GABA Neuronal Deletion of Shank3 Exons 14-16 in Mice Suppresses Striatal Excitatory Synaptic Input and Induces Social and Locomotor Abnormalities.
Shank3 is an excitatory postsynaptic scaffolding protein implicated in multiple brain disorders, including autism spectrum disorders (ASD) and Phelan-McDermid syndrome (PMS). Although previous neurobiological studies on Shank3 and Shank3-mutant mice have revealed diverse roles of Shank3 in the regulation of synaptic, neuronal and brain functions, whether Shank3 expression in specific cell types distinctly contributes to mouse phenotypes remains largely unclear. In the present study, we generated two Shank3-mutant mouse lines (exons 14-16) carrying global and GABA neuron-specific deletions and characterized their electrophysiological and behavioral phenotypes. These mouse lines show similar decreases in excitatory synaptic input onto dorsolateral striatal neurons. In addition, the abnormal social and locomotor behaviors observed in global Shank3-mutant mice are strongly mimicked by GABA neuron-specific Shank3-mutant mice, whereas the repetitive and anxiety-like behaviors are only partially mimicked. These results suggest that GABAergic Shank3 (exons 14-16) deletion has strong influences on striatal excitatory synaptic transmission and social and locomotor behaviors in mice.


Blood pressure and calf muscle oxygen extraction during plantar flexion exercise in peripheral artery disease.
Peripheral artery disease (PAD) is an atherosclerotic vascular disease that affects 200 million people worldwide. Although PAD primarily affects large arteries, it is also associated with microvascular dysfunction, an exaggerated blood pressure (BP) response to exercise, and high cardiovascular mortality. We hypothesized that fatiguing plantar flexion exercise that evokes claudication elicits a greater reduction in skeletal muscle oxygenation (SmO2) and a higher rise in BP in PAD compared with age-matched healthy subjects, but low-intensity steady-state plantar flexion elicits similar responses between groups. In the first experiment, eight patients with PAD and eight healthy controls performed fatiguing plantar flexion exercise (from 0.5 to 7 kg for up to 14 min). In the second experiment, seven patients with PAD and seven healthy controls performed low-intensity plantar flexion exercise (2.0 kg for 14 min). BP, heart rate (HR), and SmO2 were measured continuously using near-infrared spectroscopy (NIRS). SmO2 is the ratio of oxygenated hemoglobin to total hemoglobin, expressed as a percent. At fatigue, patients with PAD had a greater increase in mean arterial BP (18 ± 2 vs. vs. 10 ± 2 mmHg, P = 0.029) and HR (14 ± 2 vs. 6 ± 2 beats/min, P = 0.033) and a greater reduction in SmO2 (-54 ± 10 vs. -12 ± 4%, P = 0.001). However, both groups had similar physiological responses to low-intensity, nonpainful plantar flexion exercise. These data suggest that patients with PAD have altered oxygen uptake and/or utilization during fatiguing exercise coincident with an augmented BP response.NEW & NOTEWORTHY In this laboratory study, patients with peripheral artery disease performed plantar flexion exercise in the supine posture until symptoms of claudication occurred. Relative to age- and sex-matched healthy subjects we found that patients had a higher blood pressure response, a higher heart rate response, and a greater reduction in skeletal muscle oxygenation as determined by near-infrared spectroscopy. Our data suggest that muscle ischemia contributes to the augmented exercise pressor reflex in peripheral artery disease.


Effects of transcranial direct current stimulation over the posterior parietal cortex on novice X-ray screening performance.
Existing theories of visual search are generally deduced from lab-based studies involving the identification of a target object among similar distractors. The role of the right parietal cortex in visual search is well-established. However, less is known about real-world visual search tasks, such as X-ray screening, which require targets to be disembedded from their background. Research has shown variations in the cognitive abilities required for these tasks and typical lab-based visual search tasks. Thus, the findings of traditional visual search studies do not always transfer into the applied domain. Although brain imaging studies have offered insights into visual search tasks involving disembedding, highlighting an association between the left parietal cortex and disembedding performance, no causal link has yet been established. To this end, we carried out a pilot study (n = 34, between-subjects) administering non-invasive brain stimulation over the posterior parietal cortex (PPC) prior to completing a security X-ray screening task. The findings suggested that anodal left PPC tDCS enhanced novice performance in X-ray screening over that of sham stimulation, in line with brain imaging findings. However, the efficacy of tDCS is under question, with a growing number of failed replications. With this in mind, this study aims to re-test our original hypothesis by examining the effects of left-side parietal stimulation on novice X-ray screener performance and comparing them to those of sham stimulation and of stimulation on a control site (right PPC). As such, this within-subjects study comprised three sessions (2 mA left PPC, 2 mA right PPC, low-intensity sham stimulation left PPC), to investigate effects of anodal tDCS on X-ray screening performance. The pre-registered analysis did not detect any significant differences between left PPC tDCS and sham tDCS or left PPC tDCS and right PPC tDCS on novice performance (d') in X-ray screening. Further exploratory analyses detected no effects of left PPC tDCS on any other indices of performance in the X-ray security screening task (c, RTs and accuracy), or a disembedding control task (RTs and accuracy). The use of alternative stimulation techniques, with replicable behavioural effects on the parietal lobe (or a multi-technique approach), and well-powered studies with a systematic variation of stimulation parameters, could help to choose between two possible interpretations: that neither left nor right PPC are causally related to either tasks or that tDCS was ineffective. Finally, low-intensity sham stimulation (.016 mA), previously shown to outperform other sham conditions in between-subjects designs, was found to be ineffective for blinding participants in a within-subjects design. Our findings raise concerns for the current lack of optimal control conditions and add to the growing literature highlighting the need for replication in the field.


The role of neutrophils in rheumatic disease-associated vascular inflammation.
Vascular pathologies underpin and intertwine autoimmune rheumatic diseases and cardiovascular conditions, and atherosclerosis is increasingly recognized as the leading cause of morbidity in conditions such as systemic lupus erythematosus (SLE), rheumatoid arthritis and antineutrophil cytoplasmic antibody-associated vasculitis. Neutrophils, important cells in the innate immune system, exert their functional effects in tissues via a variety of mechanisms, including the generation of neutrophil extracellular traps and the production of reactive oxygen species. Neutrophils have been implicated in the pathogenesis of several rheumatic diseases, and can also intimately interact with the vascular system, either through modulating endothelial barriers at the blood-vessel interface, or through associations with platelets. Emerging data suggest that neutrophils also have an important role maintaining homeostasis in individual organs and can protect the vascular system. Furthermore, studies using high-dimensional omics technologies have advanced our understanding of neutrophil diversity, and immature neutrophils are receiving new attention in rheumatic diseases including SLE and systemic vasculitis. Developments in genomic, imaging and organoid technologies are beginning to enable more in-depth investigations into the pathophysiology of vascular inflammation in rheumatic diseases, making now a good time to re-examine the full scope of roles of neutrophils in these processes.


Effect of the novel functional fibre, polyglycoplex (PGX), on body weight and metabolic parameters: A systematic review of randomized clinical trials.
Hundreds of dietary supplements are marketed as weight loss pills, but the evidence for effectiveness for most is unproven. The objective of this review was to critically appraise and evaluate the evidence from published randomized clinical trials (RCTs) examining the effectiveness of polyglycoplex (PGX), a novel functional fibre, on body weight and metabolic parameters. We conducted electronic searches in Medline, Embase, Amed, Cinahl and The Cochrane Library. Only double-blinded RCTs were considered for inclusion. The reporting quality of included studies was assessed using the Cochrane criteria. Two reviewers independently determined eligibility, assessed the quality of reporting, and extracted the data. We included four RCTs with a total of 217 participants. The RCTs varied in the quality of their reporting. The evidence from the RCTs suggested that PGX has no significant effects on body weight; however, significant reductions were noted for total and LDL cholesterol. Adverse events reported included diarrhea and abdominal bloating. The evidence from available RCTs does not indicate that PGX intake causes reductions in body weight. PGX may cause reductions in total and LDL cholesterol. Few trials examining the effects of PGX have been conducted; they are characterized by small sample sizes, deficiencies in reporting quality, and are funded by a single manufacturer. Future clinical trials evaluating its effect should be adequately powered and better reported.


A Phase II Trial of First-Line Combination Chemotherapy With Cisplatin, Pemetrexed, and Nivolumab for Unresectable Malignant Pleural Mesothelioma: A Study Protocol.
The purpose of this study is to assess the efficacy and safety of combination chemotherapy with cisplatin, pemetrexed, and nivolumab for unresectable malignant pleural mesothelioma (MPM). Patients with untreated, advanced, or metastatic MPM who meet the inclusion and exclusion criteria will be included. A total of 18 patients will be enrolled from 4 Japanese institutions within 1 year. Combination chemotherapy with cisplatin (75 mg/m2), pemetrexed (500 mg/m2), and nivolumab (360 mg/person) is administered every 3 weeks for a total of 4 to 6 cycles. Then, maintenance therapy with nivolumab will be administered until disease progression, unacceptable toxicities, or the patient's condition meets the withdrawal criteria. The primary end point is the centrally reviewed overall response rate. The secondary end points include the disease control rate, overall survival, progression-free survival, and adverse events. This phase II trial evaluating first-line combination chemotherapy for unresectable MPM commenced in January 2018. This is the first prospective trial to evaluate the effect of an anti-programmed death-1 antibody combined with cisplatin and pemetrexed for unresectable MPM.


Circulating excretory-secretory antigen levels and specific antibody responses in mice infected with Toxocara canis.
Circulatory excretory-secretory antigen levels and IgM and IgG responses to larval antigens were monitored in the serum of 20 BALB/c mice that had been given approximately 500 infective eggs of Toxocara canis by stomach tube. Other groups of mice received different doses of infective eggs, ranging from 5 to 1,250 eggs. Excretory-secretory antigens were collected from culture fluid in which mechanically hatched larvae of T. canis were maintained. An indirect enzyme-linked immunosorbent assay was used to monitor specific antibody responses. Circulating antigen levels were monitored using a direct ELISA which incorporated an IgG fraction of a rabbit antiserum to the excretory-secretory antigens as a capture antibody and a biotin-conjugated form of the same rabbit IgG as the second antibody. The antigen-specific IgM response was evident the first week of infection and peaked 3 to 6 weeks post-infection. The antigen-specific IgG response first appeared the second week of infection and peaked at 6 to 8 weeks post-infection. Both isotype levels stayed near their peak values for the remainder of the study. In the untreated sera, circulating antigen was initially evident and highest the first week of infection; the antigen concentrations dropped by the third month of infection to low, but significant, levels that persisted for the duration of the study. The administration of greater than 25 eggs produced antigenemias. There appeared to be a positive linear trend between the number of eggs given and the amount of antigen in the circulation.


