Argyrophilic nucleolar organizer region in proliferating cell has a predictive value for local recurrence in superficial bladder tumor.
It has been shown in many carcinomas that the proliferation rate and number of argyrophilic nucleolar organizer regions (AgNOR) are associated with tumor aggressiveness. However, in bladder tumor the significance of the correlation between the number of AgNOR and tumor behavior remains controversial. Therefore, it would be helpful if a new technique could be developed that would allow for more accurate AgNOR counting in association with tumor behavior. We established the simultaneous staining technique of AgNOR with Ki-67 labeling to reveal the significance of AgNOR count in superficial bladder tumor. A total of 50 paraffin sections of superficial bladder tumor were stained with AgNOR and Ki-67 (MIB-1). The numbers of AgNORs in proliferating (MIB-1 positive) or resting (MIB-1 negative) cells were counted from a total of 100 nuclei. Correlations between MIB-1 associated AgNOR count and clinicopathological parameters were statistically analyzed. The AgNOR count in proliferating cells was significantly higher than that in resting cells (p<0.01), and the count significantly increased with tumor grade (p<0.01). Based on recurrence-free survival analyses the local recurrence rate was significantly higher in patients with high proliferating cell NOR but not for those with resting or whole cells. However, no AgNOR score helped to select patients at high risk for disease progression. Proliferating cell NOR had a predictive value for local recurrence in patients with superficial bladder tumor.


Rethinking 'rational imitation' in 14-month-old infants: a perceptual distraction approach.
In their widely noticed study, Gergely, Bekkering, and Király (2002) showed that 14-month-old infants imitated an unusual action only if the model freely chose to perform this action and not if the choice of the action could be ascribed to external constraints. They attributed this kind of selective imitation to the infants' capacity of understanding the principle of rational action. In the current paper, we present evidence that a simpler approach of perceptual distraction may be more appropriate to explain their results. When we manipulated the saliency of context stimuli in the two original conditions, the results were exactly opposite to what rational imitation predicts. Based on these findings, we reject the claim that the notion of rational action plays a key role in selective imitation in 14-month-olds.


Emerging from the database shadows: characterizing undocumented immigrants in a large cohort of HIV-infected persons.
Little is known about how HIV affects undocumented immigrants despite social and structural factors that may place them at risk of poor HIV outcomes. Our understanding of the clinical epidemiology of HIV-infected undocumented immigrants is limited by the challenges of determining undocumented immigration status in large data sets. We developed an algorithm to predict undocumented status using social security number (SSN) and insurance data. We retrospectively applied this algorithm to a cohort of HIV-infected adults receiving care at a large urban healthcare system who attended at least one HIV-related outpatient visit from 1997 to 2013, classifying patients as "screened undocumented" or "documented". We then reviewed the medical records of screened undocumented patients, classifying those whose records contained evidence of undocumented status as "undocumented per medical chart" (charted undocumented). Bivariate measures of association were used to identify demographic and clinical characteristics associated with undocumented immigrant status. Of 7593 patients, 205 (2.7%) were classified as undocumented by the algorithm. Compared to documented patients, undocumented patients were younger at entry to care (mean 38.5 years vs. 40.6 years, p < 0.05), less likely to be female (33.2% vs. 43.1%, p < 0.01), less likely to report injection drug use as their primary HIV risk factor (3.4% vs. 18.0%, p < 0.001), and had lower median CD4 count at entry to care (288 vs. 339 cells/mm3, p < 0.01). After medical record review, we re-classified 104 patients (50.7%) as charted undocumented. Demographic and clinical characteristics of charted undocumented did not differ substantially from screened undocumented. Our algorithm allowed us to identify and clinically characterize undocumented immigrants within an HIV-infected population, though it overestimated the prevalence of patients who were undocumented.


Skin-interfaced microfluidic system with personalized sweating rate and sweat chloride analytics for sports science applications.
Advanced capabilities in noninvasive, in situ monitoring of sweating rate and sweat electrolyte losses could enable real-time personalized fluid-electrolyte intake recommendations. Established sweat analysis techniques using absorbent patches require post-collection harvesting and benchtop analysis of sweat and are thus impractical for ambulatory use. Here, we introduce a skin-interfaced wearable microfluidic device and smartphone image processing platform that enable analysis of regional sweating rate and sweat chloride concentration ([Cl-]). Systematic studies (n = 312 athletes) establish significant correlations for regional sweating rate and sweat [Cl-] in a controlled environment and during competitive sports under varying environmental conditions. The regional sweating rate and sweat [Cl-] results serve as inputs to algorithms implemented on a smartphone software application that predicts whole-body sweating rate and sweat [Cl-]. This low-cost wearable sensing approach could improve the accessibility of physiological insights available to sports scientists, practitioners, and athletes to inform hydration strategies in real-world ambulatory settings.


