Bevacizumab for the treatment of high-grade glioma.
Gliomas are highly vascular and rich in vascular endothelial growth factor (VEGF) that promotes angiogenesis. Bevacizumab is a monoclonal antibody against VEGF inhibiting angiogenesis by preventing receptor activation. Phase II clinical trials using bevacizumab in both newly diagnosed and recurrent high-grade glioma (HGG) showed promising results. This is a review of clinical trials investigating bevacizumab in newly diagnosed and recurrent HGGs with a focus on outcome results. A future perspective about the expected role of bevacizumab is given. Bevacizumab efficacy, safety and tolerability, the combination of radiation and bevacizumab as well as the use of bevacizumab to treat pseudoprogression are discussed. Further criteria of response evaluation needed to be adjusted in the age of anti-angiogenic therapy and this will be discussed. Bevacizumab has been shown to be safe and tolerable in HGG. In the recurrent disease setting, bevacizumab alone might be sufficient for a clinical benefit and is currently approved as a single agent for this indication. While clinical trials demonstrate a prolonged progression-free survival in bevacizumab-treated HGG, a benefit on OS has not been demonstrated yet. Bevacizumab has also been introduced into other settings in neuro-oncology including concurrent administration with re-irradiation for recurrent HGG.


Escitalopram increases cortical nitric oxide synthase (NOS) in rat brain during ethanol withdrawal.
The effect of escitalopram on ethanol withdrawal syndrome (EWS) and involvement of nitric oxide system in rats was investigated. Male Wistar rats divided into five experimental groups of eight animals each: (a) control group; (b) EWS (saline) group; (c) escitalopram 2.5 mg group; (d) escitalopram 5mg group and (e) escitalopram 10 mg group. Ethanol dependence was induced in rats by ethanol-containing liquid diet and ethanol withdrawal was precipitated by replacing ethanol free diet. Ethanol receiving rats in individual groups were decapitated on 21st day of ethanol ingestion and at sixth hour of ethanol withdrawal. Brains were removed and dissected. Five regions of the brain were dissected: the frontal cortex, cerebellum, striatum, hippocampus and hypothalamus. Immunohistochemical NOS staining was performed. The NOS staining intensity in cortex and hypothalamus regions were significantly lower in EWS group than control group. During EWS period, in rats given 2.5 and 10 mg/kg escitalopram, the staining intensity in cortex, striatum and hippocampus were found to be 11.492, 8.519 and 11.234, respectively, and was statistically different than the control group. The hippocampal NOS staining intensity was found to be significantly decreased with 2.5 mg/kg escitalopram, whereas the cortex, striatum and hippocampal staining intensity were increased significantly with 5 mg/kg. In 10 mg/kg escitalopram group, staining properties were not different than those of the control group. Our results suggest that NOS decreases during ethanol withdrawal in cortex and hypothalamus of rat brain and treatment with escitalopram reverses the enzyme density in cortex but not hypothalamus.


[The place of ultrasound diagnosis in urology].
Ultrasonic diagnosis is used in urology, as in other fields, for the differentiation between cystic and solid tumours and in the planning of radiotherapeutic management. A particularly valuable application of ultrasound is in the determination of prostatic size, which provides essential information of which the preoperative decision as to surgical procedure is based. Ultrasonic tomography also has a limited application in the examination of the scrotum.


Studies on the mechanism of polyethylene glycol-mediated cell fusion using fluorescent membrane and cytoplasmic probes.
The mechanism by which polyethylene glycol (PEG) mediates cell fusion has been studied by examining the movements of membrane lipids and proteins, as well as cytoplasmic markers, from erythrocytes to monolayers of cultured cells to which they have been fused. Fluorescence and freeze-fracture electron microscopy and fluorescence recovery after photobleaching have yielded the following results: (a) In the presence of both fusogenic and nonfusogenic PEG membranes are brought together at closely apposed contact regions. (b) Fluorescent lipid probes quickly spread from the membranes of erythrocytes to cultured cells in the presence of both fusogenic and nonfusogenic PEG. (c) Proteins of the erythrocyte membranes were never observed to diffuse into the cultured cell membrane. (d) Water-soluble proteins did not diffuse from the erythrocyte interior into the target cell cytoplasm until the PEG was removed. These data suggest that the coordinate action of two distinct components is necessary for fusion as mediated by PEG. Presumably, the polymer itself promotes close apposition of the adjacent cell membranes but the fusion stimulus is provided by the additives contained in commercial PEG.


Widespread occurrence of ranavirus in pond-breeding amphibian populations.
Ranaviruses are an emerging threat for many amphibian populations, yet their distribution in amphibian communities and the association of infection with possible stressors and species is not fully understood due to historically sparse surveillance. Agricultural practices that reduce the water quality of amphibian breeding habitats (e.g., cattle access to wetlands) and environmental stressors (e.g., lower temperatures) may contribute to ranavirus emergence. We tested larval amphibians for ranavirus infection across four seasons in farm ponds (n = 40) located in Tennessee, USA. Cattle at various densities were allowed access to half of the sampled ponds. Ranavirus infections were detected in nine species and in 33 of the sampled ponds (83%), illustrating widespread occurrence of the pathogen. Species within the family Ranidae were the most frequently infected. In 13 of the ponds containing infected individuals, prevalence exceeded 40% during at least one season. Infections were detected in multiple seasons in 20 of the sampled ponds containing infections, suggesting that ranaviruses are relatively persistent in these systems. Cattle had negative effects on water quality (turbidity and ammonia) and there was a positive association between cattle abundance and ranavirus prevalence in the summer. Counter to previous field studies in North America, we found a significant positive association between water temperature and ranavirus prevalence in the fall sampling events. Despite these findings, the influences of cattle and temperature on ranavirus prevalence were not consistent across seasons. As such, the mechanisms driving high ranavirus prevalence across the landscape and over time remain unclear. Given the widespread occurrence of ranaviruses in wild amphibians, we encourage the implementation of surveillance programs to help identify potential drivers of emergence. Sites with high ranavirus prevalence should be monitored annually for outbreaks, and the long-term effects on population size determined.


The evolution of Queensland spiny mountain crayfish of the genus Euastacus. I. Testing vicariance and dispersal with interspecific mitochondrial DNA.
The upland mesic rainforests of eastern Australia have been described as a "mesothermal archipelago" where a chain of cool mountain "islands" arise from a warm "sea" of tropical and subtropical lowlands. An endemic freshwater crayfish belonging to the genus Euastacus is found on each of these mountain "islands." The Euastacus are particularly suitable for the study of evolution because each mountain harbors a unique species, there are many taxa present providing replication within the group and, most importantly, their distribution is linear, extending along a south-north axis. This group could have evolved by "simultaneous vicariance" where there was one vicariant separation event of a widespread ancestor, or by "south to north stepping stone dispersal" where there were long distance dispersal events from neighboring mountain islands, starting in the south and proceeding north in a dispersal-colonization wave. We used pairwise genetic distances between nearest geographic neighbors as a novel way to test the two hypotheses. If diversification was due to "south to north stepping stone dispersal," then pairwise genetic distances between nearest geographic neighbors should decrease progressively the farther north the taxon pairs are found, reflecting the decreasing periods of isolation. In this case there should be a negative correlation between the south to north rank order of nearest neighbors and pairwise genetic distances. A Spearman's correlation on 16S mtDNA pairwise genetic distances and geographic rank order was not significant, indicating there was no support for the south to north stepping stone dispersal hypothesis. If simultaneous vicariance was responsible for diversification then all nearest geographic neighbor taxon pairs should have similar genetic distances and, therefore, the variance in nearest neighbor distances should be zero, or close to it. To test if the observed variance was tending towards zero we developed a randomization test where nearest neighbor taxon pairs were assigned random genetic distances and the variances calculated. The observed variance lay in the < 0.05 range of the simulated variances, providing support for the simultaneous vicariance hypothesis. The data also suggest there was simultaneous vicariance of at least two ancestral Queensland lineages. The timing of this vicariant event was probably in the Pliocene, which is consistent with the divergence times reported for other Australian mesic rainforest restricted taxa.


Anti-Candida albicans IgG Antibodies in Children With Autism Spectrum Disorders.
The gut microbiota are known to have a profound influence on both mucosal and systemic immunity and are important for gastrointestinal (GI) function. In addition, new evidence shows that the microbiota significantly influence neurodevelopment and behavior. Immune dysfunction and GI distress are extremely common in individuals with autism spectrum disorders (ASD). A growing body of evidence suggests that individuals with ASD have significant aberrations in the composition of their gut microbiota, known as dysbiosis. However, these studies have focused on the bacterial components of the microbiota, leaving the fungal microbiota in ASD poorly studied. Increases in fungal species such as Candida albicans are associated with inflammatory bowel disorders, and have recently been implicated in several neurological disorders including schizophrenia. We aimed to determine if children with ASD exhibit elevations in antibodies that target C. albicans, indicating current or previous overgrowth of this fungal species. We measured anti-C. albicans immunoglobulin (IgG) in plasma from 80 children enrolled in the UC Davis MIND Institute CHARGE study. Measurements were acquired using a commercial ELISA kit. Plasma anti-C. albicans antibody positivity was found in 36.5% (19/52) of children with ASD. Anti-C. albicans antibodies in typically developing controls was (14.3%; 4/28). Overall, ASD children had a higher rate of high-positive values compared to typically developed children with an unadjusted odds ratio of 3.45 (95% confidence interval, 1.0409 to 11.4650; p = 0.041, two-tailed). GI dysfunction was found in about half of the ASD children who were positive for anti-Candida IgG. This study provides evidence of a new microbial risk factor for ASD.