Phasorial differential pulse-width pair technique for long-range Brillouin optical time-domain analysis sensors.
We introduce a novel phasorial differential pulse-width pair (PDPP) method for Brillouin optical time-domain analysis (BOTDA) sensors that combines spatial resolution enhancement with increased tolerance to non-local effects. It is based on the subtraction of the complex time-domain traces supplied by a sensor configuration that uses a phase-modulated probe wave and RF demodulation. The fundamentals of the technique are first described theoretically and using numerical simulation of the propagating waves. Then, proof-of-concept experiments demonstrate the measurement of the Brillouin frequency shift distribution over 50-km. The system is shown to withstand large variations of the pump power generated by its interaction with a powerful probe wave along the fiber; hence, highlighting the potential of the PDPP technique to increase the detected signal-to-noise ratio in long-range BOTDA. Moreover, the PDPP is also shown to increase the measurement contrast by allowing the use of relatively long duration pulses while retaining 1-m spatial resolution.


L-arginine supplementation improves responses to injury and inflammation in dextran sulfate sodium colitis.
Inflammatory bowel disease (IBD), consisting of Crohn's disease and ulcerative colitis (UC), results in substantial morbidity and is difficult to treat. New strategies for adjunct therapies are needed. One candidate is the semi-essential amino acid, L-arginine (L-Arg), a complementary medicine purported to be an enhancer of immunity and vitality in the lay media. Using dextran sulfate sodium (DSS) as a murine colonic injury and repair model with similarities to human UC, we assessed the effect of L-Arg, as DSS induced increases in colonic expression of the y(+) cationic amino acid transporter 2 (CAT2) and L-Arg uptake. L-Arg supplementation improved the clinical parameters of survival, body weight loss, and colon weight, and reduced colonic permeability and the number of myeloperoxidase-positive neutrophils in DSS colitis. Luminex-based multi-analyte profiling demonstrated that there was a marked reduction in proinflammatory cytokine and chemokine expression with L-Arg treatment. Genomic analysis by microarray demonstrated that DSS-treated mice supplemented with L-Arg clustered more closely with mice not exposed to DSS than to those receiving DSS alone, and revealed that multiple genes that were upregulated or downregulated with DSS alone exhibited normalization of expression with L-Arg supplementation. Additionally, L-Arg treatment of mice with DSS colitis resulted in increased ex vivo migration of colonic epithelial cells, suggestive of increased capacity for wound repair. Because CAT2 induction was sustained during L-Arg treatment and inducible nitric oxide (NO) synthase (iNOS) requires uptake of L-Arg for generation of NO, we tested the effect of L-Arg in iNOS(-/-) mice and found that its benefits in DSS colitis were eliminated. These preclinical studies indicate that L-Arg supplementation could be a potential therapy for IBD, and that one mechanism of action may be functional enhancement of iNOS activity.


Central clot score at computed tomography as a predictor of 30-day mortality after acute pulmonary embolism.
The severity of acute pulmonary embolism can be assessed with computed tomography (CT) using clot burden estimation. We compared the existing CT obstruction scores with an in-house developed central clot score for the prediction of 30-day pulmonary embolism (PE)-related mortality. In 125 consecutive patients [47 men, 78 women; mean age +/- standard deviation (SD, 60.4 years +/- 16.6] with acute PE, 2 readers in consensus assessed the severity of PE with 2 existing clot scoring systems (Mastora and Qanadli) and central clot score. The right ventricular dysfunction was assessed by right ventricular diameter (RVD), left ventricular diameter (LVD), ventricular ratio (VR) and septal deviation. Univariate and multivariate regression analysis were performed to correlate these parameters and 30-day PE-related mortality. Ten patients (8%) died of PE within 30 days following CT and 115 patients did not have PE-related death outcome. There was a significant difference in all 3 clot scores, LVD and VR between patients with 30-day PE-related death and those without (P < or =0.001-0.02). Univariate regression analysis showed that all three clot scores and LVD were predictors of PE death, however with multivariate analysis, only central clot score showed significant correlation with 30-day PE death [Odds ratio (OR), 1.1; 96% CI, 1-1.16; P <0.003]. A central clot index of 53% had 100% sensitivity, 76.5% specificity, 23.5% positive predictive value and 98% negative predictive value for 30-day PE death. Central clot score is a strong predictor of 30-day PE death and may therefore allow therapy and risk stratification in patients with acute PE.


The short-term effect of whole-body vibration training on vertical jump, sprint, and agility performance.
Previous studies have suggested that short-term whole-body vibration (WBV) training produces neuromuscular improvement similar to that of power and strength training. However, it is yet to be determined whether short-term WBV exposure produces neurogenic enhancement for power, speed, and agility. The purpose of this study was to investigate the effect short-term WBV training had on vertical jump, sprint, and agility performance in nonelite athletes. Twenty-four sport science students (16 men and 8 women) were randomly assigned to 2 groups: WBV training or control. Each group included 8 men and 4 women. Countermovement jump (CMJ) height, squat jump (SJ) height, sprint speed over 5, 10, and 20 m, and agility (505, up and back) were performed by each participant before and after 9 days of either no training (control) or WBV training. Perceived discomfort of every participant was recorded after daily WBV exposure and nonexposure. There were no significant differences between WBV and control groups for CMJ, SJ, sprints, and agility. Perceived discomfort differed between the first and subsequent days of WBV training (p < 0.05); however, there was no difference between the WBV and control groups. It is concluded that short-term WBV training did not enhance performance in nonelite athletes.


Two-photon and time-resolved fluorescence spectroscopy as probes for structural determination in amyloid-β peptides and aggregates.
The development of new sensitive methods for the detailed collection of conformational and morphological information about amyloids is crucial for the elucidation of critical questions regarding aggregation processes in neurodegenerative diseases. The combined approach of two-photon and time-resolved fluorescence spectroscopy described in this report interrogates the early conformational dynamics seen in soluble oligomers of amyloid-β(1-42). Concentration-dependent aggregation studies using two-photon absorption show enhanced sensitivity toward conformational changes taking place in the secondary structure of the amyloid peptide as aggregation proceeds. Fluorescence lifetimes and changes in anisotropy values indicate Förster-type energy transfer occurring as a function of aggregation state. The sensitivity of our two-photon methodology is compared to that of circular dichroism (CD) spectroscopy, and the results indicate that the two-photon absorption cross-section method exhibits superior sensitivity. A theoretical model is developed that, together with electronic structure calculations, explains the change in cross section as a function of aggregation in terms of interacting transition dipoles for aggregates showing stacked or parallel structures. This suggests that the two-photon method provides a sensitive alternative to CD spectroscopy while avoiding many of the inherent challenges particular to CD data collection. The implication of this finding is significant, as it indicates that a two-photon-based technique used in conjunction with time-resolved fluorescence might be able to reveal answers to conformational questions about amyloid-β(1-42) that are presently inaccessible with other techniques.


Gonadal and Endocrine Analysis of a Gynandromorphic Chicken.
Birds have a ZZ male and ZW female sex chromosome system. The relative roles of genetics and hormones in regulating avian sexual development have been revealed by studies on gynandromorphs. Gynandromorphs are rare bilateral sex chimeras, male on one side of the body and female on the other. We examined a naturally occurring gynandromorphic chicken that was externally male on the right side of the body and female on the left. The bird was diploid but with a mix of ZZ and ZW cells that correlated with the asymmetric sexual phenotype. The male side was 96% ZZ, and the female side was 77% ZZ and 23% ZW. The gonads of this bird at sexual maturity were largely testicular. The right gonad was a testis, with SOX9+ Sertoli cells, DMRT1+ germ cells, and active spermatogenesis. The left gonad was primarily testicular, but with some peripheral aromatase-expressing follicles. The bird had low levels of serum estradiol and high levels of testosterone, as expected for a male. Despite the low percentage of ZW cells on that side, the left side had female sex-linked feathering, smaller muscle mass, smaller leg and spur, and smaller wattle than the male side. This indicates that these sexually dimorphic structures must be at least partly independent of sex steroid effects. Even a small percentage of ZW cells appears sufficient to support female sexual differentiation. Given the lack of chromosome-wide dosage compensation in birds, various sexually dimorphic features may arise due to Z-gene dosage differences between the sexes.


Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors.
Most haematopoietic cells renew from adult haematopoietic stem cells (HSCs), however, macrophages in adult tissues can self-maintain independently of HSCs. Progenitors with macrophage potential in vitro have been described in the yolk sac before emergence of HSCs, and fetal macrophages can develop independently of Myb, a transcription factor required for HSC, and can persist in adult tissues. Nevertheless, the origin of adult macrophages and the qualitative and quantitative contributions of HSC and putative non-HSC-derived progenitors are still unclear. Here we show in mice that the vast majority of adult tissue-resident macrophages in liver (Kupffer cells), brain (microglia), epidermis (Langerhans cells) and lung (alveolar macrophages) originate from a Tie2(+) (also known as Tek) cellular pathway generating Csf1r(+) erythro-myeloid progenitors (EMPs) distinct from HSCs. EMPs develop in the yolk sac at embryonic day (E) 8.5, migrate and colonize the nascent fetal liver before E10.5, and give rise to fetal erythrocytes, macrophages, granulocytes and monocytes until at least E16.5. Subsequently, HSC-derived cells replace erythrocytes, granulocytes and monocytes. Kupffer cells, microglia and Langerhans cells are only marginally replaced in one-year-old mice, whereas alveolar macrophages may be progressively replaced in ageing mice. Our fate-mapping experiments identify, in the fetal liver, a sequence of yolk sac EMP-derived and HSC-derived haematopoiesis, and identify yolk sac EMPs as a common origin for tissue macrophages.