The Role and Effects of ANXA1 in Temporal Lobe Epilepsy: A Protection Mechanism?
BACKGROUND The endogenous protein annexin A1 (ANXA1) is an anti-inflammatory mediator in the brain that is thought to contribute to the progression of many neurological conditions. However, its exact role in temporal lobe epilepsy (TLE) remains unclear. We hypothesized that ANXA1 exerts negative actions on TLE by alleviating inflammatory damage in neurons. To identify the potential mechanism of TLE by assessing ANXA1 expression in TLE rats. MATERIAL AND METHODS TLE was induced in rats (n=70) via an intraperitoneal injection of lithium chloride (LiCl) and pilocarpine (PILO). The control group (n=10) received an injection of the equivalent amount of saline. ANXA1 expression was detected via immunohistochemistry and Western blotting. RESULTS Successful establishment of the TLE model in rats resulted in epileptic seizures. ANXA1 was immunohistochemically detected as brownish yellow particles in the dentate gyrus and the CA1 region of the door zone; this expression was predominantly localized to the cytoplasm of glia rather than neurons. ANXA1 expression was stronger in TLE rats compared with the control group. ANXA1 expression in TLE was also assessed via Western blotting, and compared between groups at various time points. ANXA1 expression was significantly increased in the acute (the first 24 h) and chronic (after 1 month) phases (P<0.001) but significantly decreased during the recovery phase (72 h, 1 week, and 2 weeks) (P<0.001). These findings suggest that ANXA1 expression is correlated with TLE activity. CONCLUSIONS Our data suggest that ANXA1 plays an important role in TLE by alleviating inflammatory damage and protecting neurons.


Cultivated autologous limbal epithelial transplantation for symptomatic bullous keratopathy.
We report the clinical outcome of cultivated autologous limbal epithelial transplantation for symptomatic bullous keratopathy in an 87-year-old female patient presenting with recurrent corneal erosion and intractable ocular pain. The patient refused corneal transplantation, and transplantation of a cultivated autologous limbal epithelial sheet was carried out to free her of the symptoms associated with chronic pseudophakic bullous keratopathy. Both integrity of transplanted epithelium and relief from intolerable ocular pain were achieved and maintained immediately after surgery. No complications were noted in either the transplanted eye or donor eye during the 43-month follow-up period. Our findings support that patients with bullous keratopathy not consenting to procedures to improve visual acuity in whom the primary aim is relief of symptoms may benefit from cultivated autologous limbal epithelial transplantation.


Field evaluation of two novel sampling devices for collecting wild oviposition site seeking malaria vector mosquitoes: OviART gravid traps and squares of electrocuting nets.
New sampling tools are needed for collecting exophilic malaria mosquitoes in sub-Saharan Africa to monitor the impact of vector control interventions. The OviART gravid trap and squares of electrocuting nets (e-nets) were recently developed under semi-field conditions for collecting oviposition site seeking Anopheles gambiae (sensu stricto) (s.s.). This study was designed to evaluate the efficacy of these traps for sampling malaria vectors under field conditions. Prior to field testing, two modifications to the prototype OviART gravid trap were evaluated by (i) increasing the surface area and volume of water in the artificial pond which forms part of the trap, and (ii) increasing the strength of the suction fan. Six sampling tools targeting gravid females (Box gravid trap, detergent-treated ponds, e-nets insect glue-treated ponds, sticky boards and sticky floating-acetate sheets) were compared under field conditions to evaluate their relative catching performance and to select a method for comparison with the OviART gravid trap. Finally, the trapping efficacy of the OviART gravid trap and the square of e-nets were compared with a Box gravid trap during the long rainy season in three household clusters in western Kenya. The OviART gravid trap prototype's catch size was doubled by increasing the pond size [rate ratio (RR) 1.9; 95 % confidence interval (CI) 1.1-3.4] but a stronger fan did not improve the catch. The square of e-nets performed better than the other devices, collecting three times more gravid Anopheles spp. than the Box gravid trap (RR 3.3; 95 % CI 1.4-7.6). The OviART gravid trap collections were comparable to those from the e-nets and 3.3 (95 % CI 1.5-7.0) times higher than the number of An. gambiae senso lato (s.l.) collected by the Box gravid trap. Both OviART gravid trap and squares of e-nets collected wild gravid Anopheles gambiae (s.l.) where natural habitats were within 200-400 m of the trap. Whilst the e-nets are difficult to handle and might therefore only be useful as a research device, the OviART gravid trap presents a promising new surveillance tool. Further field testing is needed in different eco-epidemiological settings to provide recommendations for its use.