Right atrial pressure-volume relationships in tricuspid regurgitation.
Pressure-volume relationships in the right atrium were examined before and after the creation of acute experimental tricuspid regurgitation in pigs. A 1.3 kHz multielectrode impedance catheter with a measuring current of 4 mA was used to determine instantaneous right atrial pressure and relative blood volume; right atrial dimension was assessed simultaneously with ultrasonic crystals attached to the atrial walls. Impedance volume waveforms and ultrasonic crystal dimensions closely paralleled each other at baseline and after the induction of tricuspid regurgitation. The normal right atrial pressure-volume plot exhibited a figure-of-eight configuration, with an "a-loop" and a "v-loop" corresponding to the a-wave and v-wave of the right atrial pressure tracing. With severe tricuspid regurgitation, atrial pump function was abolished, and the pressure-volume plot exhibited a single clockwise loop, consistent with complete ventricularization of the right atrium. Intermediate degrees of tricuspid regurgitation preserved the figure-of-eight loop, but the size of both the a-loop and the v-loop were increased, consistent with a Starling-type load imposed on the atrium by the regurgitant blood volume. Increased right ventricular afterload mediated by constriction of the pulmonary artery and infusion of methoxamine reversibly converted the right atrial pressure-volume loop from that of mild to that of severe tricuspid regurgitation. Alternatively, constriction of the inferior vena cava and infusion of nitroprusside changed the right atrial pressure-volume loop from that of a severe pattern of tricuspid regurgitation to a less severe type of pattern. Infusion of dobutamine increased the size of the a-loop relative to the v-loop both at baseline and after induction of tricuspid regurgitation. We conclude that tricuspid regurgitation induces changes in right atrial mechanics that can be detected and quantified with an impedance catheter.


Clinical and molecular characteristics of non-small-cell lung cancer (NSCLC) harboring EGFR mutation: results of the nationwide French Cooperative Thoracic Intergroup (IFCT) program.
EGFR mutations cause inconsistent response to EGFR tyrosine-kinase inhibitors (TKI). To better understand these features, we reviewed all cases of EGFR-mutated non-small-cell lung cancer collected in the Biomarkers France database. Of 17 664 patients, 1837 (11%) with EGFR-mutated non-small-cell lung cancer were retrospectively analyzed for clinical and molecular characteristics. Results were correlated with survival and treatment response for the 848 stage IV patients. EGFR exon 18, 19, 20 and 21 mutations were found in 102 (5.5%), 931 (51%), 102 (5.5%) and 702 (38%) patients, respectively. Over 50% of exon 18 and 20 mutated patients were smokers. The median follow-up was 51.7 months. EGFR mutation type was prognostic of overall survival (OS) versus wild-type {exon 19: hazard ratio (HR)=0.51 [95% confidence interval (CI): 0.41-0.64], P < 0.0001; exon 21: HR = 0.76 (95% CI: 0.61-0.95), P = 0.002; exon 20: HR = 1.56 (95% CI: 1.02-2.38), P = 0.004}. EGFR mutation type was prognostic of progression-free survival versus wild-type [exon 19: HR = 0.62 (95% CI: 0.49-0.78), P < 0.0001; exon 20: HR = 1.46 (95% CI: 0.96-2.21), P = 0.07]. First-line treatment choice did not influence OS in multivariate analysis. First-line TKI predicted improved progression-free survival versus chemotherapy [HR = 0.67 (95% CI: 0.53-0.85), P = 0.001]. OS was longer for del19 versus L858R, which was associated with better OS compared with other exon 21 mutations, including L861Q. TKI improved survival in patients with exon 18 mutations, while chemotherapy was more beneficial for exon 20-mutated patients. EGFR mutation type can inform the most appropriate treatment. Therapeutic schedule had no impact on OS in our study, although TKI should be prescribed in first-line considering the risk of missing the opportunity to use this treatment.


EPOS2020: a major step forward.
Together with this issue of Rhinology the new European Position Paper on Rhinosinusitis and Nasal Polyps will see the light. The European Position Paper on Rhinosinusitis and Nasal Polyps 2020 is the update of similar evidence based position papers published in 2005 and 2007 and 2012. The core objective of the EPOS2020 guideline is to provide revised, up-to-date and clear evidence-based recommendations and integrated care pathways in ARS and CRS. EPOS2020 provides an update on the literature published and studies undertaken in the eight years since the EPOS2012 position paper was published and addresses areas not extensively covered in EPOS2012 like paediatric CRS, sinus surgery, exacerbations of CRS and the prevention of CRS. EPOS2020 also involved new stakeholders, like neurologists, immunologists, pharmacists and patients and address new target users who have become more involved in the management and treatment of rhinosinusitis since the publication of the last EPOS document, including pharmacists, nurses, specialised care givers and indeed patients themselves, who employ increasing self-management of their condition using over the counter treatments. The document provides suggestion for future research in this area and offer updated guidance for definitions and outcome measurements in research in different settings.


Relation between cognitive impairment and treatment adherence in elderly hypertensive patients.
Nonadherence to medical treatment and lack of cooperation in hypertensive patients >65 years of age are believed to be caused by a number of age-related problems, such as cognitive impairment. Numerous epidemiological and prospective studies have demonstrated that hypertension that remains untreated for many years or is unsuccessfully treated for reasons such as poor compliance and adherence of the patient may lead to cognitive impairment. The objective of this study was to investigate the occurrence of cognitive impairment and its effect on treatment compliance and adherence in elderly hypertensive patients. This study was an analytical cross-sectional study. The study was conducted on 300 patients aged 65-91 years (mean age=71.8 years, SD=7.8 years) diagnosed with hypertension. The following research tools were used: 1) Hill-Bone High Blood Pressure Compliance Scale (HBCS) and 2) Mini-Mental State Examination (MMSE). We also analyzed medical documentation to obtain basic sociodemographic and clinical data. The study was approved by the Bioethics Committee of the Medical University of Wrocław (no KB-144/2016). Cognitive impairment occurred in 60% of the patients. A group of 63% patients complied with antihypertensive therapy, with the mean score of 20.8 points. Cognitive impairment was strongly correlated with the total score of the HBCS questionnaire (p<0.001) and two of its subscales: "appointment keeping" (p<0.001) and "medication taking" (p<0.001). Compliance and adherence levels are higher in patients with a higher educational level, whereas male sex adversely affects treatment adherence in elderly hypertensive patients.


Acute Citalopram administration alters zebrafish social dynamics in a behavioral teleporting experiment.
In the Star Trek universe, Scotty can "beam you up" and teleport you to a location. Even though science fiction is yet to be reality, is teleporting still possible? Recently, we have developed a robotic platform that is capable of teleporting the behavior of fish from one tank to another through biologically-inspired robotic replicas which map the behavior of a fish from a tank to the other one. Our results indicate that behavioral teleporting is a promising tool to study the biological determinants of behavior. Here we demonstrate its use in pharmacological research, by investigating the effects of selective serotonin reuptake inhibitors of social interactions.


Vaginal dilators for prevention of dyspareunia after prolapse surgery: a randomized controlled trial.
To compare rates of de novo dyspareunia in women with and without vaginal dilator use after posterior colporrhaphy. This randomized controlled trial included sexually active patients with prolapse and no bothersome baseline dyspareunia undergoing posterior colporrhaphy. Patients were randomized to daily vaginal dilator use from postoperative weeks 4 through 8 or to no dilator use. Pelvic organ prolapse quantification examination and vaginal caliber were measured at baseline, 8 weeks, and 6 months postoperatively. Sexual function was evaluated at baseline, 3 months, and 6 months postoperatively using the Pelvic Organ Prolapse/Urinary Incontinence Sexual Function Questionnaire-12. Participants completed a Patient Global Impression of Improvement at 3 months and 6 months postoperatively. Sixty patients were randomized: 30 in the dilator group and 30 in the control group. There were no differences in baseline characteristics and postoperative vaginal caliber between groups. At 3 months, 9.5% of patients reported de novo dyspareunia in the dilator group compared with 19.2% of control patients (P=.44). At 6 months, 12.5% of patients in the dilator group reported de novo dyspareunia compared with 3.8% of control patients (P=.34). There was a 13% loss-to-follow-up rate, and therefore we did not meet appropriate power to detect a difference. There were no differences in overall sexual function or Patient Global Impression of Improvement scores between groups at 3 months and 6 months. There were no significant differences in de novo dyspareunia rates, overall postoperative sexual function scores, or global improvement scores between those using vaginal dilators compared with control patients.


Dynamics of bacterial communities before and after distribution in a full-scale drinking water network.
Understanding the biological stability of drinking water distribution systems is imperative in the framework of process control and risk management. The objective of this research was to examine the dynamics of the bacterial community during drinking water distribution at high temporal resolution. Water samples (156 in total) were collected over short time-scales (minutes/hours/days) from the outlet of a treatment plant and a location in its corresponding distribution network. The drinking water is treated by biofiltration and disinfectant residuals are absent during distribution. The community was analyzed by 16S rRNA gene pyrosequencing and flow cytometry as well as conventional, culture-based methods. Despite a random dramatic event (detected with pyrosequencing and flow cytometry but not with plate counts), the bacterial community profile at the two locations did not vary significantly over time. A diverse core microbiome was shared between the two locations (58-65% of the taxa and 86-91% of the sequences) and found to be dependent on the treatment strategy. The bacterial community structure changed during distribution, with greater richness detected in the network and phyla such as Acidobacteria and Gemmatimonadetes becoming abundant. The rare taxa displayed the highest dynamicity, causing the major change during water distribution. This change did not have hygienic implications and is contingent on the sensitivity of the applied methods. The concept of biological stability therefore needs to be revised. Biostability is generally desired in drinking water guidelines but may be difficult to achieve in large-scale complex distribution systems that are inherently dynamic.


Standardized Polyalthia longifolia leaf extract induces the apoptotic HeLa cells death via microRNA regulation: identification, validation, and therapeutic potential.
Polyalthia longifolia var. angustifolia Thw. (Annonaceae), is a famous traditional medicinal plant in Asia. Ample data specifies that the medicinal plant P. longifolia has anticancer activity; however, the detailed mechanisms of action still need to be well studied. Recent studies have revealed the cytotoxicity potential of P. longifolia leaf against HeLa cells. Therefore, the current study was conducted to examine the regulation of miRNAs in HeLa cancer cells treated with the standardized P. longifolia methanolic leaf extract (PLME). The regulation of miRNAs in HeLa cancer cells treated with the standardized PLME extract was studied through Illumina, Hi-Seq. 2000 platform of Next-Generation Sequencing (NGS) and various in silico bioinformatics tools. The PLME treatment regulated a subset of miRNAs in HeLa cells. Interestingly, the PLME treatment against HeLa cancer cells identified 10 upregulated and 43 downregulated (p < 0.05) miRNAs associated with apoptosis induction. Gene ontology (GO) term analysis indicated that PLME induces cell death in HeLa cells by inducing the pro-apoptotic genes. Moreover, the downregulated oncomiRs modulated by PLME treatment in HeLa cells were identified, targeting apoptosis-related genes through gene ontology and pathway analysis. The LC-ESI-MS/MS analysis identified the presence of Vidarabine and Anandamide compounds that were previously reported to exhibit anticancer activity. The findings of this study obviously linked the cell cytotoxicity effect of PLME treatment against the HeLa cells with regulating various miRNAs expression related to apoptosis induction in the HeLa cells. PLME treatment induced apoptotic HeLa cell death mechanism by regulating multiple miRNAs. The identified miRNAs regulated by PLME may provide further insight into the mechanisms that play a critical role in cervical cancer, as well as novel ideas regarding gene therapeutic strategies.