Genetics and biology of pancreatic ductal adenocarcinoma.
With 5-year survival rates remaining constant at 6% and rising incidences associated with an epidemic in obesity and metabolic syndrome, pancreatic ductal adenocarcinoma (PDAC) is on track to become the second most common cause of cancer-related deaths by 2030. The high mortality rate of PDAC stems primarily from the lack of early diagnosis and ineffective treatment for advanced tumors. During the past decade, the comprehensive atlas of genomic alterations, the prominence of specific pathways, the preclinical validation of such emerging targets, sophisticated preclinical model systems, and the molecular classification of PDAC into specific disease subtypes have all converged to illuminate drug discovery programs with clearer clinical path hypotheses. A deeper understanding of cancer cell biology, particularly altered cancer cell metabolism and impaired DNA repair processes, is providing novel therapeutic strategies that show strong preclinical activity. Elucidation of tumor biology principles, most notably a deeper understanding of the complexity of immune regulation in the tumor microenvironment, has provided an exciting framework to reawaken the immune system to attack PDAC cancer cells. While the long road of translation lies ahead, the path to meaningful clinical progress has never been clearer to improve PDAC patient survival.


Timing of stroke in patients undergoing total hip replacement and matched controls: a nationwide cohort study.
Stroke is a potentially fatal complication of total hip replacements (THR). However, timing of stroke in THR patients compared with matched controls and influence of drug use remain unknown. The objective of this study was to determine timing of stroke in patients with THR compared with matched control subjects. A nationwide cohort study was conducted within the Danish registers (1998-2007). Included patients were those with a primary THR in the study period (n=66,583) and were matched by age, sex, and region to three referent subjects without THR or total knee replacements. Time-dependent Cox models were used to derive hazard ratios and were adjusted for disease history and drug use. A 4.7-fold increased risk of ischemic stroke (adjusted hazard ratio, 4.69; 95% CI, 3.12-7.06), and a 4.4-fold increased risk of hemorrhagic stroke (adjusted hazard ratio, 4.40; 95% CI, 2.01-9.62) were found within 2 weeks following THR, compared with matched controls. The risk remained elevated during the first 6 postoperative weeks for ischemic stroke, and the first 12 weeks for hemorrhagic stroke. Outpatient antiplatelet drug use lowered the 6-week hazard ratios for ischemic stroke by 70%, although not affecting risk of hemorrhagic stroke. This study shows, that THR patients have a 4.7-fold increased risk of ischemic stroke, and a 4.4-fold increased risk of hemorrhagic stroke during the first 2 weeks postsurgery. Risk assessment of stroke in individual patients undergoing THR (ie, evaluate other risk factors for stroke) should be considered during the first 6 to 12 weeks.


Calibration and prediction of removal function in magnetorheological finishing.
A calibrated and predictive model of the removal function has been established based on the analysis of a magnetorheological finishing (MRF) process. By introducing an efficiency coefficient of the removal function, the model can be used to calibrate the removal function in a MRF figuring process and to accurately predict the removal function of a workpiece to be polished whose material is different from the spot part. Its correctness and feasibility have been validated by simulations. Furthermore, applying this model to the MRF figuring experiments, the efficiency coefficient of the removal function can be identified accurately to make the MRF figuring process deterministic and controllable. Therefore, all the results indicate that the calibrated and predictive model of the removal function can improve the finishing determinacy and increase the model applicability in a MRF process.


Reasons for nonadherence to the dapivirine vaginal ring: narrative explanations of objective drug-level results.
MTN-020/ASPIRE trial and IPM-027/Ring Study recently proved the dapivirine vaginal ring was safe and effective with consistent use. To optimize the ring's impact, the barriers and facilitators to ring adherence must be understood and addressed. Former ASPIRE participants were stratified by age group (18-21; 22-45) and randomly selected at seven sites in Malawi, South Africa, Uganda and Zimbabwe, 12-17 months after trial exit. Using in-depth interviews or focus group discussions, ring use barriers were explored using structured guides and visual tools including individual-level depictions of dapivirine levels detected in plasma and returned rings. A total of 187 were enrolled; 37% were 18-21 years when they began ASPIRE. Most (75%) had drug-level results, suggesting inconsistent ring use throughout ASPIRE. Participants viewed themselves as adherent, while simultaneously describing regular instances and reasons for ring removal (e.g. for sex or menses). Less adherent women reported fears that partners would oppose the ring or feel it during sex. High adherers expressed altruistic motivations for ring use. Women of all ages attributed young women's nonadherence to their tendency to be less 'serious' about the future, HIV prevention and the study; motivated predominantly by benefits; more fearful of fertility-related consequences; and to having less relationship control. When presented with objective adherence data, participants provided reasons for intermittent ring use, while simultaneously portraying themselves as consistent ring users. Further research is needed to understand how women could use the ring in a way that fits into the context of their relationships and their lives while still conferring adequate HIV prophylaxis.


Intrinsic motivation and amotivation in first episode and prolonged psychosis.
The deleterious functional implications of motivation deficits in psychosis have generated interest in examining dimensions of the construct. However, there remains a paucity of data regarding whether dimensions of motivation differ over the course of psychosis. Therefore, this study examined two motivation dimensions, trait-like intrinsic motivation, and the negative symptom of amotivation, and tested the impact of illness phase on the 1) levels of these dimensions and 2) relationship between these dimensions. Participants with first episode psychosis (FEP; n=40) and prolonged psychosis (n=66) completed clinician-rated measures of intrinsic motivation and amotivation. Analyses revealed that when controlling for group differences in gender and education, the FEP group had significantly more intrinsic motivation and lower amotivation than the prolonged psychosis group. Moreover, intrinsic motivation was negatively correlated with amotivation in both FEP and prolonged psychosis, but the magnitude of the relationship did not statistically differ between groups. These findings suggest that motivation deficits are more severe later in the course of psychosis and that low intrinsic motivation may be partially independent of amotivation in both first episode and prolonged psychosis. Clinically, these results highlight the importance of targeting motivation in early intervention services.


Assessing the publishing priorities and preferences among STEM researchers at a large R1 institution.
The cost of academic publishing has increased substantially despite the ease with which information can be shared on the web. Open Access publishing is a key mechanism for amplifying research access, inclusivity, and impact. Despite this, shifting to a free-to-read publishing environment requires navigating complex barriers that vary by career status and publishing expectations. In this article, we investigate the motivations and preferences of researchers situated within our large research institution as a case study for publishing attitudes at similar institutions. We surveyed the publishing priorities and preferences of researchers at various career stages in STEM fields as they relate to openness, data practices, and assessment of research impact. Our results indicate that publishing preferences, data management experience and research impact assessment vary by career status and departmental approaches to promotion. We find that open access publishing is widely appreciated regardless of career status, but financial limitations and publishing expectations were common barriers to publishing in Open Access journals. Our findings shed light on publishing attitudes and preferences among researchers at a major R1 research institution, and offer insight into advocacy strategies that incentivize open access publishing.


Activities of ceftazidime and avibactam against β-lactamase-producing Enterobacteriaceae in a hollow-fiber pharmacodynamic model.
Avibactam is a novel non-β-lactam β-lactamase inhibitor that is currently undergoing phase 3 clinical trials in combination with ceftazidime. Ceftazidime is hydrolyzed by a broad range of β-lactamases, but avibactam is able to inhibit the majority of these enzymes. The studies described here attempt to provide insight into the amount of avibactam required to suppress bacterial growth in an environment where the concentrations of both agents are varying as they would when administered to humans. Following the simulation of a single intravenous dose of the drug, ceftazidime alone had no effect on any test organism, but a ceftazidime-avibactam combination resulted in rapid killing of all of the strains, with growth suppressed for the 8 h of the study. For seven of eight strains, this was achieved with a 1-g-250-mg profile, but a 2-g-500-mg profile was necessary to completely suppress a high-level-AmpC-producing isolate. When ceftazidime was infused continuously for 24 h with a single bolus dose of avibactam, rapid killing of all of the strains was again observed, with growth suppressed for 10 to >24 h. Regrowth appeared to commence once the avibactam concentration dropped below a critical concentration of approximately 0.3 μg/ml. In a third series of studies, ceftazidime was administered every 8 h for 24 h with avibactam administered at fixed concentrations for short periods during each ceftazidime dose profile. Simulating a 1-g dose of ceftazidime, an avibactam pulse of >0.25 and <0.5 μg/ml was required to suppress growth for 24 h.


Autoreactive cytotoxic T lymphocytes in human immunodeficiency virus type 1-infected subjects.
A subtractive analysis of peptides eluted from major histocompatibility complex (MHC) class I human histocompatibility leukocyte antigen (HLA)-A2.1 molecules purified from either human immunodeficiency virus type-1 (HIV-1)-infected or uninfected cells was performed using micro high-performance liquid chromatography and mass spectrometry. Three peptides unique to infected cells were identified and found to derive from a single protein, human vinculin, a structural protein not known to be involved in viral pathogenesis. Molecular and cytofluorometric analyses revealed vinculin mRNA and vinculin protein overexpression in B and T lymphocytes from HIV-1-infected individuals. Vinculin peptide-specific CTL activity was readily elicited from peripheral blood lymphocytes of the majority of HLA-A2.1+, HIV+ patients tested. Our observations suggest that atypical vinculin expression and MHC class I-mediated presentation of vinculin-derived peptides accompany HIV infection of lymphoid cells in vivo, with a resultant induction of antivinculin CTL in a significant portion of HIV+ (HLA-A2.1+) individuals.


Parent and peer social support compensation and internalizing problems in adolescence.
The general benefits of social support are well-documented; however, little is understood about the unique contributions of social support from specific sources, such as parents and peers. In addition, it is unknown whether social support from some sources might buffer against a lack of social support from another source for the outcome of internalizing problems. The current study investigated two research questions: (a) What is the association between social support from mothers, fathers, classmates, and close friends and internalizing problems for adolescent boys and girls? and (b) Can social support from one source (e.g., mothers) buffer against internalizing problems when social support from another source (e.g., fathers) may be lacking? Do these associations vary by gender? These research questions were examined with a sample of 364 middle school students (61.3% female). Students completed a series of self-report questionnaires assessing social support and internalizing symptoms. Multiple group structural equation models indicated that social support provided general benefits from all sources for early adolescent boys and girls. Regarding stress-buffering with low support as a stressor, no evidence was found for compensation between mothers and fathers. Mother support compensated for low classmate support for both boys and girls and father support compensated for low classmate support for boys. Classmate support compensated for low close friend support for boys. Implications in the context of the school environment for both boys and girls are discussed.