The Impact of Saccharomyces cerevisiae on a Wine Yeast Consortium in Natural and Inoculated Fermentations.
Natural, also referred to as spontaneous wine fermentations, are carried out by the native microbiota of the grape juice, without inoculation of selected, industrially produced yeast or bacterial strains. Such fermentations are commonly initiated by non-Saccharomyces yeast species that numerically dominate the must. Community composition and numerical dominance of species vary significantly between individual musts, but Saccharomyces cerevisiae will in most cases dominate the late stages of the fermentation and complete the process. Nevertheless, non-Saccharomyces species contribute significantly, positively or negatively, to the character and quality of the final product. The contribution is species and strain dependent and will depend on each species or strain's absolute and relative contribution to total metabolically active biomass, and will therefore, be a function of its relative fitness within the microbial ecosystem. However, the population dynamics of multispecies fermentations are not well understood. Consequently, the oenological potential of the microbiome in any given grape must, can currently not be evaluated or predicted. To better characterize the rules that govern the complex wine microbial ecosystem, a model yeast consortium comprising eight species commonly encountered in South African grape musts and an ARISA based method to monitor their dynamics were developed and validated. The dynamics of these species were evaluated in synthetic must in the presence or absence of S. cerevisiae using direct viable counts and ARISA. The data show that S. cerevisiae specifically suppresses certain species while appearing to favor the persistence of other species. Growth dynamics in Chenin blanc grape must fermentation was monitored only through viable counts. The interactions observed in the synthetic must, were upheld in the natural must fermentations, suggesting the broad applicability of the observed ecosystem dynamics. Importantly, the presence of indigenous yeast populations did not appear to affect the broad interaction patterns between the consortium species. The data show that the wine ecosystem is characterized by both mutually supportive and inhibitory species. The current study presents a first step in the development of a model to predict the oenological potential of any given wine mycobiome.


Progress and challenges of the rural cooperative medical scheme in China.
During China's transition to a market economy in the 1980s and 1990s, the rural population faced substantial barriers to accessing health care and encountered heavier financial burdens than urban residents in paying for necessary health services. In 2003, China started to implement a rural cooperative medical scheme (RCMS), mainly through government subsidies. The scheme operates at the county level and offers a modest benefit package. In spite of rapid economic growth since the early 1980s, income disparities in China have increased, particularly between rural and urban populations. In response, the government has put greater emphasis on social development, including health system development. Examples are the prioritization of improved access to health services and the reduction of the burden of payment for necessary services. After 10 years of implementation, the RCMS now provides coverage to the entire rural population and has substantially improved access to health care. Yet despite a drop in out-of-pocket payments as a proportion of total health expenditure, paying for necessary services continues to cause financial hardship for many rural residents. In its first decade, the RCMS made progress through political mobilization, government subsidies, the readiness of the health-care delivery system, and the availability of a monitoring and evaluation system. Further improving the RCMS will require a focus on cost containment, quality improvement and making the scheme portable.


