable neurons in the network. The degree of and participation in coordination was highly correlated across all pair types. Our approach provides a unique model for studying the development of novel, coordinated reach-to-grasp movement at the behavioral and cortical levels. NEW & NOTEWORTHY Given that motor cortex undergoes reorganization after amputation, our work focuses on training nonhuman primates with chronic amputations to use neurons that are not reach or grasp related to control a robotic arm to reach to grasp through the use of operant conditioning, mimicking early development. We studied the development of a novel, coordinated behavior at the behavioral and cortical level, and the neural plasticity in M1 associated with learning to use a brain-machine interface.


Identifying effective behavioural components of Intervention and Comparison group support provided in SMOKing cEssation (IC-SMOKE) interventions: a systematic review protocol.
Systematic reviews of behaviour change interventions for smoking cessation vary in scope, quality, and applicability. The current review aims to generate more accurate and useful findings by (1) a detailed analysis of intervention elements that change behaviour (i.e. behaviour change techniques (BCTs)) and potential moderators of behaviour change (i.e. other intervention and sample characteristics) and (2) assessing and controlling for variability in support provided to comparison groups in smoking cessation trials. A systematic review will be conducted of randomized controlled trials of behaviour change interventions for smoking cessation in adults (with or without pharmacological support), with a minimum follow-up of 6 months, published after 1995. Eligible articles will be identified through the Cochrane Tobacco Addiction Group Specialized Register. Study authors will be asked for detailed descriptions of smoking cessation support provided to intervention and comparison groups. All data will be independently coded by two researchers. The BCT taxonomy v1 (tailored to smoking cessation interventions) and template for intervention description and replication criteria will be used to code intervention characteristics. Data collection will further include sample and trial characteristics and outcome data (smoking cessation rates). Multilevel mixed-effects meta-regression models will be used to examine which BCTs and/or BCT clusters delivered to intervention and comparison groups explain smoking cessation rates in treatment arms (and effect sizes) and what key moderators of behaviour change are. Predicted effect sizes of each intervention will be computed assuming all interventions are compared against comparison groups receiving the same levels of behavioural support (i.e. low, medium, and high levels). Multi-disciplinary advisory board members (policymakers, health care providers, and (ex-)smokers) will provide strategic input throughout the project to ensure the review's applicability to policy and practice. By capturing BCTs in intervention and comparison groups, this systematic review will provide more accurate estimates of the effectiveness of smoking cessation interventions, the most promising BCTs and/or BCT clusters associated with smoking cessation rates in intervention and comparison arms, and important moderators of behaviour change. The results could set new standards for conducting meta-analyses of behaviour change interventions and improve research, service delivery, and training in the area of smoking cessation. PROSPERO CRD42015025251.


Application of Machine Learning in Modeling the Relationship between Catchment Attributes and Instream Water Quality in Data-Scarce Regions.
This research delves into the efficacy of machine learning models in predicting water quality parameters within a catchment area, focusing on unraveling the significance of individual input variables. In order to manage water quality, it is necessary to determine the relationship between the physical attributes of the catchment, such as geological permeability and hydrologic soil groups, and in-stream water quality parameters. Water quality data were acquired from the Iran Water Resource Management Company (WRMC) through monthly sampling. For statistical analysis, the study utilized 5-year means (1998-2002) of water quality data. A total of 88 final stations were included in the analysis. Using machine learning methods, the paper gives relations for 11 in-stream water quality parameters: Sodium Adsorption Ratio (SAR), Na+, Mg2+, Ca2+, SO42-, Cl-, HCO3-, K+, pH, conductivity (EC), and Total Dissolved Solids (TDS). To comprehensively evaluate model performance, the study employs diverse metrics, including Pearson's Linear Correlation Coefficient (R) and the mean absolute percentage error (MAPE). Notably, the Random Forest (RF) model emerges as the standout model across various water parameters. Integrating research outcomes enables targeted strategies for fostering environmental sustainability, contributing to the broader goal of cultivating resilient water ecosystems. As a practical pathway toward achieving a delicate balance between human activities and environmental preservation, this research actively contributes to sustainable water ecosystems.