Performance of an adipokine pathway-based multilocus genetic risk score for prostate cancer risk prediction.
Few biomarkers are available to predict prostate cancer risk. Single nucleotide polymorphisms (SNPs) tend to have weak individual effects but, in combination, they have stronger predictive value. Adipokine pathways have been implicated in the pathogenesis. We used a candidate pathway approach to investigate 29 functional SNPs in key genes from relevant adipokine pathways in a sample of 1006 men eligible for prostate biopsy. We used stepwise multivariate logistic regression and bootstrapping to develop a multilocus genetic risk score by weighting each risk SNP empirically based on its association with disease. Seven common functional polymorphisms were associated with overall and high-grade prostate cancer (Gleason≥7), whereas three variants were associated with high metastatic-risk prostate cancer (PSA≥20 ng/mL and/or Gleason≥8). The addition of genetic variants to age and PSA improved the predictive accuracy for overall and high-grade prostate cancer, using either the area under the receiver-operating characteristics curves (P<0.02), the net reclassification improvement (P<0.001) and integrated discrimination improvement (P<0.001) measures. These results suggest that functional polymorphisms in adipokine pathways may act individually and cumulatively to affect risk and severity of prostate cancer, supporting the influence of adipokine pathways in the pathogenesis of prostate cancer. Use of such adipokine multilocus genetic risk score can enhance the predictive value of PSA and age in estimating absolute risk, which supports further evaluation of its clinical significance.


Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast.
The dengue virus type 2 genomic RNA was amplified by reverse transcription-PCR and cloned as four cDNA fragments. We could not assemble these four fragments into full-length cDNA in Escherichia coli. The full-length dengue virus cDNA was constructed by homologous recombination in yeast, either as part of a yeast artificial chromosome or in a yeast-E. coli shuttle vector. Full-length cDNA clones were propagated once in E. coli to prepare useful quantities of DNA. In vitro transcription of these clones produced full-length RNA transcripts. Introduction of these transcripts into LLC-MK2 cells produced typical dengue infection, as judged by cytopathic effects and indirect immunofluorescence. Infectivity was sensitive to RNase digestion and was dependent on the presence of cap analog in the transcription reaction mixture. Virus in the medium was passaged on C6-36 cells to produce stocks, and these stocks had titers and plaque morphologies similar to those of the parental dengue virus type 2. Intracellular dengue virus RNA from cells infected with transcript-derived virus contained an introduced BstEII site, proving that infectivity was derived from RNA transcripts and not from contamination with parental dengue virus. Transcript-derived virus was comparable to dengue virus type 2 for growth and protein expression in tissue culture cells. Sequence analysis of the dengue virus cDNA in one full-length clone revealed only one unexpected silent mutation. By using yeast technology, it will be easy to introduce specific mutations into the dengue virus cDNA, allowing analysis of the virus phenotype in cells transfected with mutant transcripts.


Certolizumab pegol (CDP870) for rheumatoid arthritis in adults.
Tumour necrosis factor (TNF)-alpha inhibitors are beneficial for the treatment of rheumatoid arthritis (RA) for reducing the risk of joint damage, improving physical function and improving the quality of life. This review is an update of the 2014 Cochrane Review of the treatment of RA with certolizumab pegol. To assess the clinical benefits and harms of certolizumab pegol (CZP) in people with RA who have not responded well to conventional disease-modifying anti-rheumatic drugs (DMARDs). We searched the Cochrane Central Register of Controlled Trials (CENTRAL: Cochrane Library 2016, Issue 9), MEDLINE, Embase, Web of Knowledge, reference lists of articles, clinicaltrials.gov and ICTRP of WHO. The searches were updated from 2014 (date of the last search for the previous version) to 26 September 2016. Randomised controlled trials that compared certolizumab pegol with any other agent, including placebo or methotrexate (MTX), in adults with active RA, regardless of current or prior treatment with conventional disease-modifying anti-rheumatic drugs (DMARDs), such as MTX. Two review authors independently checked search results, extracted data and assessed trial quality. We resolved disagreements by discussion or referral to a third review author. We included 14 trials in this update, three more than previously. Twelve trials (5422 participants) included measures of benefit. We pooled 11 of them, two more than previously. Thirteen trials included information on harms, (5273 participants). The duration of follow-up varied from 12 to 52 weeks and the range of doses of certolizumab pegol varied from 50 to 400 mg given subcutaneously. In Phase III trials, the comparator was placebo plus MTX in seven trials and placebo in five. In the two Phase II trials the comparator was only placebo.The approved dose of certolizumab pegol, 200 mg every other week, produced clinically important improvements at 24 weeks for the following outcomes:- American College of Rheumatology (ACR) 50% improvement (pain, function and other symptoms of RA): 25% absolute improvement (95% confidence interval (CI) 20% to 33%); number need to treat for an additional beneficial outcome (NNTB) of 4 (95% CI 3 to 5); risk ratio (RR) 3.80 (95% CI 2.42 to 5.95), 1445 participants, 5 studies.- The Health Assessment Questionnaire (HAQ): -12% absolute improvement (95% CI -9% to -14%); NNTB of 8 (95% CI 7 to 11); mean difference (MD) - 0.35 (95% CI -0.43 to -0.26; 1268 participants, 4 studies) (scale 0 to 3; lower scores mean better function).- Proportion of participants achieving remission (Disease Activity Score (DAS) < 2.6) absolute improvement 10% (95% CI 8% to 16%); NNTB of 8 (95% CI 6 to 12); risk ratio (RR) 2.94 (95% CI 1.64 to 5.28), 2420 participants, six studies.- Radiological changes: erosion score (ES) absolute improvement -0.29% (95% CI -0.42% to -0.17%); NNTB of 6 (95% CI 4 to 10); MD -0.67 (95% CI -0.96 to -0.38); 714 participants, two studies (scale 0 to 230), but not a clinically important difference.-Serious adverse events (SAEs) were statistically but not clinically significantly more frequent for certolizumab pegol (200 mg every other week) with an absolute rate difference of 3% (95% CI 1% to 4%); number needed to treat for an additional harmful outcome (NNTH) of 33 (95% CI 25 to 100); Peto odds ratio (OR) 1.47 (95% CI 1.13 to 1.91); 3927 participants, nine studies.There was a clinically significant increase in all withdrawals in the placebo groups (for all doses and at all follow-ups) with an absolute rate difference of -29% (95% CI -16% to -42%), NNTH of 3 (95% CI 2 to 6), RR 0.47 (95% CI 0.39 to 0.56); and there was a clinically significant increase in withdrawals due to adverse events in the certolizumab groups (for all doses and at all follow-ups) with an absolute rate difference of 2% (95% CI 0% to 3%); NNTH of 58 (95% CI 28 to 329); Peto OR 1.45 (95% CI 1.09 to 1.94) 5236 participants Twelve studies.We judged the quality of evidence to be high for ACR50, DAS remission, SAEs and withdrawals due to adverse events, and moderate for HAQ and radiological changes, due to concerns about attrition bias. For all withdrawals we judged the quality of evidence to be moderate, due to inconsistency. The results and conclusions did not change from the previous review. There is a moderate to high certainty of evidence from randomised controlled trials that certolizumab pegol, alone or combined with methotrexate, is beneficial in the treatment of RA for improved ACR50 and health-related quality of life, an increased chance of remission of RA, and reduced joint damage as seen on x-ray. Fewer people stopped taking their treatment, but most of these who did stopped due to serious adverse events. Adverse events were more frequent with active treatment. We found a clinically but not statistically significant risk of serious adverse events.


Using mirror visual feedback and virtual reality to treat fibromyalgia.
Fibromyalgia is a condition characterized by long term body-wide pain and tender points in joints, muscles and soft tissues. Other symptoms include chronic fatigue, morning stiffness, and depression. It is well known that these symptoms are exacerbated under periods of high stress. When pain becomes severe enough, the mind can enter what is known as a dissociative state, characterized by depersonalization - the feeling of detachment from one's physical body and the illusion of watching one's physical body from outside. In evolutionary terms, dissociative states are thought to be an adaptive mechanism to mentally distance oneself from pain, often during trauma. Similar dissociative experiences are reported by subjects who have used psychoactive drugs such as ketamine. We have previously used non-invasive mirror visual feedback to treat subjects with chronic pain from phantom limbs and suggested its use for complex regional pain syndrome: once considered intractable pain. We wondered whether such methods would work to alleviate the chronic pain of fibromyalgia. We tested mirror visual feedback on one fibromyalgia patient. On 15 trials, the patient's lower limb pain rating (on a scale from 1 to 10) decreased significantly. These preliminary results suggest that non-invasive dissociative anesthetics such as VR goggles, ketamine, and mirror visual feedback could be used to alleviate chronic pain from fibromyalgia. This would furnish us with a better understanding of the mechanism by which external visual feedback interacts with the internal physical manifestation of pain.


Maternal alcohol ingestion reduces surfactant protein A expression by preterm fetal lung epithelia.
In addition to neurodevelopmental effects, alcohol consumption at high levels during pregnancy is associated with immunomodulation and premature birth. Premature birth, in turn, is associated with increased susceptibility to various infectious agents such as respiratory syncytial virus (RSV). The initial line of pulmonary innate defense includes the mucociliary apparatus, which expels microorganisms trapped within the airway secretions. Surfactant proteins A and D (SP-A and SP-D, respectively) are additional components of pulmonary innate immunity and have an important role in pulmonary defense against inhaled pathogens. The purpose of this study was to determine if chronic alcohol consumption during the third trimester of pregnancy alters the function of the mucociliary apparatus and expression of SP-A and SP-D of fetal lung epithelia. Sixteen, date-mated ewes were assigned to two different groups; an ethanol-exposed group in which ewes received ethanol through surgically implanted intra-abomasal cannula during the third trimester of pregnancy, and a control group in which ewes received the equivalent amount of water instead of ethanol. Within these two groups, ewes were further randomly assigned to a full-term group in which the lambs were naturally delivered, and a preterm group in which the lambs were delivered prematurely via an abdominal incision and uterotomy. Ethanol was administered five times a week as a 40% solution at 1g/kg of body weight. The mean maternal serum alcohol concentration measured 6h postadministration was 16.3+/-4.36 mg/dl. Tracheas from six full-term lambs were collected to assess ciliary beat frequency (CBF). The lung tissue from all (24) lambs was collected for immunohistochemistry analysis of SP-A and SP-D protein production and fluorogenic real-time quantitative polymerase chain reaction analysis of SP-A and SP-D mRNA levels. Exposure to ethanol during pregnancy significantly blocked stimulated increase in CBF through ethanol-mediated desensitization of cAMP-dependent protein kinase. In addition, preterm born/ethanol-exposed lambs showed significantly decreased SP-A mRNA expression when compared with the preterm born/control group (P=.004); no significant changes were seen with SP-D. The full-term/ethanol-exposed lambs had no significant alterations in mRNA levels, but had significantly less detectable SP-A protein when compared with the full-term/control lambs (P=.02). These findings suggest that chronic maternal ethanol consumption during the third trimester of pregnancy alters innate immune gene expression in fetal lung. These alterations may underlie increased susceptibility of preterm infants, exposed to ethanol in utero, to RSV and other microbial agents.