Access to healthcare, HIV/STI testing, and preferred pre-exposure prophylaxis providers among men who have sex with men and men who engage in street-based sex work in the US.
Pre-exposure prophylaxis (PrEP) is a promising strategy for HIV prevention among men who have sex with men (MSM) and men who engage in sex work. But access will require routine HIV testing and contacts with healthcare providers. This study investigated men's healthcare and HIV testing experiences to inform PrEP implementation. We conducted 8 focus groups (n = 38) in 2012 and 56 in-depth qualitative interviews in 2013-14 with male sex workers (MSWs) (n = 31) and other MSM (n = 25) in Providence, RI. MSWs primarily met clients in street-based sex work venues. Facilitators asked participants about access to healthcare and HIV/STI testing, healthcare needs, and preferred PrEP providers. MSWs primarily accessed care in emergency rooms (ERs), substance use clinics, correctional institutions, and walk-in clinics. Rates of HIV testing were high, but MSWs reported low access to other STI testing, low insurance coverage, and unmet healthcare needs including primary care, substance use treatment, and mental health services. MSM not engaging in sex work were more likely to report access to primary and specialist care. Rates of HIV testing among these MSM were slightly lower, but they reported more STI testing, more insurance coverage, and fewer unmet needs. Preferred PrEP providers for both groups included primary care physicians, infectious disease specialists, and psychiatrists. MSWs were also willing to access PrEP in substance use treatment and ER settings. PrEP outreach efforts for MSWs and other MSM should engage diverse providers in many settings, including mental health and substance use treatment, ERs, needle exchanges, correctional institutions, and HIV testing centers. Access to PrEP will require financial assistance, but can build on existing healthcare contacts for both populations.


Sex and death in the male pea aphid, Acyrthosiphon pisum: The life-history effects of a wing dimorphism.
Insect dispersal dimorphisms, in which both flight-capable and flightless individuals occur in the same species, are thought to reflect a balance between the benefits and costs of dispersal. Fitness costs and benefits associated with wing dimorphism were investigated in the male pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). In one-on-one mating competitions in small arenas between winged and wingless males, the winged aphids obtained most of the matings with virgin females. In contrast, during competition experiments in larger cages with multiple individuals of each morph, the winged males no longer had a clear mating advantage over wingless males. In the absence of competition, wingless males had marginally higher lifetime reproductive success than winged males, probably because mating winged males tended to die faster than wingless males. In the absence of females, winged males survived longer than wingless males and this difference disappeared under starvation conditions. Mating males of both morphs died significantly faster than males without access to females. There does not appear to be a direct tradeoff of dispersal ability with life history characteristics in pea aphid males, suggesting that the advantages of producing winged males may result from outbreeding.


An Evaluation of Publication Bias in High-Impact Orthopaedic Literature.
Statistical analysis of systematic reviews allows the results of previous studies to be combined and synthesized to assess the overall health effect of the intervention in question. Systematic reviews can also be used to guide the creation of clinical practice guidelines and are considered to have a high level of evidence. Thus, it is important that their methodological quality is of the highest standard. Publication bias presents 2 problems: (1) studies with significant results may be overrepresented in systematic reviews and meta-analyses ("false positives") and (2) studies without significant results may not be included in systematic reviews and meta-analyses ("false negatives") because each study, on its own, was underpowered, meaning that some treatment options that may have clinical benefit will not be adopted. We performed a study to evaluate the techniques used by authors to report and evaluate publication bias in the top 10 orthopaedic journals as well as 3 orthopaedic-related Cochrane groups. Two authors independently screened the titles and abstracts to identify systematic reviews and meta-analyses. We assessed publication bias in the systematic reviews that did not assess publication bias themselves. Our final sample included 694 systematic reviews or meta-analyses that met our inclusion criteria. Our review included 502 studies (72%) that focused on clinical outcomes, with the majority of the remaining studies focused on predictive and prognostic accuracy (20%) or diagnostic accuracy (5%). Publication bias was discussed in 295 (42.5%) of the included studies and was assessed in 135 (19.5%). Of the studies that assessed publication bias, 31.9% demonstrated evidence of publication bias. Only 43% and 22% of studies that involved use of the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines discussed and assessed publication bias, respectively. Publication bias is infrequently discussed and assessed in the high-impact orthopaedic literature. Furthermore, nearly one-third of the studies that assessed for publication bias demonstrated evidence of publication bias. In addition to these shortcomings, fewer than half of these studies involved use of the PRISMA guidelines and yet only one-fourth of the studies assessed for publication bias. By understanding the degree to which publication bias is discussed and presented in high-impact orthopaedic literature, changes can be made by journals and researchers alike to improve the overall quality of research produced and reported.