Degradation of a cytosolic protein requires endoplasmic reticulum-associated degradation machinery.
Protein misfolding is monitored by a variety of cellular "quality control" systems. Endoplasmic reticulum (ER) quality control handles misfolded secretory and membrane proteins and is well characterized. However, less is known about the quality control of misfolded cytosolic proteins (CytoQC). To study CytoQC, we have employed a genetic system in Saccharomyces cerevisiae using a transplantable degron, CL1 (1). Attachment of CL1 to the cytosolic protein Ura3p destabilizes Ura3p, targeting it for rapid proteasomal degradation. We have performed a comprehensive analysis of Ura3p-CL1 degradation requirements. As shown previously, we observe that the ER-localized ubiquitin E2 (Ubc6p, Ubc7p, and Cue1p) and E3 (Doa10p) machinery involved in ER-associated degradation (ERAD) are also responsible for the degradation of the cytosolic substrate Ura3p-CL1. Importantly, we find that the cytosol/ER membrane-localized chaperones Ydj1p and Ssa1p, known to be necessary for the ERAD of membrane proteins with misfolded cytosolic domains, are also required for the ubiquitination and degradation of Ura3p-CL1. In addition, we show a role for the Cdc48p-Npl4p-Ufd1p complex in the degradation of Ura3p-CL1. When ubiquitination is blocked, a portion of Ura3p-CL1 is ER membrane-localized. Furthermore, access to the cytosolic face of the ER is required for the degradation of CL1 degron-containing proteins. The ER is distributed throughout the cytosol, and our data, together with previous studies, suggest that the cytosolic face of the ER membrane serves as a "platform" for the degradation of Ura3p-CL1, which may also be the case for other CytoQC substrates.


Evaluating the safety of very short-term (10 days) dual antiplatelet therapy after Genous™ bio-engineered R stent™ implantation: the multicentre pilot Genous trial.
Percutaneous coronary stenting is synonymous with dual antiplatelet therapy, ranging from four weeks to lifelong. However, even short-term (four weeks) therapy with aspirin and thienopyridines is occasionally contraindicated. No study has ever appraised very short-term dual antiplatelet therapy after stenting. We thus aimed to exploit the pro-healing features of the Genous™ Bio-engineered R stent™ (Genous) (OrbusNeich Medical Technologies, Hong Kong, People's Republic of China) and evaluate the safety of a 10-day dual antiplatelet regimen after its implantation in up to 50 patients. Forty-nine consecutive patients with de novo lesions located in vessels able to receive a 2.5 mm Genous stent were included. After stenting, they received lifelong aspirin plus clopidogrel for 10 days. The primary endpoint of the study was sudden cardiac death, myocardial infarction or angiographic evidence of stent thrombosis ascribable to the study stent. Almost 70% of patients effectively discontinued clopidogrel nine to 11 days after stenting. At three-month clinical follow-up, no patient had died or reached the primary endpoint (95%; confidence interval 0-7.3%). Repeat revascularisation occurred instead in three (6.1%[2.1-16.5%]), with target lesion revascularisation in two (4.1%[1.1-13.7%]). Even very short-term dual antiplatelet therapy seems safe after coronary stenting with Genous in de novo coronary artery lesions located in secondary branch vessels. This preliminary exploratory study gives some support to planning a large trial to test the hypothesis of short dual antiplatelet therapy following Genous stent implantation.


Intramolecular coupling of active sites in the pyruvate dehydrogenase multienzyme complex of Escherichia coli.
The intramolecular passage of substrate between the component enzymes of the pyruvate dehydrogenase multienzyme complex of Escherichia coli was examined. A series of partly reassembled complexes, varying only in their E1 (pyruvate decarboxylase, EC 184.108.40.206) content, was incubated with pyruvate in the absence of CoA, conditions under which the lipoic acid residues covalently bound to the E2 (lipoate acetyltransferase, EC22.214.171.124) chains of the complex become reductively acetylated, and the reaction then ceases. The fraction of E2 chains thus acetylated was estimated by specific reaction of the thiol groups in the acetyl-lipoic acid moieties with N-ethyl[2,3-14C]maleimide. The simplest interpretation of the results was that a single E1 dimer is capable of catalysing the rapid acetylation of 8-12 E2 chains, in good agreement with the results of Bates, Danson, Hale, Hooper & Perham [(1977) Nature (London) 268, 313-316]. This novel functional connexion of active sites must be brought about by transacetylation reactions between lipoic acid residues of neighbouring E2 chains in the enzyme complex. There was also a slow transacylation process between the rapidly acetylated lipoic acid residues and those that did not react in the initial, faster phase. This interaction was not investigated in detail, since it is too slow to be of kinetic significance in the normal enzymic reaction.


Baseline characteristics and the effect of selenium supplementation on cancer incidence in a randomized clinical trial: a summary report of the Nutritional Prevention of Cancer Trial.
The Nutritional Prevention of Cancer Trial was a randomized, clinical trial designed to evaluate the efficacy of selenium as selenized yeast (200 microg daily) in preventing the recurrence of nonmelanoma skin cancer among 1312 residents of the Eastern United States. Original secondary analyses through December 31, 1993 showed striking inverse associations between treatment and the incidence of total [hazard ratio (HR) = 0.61, 95% confidence interval (CI) = 0.46-0.82], lung, prostate, and colorectal cancer and total cancer mortality. This report presents results through February 1, 1996, the end of blinded treatment. Effect modification by baseline characteristics is also evaluated. The effects of treatment overall and within subgroups of baseline age, gender, smoking status, and plasma selenium were examined using incidence rate ratios and Cox proportional hazards models. Selenium supplementation reduced total (HR = 0.75, 95% CI = 0.58-0.97) and prostate (HR = 0.48, 95% CI = 0.28-0.80) cancer incidence but was not significantly associated with lung (HR = 0.74, 95% CI = 0.44-1.24) and colorectal (HR = 0.46, 95% CI = 0.21-1.02) cancer incidence. The effects of treatment on other site-specific cancers are also described. The protective effect of selenium was confined to males (HR = 0.67, 95% CI = 0.50-0.89) and was most pronounced in former smokers. Participants with baseline plasma selenium concentrations in the lowest two tertiles (<121.6 ng/ml) experienced reductions in total cancer incidence, whereas those in the highest tertile showed an elevated incidence (HR = 1.20, 95% CI = 0.77-1.86). The Nutritional Prevention of Cancer trial continues to show a protective effect of selenium on cancer incidence, although not all site-specific cancers exhibited a reduction in incidence. This treatment effect was restricted to males and to those with lower baseline plasma selenium concentrations.


Age-related increase in alanine aminotransferase correlates with elevated levels of plasma amino acids, decanoylcarnitine, Lp-PLA2 Activity, oxidative stress, and arterial stiffness.
We investigated plasma metabolite profiles that correlated with age-related serum alanine aminotransferase (ALT) levels. The study included 602 healthy, nondiabetic subjects (aged 30-65 years); 393 individuals had normal ALT levels at baseline. Fifty-three (13.5%) individuals developed elevated ALT levels after 3 years. The remaining 340 subjects with normal ALT were matched to the elevated-ALT group (n = 53) for age, gender, BMI, fasting glucose, and ALT to form the control group (n = 53). At the 3-year follow-up, the elevated-ALT group exhibited greater increases in waist circumference, serum free fatty acid, ALT, aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), bilirubin, plasma oxidized LDL, Lp-PLA2 activity, urinary 8-epi-prostaglandin F2α (8-epi-PGF2α), and brachial-ankle pulse-wave velocity (ba-PWV) compared to the control group after baseline adjustment. The elevated-ALT group exhibited greater increases in plasma l-valine (q = 0.036), l-leucine (q = 0.012), l-phenylalanine (q = 0.012), and decanoylcarnitine (q = 0.002). Mean ALT levels positively correlated with changes in these four metabolites, which correlated with changes in AST, GGT, Lp-PLA2 activity, urinary 8-epi-PGF2α, and ba-PWV. Mean ALT changes did not significantly correlate with HOMA-insulin resistance. These results suggest that increased plasma levels of l-valine, l-leucine, l-phenylalanine, and decanoylcarnitine precede insulin resistance during periods of elevated ALT. This metabolic disturbance coincides with enhanced risk factors for cardiovascular disease.


Cesarean section. Assessment of the convenience factor.
To assess the potential input of physician convenience on the performance of cesarean sections (C-sections), analysis of C-sections by time of day and day of the week was undertaken at four Chicago-area hospitals. The primary C-section rate at the University of Chicago Chicago Lying-In Hospital, with a relatively high-risk patient population and a full-time salaried resident and faculty staff, was lower than at three other hospitals staffed predominantly by private practitioners. Indications for primary C-sections were classified as "acute," "semiacute" or "nonacute." Acute and semiacute C-sections were performed without demonstrated time biases in regard to the time of day or the day of the week at all four hospitals. Nonacute C-sections (70% cephalopelvic disproportion) were not performed as frequently at night (12-8 A.M.) as at other times at three of the four hospitals, but there were no differences in the individual characteristics of the outcomes of such deliveries between day and night. There was also no Friday afternoon or Monday morning frequency increase. The data failed to reveal significant variations in the performance of acute C-sections but did show day-night variability in C-sections done for non-acute indications.


Alopecia: botanical approaches in review.
The use of herbal medications in dermatologic disease has become common practice among consumers. In this paper, the authors review and discuss the existing evidence-based botanical modalities in the peer-reviewed literature with a particular focus on various presentations of alopecia. To maximize potential clinical application, this review has been limited human studies. The goal of the study was to provide a thorough evaluation of the current understanding of the use of non-pharmaceutical botanical products in the management of hair loss.