1H- and 31P-NMR studies of ditercalinium binding to a d(GCGC)2 and d(CCTATAGG)2 minihelices: a sequence specificity study.
The structures of the complexes formed in aqueous solution between ditercalinium, a bis-intercalating drug, and both the self-complementary tetranucleotide d(GCGC)2 and octanucleotide d(CCTATAGG)2, have been investigated by 400-MHz 1H-nmr and 162-MHz 31P-nmr. All the nonexchangeable protons, as well as the exchangeable imino protons and the phosphorus signals, have been assigned. Both oligonucleotides have been shown to adopt a right-handed B-DNA type structure. The addition of ditercalinium to the oligonucleotides lead to the formation of complexes in slow exchange at the nmr time scale with the free helices. At all drug-to-helix ratios studied, the ditercalinium was found in the bound form, whereas free and complexed oligonucleotides were in slow exchange, allowing resonance assignments through two-dimensional chemical exchange experiments. for d(GCGC)2 the strong upfield shifts induced on all aromatic protons of both the bases and the drug by complexation with ditercalinium suggest an interaction by intercalation of the two rings. However, the loss of twofold symmetry upon binding, as well as the chemical shift variation of the drug proton signals of one of the chromophores with temperature and concentration, favor a model in which the drug-nucleotide complexes have one ring of the drug intercalated and the other stacked on top of the external base pair. The intermolecular contacts between drug protons and nucleotide protons give a defined geometry for complexation that is consistent with the proposed model. In contrast, with d(CCTATAGG)2 several drug-nucleotide complexes were formed and a large increase in line broadening was observed at high drug-to-DNA ratios, precluding a detailed analysis of these complexes. However, the large upfield shift in the imino proton resonances together with the shielding of the ditercalinium ring protons favor a model with bis-intercalation of ditercalinium. This model is supported by the downfield shift of at least 4 out of 14 phosphorus signals. The results are compared with those obtained on ditercalinium binding to the homologous sequences d(CGCG)2 and d(TTCGCGAA)2, and discussed in terms of sequence specificity.


Estimation of the In Vivo MIC of Cipargamin in Uncomplicated Plasmodium falciparum Malaria.
The MIC of an antimalarial drug for a particular infection is the drug level associated with a net parasite multiplication rate of one per asexual cycle. To ensure the cure of malaria, the MIC must be exceeded until all parasites have been eliminated. The development of highly sensitive and accurate PCR quantitation of low-density malaria parasitemia enables the prospective pharmacokinetic-pharmacodynamic (PK-PD) characterization of antimalarial drug effects and now allows identification of the in vivo MIC. An adaptive design and a PK-PD modeling approach were used to determine prospectively the MIC of the new antimalarial cipargamin (KAE609) in adults with uncomplicated Plasmodium falciparum malaria in an open-label, dose-ranging phase 2a study. Vietnamese adults with acute P. falciparum malaria were allocated sequentially to treatment with a single 30-mg (n = 6), 20-mg (n = 5), 10-mg (n = 7), or 15-mg (n = 7) dose of cipargamin. Artemisinin-based combination therapy was given after parasite densities had fallen and then risen as cipargamin levels declined below the MIC but before a return of signs or symptoms. The rates of parasite clearance were dose dependent, with near saturation of the effect being seen at an adult dose of 30 mg. The developed PK-PD model accurately predicted the therapeutic responses in 23/25 patients. The predicted median in vivo MIC was 0.126 ng/ml (range, 0.038 to 0.803 ng/ml). Pharmacometric characterization of the relationship between antimalarial drug concentrations and parasite clearance rates following graded subtherapeutic antimalarial drug dosing is safe and provides a rational framework for dose finding in antimalarial drug development. (This study has been registered at ClinicalTrials.gov under identifier NCT01836458.).


Disability and health outcomes in geospatial analyses of Southeastern U.S. county health data.
People with disabilities tend to be at risk for secondary conditions. There is a need for comprehensive disability and health databases, including geographic information systems to evaluate trends in health, functioning, and employment. We evaluated county levels in morbidity and mortality across the Southeastern United States using spatial regression, examining 2015 trends in accordance with Healthy People 2020 objectives. We merged 2015 National County Health Rankings and the 2015 Social Security Administration's Report on SSDI Beneficiaries, all for n = 1387 Southeastern U.S. county units. We used GeoDa to regress health and disability multivariable models for the dependent variable, age-adjusted Years of Potential Life Lost (YPLL) per 100,000 population. The principal Health/Demographic multivariable model of factors impacting YPLL yielded an adjusted R2 = 0.743 (F = 188.3, p < 0.001) with percentage physically inactive, preventable hospital stays, percentage diabetics, and low college attendance figuring prominently. A Socioeconomic/Demographic multivariable model impacting YPLL yielded R2 = 0.631 (F = 156.0, p < 0.001), with disability and percentage unemployment being major associated variables. For the Southeastern U.S., counties with higher prevalence of SSDI disability workers correlated with significantly higher YPLL and poorer health outcomes. The research augments CDC Disability and Health GIS systems to measure Healthy People 2020 outcomes for persons with disabilities nationwide. Spatial regression represents a robust approach for improved analysis of geographic data for population health measures.


Slit Injection of 1,3-Dichloropropene for Management of Belonolaimus longicaudatus on Established Bermudagrass.
Belonolaimus longicaudatus is a serious problem on bermudagrass, a common warm-season turfgrass, in Florida. The cancellation of organophosphate nematicides necessitates that new management tools be identified for use on sports turf. Postplant application of 1,3-dichloropropene (1,3-D) on bermudagrass was evaluated for management of B. longicaudatus on golf course fairways and driving ranges. A series of 10 experiments were conducted to evaluate the effectiveness of 1,3-D in reducing population densities of B. longicaudatus and enhancing bermudagrass recovery from nematode damage. In 5 of 10 experiments, 1,3-D injected at 46.8 liters/ha was effective in reducing population densities of B. longicaudatus (P < 0.05) compared to untreated plots 2 to 4 weeks after treatment. One month after treatment, population densities of B. longicaudatus ranged from 59% to 97% of those in untreated plots. Nematode suppression generally lasted 2 months or less. Turf visual performance was improved following injection with 1,3-D (P < 0.05) over untreated plots when other factors were not limiting. Turf root development also was enhanced following injection with 1,3-D. Postplant injection of 1,3-D could be a useful nematode management tool for certain sports turf applications.


Dynamics of gene expression patterns during early development of the European seabass (Dicentrarchus labrax).
Larval and embryonic stages are the most critical period in the life cycle of marine fish. Key developmental events occur early in development and are influenced by external parameters like stress, temperature, salinity, and photoperiodism. Any failure may cause malformations, developmental delays, poor growth, and massive mortalities. Advanced understanding of molecular processes underlying marine larval development may lead to superior larval rearing conditions. Today, the new sequencing and bioinformatic methods allow transcriptome screens comprising messenger (mRNA) and microRNA (miRNA) with the scope of detecting differential expression for any species of interest. In the present study, we applied Illumina technology to investigate the transcriptome of early developmental stages of the European seabass (Dicentrarchus labrax). The European seabass, in its natural environment, is a euryhaline species and has shown high adaptation processes in early life phases. During its embryonic and larval phases the European seabass lives in a marine environment and as a juvenile it migrates to coastal zones, estuaries, and lagoons. Investigating the dynamics of gene expression in its early development may shed light on factors promoting phenotypic plasticity and may also contribute to the improvement and advancement of rearing methods of the European seabass, a species of high economic importance in European and Mediterranean aquaculture. We present the identification, characterization, and expression of mRNA and miRNA, comprising paralogous genes and differentially spliced transcripts from early developmental stages of the European seabass. We further investigated the detection of possible interactions of miRNA with mRNA.


Antimalarial and antitubercular nostocarboline and eudistomin derivatives: synthesis, in vitro and in vivo biological evaluation.
The synthesis of nine nostocarboline derivatives with substitutions of the 2-methyl group by alkyl, aryl and functionalized residues, 10 symmetrical bis cationic dimers linking 6-Cl-norharmane through the 2-position and fifteen derivatives of the marine alkaloids eudistomin N and O is reported. These compounds were evaluated in vitro against four parasites (Trypanosoma brucei rhodesiense STIB 900, Trypanosoma cruzi Tulahuen C2C4, Leishmania donovani MHOM-ET-67/L82 axenic amastigotes, and Plasmodium falciparum K1 strain), against Mycobacterium tuberculosis H37Rv, Mycobacterium smegmatis mc(2)155 and Corynebacterium glutamicum ATCC13032, and cytotoxicity was determined against L6 rat myoblast cells. Nostocarboline and derivatives displayed potent and selective in vitro inhibition of P. falciparum with weak cytotoxicity. The dimers displayed submicromolar inhibition of L. donovani and T. brucei, and nanomolar activity against P. falciparum, albeit with pronounced cytotoxicity. One dimer showed a MIC(99) value against M. tuberculosis of 2.5 microg/ml. The alkylated eudistomin N and O derivatives displayed activities down to 18 nM against P. falciparum for N-Me Eudistomin N. Four dimers, nostocarboline and three eudostomin derivatives were evaluated in an in vivo Plasmodium berghei mouse model. No significant activity was observed for the dimers, but a 50% reduction in parasitaemia was observed at 4 x 50 mg/kg ip for nostocarboline.