Dose-dependent effects of diallyl disulfide on plasma glucose and free fatty acid levels in rats.
The acute effects of oral administration of diallyl disulfide (DADS), the major organosulfur compound of garlic, on plasma glucose and free fatty acid (FFA) concentrations were examined in rats. Male, 10-week-old Sprague-Dawley rats were divided into DADS-free and DADS-administered (dose = 10, 20, and 40 mg/kg body weight [BW]) groups. Plasma samples were prepared from whole blood drawn from the tail vein 0, 1, 2, 4, and 6 hr after administration. The stomachs were isolated, and the contents were measured 8 hr after administration. In DADS-administered groups, plasma glucose concentrations were increased in a dose-dependent manner 1 hr after the administration. The increase was transient, except in groups administered 40 mg/kg BW of DADS, in which plasma glucose levels remained significantly higher than the DADS-free levels throughout the experimental period. Similar patterns were observed in the plasma FFA concentrations, although the significant differences were lower than those observed in the plasma glucose concentrations. The gastric contents were dose-dependently elevated after DADS administration. The increase was significant when 20 or 40 mg/kg BW of DADS was administered. These results suggest that oral administration of DADS can mobilize energy substrates into the blood, although a higher dose of DADS slows gastric emptying.


Do associations between objectively-assessed physical activity and neighbourhood environment attributes vary by time of the day and day of the week? IPEN adult study.
To more accurately quantify the potential impact of the neighbourhood environment on adults' physical activity (PA), it is important to compare environment-PA associations between periods of the day or week when adults are more versus less likely to be in their neighbourhood and utilise its PA resources. We examined whether, among adults from 10 countries, associations between objectively-assessed neighbourhood environment attributes and moderate-to-vigorous physical activity (MVPA) varied by time of the day and day of the week. The secondary aim was to examine whether such associations varied by employment status, gender and city. This cross-sectional study included 6,712 adults from 14 cities across 10 countries with ≥1 day of valid accelerometer-assessed MVPA and complete information on socio-demographic and objectively-assessed environmental characteristics within 0.5 and 1 km street-network buffers around the home. Accelerometer measures (MVPA min/h) were created for six time periods from early morning until late evening/night, for weekdays and weekend days separately. Associations were estimated using generalized additive mixed models. Time of the day, day of week, gender and employment status were significant moderators of environment-MVPA associations. Land use mix was positively associated with MVPA in women who were employed and in men irrespective of their employment status. The positive associations between MVPA and net residential density, intersection density and land use mix were stronger in the mornings of weekdays and the afternoon/evening periods of both weekdays and weekend days. Associations between number of parks and MVPA were stronger in the mornings and afternoon/evenings irrespective of day of the week. Public transport density showed consistent positive associations with MVPA during weekends, while stronger effects on weekdays were observed in the morning and early evenings. This study suggests that space and time constraints in adults' daily activities are important factors that determine the impact of neighbourhood attributes on PA. Consideration of time-specific associations is important to better characterise the magnitude of the effects of the neighbourhood environment on PA. Future research will need to examine the contribution of built environment characteristics of areas surrounding other types of daily life centres (e.g., workplaces) to explaining adults' PA at specific times of the day.


Radiating despite a Lack of Character: Ecological Divergence among Closely Related, Morphologically Similar Honeyeaters (Aves: Meliphagidae) Co-occurring in Arid Australian Environments.
Quantifying the relationship between form and function can inform use of morphology as a surrogate for ecology. How the strength of this relationship varies continentally can inform understanding of evolutionary radiations; for example, does the relationship break down when certain lineages invade and diversify in novel habitats? The 75 species of Australian honeyeaters (Meliphagidae) are morphologically and ecologically diverse, with species feeding on nectar, insects, fruit, and other resources. We investigated Meliphagidae ecomorphology and community structure by (1) quantifying the concordance between morphology and ecology (foraging behavior), (2) estimating rates of trait evolution in relation to the packing of ecological space, and (3) comparing phylogenetic and trait community structure across the broad environmental gradients of the continent. We found that morphology explained 37% of the variance in ecology (and 62% vice versa), and we uncovered well-known bivariate relationships among the multivariate ecomorphological data. Ecological trait diversity declined less rapidly than phylogenetic diversity along a gradient of decreasing precipitation. We employ a new method (trait fields) and extend another (phylogenetic fields) to show that while species in phylogenetically clustered, arid-environment assemblages are similar morphologically, they are as varied in foraging behavior as those from more diverse assemblages. Thus, although closely related and similar morphologically, these arid-adapted species have diverged in ecological space to a similar degree as their mesic counterparts.