[Uterine response of golden hamsters to estrogenic stimulation in postnatal ontogeny].
The proliferative activity of the cells of uterine epithelium and stroma of the developing golden hamsters and the activity of acid phosphatase isozymes in these tissues was studied. The dynamics of changes of these indices during the normal development and upon estrogenic stimulation was followed. The uterine cells were shown to react to estrogenic stimulation was followed. The uterine cells were shown to react to estrogenic stimulation by the increase of proliferation from the first days of postnatal development. Characteristic changes in the ratio of the acid phosphatase isozymes in the epithelial cells in response to estrogenic stimulation appeared in the 2nd week of postnatal development only. The definite level of differentiation of cells-targets appears to be indispensable for the realization of certain specific effects of the hormone.


Molecular Dynamics and Docking Simulations of Homologous RsmE Methyltransferases Hints at a General Mechanism for Substrate Release upon Uridine Methylation on 16S rRNA.
In this study, molecular dynamics (MD) and docking simulations were carried out on the crystal structure of Neisseria Gonorrhoeae RsmE aiming at free energy of binding estimation (ΔGbinding) of the methyl transfer substrate S-adenosylmethionine (SAM), as well as its homocysteine precursor S-adenosylhomocysteine (SAH). The mechanistic insight gained was generalized in view of existing homology to two other crystal structures of RsmE from Escherichia coli and Aquifex aeolicus. As a proof of concept, the crystal poses of SAM and SAH were reproduced reflecting a more general pattern of molecular interaction for bacterial RsmEs. Our results suggest that a distinct set of conserved residues on loop segments between β12, α6, and Met169 are interacting with SAM and SAH across these bacterial methyltransferases. Comparing molecular movements over time (MD trajectories) between Neisseria gonorrhoeae RsmE alone or in the presence of SAH revealed a hitherto unknown gatekeeper mechanism by two isoleucine residues, Ile171 and Ile219. The proposed gating allows switching from an open to a closed state, mimicking a double latch lock. Additionally, two key residues, Arg221 and Thr222, were identified to assist the exit mechanism of SAH, which could not be observed in the crystal structures. To the best of our knowledge, this study describes for the first time a general catalytic mechanism of bacterial RsmE on theoretical ground.


Game Transfer Phenomena and Problematic Interactive Media Use: Dispositional and Media Habit Factors.
The study of the effects of interactive media has mainly focused on dysregulated behaviors, the conceptualization of which is supported by the paradigms of addiction. Research into Game Transfer Phenomena (GTP) examines the interplay between video game features, events while playing, and the manipulation of hardware, which can lead to sensory-perceptual and cognitive intrusions (e.g., hallucinations and recurrent thoughts) and self-agency transient changes (e.g., automatic behaviors) related to video games. GTP can influence the interpretation of stimuli and everyday interactions and, in contrast to gaming disorder, are relatively common and not necessarily negative. However, some players have reported feeling distress due to their GTP. This study focuses on how dispositional and interactive media habit factors are related to GTP and two forms of problematic interactive media [problematic video game playing (PVG) and problematic social media use (PSMU)]. A sample of 343 university students who played video games completed an online survey (58.7% male, 19-25 years old). Not all who had experienced GTP were identified as exhibiting PVG or PSMU, but all of those in the PVG group had experienced GTP. Overall, the profiles of the groups, including GTP (91.4%), PVG (28.5%), and PSMU (24.8%), were in accordance with previous findings. Those in the GTP and the PVG groups were characterized by being male, being highly engaged in the game (either while playing or via game-related activities), and showed preferences for game-related activities. However, while those in the GTP group were significantly more likely to be fantasy-prone, those with PVG were the ones who played most per day. Those in the PSMU group were characterized by being female and/or extroverted, frequently using social/sharing platforms, and seldom playing video games. A hierarchical binary logistic regression revealed that males were more likely to experience GTP. Increases in PVG, fantasy proneness, and neuroticism increased the odds of GTP. Future work can benefit from considering the role of GTP in gaming disorder, since intrusive thoughts, cognitive biases, and poor impulse control are pivotal in the initiation and maintenance of dysfunctional playing behaviors.


Evaluation of a glucose oxidase procedure.
A commerical adaptation of Trinder's glucose oxidase procedure for glucose determination was evaluated by regression analysis. Excellent correlation was found between the glucose oxidase procedure and a hexokinase procedure. Good correlation was observed between an AutoAnalyzer I, the SMA 12/60, and a Beckman Glucose Analyzer, all utilizing variations of the glucose oxidase procedure. This combination provides a system that can handle screening, routine, and emergency requests in essentially the same manner and with comparable true glucose results. No interference was found with common anticoagulants and preservatives. Boehringer Mannheim Corporation's kit for glucose determinations appears to be of equivalent quality to the original method as outlined by Trinder and evaluated by others.


Association of urinary bisphenol A concentration with medical disorders and laboratory abnormalities in adults.
Bisphenol A (BPA) is widely used in epoxy resins lining food and beverage containers. Evidence of effects in animals has generated concern over low-level chronic exposures in humans. To examine associations between urinary BPA concentrations and adult health status. Cross-sectional analysis of BPA concentrations and health status in the general adult population of the United States, using data from the National Health and Nutrition Examination Survey 2003-2004. Participants were 1455 adults aged 18 through 74 years with measured urinary BPA and urine creatinine concentrations. Regression models were adjusted for age, sex, race/ethnicity, education, income, smoking, body mass index, waist circumference, and urinary creatinine concentration. The sample provided 80% power to detect unadjusted odds ratios (ORs) of 1.4 for diagnoses of 5% prevalence per 1-SD change in BPA concentration, or standardized regression coefficients of 0.075 for liver enzyme concentrations, at a significance level of P < .05. Chronic disease diagnoses plus blood markers of liver function, glucose homeostasis, inflammation, and lipid changes. Higher urinary BPA concentrations were associated with cardiovascular diagnoses in age-, sex-, and fully adjusted models (OR per 1-SD increase in BPA concentration, 1.39; 95% confidence interval [CI], 1.18-1.63; P = .001 with full adjustment). Higher BPA concentrations were also associated with diabetes (OR per 1-SD increase in BPA concentration, 1.39; 95% confidence interval [CI], 1.21-1.60; P < .001) but not with other studied common diseases. In addition, higher BPA concentrations were associated with clinically abnormal concentrations of the liver enzymes gamma-glutamyltransferase (OR per 1-SD increase in BPA concentration, 1.29; 95% CI, 1.14-1.46; P < .001) and alkaline phosphatase (OR per 1-SD increase in BPA concentration, 1.48; 95% CI, 1.18-1.85; P = .002). Higher BPA exposure, reflected in higher urinary concentrations of BPA, may be associated with avoidable morbidity in the community-dwelling adult population.


Establishment of a human induced pluripotent stem cell neuronal model for identification of modulators of A53T α-synuclein levels and aggregation.
Inhibiting formation or promoting degradation of α-synuclein aggregates are among the therapeutical approaches under investigation as disease-modifying treatment strategies for Parkinson's disease. To support these developments, several in vitro models based on seeded α-synuclein aggregation have been established in immortalized cell lines and murine primary neurons. Here, we report on a humanized model with a reproducibility and throughput that enables its use in supporting target identification and validation in pharmacological research. A human induced pluripotent stem cell (iPSC) line was genetically modified to express HA-tagged α-synuclein with the point mutation in position 53 from Alanine to Threonine (A53T) under an inducible system and differentiated into cortical neurons expressing neuronal markers and exhibiting spontaneous activity. Intracellular α-synuclein aggregation was triggered by exposure to exogenous added fibrillated recombinant wild-type human α-synuclein fibrils91 and demonstrated by several endpoints; the formation of Triton-insoluble SDS-soluble α-synuclein, biochemically in a fluorescence resonance energy transfer based aggregation assay and by immunocytochemistry of phosphorylated α-synuclein positive puncta. We demonstrate the feasibility of upscaling the iPSC neuron production for drug discovery and that the model has a suitable dynamic range allowing for both detection of increased and decreased α-synuclein aggregation. Moreover, gene modulation is feasible using siRNAs, making the model suitable for genetic screening for modulators of α-synuclein aggregation. Data on effects of USP8, USP13 and USP9X knockdown on α-synuclein expression and aggregation contradicts published data from immortalized cell lines and murine systems. This highlight the importance of including humanized neuronal models in the confirmation of biological mechanisms in specific variations of Parkinson's disease.


Bcl-xL inhibits tBid and Bax via distinct mechanisms.
The proteins of the Bcl-2 family are key regulators of apoptosis. They form a complex interaction network in the cytosol and in cellular membranes, whose outcome determines mitochondrial permeabilization and commitment to death. However, we still do not understand how the action of the different family members is orchestrated to regulate apoptosis. Here, we combined quantitative analysis of the interactions and the localization dynamics of the family representatives Bcl-xL, Bax and tBid, in living cells. We discovered that Bax and tBid are able to constitutively shuttle between cytosol and mitochondria in the absence of other Bcl-2 proteins. Bcl-xL clearly stabilized tBid at mitochondria, where they formed tight complexes. In contrast, Bcl-xL promoted Bax retrotranslocation to the cytosol without affecting its shuttling rate, but by forming weak inhibitory mitochondrial complexes. Furthermore, analysis of phospho-mimetics of Bcl-xL suggested that phosphorylation regulates the function of Bcl-xL via multiple mechanisms. Altogether, our findings support a model in which the Bcl-2 network not only modulates protein/protein interactions among the family members, but also their respective intracellular localization dynamics, to regulate apoptosis.


Child mortality associated with reasons for non-breastfeeding and weaning: is breastfeeding best for HIV-positive mothers?
To estimate child mortality associated with reasons for the non-initiation of breastfeeding and weaning caused by preceding morbidity, compared with voluntary weaning as a result of maternal choice. Demographic and Health Surveys were analysed from 14 developing countries. Women reported whether they initiated lactation or weaned, and if so, their reasons for non-initiation or stopping breastfeeding were classified as voluntary choice or as a result of preceding maternal/infant illness. Rates of child mortality and survival analyses were estimated, by reasons for non-breastfeeding or weaning. Mortality was highest among never-breastfed children. Child mortality among women who never initiated breastfeeding was significantly higher than among women who weaned. Preceding maternal/infant morbidity was the most common reason for not breastfeeding (63.9%), and the mortality of children never breastfed because of preceding morbidity was higher than in children not breastfed as a result of maternal choice; 326.8 per 1000 versus 34.8 per 1000, respectively. Mortality among breastfed children who were weaned because of preceding morbidity was higher than among those weaned voluntarily; 19.2 per 1000 versus 9.3 per 1000, respectively. Failure to initiate lactation was significantly more frequent among women reporting complications of delivery and with low birthweight infants. Child mortality as a result of the voluntary non-initiation of breastfeeding or voluntary weaning was lower than previously estimated, and this should be used as a benchmark when counselling HIV-positive mothers on the risks of non-breastfeeding or weaning to prevent mother-to-child transmission of HIV.