P2X7 receptor antagonism increases regulatory T cells and reduces clinical and histological graft-versus-host disease in a humanised mouse model.
Graft-versus-host disease (GVHD) is a severe inflammatory response arising from allogeneic haematopoietic stem cell transplantation. Previous studies revealed that antagonism of the P2X7 receptor with Brilliant Blue G (BBG) reduced liver GVHD but did not alter clinical GVHD in a humanised mouse model. Therefore, the present study aimed to trial a modified injection regime using more frequent dosing of BBG to improve outcomes in this model of GVHD. NOD-scid IL2Rγnull (NSG) mice were injected intraperitoneally (i.p.) with 10 × 106 human peripheral blood mononuclear cells (hPBMCs) (day 0), then daily with BBG (50 mg/kg) or saline (days 0-10). BBG significantly reduced clinical score, mortality and histological GVHD compared with saline treatment (endpoint). BBG significantly increased proportions of human regulatory T cells (Tregs) and human B cells and reduced serum human interferon-γ compared with saline treatment prior to development of clinical GVHD (day 21). To confirm the therapeutic benefit of P2X7 antagonism, NSG mice were injected i.p. with 10 × 106 hPBMCs (day 0), then daily with pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) (300 mg/kg) or saline (days 0-10). PPADS increased human Treg proportions compared with saline treatment (day 21), but potential clinical benefits were confounded by increased weight loss with this antagonist. To investigate the role of P2X7 antagonism on Treg survival, hPBMCs were cultured in reduced serum conditions to promote cell death. BBG increased proportions of Tregs (and B cells) compared with saline under these conditions. In conclusion, P2X7 antagonism reduces clinical and histological GVHD in a humanised mouse model corresponding to an increase in human Tregs.


A modified method for whole exome resequencing from minimal amounts of starting DNA.
Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes.


A novel composite of molecularly imprinted polymer-coated PdNPs for electrochemical sensing norepinephrine.
A novel composite of molecularly imprinted polymer-coated palladium nanoparticles (MIP-coated PdNPs) was synthesized by sol-gel method using norepinephrine as template, phenyl trimethoxysilane as functional monomer and tetramethoxysilane as crosslinker. The combination of PdNPs and silica-based MIP endowed the composite with good electrochemical catalytic property, large surface area and template selectivity. MIP-coated PdNPs were characterized by Fourier transform infrared spectroscopy and Transmission electron microscopy. Then MIP-coated PdNPs composite was used as a recognition element in the construction of an electrochemical sensor for norepinephrine. The properties of MIP-coated PdNPs sensor such as special binding, adsorption dynamics and selective recognition ability were evaluated by differential pulse voltammetry. The results demonstrated that MIP-coated PdNPs sensor not only possessed a short response time, but also high binding capacity for norepinephrine, which enabled the imprinted sensor with higher current response than that of non-imprinted material and MIP without PdNPs. In addition, the MIP-coated PdNPs sensor exhibited selectivity for norepinephrine in comparison to other analogs. The MIP-coated PdNPs sensor had a wide linear range over norepinephrine concentration from 0.5 to 80.0μM with a detection limit of 0.1μM. The MIP-coated PdNPs sensor was proved to be a suitable sensing tool for the fast, sensitive and selective determination of norepinephrine in injection and urine samples.


Quality indicators for enteral and parenteral nutrition therapy: application in critically ill patients "at nutritional risk".
Quality Indicators for Nutritional Therapy (QINT) allow a practical assessment of nutritional therapy (NT) quality. To apply and monitor QINT for critically ill patients at nutritional risk. Cross sectional study including critically ill patients > 18 years old, at nutritional risk, on exclusive enteral (ENT) or parenteral nutritional therapy (PNT) for > 72 hours. After three consecutive years, 9 QINT were applied and monitored. Statistical analysis was performed with SPSS version 17.0. A total of 145 patients were included, 93 patients were receiving ENT, among then 65% were male and the mean age was 55.7 years (± 17.4); 52 patients were receiving PNT, 67% were male and the mean age was 58.1 years (± 17.4). All patients (ENT and PNT) were nutritionally screened at admission and their energy and protein needs were individually estimated. Only ENT was early initiated, more than 70% of the prescribed ENT volume was infused and there was a reduced withdrawal of enteral feeding tube. The frequency of diarrhea episodes and digestive fasting were not adequate in ENT patients. The proper supply of energy was contemplated only for PNT patients and there was an expressive rate of oral intake recovery in ENT patients. After three years of research, the percentage of QINT adequacy varied between 55%-77% for ENT and 60%-80% for PNT. The results were only made possible by the efforts of a multidisciplinary team and the continuous re-evaluation of the procedures in order to maintain the nutritional assistance for patients at nutritional risk.


Socio-demographic and clinical profile of substance abuse patients admitted to an emergency department in Ankara, Turkey.
Illicit drug abuse is an important health problem around the world. Cannabis is the most commonly used illicit drug, followed by amphetamines, opioids, and cocaine. In this study, we searched the socio-demographic features of drug abusers admitted to an emergency department. The study was performed in the emergency department of an education and training hospital in Ankara, Turkey. The socio-demographic features, substances commonly consumed, and the reason for the patients' presentation were recorded. The mean age of patients was 28.3±9.2. Of those under study, 93.3% were male; 68.7% were discharged from the emergency department after a short observation period; and 10.4% were hospitalized. The median age of the patients' first experience with illicit substances was 19 (4-56). Illicit drug use remains a significant medical health problem, not only in Turkey but also around the world. Abusers frequently seek emergency services, complaining of various symptoms, but generally do not mention their substance abuse. Emergency physicians must ask patients if they use abuse drugs.


Surface lymphotoxin alpha/beta complex is required for the development of peripheral lymphoid organs.
For more than a decade, the biological roles and the apparent redundancy of the cytokines tumor necrosis factor (TNF) and lymphotoxin (LT) have been debated. LT alpha exists in its soluble form as a homotrimer, which like TNF only binds the TNF receptors, TNF-R55 or TNF-R75. The cell surface form of LT exists as a heteromer of LT alpha and LT beta subunits and this complex specifically binds the LT beta receptor (LT beta-R). To discriminate the functions of the LT and TNF systems, soluble LT beta-R-immunoglobulin (Ig) or TNF-R-Ig fusion proteins were introduced into embryonic circulation by injecting pregnant mice. Exposure to LT beta-R-Ig during gestation disrupted lymph node development and splenic architecture in the progeny indicating that both effects are mediated by the surface LT alpha/beta complex. These data are the first to identify a cell surface ligand involved in immune organ morphogenesis. Moreover, they unambiguously discriminate the functions of the various TNF/LT ligands, provide a unique model to study compartmentalization of immune responses and illustrate the generic utility of receptor-Ig fusion proteins for dissecting/ordering ontogenetic events in the absence of genetic modifications.


B-type natriuretic peptide and adiponectin releases in rat model of myocardial damage induced by isoproterenol administration.
B-type natriuretic peptide (BNP) and adiponectin play important role in the cardiovascular homeostasis regulation. We investigated BNP and adiponectin serum levels followed by isoproterenol (ISO) administration to rats and explored the relationship between them. Cardiac troponin I (cTnI) blood level was used as biochemical evidence of myocardial damage development. Adult male Wistar rats (average body weight 273.33 ± 21.63 g) were distributed into groups: control group received saline (n=6) and ISO groups (n=12) treated with ISO (subcutaneous single dose 100 mg/kg of rat body weight). ISO group was divided into two groups according to the time of BNP, adiponectin and cTnI determination: ISO I (n=6; 2 hours after ISO administration); ISO II (n=6; 4 hours after ISO administration). Blood for determination of parameters was taken from rat abdominal aorta. BNP, adiponectin and cTnI were determined by ELISA method. Data were statistically analysed by using SPSS version 13 computer program. P value less 0.05 was considered statistically significant. Blood BNP and adiponectin were lower at 2 hours after ISO administration in comparison with control group (p=0.004 for BNP and p=0.174 for adiponectin). Four hours after ISO administration, we have noted significant elevation of both parameters compared to ISO I group (p=0.004 for BNP; p=0.02 for adiponectin). Test of correlation have showed significant relation between their blood levels during experimental period (rho=0.577; p=0.01). BNP and adiponectin are not simple indicators of myocardial damage development. They have possible associated and additive effects in cardiovascular homeostasis regulation.


Integrated side-stream reactor for biological nutrient removal and minimization of sludge production.
Integrated processes to reduce in situ the sludge production in wastewater treatment plants are gaining attention in order to facilitate excess sludge management. In contrast to post-treatments, such as anaerobic digestion which is placed between the activated sludge system and dewatering processes, integrated technologies are placed in the sludge return line. This study evaluates the application of an anoxic side-stream reactor (SSR) which creates a physiological shock and uncouples the biomass metabolism and diverts the activity from assimilation for biosynthesis to non-growth activities. The effect of this system in biological nutrient removal for both nitrogen and phosphorus was evaluated for the anaerobic, anoxic and aerobic reactors. The RedOx potential within the SSR was maintained at -150 mV while the sludge loading rate was modified by increasing the percentage of recycled activated sludge feed to the SSR (0 and 40% at laboratory scale and 0, 10, 50 and 100% at pilot scale). The use of the SSR presented a slight reduction of phosphorus removal but maintained the effluent quality to the required discharge values. Nitrogen removal efficiency increased from 75 to 86% while reducing the sludge production rate by 18.3%.


Tuberous sclerosis complex and epilepsy: prognostic significance of electroencephalography and magnetic resonance imaging.
Tuberous sclerosis complex is a disease that affects many organs, including the central nervous system. Nervous system involvement in the form of hamartomas often results in seizures. In this study we wanted to determine the outcome of epilepsy in tuberous sclerosis complex and determine whether interictal electroencephalograms (EEGs) and hamartoma burden as seen with magnetic resonance imaging (MRI) are predictive of degree of seizure control. The study population consisted of 30 patients. For each patient two sets of EEG and MRI data, separated by at least 12 months, and information on seizure frequency at time of data collection were obtained. Sensitivity, specificity, and positive and negative predictive values of various EEG and MRI findings were determined. Seizure control improved in 20 and worsened in 10 patients. In relation to seizure control, the specificity of an abnormal sleep EEG and the positive predictive value of normal sleep EEG were 100%. MRI and EEG background were neither sensitive nor specific for predicting seizure control. A majority of children with tuberous sclerosis complex can achieve good seizure control. The sleep EEG is helpful in predicting eventual seizure control.