Deficits in the management of patients with adrenocortical carcinoma in Germany.
Adrenocortical carcinoma (ACC) is a rare tumor with a poor prognosis. Often, the physicians who first treat patients with ACC have no prior experience with the disease. The aim of our study was to evaluate the quality of medical care for patients with ACC in Germany. Data from the German ACC registry were analyzed with regard to the patients' preoperative diagnostic evaluation, histopathological reporting, and clinical follow-up. The findings were compared with the recommendations of the European Network for the Study of Adrenal Tumors (ENSAT). Data were analyzed from 387 patients who had been given an initial diagnosis of ACC in the years 1998 to 2009. 21% of them underwent no hormonal evaluation before surgery, and 59% underwent an inadequate hormonal evaluation. This exposed the patients to unnecessary perioperative risks and impaired their follow-up. 48% did not undergo CT scanning of the chest, even though the lungs are the most frequent site of metastases of ACC. For 13% of the patients, the diagnosis of ACC was later revised by a reference pathologist. For 11% of the patients, the histopathology report contained no information about resection status, even though this is an important determinant of further treatment and prognosis. Optimal management requires re-staging at three-month intervals, yet some patients underwent re-staging only after a longer delay, or not at all. We have identified significant deficits in the care of patients with ACC in Germany. We suspect that the situation is similar for other rare diseases. The prerequisite to better care is close and early cooperation of the treating physicians with specialized centers.


Enhancing activity and thermostability of lipase A from Serratia marcescens by site-directed mutagenesis.
Lipases as significant biocatalysts had been widely employed to catalyze various chemical reactions such as ester hydrolysis, ester synthesis, and transesterification. Improving the activity and thermostability of enzymes is desirable for industrial applications. The lipase of Serratia marcescens belonging to family I.3 lipase has a very important pharmaceutical application in production of chiral precursors. In the present study, to achieve improved lipase activity and thermostability, using computational predictions of protein, four mutant lipases of SML (MutG2P, MutG59P, Mut H279K and MutL613WA614P) were constructed by site-directed mutagenesis. The recombinant mutant proteins were over-expressed in E. coli and purified by affinity chromatography on the Ni-NTA system. Circular dichroism spectroscopy, differential scanning calorimetry and kinetic parameters (Km and kcat) were determined. Our results have shown that the secondary structure of all lipases was approximately similar to one another. The MutG2P and MutG59P were more stable than wild type by approximately 2.3 and 2.9 in T1/2, respectively. The catalytic efficiency (kcat/Km) of MutH279K was enhanced by 2-fold as compared with the wild type (p<0.05). These results indicate that using protein modeling program and creating mutation, can enhance lipase activity and/or thermostability of SML and it also could be used for improving other properties of enzyme to the desired requirements as well as further mutations.


Al13H-: hydrogen atom site selectivity and the shell model.
Using a combination of anion photoelectron spectroscopy and density functional theory calculations, we explored the influence of the shell model on H atom site selectivity in Al(13)H(-). Photoelectron spectra revealed that Al(13)H(-) has two anionic isomers and for both of them provided vertical detachment energies (VDEs). Theoretical calculations found that the structures of these anionic isomers differ by the position of the hydrogen atom. In one, the hydrogen atom is radially bonded, while in the other, hydrogen caps a triangular face. VDEs for both anionic isomers as well as other energetic relationships were also calculated. Comparison of the measured versus calculated VDE values permitted the structure of each isomer to be confirmed and correlated with its observed photoelectron spectrum. Shell model, electron-counting considerations correctly predicted the relative stabilities of the anionic isomers and identified the stable structure of neutral Al(13)H.


DeepScratch: Single-cell based topological metrics of scratch wound assays.
Changes in tissue architecture and multicellular organisation contribute to many diseases, including cancer and cardiovascular diseases. Scratch wound assay is a commonly used tool that assesses cells' migratory ability based on the area of a wound they cover over a certain time. However, analysis of changes in the organisational patterns formed by migrating cells following genetic or pharmacological perturbations are not well explored in these assays, in part because analysing the resulting imaging data is challenging. Here we present DeepScratch, a neural network that accurately detects the cells in scratch assays based on a heterogeneous set of markers. We demonstrate the utility of DeepScratch by analysing images of more than 232,000 lymphatic endothelial cells. In addition, we propose various topological measures of cell connectivity and local cell density (LCD) to characterise tissue remodelling during wound healing. We show that LCD-based metrics allow classification of CDH5 and CDC42 genetic perturbations that are known to affect cell migration through different biological mechanisms. Such differences cannot be captured when considering only the wound area. Taken together, single-cell detection using DeepScratch allows more detailed investigation of the roles of various genetic components in tissue topology and the biological mechanisms underlying their effects on collective cell migration.