Responsiveness to nicotine of neurons of the caudal nucleus of the solitary tract correlates with the neuronal projection target.
The caudal nucleus of the solitary tract (NTS) is the key integrating center of visceral sensory-motor signaling supporting autonomic homeostasis. Two key projections of this nucleus are the parabrachial nucleus (PbN) and the dorsal motor nucleus of the vagus (DMV). The PbN integrates and relays viscerosensory information primarily to the forebrain, supporting behavioral, emotional, and endocrine responses to visceral events, while the DMV contains parasympathetic preganglionic cholinergic motoneurons that support primarily gastrointestinal reflexes. Subsets of caudal NTS neurons express presynaptic and somatodendritic nicotinic acetylcholine receptors (nAChRs). However, the anatomical identification of nicotine-responsive caudal NTS neurons has not been determined. This study used in vivo and ex vivo fluorescent tracing and slice patch-clamp electrophysiological recordings from anatomically identified caudal NTS neurons to test the hypothesis that the responsiveness of these cells to nicotine correlates with the target of their axonal projections. The results demonstrate that the majority of glutamatergic terminals that synapse on PbN-projecting caudal NTS neurons are unaffected by nicotine. Moreover, only a fraction of these cells express somatodendritic nAChRs. In contrast, the majority of DMV-projecting caudal NTS neurons exhibit robust presynaptic and somatodendritic responsiveness to nicotine. However, PbN-projecting neurons also exhibit significantly lower background frequencies of glutamatergic miniature postsynaptic currents than DMV-projecting neurons. Therefore, presynaptic unresponsiveness to nicotine may result from deficient glutamatergic innervation of PbN-projecting neurons. Nevertheless, the caudal NTS contains function-specific subsets of cells with target-specific responsiveness to nicotine. These results may support development of therapeutic strategies for selective targeting of specific autonomic pathways and impaired autonomic homeostasis.


Synthetic Notch-Receptor-Mediated Transmission of a Transient Signal into Permanent Information via CRISPR/Cas9-Based Genome Editing.
Synthetic receptor biology and genome editing are emerging techniques, both of which are currently beginning to be used in preclinical and clinical applications. We were interested in whether a combination of these techniques approaches would allow for the generation of a novel type of reporter cell that would recognize transient cellular events through specifically designed synthetic receptors and would permanently store information about these events via associated gene editing. Reporting cells could be used in the future to detect alterations in the cellular microenvironment, including degenerative processes or malignant transformation into cancer cells. Here, we explored synthetic Notch (synNotch) receptors expressed in human embryonic kidney cells to investigate the efficacy of antigen recognition events in a time- and dose-dependent manner. First, we evaluated the most suitable conditions for synNotch expression based on dsRed-Express fluorophore expression. Then, we used a synNotch receptor coupled to transcriptional activators to induce the expression of a Cas9 nuclease targeted to a specific genomic DNA site. Our data demonstrate that recognition of various specific antigens via synNotch receptors robustly induced Cas9 expression and resulted in an indel formation frequency of 34.5%-45.5% at the targeted CXCR4 locus. These results provide proof of concept that reporter cells can be designed to recognize a given event and to store transient information permanently in their genomes.


Advances in spatiotemporal models for non-communicable disease surveillance.
Surveillance systems are commonly used to provide early warning detection or to assess an impact of an intervention/policy. Traditionally, the methodological and conceptual frameworks for surveillance have been designed for infectious diseases, but the rising burden of non-communicable diseases (NCDs) worldwide suggests a pressing need for surveillance strategies to detect unusual patterns in the data and to help unveil important risk factors in this setting. Surveillance methods need to be able to detect meaningful departures from expectation and exploit dependencies within such data to produce unbiased estimates of risk as well as future forecasts. This has led to the increasing development of a range of space-time methods specifically designed for NCD surveillance. We present an overview of recent advances in spatiotemporal disease surveillance for NCDs, using hierarchically specified models. This provides a coherent framework for modelling complex data structures, dealing with data sparsity, exploiting dependencies between data sources and propagating the inherent uncertainties present in both the data and the modelling process. We then focus on three commonly used models within the Bayesian Hierarchical Model (BHM) framework and, through a simulation study, we compare their performance. We also discuss some challenges faced by researchers when dealing with NCD surveillance, including how to account for false detection and the modifiable areal unit problem. Finally, we consider how to use and interpret the complex models, how model selection may vary depending on the intended user group and how best to communicate results to stakeholders and the general public.


Evolution of surgical management of gallbladder carcinoma and impact on outcome: results from two decades at a single-institution.
The surgical approach to gallbladder cancer (GBCA) has evolved in recent years, but the impact on outcomes is unknown. This study describes differences in presentation, surgery, chemotherapy strategy, and survival for patients with GBCA over two decades at a tertiary referral center. A single-institution database was queried for patients with GBCA who underwent surgical evaluation and exploration and was studied retrospectively. Univariate logistic regression was used to assess the relationship between time and treatment. Univariate Cox proportional hazard regression assessed the association between year of diagnosis and survival. From 1992 to 2015, 675 patients with GBCA were evaluated and 437 underwent exploration. Complete resection rates increased over time (p < 0.001). In those submitted to complete resection (n = 255, 58.4%), more recent years were associated with lower likelihood of bile duct resection and major hepatectomy but greater odds of neoadjuvant and adjuvant chemotherapy (p < 0.05). No significant association was found between year of diagnosis and OS or RFS (p > 0.05) for patients with complete resection. Over the study period, GBCA treatment evolved to include fewer biliary and major hepatic resections with no apparent adverse impact on outcome. Further prospective trials, specifically limited to GBCA, are needed to determine the impact of adjuvant chemotherapy.


TRPV4-mediated channelopathies.
Transient receptor potential vanilloid sub type 4 (TRPV4) is a member of non-selective cation channel that is important for sensation of several physical and chemical stimuli and also involved in multiple physiological functions. Recently it gained immense medical and clinical interest as several independent studies have demonstrated that mutations in the TRPV4 gene can results in genetic disorders like Brachyolmia, Charcot-Marie-Tooth disease type 2C, Spinal Muscular Atrophy and Hereditary Motor and Sensory Neuropathy type 2. Close analysis of the data obtained from these naturally occurring as well as other TRPV4 mutants suggest that it is not the altered channel activity of these mutants per se, but the involvement and interaction of other factors that seem to modulate oligomerization, trafficking and degradation of TRPV4 channels. Also, these factors can either enhance or reduce the activity of TRPV4. In addition, there are some potential signaling events that can also be involved in these genetic disorders. In this review, we analyzed how and what extent certain cellular and molecular functions like oligomerization, surface expression, ubiquitination and functional interactions might be affected by these mutations.


Oligonol, a Low Molecular Weight Polyphenol, Enhances Apoptotic Cell Death in Ovarian Cancer Cells via Suppressing NF-κB Activation.
Oligonol, a low molecular weight polyphenol derived from lychee fruit, not only has anti-inflammatory effects in various disease conditions but also has antitumor-promoting effects. We evaluate the nuclear factor-kappa B (NF-κB)-related anticancer effect of oligonol in ovarian cancer using SKOV-3 cells. Cell viability was examined after oligonol treatment using MTT assay and reactive oxygen species (ROS) production measurement. Subsequently, apoptotic cell death was visualized by the TdT-mediated dUTP nick-end labeling (TUNEL) method. The effect of oligonol on the NF-κB signaling pathway was evaluated using western blot analysis and luciferase activity measurement of p65, an NF-κB subunit. Cell viability significantly decreased after oligonol treatment of 72 h. Apoptosis-related markers were highly expressed in oligonol-treated cells, and increased apoptosis after oligonol treatment was also confirmed using the TUNEL assay. Western blotting results showed the expression of NF-κB signaling pathway factors, p-ERK, TRAF2, and p-IκBα, increased following treatment with oligonol, whereas p65 and COX-2 expression decreased. Immunofluorescence imaging results showed p65 luciferase activity in the nucleus as well as a shift to cytoplasmic expression. Oligonol treatment significantly enhances apoptotic cell death in SKOV-3 cells, with the suppression of NF-κB activation, which plays an essential role in this anticancer effect.


Adaptive clinical trials: progress and challenges.
Adaptive designs promise the flexibility to redesign clinical trials at interim stages. This flexibility would provide greater efficiency in drug development. However, despite this promise, many hesitate to implement such designs. Here we explore three possible reasons for the hesitation: (i) confusion with respect to the definition of an 'adaptive design'; (ii) controversy surrounding the use of sample size re-estimation methods; and (iii) logistical barriers that must be overcome in order to use adaptive designs within existing trial frameworks.The large volume of recent work has created confusion with respect to the definition of an 'adaptive design'. Unfortunately, this has resulted in reduced usage of many acceptable methods because of guilt by association with the more controversial methods. This review attempts to clarify the differences among many common types of proposed adaptive designs. Once the differences are noted, it becomes apparent that some adaptive designs are well accepted while others remain very controversial. In fact, much of the controversy and criticism surrounding adaptive designs has focused on their use for sample size re-estimation. Hence, this review also examines the different types of adaptive designs for sample size re-estimation in order to clarify the controversy surrounding the use of these methods. Specifically, separating the controversial from good practice requires clarifying differences between adaptive designs with sample size re-estimation based on a revised treatment effect and re-estimation based only on nuisance parameters (internal pilot designs). Finally, many logistical barriers must be overcome in order to use adaptive designs within existing trial frameworks.If the promise of adaptive designs is to be achieved, it will be important to bring together large groups of individuals from funding sources and regulatory agencies to address these limitations. Very few discussions of these issues have appeared in journals that are targeted to clinical audiences. In fact, current use of adaptive designs is not really hindered by the lack of statistical methods to accommodate the adaptations. Rather, there is a need for education as to which adaptive designs are acceptable and which are not acceptable. These discussions will require the involvement of many individuals outside the statistical community. In this review, we summarize the existing methods and current controversies with the intent of providing a clarification that will enable these individuals to participate in these much-needed discussions.