Pregnancy outcome in thoracic aortic disease data from the Registry Of Pregnancy And Cardiac disease.
Cardiovascular disease is the leading cause of death during pregnancy with thoracic aortic dissection being one of the main causes. Thoracic aortic disease is commonly related to hereditary disorders and congenital heart malformations such as bicuspid aortic valve (BAV). Pregnancy is considered a high risk period in women with underlying aortopathy. The ESC EORP Registry Of Pregnancy And Cardiac disease (ROPAC) is a prospective global registry that enrolled 5739 women with pre-existing cardiac disease. With this analysis, we aim to study the maternal and fetal outcome of pregnancy in women with thoracic aortic disease. Thoracic aortic disease was reported in 189 women (3.3%). Half of them were patients with Marfan syndrome (MFS), 26% had a BAV, 8% Turner syndrome, 2% vascular Ehlers-Danlos syndrome and 11% had no underlying genetic defect or associated congenital heart defect. Aortic dilatation was reported in 58% of patients and 6% had a history of aortic dissection. Four patients, of whom three were patients with MFS, had an acute aortic dissection (three type A and one type B aortic dissection) without maternal or fetal mortality. No complications occurred in women with a history of aortic dissection. There was no significant difference in median fetal birth weight if treated with a beta-blocker or not (2960 g (2358-3390 g) vs 3270 g (2750-3570 g), p value 0.25). This ancillary analysis provides the largest prospective data review on pregnancy risk for patients with thoracic aortic disease. Overall pregnancy outcomes in women with thoracic aortic disease followed according to current guidelines are good.


Central Sensitization Phenotypes in Post Acute Sequelae of SARS-CoV-2 Infection (PASC): Defining the Post COVID Syndrome.
To develop and implement criteria for description of post COVID syndrome based on analysis of patients presenting for evaluation at Mayo Clinic Rochester between November 2019 and August 2020. A total of 465 patients with a history of testing positive for COVID-19 were identified and their medical records reviewed. After a thorough review, utilizing the DELPHI methods by an expert panel, 42 (9%) cases were identified with persistent central sensitization (CS) symptoms persisting after the resolution of acute COVID-19, herein referred to as Post COVID syndrome (PoCoS). In this report we describe the baseline characteristics of these PoCoS patients. Among these 42 PoCoS patients, the mean age was 46.2 years (median age was 46.5 years). Pain (90%), fatigue (74%), dyspnea (43%), and orthostatic intolerance (38%) were the most common symptoms. The characteristics of an initial 14 patients were utilized for the development of clinical criteria via a modified Delphi Method by a panel of experts in central sensitization disorders. These criteria were subsequently applied in the identification of 28 additional cases of suspected PoCoS. A 2-reviewer system was used to analyze agreement with using the criteria, with all 28 cases determined to be either probable or possible cases by the reviewers. Inter-reviewer agreement using these proposed defining criteria was high with a Cohen's alpha of .88. Here we present what we believe to be the first definitional criteria for Post COVID syndrome. These may be useful in clinical phenotyping of these patients for targeted treatment and future research.


Comparison of fluorescent in situ hybridization and conventional culturing for detection of Helicobacter pylori in gastric biopsy specimens.
In this study, we have investigated 201 gastric biopsy specimens obtained from dyspeptic patients for the presence of Helicobacter pylori. By means of fluorescent in situ hybridization (FISH) with rRNA-targeted fluorescence-labeled oligonucleotide probes specific for H. pylori, this pathogen was detected in 63 biopsy specimens. By using conventional culturing, H. pylori was isolated from 49 of these 63 gastric biopsy specimens. In contrast, FISH failed to identify H. pylori in four samples from which the pathogen was cultured. The lowest sensitivity was obtained by using the urease test. H. pylori was detected indirectly by this method in 43 of 67 biopsy specimens, which were positive for the pathogen as determined by FISH and/or culturing. All 49 H. pylori isolates that were detected by FISH and culturing underwent antimicrobial susceptibility testing for clarithromycin, a macrolide drug that is a key component in the therapy of peptic ulcer disease caused by this pathogen. Clarithromycin susceptibility testing of cultured isolates was carried out by the E-test, whereas FISH was used on biopsy specimens to detect clarithromycin-resistant mutant strains. No discrepancies were found between these two methods. Thirty-seven strains were clarithromycin sensitive, and eight H. pylori isolates were resistant to the macrolide. From another four biopsy specimens, a mixture of clarithromycin-sensitive and -resistant strains was identified by both methods. Thus, FISH is a reliable technique for determining the clarithromycin susceptibility of this pathogen. Taken together, FISH is a more sensitive and rapid technique than culturing for detection of H. pylori in gastric biopsy specimens. However, in the microbiology routine diagnostic laboratory, the combination of both FISH and conventional culturing significantly increases the sensitivity in detection of H. pylori.


Enhanced degradation of softwood versus hardwood by the white-rot fungus Pycnoporus coccineus.
White-rot basidiomycete fungi are potent degraders of plant biomass, with the ability to mineralize all lignocellulose components. Recent comparative genomics studies showed that these fungi use a wide diversity of enzymes for wood degradation. Deeper functional analyses are however necessary to understand the enzymatic mechanisms leading to lignocellulose breakdown. The Polyporale fungus Pycnoporus coccineus BRFM310 grows well on both coniferous and deciduous wood. In the present study, we analyzed the early response of the fungus to softwood (pine) and hardwood (aspen) feedstocks and tested the effect of the secreted enzymes on lignocellulose deconstruction. Transcriptomic and proteomic analyses revealed that P. coccineus grown separately on pine and aspen displayed similar sets of transcripts and enzymes implicated in lignin and polysaccharide degradation. In particular, the expression of lignin-targeting oxidoreductases, such as manganese peroxidases, increased upon cultivation on both woods. The sets of enzymes secreted during growth on both pine and aspen were more efficient in saccharide release from pine than from aspen, and characterization of the residual solids revealed polysaccharide conversion on both pine and aspen fiber surfaces. The combined analysis of soluble sugars and solid residues showed the suitability of P. coccineus secreted enzymes for softwood degradation. Analyses of solubilized products and residual surface chemistries of enzyme-treated wood samples pointed to differences in fiber penetration by different P. coccineus secretomes. Accordingly, beyond the variety of CAZymes identified in P. coccineus genome, transcriptome and secretome, we discuss several parameters such as the abundance of manganese peroxidases and the potential role of cytochrome P450s and pectin degradation on the efficacy of fungi for softwood conversion.


ESCRT Machinery Mediates Cytokinetic Abscission in the Unicellular Red Alga Cyanidioschyzon merolae.
In many eukaryotes, cytokinesis proceeds in two successive steps: first, ingression of the cleavage furrow and second, abscission of the intercellular bridge. In animal cells, the actomyosin contractile ring is involved in the first step, while the endosomal sorting complex required for transport (ESCRT), which participates in various membrane fusion/fission events, mediates the second step. Intriguingly, in archaea, ESCRT is involved in cytokinesis, raising the hypothesis that the function of ESCRT in eukaryotic cytokinesis descended from the archaeal ancestor. In eukaryotes other than in animals, the roles of ESCRT in cytokinesis are poorly understood. To explore the primordial core mechanisms for eukaryotic cytokinesis, we investigated ESCRT functions in the unicellular red alga Cyanidioschyzon merolae that diverged early in eukaryotic evolution. C. merolae provides an excellent experimental system. The cell has a simple organelle composition. The genome (16.5 Mb, 5335 genes) has been completely sequenced, transformation methods are established, and the cell cycle is synchronized by a light and dark cycle. Similar to animal and fungal cells, C. merolae cells divide by furrowing at the division site followed by abscission of the intercellular bridge. However, they lack an actomyosin contractile ring. The proteins that comprise ESCRT-I-IV, the four subcomplexes of ESCRT, are partially conserved in C. merolae. Immunofluorescence of native or tagged proteins localized the homologs of the five ESCRT-III components [charged multivesicular body protein (CHMP) 1, 2, and 4-6], apoptosis-linked gene-2-interacting protein X (ALIX), the ESCRT-III adapter, and the main ESCRT-IV player vacuolar protein sorting (VPS) 4, to the intercellular bridge. In addition, ALIX was enriched around the cleavage furrow early in cytokinesis. When the ESCRT function was perturbed by expressing dominant-negative VPS4, cells with an elongated intercellular bridge accumulated-a phenotype resulting from abscission failure. Our results show that ESCRT mediates cytokinetic abscission in C. merolae. The fact that ESCRT plays a role in cytokinesis in archaea, animals, and early diverged alga C. merolae supports the hypothesis that the function of ESCRT in cytokinesis descended from archaea to a common ancestor of eukaryotes.


Paeoniflorin in experimental BALB/c mansoniasis: A novel anti-angiogenic therapy.
Chronic hepatic schistosomiasis causes portal hypertension, fibrosis and lethal hepatosplenic complications. Previous studies focused mainly on schistosomicidal drugs and neglected the therapeutic approaches against the vascular complications after portal hypertension. Investigating a novel anti-angiogenic therapy is an urgent. The current study is to evaluate the performance of Paeoniflorin (PAE) as an anti-angiogenic therapy, being a powerful anti-fibrotic, compared to artemether (ART) and praziqantel (PZQ) in schistosomiasis mansoni BALB/c mice. Thirty two laboratory bred male BALB/c Swiss albino mice. The mice were classified into four groups (8 mice each), control infected (CI), PZQ (300 mg/kg/12 h), ART (0.1 ml/mg/d) and PAE (50 mg/kg/d) treated groups for one month. All mice groups were sacrificed 15 weeks post infection for assessment of the drugs' efficacy by parasitological, histopathological and immunohistochemical studies. Our results in PAE group showed marked reduction in the mean egg count/gram stool, worm burden, egg count/gram liver tissue, granuloma diameter and pro-angiogenic factors as vascular endothelial growth factor (VEGF), Proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (α-SMA) and CD34; conversely, there was an augmentation of the tissue inhibitor metalloproteinases-2 (TIMP-2) as an anti-angiogenic expression that was exceeded ART and PZQ treated groups compared to CI group (p˂0.001). Conclusively, PAE has an anti-angiogenic impact with no vascular proliferative activity or recanalization, no micro-vessel density (MVD) changes, granuloma resolution and fibrosis regression. PAE is predicted to be a potential therapy for chronic hepatic diseases associated with fibrosis and angiogenesis, hopeful in protecting from advanced serious complications; cancer and metastasis.