Molecular, metabolic, and genetic control: An introduction.
The living cell is a miniature, self-reproducing, biochemical machine. Like all machines, it has a power supply, a set of working components that carry out its necessary tasks, and control systems that ensure the proper coordination of these tasks. In this Special Issue, we focus on the molecular regulatory systems that control cell metabolism, gene expression, environmental responses, development, and reproduction. As for the control systems in human-engineered machines, these regulatory networks can be described by nonlinear dynamical equations, for example, ordinary differential equations, reaction-diffusion equations, stochastic differential equations, or cellular automata. The articles collected here illustrate (i) a range of theoretical problems presented by modern concepts of cellular regulation, (ii) some strategies for converting molecular mechanisms into dynamical systems, (iii) some useful mathematical tools for analyzing and simulating these systems, and (iv) the sort of results that derive from serious interplay between theory and experiment. (c) 2001 American Institute of Physics.


Pacific Peoples, Violence, and the Power and Control Wheel.
This qualitative project was the first to study values and practices about sexual assault among migrant communities from the Cook Islands, Fiji, Niue, Samoa, Tokelau, Tonga, and Tuvalu in New Zealand. It aimed to identify customs, beliefs, and practices among these ethnic groups that were protective and preventive factors against sexual violence. Researchers were ethnically matched with 78 participants from the seven ethnic communities, and conducted individual interviews and one female focus group using protocols that were culturally appropriate for each ethnic group. Interviews were thematically analyzed. The study identified the brother-sister covenant and the sanctity of women as strong protective and preventive factors against sexual violence, expressed differently in each culture. Most participants viewed sexual violence as involving their extended families, village, and church communities, rather than solely the individuals concerned. However, the communal values and practices of these seven Pacific cultures raise questions about the individualistic assumptions and the meaning of violence underlying the Power and Control Wheel and the Duluth Model of domestic violence. It also raises questions about how such an individualized model can help services effectively support women in these collective societies who are experiencing violence, and how it can contribute to Pacific community prevention of violence. This study is therefore relevant to countries with significant populations of Pacific peoples and other collective cultures.


Is medical students' moral orientation changeable after preclinical medical education?
Moral orientation can affect ethical decision-making. Very few studies have focused on whether medical education can change the moral orientation of the students. The purpose of the present study was to document the types of moral orientation exhibited by medical students, and to study if their moral orientation was changed after preclinical education. From 2007 to 2009, the Mojac scale was used to measure the moral orientation of Taiwan medical students. The students included 271 first-year and 109 third-year students. They were rated as a communitarian, dual, or libertarian group and followed for 2 years to monitor the changes in their Mojac scores. In both first and third-year students, the dual group after 2 years of preclinical medical education did not show any significant change. In the libertarian group, first and third-year students showed a statistically significant increase from a score of 99.4 and 101.3 to 103.0 and 105.7, respectively. In the communitarian group, first and third-year students showed a significant decline from 122.8 and 126.1 to 116.0 and 121.5, respectively. During the preclinical medical education years, students with communitarian orientation and libertarian orientation had changed in their moral orientation to become closer to dual orientation. These findings provide valuable hints to medical educators regarding bioethics education and the selection criteria of medical students for admission.


Constituents of osmeterial secretion of pre-final instar larvae of citrus swallowtail,Papilio demodocus (Esper) (Lepidoptera: Papilionidae).
The defensive osmeterial secretion of pre-final instar larvae of the citrus swallowtail,Papilio demodocus (Lepidoptera: Papilionidae) was found to contain methyl 3-hydroxybutanoate, 3-hydroxybutanoic acid, α-pinene, myrcene, limonene, β-phellandrene, (Z)-ocimene, (E)-ocimene, β-caryophyllene, (E)-β-farnesene, and germacrene-A, as well as a further number of unidentified sesquiterpenoid constituents. The presence of germacrene-A in the secretion was inferred from the formation of β-elemene under certain GC conditions. Larvae of the second, third, and fourth instars produce qualitatively similar secretions. Remarkable quantitative differences were found between the secretions of individual larvae. These variations could not be correlated with the diet on which the larvae were fed, their sex, instar, or color form. However, in a number of larvae the two prongs of the osmeterium were found to produce quantitatively different secretions.