Selective translational control of the Alzheimer amyloid precursor protein transcript by iron regulatory protein-1.
Iron influx increases the translation of the Alzheimer amyloid precursor protein (APP) via an iron-responsive element (IRE) RNA stem loop in its 5'-untranslated region. Equal modulated interaction of the iron regulatory proteins (IRP1 and IRP2) with canonical IREs controls iron-dependent translation of the ferritin subunits. However, our immunoprecipitation RT-PCR and RNA binding experiments demonstrated that IRP1, but not IRP2, selectively bound the APP IRE in human neural cells. This selective IRP1 interaction pattern was evident in human brain and blood tissue from normal and Alzheimer disease patients. We computer-predicted an optimal novel RNA stem loop structure for the human, rhesus monkey, and mouse APP IREs with reference to the canonical ferritin IREs but also the IREs encoded by erythroid heme biosynthetic aminolevulinate synthase and Hif-2α mRNAs, which preferentially bind IRP1. Selective 2'-hydroxyl acylation analyzed by primer extension analysis was consistent with a 13-base single-stranded terminal loop and a conserved GC-rich stem. Biotinylated RNA probes deleted of the conserved CAGA motif in the terminal loop did not bind to IRP1 relative to wild type probes and could no longer base pair to form a predicted AGA triloop. An AGU pseudo-triloop is key for IRP1 binding to the canonical ferritin IREs. RNA probes encoding the APP IRE stem loop exhibited the same high affinity binding to rhIRP1 as occurs for the H-ferritin IRE (35 pm). Intracellular iron chelation increased binding of IRP1 to the APP IRE, decreasing intracellular APP expression in SH-SY5Y cells. Functionally, shRNA knockdown of IRP1 caused increased expression of neural APP consistent with IRP1-APP IRE-driven translation.


Rapid identification of virulence genes in enterotoxigenic Escherichia coli isolates associated with diarrhoea in Queensland piggeries.
To identify virulence genes in enterotoxigenic E coli (ETEC) isolates associated with diarrhoea in neonatal, 1 to 3 week-old and weaned pigs in southeast Queensland. Multiplex PCR and serotyping were applied to E coli isolates obtained over a 5-year period (1998-2002) from cases diagnosed at Toowoomba Veterinary Laboratory. A total of 126 isolates from 25 different Queensland piggeries were tested for haemolytic activity on 5% sheep blood agar and by multiplex PCR for the presence of five commonly recognised fimbrial (F4, F5, F6, F41 and F18) and three enterotoxin genes (STa, STb, LT). A subset of 62 representative isolates were serotyped by slide agglutination. For comparative purposes, multiplex PCR was also performed on the DNA of 31 ETEC isolates from 9 serotypes originating from piggeries in southern New South Wales. A total of 113 (89.7%) of the isolates from Queensland possessed ETEC virulence genes, including 14 of 15 isolates from neonatal pigs (93.3%), 18 of 23 isolates from 1 to 3 week old pigs (78.3%) and 81 of 88 isolates from weaned pigs (92.1%). F4:STa:STb:LT (serotype O149) was the most prevalent pathotype in neonatal and 1-3 week old pigs and F4:STa:STb:LT (serotype O149) and F18:STa:STb:LT (serotype O141) were most prevalent in weaned pigs. In comparison, isolates obtained from neonatal pigs from New South Wales belonged to a more diverse range of pathotypes and serotypes. Multiplex PCR was a rapid and specific method for detecting the presence of ETEC virulence genes in porcine E coli isolates. For isolates obtained from cases of suspected colibacillosis in Queensland, growth of a heavy pure culture of haemolytic E coli was a sensitive prognostic indicator of the presence of ETEC virulence genes in the isolate. ETEC pathotypes and serotypes remained stable in Queensland piggeries over the five-year study period and appear to have changed little over the last three decades.


The fitness consequences of kin-biased dispersal in a cooperatively breeding bird.
Cooperative alliances among kin may not only lead to indirect fitness benefits for group-living species, but can also provide direct benefits through access to mates or higher social rank. However, the immigrant sex in most species loses any potential benefits of living with kin unless immigrants disperse together or recruit relatives into the group in subsequent years. To look for evidence of small subgroups of related immigrants within social groups (kin substructure), we used microsatellites to assess relatedness between immigrant females of the cooperatively breeding superb starling, Lamprotornis superbus. We determined how timing of immigration led to kin subgroup formation and if being part of one influenced female fitness. Although mean relatedness in groups was higher for males than females, 26% of immigrant females were part of a kin subgroup with a sister. These immigrant sibships formed through kin recruitment across years more often than through coalitions immigrating together in the same year. Furthermore, females were more likely to breed when part of a kin subgroup than when alone, suggesting that female siblings form alliances that may positively influence their fitness. Ultimately, kin substructure should be considered when determining the role of relatedness in the evolution of animal societies.


Effects of a minimally supervised exercise program for mentally retarded adults.
Previous exercise studies that attempted to improve the cardiovascular fitness (CVF) of mentally retarded (MR) adults were flawed with methodological shortcomings that prevented conclusive results. At issue in these training studies were fitness test validity and reliability, exactness of duration and intensity of training, and an inordinate amount of supervision. Therefore, we sought to determine whether moderately MR adults (seven males, five females; IQ = 61 +/- 3, age = 25 +/- 3 yr) could improve their CVF through a minimally supervised 16-wk training program. Each subject repeated exercise tests twice on two different modes of exercise, the treadmill (TM) and Schwinn Air-Dyne ergometer (SAE), before training to ensure validity and reliability of initial CVF levels. Intensity and frequency of exercise were closely monitored. An observer was present during the training bouts, but, following initial instructions, no additional encouragement or instructions were given. Although the training program significantly increased peak VO2 (29.2 +/- 8 to 33.5 +/- 9 ml.kg-1.min-1) and peak ventilation (73 +/- 26 to 81 +/- 231.min-1) when assessed on the TM, significant changes in these same parameters were not seen when assessed on the SAE. The importance of these results was discussed.


The effect on sitting posture of a desk with a 10 degree inclination for reading and writing.
A pilot study was carried out on 10 subjects to examine the effect that a desk with a 10 degree inclination had on their sitting posture while reading and writing. A continuous recording of the subjects head and trunk positions was made. Each subject was studied during two periods of 1.5 h on different days. The subject worked at a flat desk on the first day and at a desk with a 10 degree inclination on the second. On average, the position of the head in the sagittal plane was found to be 6 degrees more erect and the position of the trunk 7 degrees more erect when working at a desk with a 10 degree inclination than when working at a flat desk. The maximal decrease in load observed on the cervical spine was 35% and on the thoracic spine 95%. The angle between the head and the trunk did not change significantly when using an inclined desk. Differences in posture in the frontal plane were not observed in this study.


Defending Comprehensive Tobacco Control Policy Implementation in Nepal From Tobacco Industry Interference (2011-2018).
Nepal passed a comprehensive tobacco control law in 2011. Tobacco control advocates successfully countered tobacco industry (TI) interference to force implementation of law. Policy documents, news stories, and key informant interviews were triangulated and interpreted using the Policy Dystopia Model (PDM). The TI tried to block and weaken the law after Parliament passed it. Tobacco control advocates used litigation to force implementation of the law while the TI used litigation in an effort to block implementation. The TI argued that tobacco was socially and economically important, and used front groups to weaken the law. Tobacco control advocates mobilized the media, launched public awareness campaigns, educated the legislature, utilized lawsuits, and monitored TI activities to successfully counter TI opposition. Both tobacco control advocates and the industry used the discursive and instrumental strategies described in the PDM. The model was helpful for understanding TI activities in Nepal and could be applied to other low- and middle-income countries. Civil society, with the help of international health groups, should continue to track TI interference and learn the lessons from other countries to proactively to counter it. The PDM provides an effective framework to understand battles over implementation of a strong tobacco control law in Nepal, a low- and middle-income country. The TI applied discursive and instrumental strategies in Nepal in its efforts to weaken and delay the implementation of the law at every stage of implementation. It is important to continuously monitor TI activities and learn lessons from other countries, as the industry often employ the same strategies globally. Tobacco control advocates utilized domestic litigation, media advocacy, and engaged with legislators, politicians, and other stakeholders to implement a strong tobacco control law. Other low- and middle-income countries can adapt these lessons from Nepal to achieve effective implementation of their laws.


Bcl-2 overexpression enhances tumor-specific T-cell survival.
Although immunotherapy based on the adoptive transfer of tumor-specific T lymphocytes has been shown to result in dramatic clinical responses in some patients, the relatively low levels of engraftment and persistence of the adoptively transferred cells may limit these responses in many patients. In an attempt to develop strategies for prolonging the survival of adoptively transferred T cells, we have carried out studies in which T cells obtained from healthy donors as well as tumor-specific T cells were transduced with a retrovirus expressing the human Bcl-2 gene. Our results indicate that these transduced T cells overexpress Bcl-2, are resistant to death, and have a survival advantage following interleukin-2 withdrawal compared with control T cells transduced with a retrovirus expressing green fluorescent protein. Tumor-specific T cells overexpressing Bcl-2 maintained their ability to specifically recognize and respond to target cells. Furthermore, we show that adoptive immunotherapy of an established B16 tumor can be significantly enhanced by overexpressing Bcl-2 in melanoma-specific T-cell receptor transgenic T cells. Our data suggest that adoptive immunotherapy approaches to the treatment of cancer patients may be enhanced using Bcl-2-modified tumor-reactive T cells.