Impact of platinum group elements on the soil invertebrate Enchytraeus crypticus.
The platinum group elements (PGE) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree over the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGE on organisms are extremely rare. Enchytraeids are ecologically relevant soil organisms, due to their activity in decomposition and bioturbation in many soil types worldwide. The experiments were carried out as described in the OECD Guideline 220 [CSN EN ISO 16387--Soil quality--Effects of pollutants on Enchytraeidae (Enchytraeus sp.)--Determination of effects on reproduction]. The reproductive effects of platinum (PtCl4), palladium (PdCl2) and rhodium (RhCl3) were examined. The concentrations of PGE tested were as follows: 5, 10, 25, 50 and 100 μmol x L(-1) PdCl2; 50, 100, 150, 200 and 250 μmol x L-1 PtCl4/RhCl3. The EC50 (medium effective concentration) was determined after 28 days of exposure. The inhibition of the reproduction of PGE-exposed enchytraeids was compared against controls. Values of 28dEC50 of PtCl4, PdCl2 and RhCl3 amounted to 161.9 μmol x L(-1), 70.0 μmol x L(-1) and 246.6 μmol x L(-1), respectively. We can confirm that the relative order of toxicities is Pd (II) >Pt(IV) >>Rh(III). To the best of our knowledge, this is the first study to use Enchytraeus crypticus as an indicator species to assess the risk of soil contamination by platinum, palladium and rhodium. Results of this study contribute important data on the ecotoxicity of a rarely studied elements.


The deception and fallacies of sponsored randomized prospective double-blinded clinical trials: the bisphosphonate research example.
The randomized prospective double-blinded clinical trial (RCT) is accepted as Level I evidence and is highly regarded. However, RCTs that gained FDA approval of drugs such as Vioxx, Fen-Phen, and oral and intravenous bisphosphonates have proven to generate misleading results and have not adequately identified serious adverse reactions. The development, research, and clinical marketing of the oral and intravenous bisphosphonates can serve as a representative example for the deteriorated value of many of today's RCTs. The expected high value of RCTs is jeopardized by: (1) sponsorship that incorporates bias; (2) randomization that can select out an expected improved result or eliminate higher-risk individuals; (3) experimental design that can avoid recognition of serious adverse reactions; (4) blinding that can easily become unblinded by the color, shape, odor, or administration requirements of a drug; (5) definitions that can define an observation as something other than what it actually represents, or fail to define it as an adverse reaction; (6) labeling of retrospective data as a prospective trial by using adjudicators prospectively to look at retrospective data; (7) change of the length of study to avoid the longer-term adverse reaction from accumulation of drug or treatment effects; (8) ghost writing, as when drug company physicians or a hired corporation either edit or write the entire protocol and/or manuscript for publication. Such corruption of the well-intended properly conducted RCT should be viewed with a sense of outrage by practitioners and requires a restructuring of the levels of evidence accepted today.


Thermoresponsive double network cryogels from dendronized copolymers showing tunable encapsulation and release of proteins.
A series of double-network cryogels (DNCs) were prepared from oligoethylene glycol (OEG)-based dendronized copolymers as the first network and polyvinyl alcohol (PVA) as the second network. The dendronized copolymers were composed of dendritic OEG units, as well as triethoxysilyl, amine or carboxyl groups. The dendritic OEGs ensured characteristic thermoresponsiveness and triethoxysilyl groups were for crosslinking, while amino and carboxyl groups were designed to provide charged species for specifically interacting with proteins. PVA behaves as a mutual support which ensures the stability and mechanical properties of the cryogels. These DNCs exhibited good thermoresponsiveness and biocompatibility. Upon increasing the temperature above their cloud points (Tcps), the cryogels became dehydrated with enhanced hydrophobicity, and thus their porous structures shrank. This property was utilized to mediate encapsulation and release of proteins. The loading and release efficiencies of different proteins were found to be dominated by hydrophilic to hydrophobic transitions of the first network, and at the same time, by the properties of proteins. At room temperature, the cryogel could efficiently capture model proteins, while above Tcps, more than 80% of lysozyme could be released in 24 h with enzyme activity remaining unchanged. These series of smart cryogels based on thermoresponsive dendronized copolymers may provide promising applications for the reversible capture of enzymes or proteins.


Defined plant extracts can protect human cells against combined xenobiotic effects.
Pollutants representative of common environmental contaminants induce intracellular toxicity in human cells, which is generally amplified in combinations. We wanted to test the common pathways of intoxication and detoxification in human embryonic and liver cell lines. We used various pollutants such as Roundup residues, Bisphenol-A and Atrazine, and five precise medicinal plant extracts called Circ1, Dig1, Dig2, Sp1, and Uro1 in order to understand whether specific molecular actions took place or not. Kidney and liver are major detoxification organs. We have studied embryonic kidney and hepatic human cell lines E293 and HepG2. The intoxication was induced on the one hand by a formulation of one of the most common herbicides worldwide, Roundup 450 GT+ (glyphosate and specific adjuvants), and on the other hand by a mixture of Bisphenol-A and Atrazine, all found in surface waters, feed and food. The prevention and curative effects of plant extracts were also measured on mitochondrial succinate dehydrogenase activity, on the entry of radiolabelled glyphosate (in Roundup) in cells, and on cytochromes P450 1A2 and 3A4 as well as glutathione-S-transferase. Clear toxicities of pollutants were observed on both cell lines at very low sub-agricultural dilutions. The prevention of such phenomena took place within 48 h with the plant extracts tested, with success rates ranging between 25-34% for the E293 intoxicated by Roundup, and surprisingly up to 71% for the HepG2. By contrast, after intoxication, no plant extract was capable of restoring E293 viability within 48 h, however, two medicinal plant combinations did restore the Bisphenol-A/Atrazine intoxicated HepG2 up to 24-28%. The analysis of underlying mechanisms revealed that plant extracts were not capable of preventing radiolabelled glyphosate from entering cells; however Dig2 did restore the CYP1A2 activity disrupted by Roundup, and had only a mild preventive effect on the CYP3A4, and no effect on the glutathione S-transferase. Environmental pollutants have intracellular effects that can be prevented, or cured in part, by precise medicinal plant extracts in two human cell lines. This appears to be mediated at least in part by the cytochromes P450 modulation.


Purpose in life predicts better emotional recovery from negative stimuli.
Purpose in life predicts both health and longevity suggesting that the ability to find meaning from life's experiences, especially when confronting life's challenges, may be a mechanism underlying resilience. Having purpose in life may motivate reframing stressful situations to deal with them more productively, thereby facilitating recovery from stress and trauma. In turn, enhanced ability to recover from negative events may allow a person to achieve or maintain a feeling of greater purpose in life over time. In a large sample of adults (aged 36-84 years) from the MIDUS study (Midlife in the U.S., http://www.midus.wisc.edu/), we tested whether purpose in life was associated with better emotional recovery following exposure to negative picture stimuli indexed by the magnitude of the eyeblink startle reflex (EBR), a measure sensitive to emotional state. We differentiated between initial emotional reactivity (during stimulus presentation) and emotional recovery (occurring after stimulus offset). Greater purpose in life, assessed over two years prior, predicted better recovery from negative stimuli indexed by a smaller eyeblink after negative pictures offset, even after controlling for initial reactivity to the stimuli during the picture presentation, gender, age, trait affect, and other well-being dimensions. These data suggest a proximal mechanism by which purpose in life may afford protection from negative events and confer resilience is through enhanced automatic emotion regulation after negative emotional provocation.


Rapid chromosome evolution in recently formed polyploids in Tragopogon (Asteraceae).
Polyploidy, frequently termed "whole genome duplication", is a major force in the evolution of many eukaryotes. Indeed, most angiosperm species have undergone at least one round of polyploidy in their evolutionary history. Despite enormous progress in our understanding of many aspects of polyploidy, we essentially have no information about the role of chromosome divergence in the establishment of young polyploid populations. Here we investigate synthetic lines and natural populations of two recently and recurrently formed allotetraploids Tragopogon mirus and T. miscellus (formed within the past 80 years) to assess the role of aberrant meiosis in generating chromosomal/genomic diversity. That diversity is likely important in the formation, establishment and survival of polyploid populations and species. Applications of fluorescence in situ hybridisation (FISH) to natural populations of T. mirus and T. miscellus suggest that chromosomal rearrangements and other chromosomal changes are common in both allotetraploids. We detected extensive chromosomal polymorphism between individuals and populations, including (i) plants monosomic and trisomic for particular chromosomes (perhaps indicating compensatory trisomy), (ii) intergenomic translocations and (iii) variable sizes and expression patterns of individual ribosomal DNA (rDNA) loci. We even observed karyotypic variation among sibling plants. Significantly, translocations, chromosome loss, and meiotic irregularities, including quadrivalent formation, were observed in synthetic (S(0) and S(1) generations) polyploid lines. Our results not only provide a mechanism for chromosomal variation in natural populations, but also indicate that chromosomal changes occur rapidly following polyploidisation. These data shed new light on previous analyses of genome and transcriptome structures in de novo and establishing polyploid species. Crucially our results highlight the necessity of studying karyotypes in young (<150 years old) polyploid species and synthetic polyploids that resemble natural species. The data also provide insight into the mechanisms that perturb inheritance patterns of genetic markers in synthetic polyploids and populations of young natural polyploid species.


Partial bit delay correlative modulation used to improve the dispersion tolerance of an optical duobinary system.
Phase effect of the optical duobinary (ODB) modulation on the chromatic dispersion (CD) tolerance is more important than the ODB modulation-induced bandwidth effect. To show this, we evaluate, respectively, the filter bandwidth effect and the phase effect of the partial bit delay correlative modulation (PBDCM) on the CD tolerance. Due to the cancellation between the CD-induced and the PBDCM-induced phase effects, the PBDCM method can increase the CD-limited transmission to 2 ??? 3 times of that using standard 1-bit delay modulation, while the optimized filter bandwidth method can increase the transmission by 50% or less, depending on the input signal format. The PBDCM can be physically realized by adjusting the delay time of the delay-and-add circuit in the conventional duobinary transmitter (without lower-pass filter). These conclusions are also valid for systems having non-negligible polarization-mode dispersion (PMD) and polarization dependent loss (PDL).