Exposure-response relationships for saquinavir, zidovudine, and zalcitabine in combination therapy.
The relationship of CD4+ cell response, level of RNA in plasma, and quantitative peripheral blood mononuclear cell (PBMC) titer to apparent drug exposure was investigated by using data from AIDS Clinical Trial Group protocol 229, a multicenter randomized study. Patients received either saquinavir, zalcitabine, or a combination of both, along with open-label zidovudine. Approximately 100 patients were enrolled in each arm, and the primary study duration was 24 weeks. Individual drug exposure, the area under the concentration-time curve, was estimated by using population-based pharmacokinetic methods. Response was defined as the maximum increase in CD4+ cell count or the maximum decrease in RNA in plasma or PBMC titer adjusted for baseline CD4+ cell count, RNA in plasma, and PBMC titer, respectively. Regression of responses on exposure demonstrated an exposure effect for saquinavir which was significant for the maximum increase in CD4+ cell count and the decrease in RNA in plasma. For the PBMC titer, no significant relationship could be demonstrated but the results suggested a trend similar to that of the other response variables. For all three response variables, the slope of the saquinavir exposure response was greater with the triple combination (saquinavir, zidovudine, and zalcitabine) than with the combination of saquinavir and zidovudine, suggesting possible synergism between saquinavir and zalcitabine.


Public health facility resource availability and provider adherence to first antenatal guidelines in a low resource setting in Accra, Ghana.
Lack of resources has been identified as a reason for non-adherence to clinical guidelines. Our aim was to describe public health facility resource availability in relation to provider adherence to first antenatal visit guidelines. A cross-sectional analysis of the baseline data of a prospective cohort study on adherence to first antenatal care visit guidelines was carried out in 11 facilities in the Greater Accra Region of Ghana. Provider adherence was studied in relation to health facility resource availability such as antenatal workload for clinical staffs, routine antenatal drugs, laboratory testing, protocols, ambulance and equipment. Eleven facilities comprising 6 hospitals (54.5 %), 4 polyclinics (36.4 %) and 1 health center were randomly sampled. Complete provider adherence to first antenatal guidelines for all the 946 participants was 48.1 % (95 % CI: 41.8-54.2 %), varying significantly amongst the types of facilities, with highest rate in the polyclinics. Average antenatal workload per month per clinical staff member was higher in polyclinics compared to the hospitals. All facility laboratories were able to conduct routine antenatal tests. Most routine antenatal drugs were available in all facilities except magnesium sulphate and sulphadoxine-pyrimethamine which were lacking in some. Antenatal service protocols and equipment were also available in all facilities. Although antenatal workload varies across different facility types in the Greater Accra region, other health facility resources that support implementation of first antenatal care guidelines are equally available in all the facilities. These factors therefore do not adequately account for the low and varying proportions of complete adherence to guidelines across facility types. Providers should be continually engaged for a better understanding of the barriers to their adherence to these guidelines.


Pleiotropic plasma membrane ATPase mutations of Saccharomyces cerevisiae.
We isolated a large number of mutations in the structural gene for the plasma membrane ATPase (PMA1) of Saccharomyces cerevisiae. These mutations were selected by their resistance to the aminoglycoside antibiotic hygromycin B. Biochemical analysis of purified membrane preparations showed that the plasma membrane ATPase activity of the mutants was reduced as much as 75%. Intragenic complementation of pma1 mutants suggested that the yeast plasma membrane ATPase was a multimeric enzyme. The pma1 mutants were apparently defective in maintaining internal pH; more than half of the mutants were unable to grow either at a low pH or in the presence of a weak acid. Most pma1 mutants were also osmotic pressure sensitive. At a very low temperature (5 degrees C) many pma1 mutants were unable to grow and were arrested as unbudded cells. The three most severely affected mutants were also unable to grow in the presence of NH4+. The most extreme mutant exhibited a severe defect in progression through the cell cycle; on synthetic medium, the cells progressively accumulated nucleus-containing small buds that generally failed to complete bud enlargement and cytokinesis. Most of the pleiotropic phenotypes of pma1 mutants could be suppressed by the addition of 50 mM KCl but not NaCl to the medium.


Teashirt 3 regulates development of neurons involved in both respiratory rhythm and airflow control.
Neonatal breathing in mammals involves multiple neuronal circuits, but its genetic basis remains unclear. Mice deficient for the zinc finger protein Teashirt 3 (TSHZ3) fail to breathe and die at birth. Tshz3 is expressed in multiple areas of the brainstem involved in respiration, including the pre-Bötzinger complex (preBötC), the embryonic parafacial respiratory group (e-pF), and cranial motoneurons that control the upper airways. Tshz3 inactivation led to pronounced cell death of motoneurons in the nucleus ambiguus and induced strong alterations of rhythmogenesis in the e-pF oscillator. In contrast, the preBötC oscillator appeared to be unaffected. These deficits result in impaired upper airway function, abnormal central respiratory rhythm generation, and altered responses to pH changes. Thus, a single gene, Tshz3, controls the development of diverse components of the circuitry required for breathing.


Molecular composition of the adenovirus type 2 virion.
The representation of the different structural polypeptides within the adenovirus virion has been accurately determined, and the particle molecular weight has been derived. A stoichiometric analysis was performed with [35S]methionine as a radiolabel, and analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate the polypeptides. The recently available sequence of the adenovirus type 2 genome was used to determine the number of methionines in each polypeptide. The resulting relative representation was placed on an absolute scale by using the known number of hexon polypeptides per virion. The analysis provides new information on the composition of the vertex region, which has been the subject of some controversy. Penton base was found to be present in 60 copies, distributed as pentamers at each of the 12 vertices. Three fiber monomers were associated with one penton base to form the penton complex. Polypeptide IX was present in 240 copies per virion and 12 copies per group-of-nine hexons, supporting a model proposed earlier for the distribution of this protein. The location of polypeptide IX explains the dissociation of the virus outer capsid into groups-of-nine hexons. The penton base was microheterogeneous, and the relative amounts suggest that the symmetry mismatch, which occurs within the penton complex between base and fiber, is resolved by the synthesis of penton base polypeptides from two closely spaced start codons.


Opiates and the gastrointestinal tract.
Opiate peptides, encephalins and endorphins, originally isolated from the brain, have been detected by the immunocytochemistry and radioimmunoassay in the digestive system and localized in the distinctive endocrine-paracrine cells of APUD series of the gastrointestinal mucosa and the nerve fibers of the myenteric plexus. The physiological role of endogenous opiates is unknown but the spectrum of biological action on the digestive system resembles that of morphine and related drugs. They strongly affect gastrointestinal motility such as gastric and intestinal contractions, gastric emptying, intestinal transit, biliary pressure and defecation. They also increase gastric secretory activity and raise gastric mucosal microcirculation without affecting gastrin release. They inhibit pancreatic bicarbonate and enzyme secretion probably via suppressing the release of intestinal hormones, secretin and cholecystokinin. The actions of opiates on the motility and secretion can be reversed by specific opiate receptor antagonists, e.g. naloxone, indicating that opiate receptors may be involved in these actions.


Language and theory-of-mind development in prelingually deafened children with cochlear implants: a preliminary investigation.
One cognitive domain in which deaf children are known to lag behind their hearing peers is that of 'theory of mind'. Impoverished early conversational experience and deprivation of discussion about intangible mental states are seen as likely contributors to this deficit. The present study explored the relation between language and theory-of-mind capabilities in prelingually deafened children aged 4 to 11 years who were cochlear implant users. The children completed theory-of-mind tasks that indexed their ability to reason about conflicting mental representations, namely the understanding of false belief and the distinction between appearance and reality. Controlling for age, language ability was found to correlate positively and reliably with theory-of-mind performance.


[Daily excretion of melatonin in patients with cancer of the stomach and large intestine].
Diurnal excretion of melatonin was studied in 37 cases of gastric and rectal cancer. In cancer patients nocturnal excretion was lower than in daytime while in healthy subjects the situation was reversed. Increased extrapineal production of melatonin as a possible cause of disorders in its excretion rhythm is discussed.


Guillotine needle biopsy as a supplement to endosonography in the diagnosis of gastric submucosal tumors.
The presumptive diagnosis of gastric submucosal tumors can be made by endoscopic ultrasonography (EUS) but histological confirmation is still required. A special guillotine biopsy device (Flexi-Temno) which enables collection of adequate submucosal samples by the endoscopic approach was therefore evaluated. After visualization by EUS the guillotine needle biopsy was performed in 21 patients with submucosal tumors of the stomach. There were 2 failures in 2 patients with leiomyomas. The diagnosis suggested by EUS was confirmed by guillotine biopsy in 17 cases. Guillotine biopsy detected 2 cases of unsuspected leiomyosarcoma which were confirmed surgically. In cases of solid submucosal tumors confirmed on EUS, the guillotine needle biopsy enables a definitive histologic diagnosis. Thus malignancies can be detected when EUS findings are not significant.


Revealing Acquired Resistance Mechanisms of Kinase-Targeted Drugs Using an on-the-Fly, Function-Site Interaction Fingerprint Approach.
Although kinase-targeted drugs have achieved significant clinical success, they are frequently subject to the limitations of drug resistance, which has become a primary vulnerability to targeted drug therapy. Therefore, deciphering resistance mechanisms is an important step in designing more efficacious, antiresistant drugs. Here we studied two FDA-approved kinase drugs: Crizotinib and Ceritinib, which are first- and second-generation anaplastic lymphoma kinase (ALK) targeted inhibitors, to unravel drug-resistance mechanisms. We used an on-the-fly, function-site interaction fingerprint (on-the-fly Fs-IFP) approach, combining binding free-energy surface calculations with the Fs-IFPs. Establishing the potentials of mean force and monitoring the atomic-scale protein-ligand interactions, before and after L1196M-induced drug resistance, revealed insights into drug-resistance/antiresistant mechanisms. Crizotinib prefers to bind the wild-type ALK kinase domain, whereas Ceritinib binds more favorably to the mutated ALK kinase domain, in agreement with experimental results. We determined that ALK kinase-drug interactions in the region of the front pocket are associated with drug resistance. Additionally, we find that the L1196M mutation does not simply alter the binding modes of inhibitors but also affects the flexibility of the entire ALK kinase domain. Our work provides an understanding of the mechanisms of ALK drug resistance, confirms the usefulness of the on-the-fly Fs-IFP approach, and provides a practical paradigm to study drug-resistance mechanisms in prospective drug discovery.


Nonparametric Bayesian density estimation on manifolds with applications to planar shapes.
Statistical analysis on landmark-based shape spaces has diverse applications in morphometrics, medical diagnostics, machine vision and other areas. These shape spaces are non-Euclidean quotient manifolds. To conduct nonparametric inferences, one may define notions of centre and spread on this manifold and work with their estimates. However, it is useful to consider full likelihood-based methods, which allow nonparametric estimation of the probability density. This article proposes a broad class of mixture models constructed using suitable kernels on a general compact metric space and then on the planar shape space in particular. Following a Bayesian approach with a nonparametric prior on the mixing distribution, conditions are obtained under which the Kullback-Leibler property holds, implying large support and weak posterior consistency. Gibbs sampling methods are developed for posterior computation, and the methods are applied to problems in density estimation and classification with shape-based predictors. Simulation studies show improved estimation performance relative to existing approaches.


Competence to stand trial should require rational understanding.
Rationality is explicit in the United States Supreme Court's Dusky standard but not in most U.S. CST standards. It is hard to imagine that the legal purposes of competency to stand trial (CST) determinations are served if a defendant's understanding of the proceedings is irrational (e.g., delusional or psychotically confused) or if the defendant cannot consult rationally with counsel. Most insanity tests include a rationality criterion. In United States v. Timmins, the Ninth Circuit emphasized the importance of rationality in CST in an opinion that also illustrated that the district court applied the federal standard, which does not mention rationality, without considering rationality. With its recent decision in Panetti v. Quarterman, the United States Supreme Court now requires rational understanding for competence to be executed. If there had been any doubt that unqualified understanding is not invariably taken to mean rational understanding by trial and appellate courts, the legal history of Panetti now dispels this misapprehension. The time is ripe for recognition of a uniform standard of CST that requires rationality.


Pervasive admixture and the spread of a large-lipped form in a cichlid fish radiation.
Adaptive radiations have proven important for understanding the mechanisms and processes underlying biological diversity. The convergence of form and function, as well as admixture and adaptive introgression, are common in adaptive radiations. However, distinguishing between these two scenarios remains a challenge for evolutionary research. The Midas cichlid species complex (Amphilophus spp.) is a prime example of adaptive radiation, with phenotypic diversification occurring at various stages of genetic differentiation. One species, A. labiatus, has large fleshy lips, is associated with rocky lake substrates, and occurs patchily within Lakes Nicaragua and Managua. By contrast, the similar, but thin-lipped, congener, A. citrinellus, is more common and widespread. We investigated the evolutionary history of the large-lipped form, specifically regarding whether the trait has evolved independently in both lakes from ancestral thin-lipped populations, or via dispersal and/or admixture events. We collected samples from distinct locations in both lakes, and assessed differences in morphology and ecology. Using RAD-seq, we genotyped thousands of SNPs to measure population structure and divergence, demographic history, and admixture. We found significant between-species differences in ecology and morphology, local intraspecific differences in body shape and trophic traits, but only limited intraspecific variation in lip shape. Despite clear ecological differences, our genomic approach uncovered pervasive admixture between the species and low genomic differentiation, with species within lakes being genetically more similar than species between lakes. Taken together, our results suggest a single origin of large-lips, followed by pervasive admixture and adaptive introgression, with morphology being driven by local ecological opportunities, despite ongoing gene-flow.


Investigating pertussis toxin and its impact on vaccination.
Whooping cough, caused by Bordetella pertussis, remains a major global health problem. Each year around 40 million of pertussis cases resulting in 200,000-400,000 annual deaths occur worldwide. Pertussis toxin is a major virulence factor of B. pertussis. Murine studies have shown its importance in bacterial colonization and in immunomodulation to evade innate or adaptive immunity. The toxin is composed of an A protomer expressing ADP-ribosyltransferase activity and a B oligomer, responsible for toxin binding to target cells. The toxin is also a major protective antigen in all currently available vaccines. However, vaccine escape mutants with altered toxin expression have recently been isolated in countries with high vaccination coverage illustrating the need for improved pertussis vaccines.


Fluorescence signaling molecularly imprinted polymers for antibiotics prepared via site-directed post-imprinting introduction of plural fluorescent reporters within the recognition cavity.
An antibiotic-imprinted cavity with two different fluorescent dyes was prepared by molecular imprinting and subsequent post-imprinting modifications (PIMs), for the readout of a specific binding event as a fluorescence signal. The fluorescent dyes were site-specifically introduced into the cavity using an orthogonal reversible bonding reaction, Schiff base formation, and a disulfide exchange reaction. The template molecule, comprising cephalexin connected to a Schiff base monomer and a disulfide monomer, was copolymerized with a crosslinker. This was followed by Schiff base hydrolysis and a disulfide exchange reaction, yielding the APO-type scaffold (PRECURSOR). Two different fluorophores, 4-formylsalicylic acid (FSA) and 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD), were introduced into the PRECURSOR by site-directed PIMs, yielding a fluorescent HOLO polymer. The obtained fluorescent polymer, HOLO(FSA)(Cys-DBD), was able to selectively readout the binding events as a fluorescence signal change. We found that the proposed strategy has the potential to open up a new class of synthetic materials possessing various desirable functions.


Toward the next quarter-century: conceptual and methodological challenges for attachment theory.
Attachment theory and research have offered fundamental insights into early sociopersonality development for the past quarter-century. As its scope expands throughout the life course with applications to developmental psychopathology, however, attachment work faces important conceptual and methodological challenges. These include (a) expanding Bowlby's theoretical formulations to address developmental changes in the nature of attachment organization beyond infancy, the converging influence of multiple attachment relationships, and the nature and development of internal working models: (b) systematically validating assessments of attachment security for older ages in the context of enhanced theoretical understanding of how attachment itself changes with age; (c) new methodological approaches to understanding the relations between attachment and later behavior in light of empirical evidence of stability and change in attachment security and the need for explicit theoretical predictions of the sequelae of attachment security; and (d) more complex conceptualizations of the associations among attachment, contextual risk, and later behavior. These are similar to the challenges facing the original pioneers of attachment theory and research, suggesting that familiar problems must now be reconsidered against the landscape of new applications of attachment work and the insights of contemporary developmental science.


Vacuolation and infection thread in root nodules of soybean.
Profuse vacuolation takes place in the soybean root nodule cells where infection threads carry rhizobia. After the rhizobia are released the disappearance of the infection thread is attributed to its degradation within large vacuoles which result from fusion of small vacuoles.


Multifunctional Micromachines Constructed by Combining Multiple Protein-Based Components with Different Functions.
Untethered mobile micromachines have considerable potential to realize more effective and minimally invasive medicine. Although diverse medical micromachines have been reported over the past few decades, these machines were developed for performing only specific tasks and the functions imparted to them were limited to a few. Hence, the methodologies for imparting a wide variety of functions to machines have not been fully explored. In this study, a novel construction strategy for the multifunctional micromachines is presented, where a specific function can be added to the machine in one step by directly combining the protein-based component, possessing the biological function of constituent proteins, to an arbitrary position of the machine by using an inkjet printing technique. As a proof-of-concept demonstration, various types of machines were constructed by combining multiple components with different functions. These constructed machines successfully performed functions as diverse as enzyme-powered self-propulsion, collection of target objects, including the bilirubin and living cells, enzyme-mediated conversion of substrate molecules to different ones, magnetic guidance, and release of anti-inflammatory drug diapocynin. The study's progressive approach as well as multifunctional and biocompatible machines composed of proteins will profoundly impact the development of intelligent machines equipped with multiplex sophisticated functionalities.


Fear of falling in robust community-dwelling older people: results of a cross-sectional study.
To identify the prevalence and risk factors for fear of falling among robust community-dwelling older people. Different models including various risk factors emerged when previous studies attempted to explore the factors' multivariate associations with fear of falling. Attempting to detect fear of falling by a single question in some previous studies may not have been sufficiently sensitive, particularly in a robust population. Although some studies used a validated instrument to detect participants' fear of falling, their self-perceived efficacy in performing various activities was seldom reported. Information to provide an insight into the planning of well-targeted interventions to reduce fear of falling has been lacking. A cross-sectional study of 445 robust community-dwelling older people aged ≥65. Besides their demographic and fall-related background, participants' fear of falling level was assessed using the Chinese Fall Efficacy Scale-International. Other instruments included the timed up and go test, the Chinese Geriatric Depression Scale-Short Form, the Chinese Anxiety Disorder Scale, the Chinese Lubben Social Network Scale and the Chinese Personal Wellbeing Index, which were used to assess different variables that may be associated with fear of falling. Of the 64·73% of participants who had fear of falling, 65·63% had no history of fall but still had fear of falling. This result was higher than in most previous studies. Participants' main concerns were walking on slippery/uneven surfaces, in crowded places and up/down a slope. Multivariate analyses showed that being female, suffering from poor vision and arthritis, poor performance in the timed up and go test, expressing more depressive and anxiety symptoms and low self-perceived well-being are multiply associated with fear of falling. Fear of falling is prevalent among robust community-dwelling older people regardless of their fall history. They showed lack of self-perceived efficacy in performing various outdoor activities. The findings can assist in developing multidimensional strategies for reducing fear of falling in robust community-dwelling older people.


Comparison of mercury concentrations in landlocked, resident, and sea-run fish (Salvelinus spp.) from Nunavut, Canada.
Mercury concentrations ([Hg]) in Arctic food fish often exceed guidelines for human subsistence consumption. Previous research on two food fish species, Arctic char (Salvelinus alpinus) and lake trout (Salvelinus namaycush), indicates that anadromous fish have lower [Hg] than nonanadromous fish, but there have been no intraregional comparisons. Also, no comparisons of [Hg] among anadromous (sea-run), resident (marine access but do not migrate), and landlocked (no marine access) life history types of Arctic char and lake trout have been published. Using intraregional data from 10 lakes in the West Kitikmeot area of Nunavut, Canada, we found that [Hg] varied significantly among species and life history types. Differences among species-life history types were best explained by age-at-size and C:N ratios (indicator of lipid); [Hg] was significantly and negatively related to both. At a standardized fork length of 500 mm, lake trout had significantly higher [Hg] (mean 0.17 µg/g wet wt) than Arctic char (0.09 µg/g). Anadromous and resident Arctic char had significantly lower [Hg] (each 0.04 µg/g) than landlocked Arctic char (0.19 µg/g). Anadromous lake trout had significantly lower [Hg] (0.12 µg/g) than resident lake trout (0.18 µg/g), but no significant difference in [Hg] was seen between landlocked lake trout (0.21 µg/g) and other life history types. Our results are relevant to human health assessments and consumption guidance and will inform models of Hg accumulation in Arctic fish.