Criteria for the diagnosis of fibromyalgia: validation of the modified 2010 preliminary American College of Rheumatology criteria and the development of alternative criteria.
To validate the 2011 modification of the 2010 American College of Rheumatology (ACR) preliminary criteria for the diagnosis of fibromyalgia (2011ModCr) and develop alternative criteria in a sample of patients with diverse pain disorders that are commonly seen in everyday practice by pain specialists, rheumatologists, and psychologists. Eight clinicians from geographically varied locations in the US evaluated patients with chronic pain and psychiatric disorders using a standard set of questions that included the 2011ModCr questions, the Symptom Impact Questionnaire (SIQR), a 28-area pain location inventory (PLI), and the Short Form 36. Alternative diagnostic criteria were developed from the same data set using logistic regression and receiver operating curve analysis. Complete data on 321 patients were evaluated; there were 135 patients with fibromyalgia (according to the 1990 ACR criteria) and 186 patients with 16 other common chronic pain problems. Comparing the 2011ModCr with the 1990 ACR criteria provided a sensitivity of 83%, a specificity of 67%, and a correct classification of 74%. Alternative criteria were derived from the 10-item symptom score from the SIQR symptoms and the 28-area PLI. Maximal diagnostic accuracy was obtained with ≥17 pain sites (range 0-28) and an SIQR symptom score of ≥21 (range 0-50). These alternative criteria had a diagnostic sensitivity of 81%, a specificity of 80%, and a correct classification of 80%. The 2011ModCr had robust operating characteristics. Alternative criteria based on symptom items from the SIQR and pain locations from the PLI had comparable operating characteristics, with somewhat better specificity and ease of use.


Comparison of three methods for beat-to-beat-interval extraction from continuous blood pressure and electrocardiogram with respect to heart rate variability analysis.
In recent years the analysis of heart rate variability (HRV) has become a suitable method for characterizing autonomous cardiovascular regulation. The aim of this study was to investigate the differences in HRV estimated from continuous blood pressure (BP) measurement by different methods in comparison to electrocardiogram (ECG) signals. The beat-to-beat intervals (BBI) were simultaneously extracted from the ECG and blood pressure of 9 cardiac patients (10 min, Colin system, 1000-Hz sampling frequency). For both data types, slope, peak, and correlation detection algorithms were applied. The short-term variability was calculated using concurrent 10-min BP and ECG segments. The root mean square errors in comparison to ECG slope detection were: 1.74 ms for ECG correlation detection; 5.42 ms for ECG peak detection; 5.45 ms for BP slope detection; 5.75 ms for BP correlation detection; and 11.96 ms for BP peak detection. Our results show that the variability obtained with ECG is the most reliable. Moreover, slope detection is superior to peak detection and slightly superior to correlation detection. In particular, for ECG signals with higher frequency characteristics, peak detection often exhibits more artificial variability. Besides measurement noise, respiratory modulation and pulse transit time play an important role in determining BBI. The slope detection method applied to ECG should be preferred, because it is more robust as regards morphological changes in the signals, as well as physiological properties. As the ECG is not recorded in most animal studies, distal pulse wave measurement in combination with correlation or slope detection may be considered an acceptable alternative.


[Observation on the efficacy of acupuncture at key acupoints combined with rehabilitation therapy for spasmodic hemiplegia after cerebral infarction].
To assess the clinical efficacy of acupuncture at key acupoints combined with the routine rehabilitation training of limb function on spasmodic hemiplegia after cerebral infarction. Eighty-six cases were randomized into an acupuncture combined with rehabilitation group (group A, 44 cases) and a rehabilitation group (group B, 42 cases). In group A, the key acupoints were selected from head, face, chest, abdomen, shoulder, back, hands, feet and ankles such as Cuanzhu (BL 2), Danzhong (CV 17), Jianyu (LI 15) and Yanglao (SI 16) were stimulated with acupuncture. In combination, the routine limb rehabilitation training was applied, once every day. In group B, the routine limb rehabilitation training was applied alone. In both groups, 10 treatments made one session and 2 sessions were required totally. Before and after treatment, Fugl-Meyer scale and functional independent measurement (FIM) scale were adopted to assess the limb motor level and the activity of daily life in the patients respectively. The modified Ashworth scale was used to assess the effect of anti-spasm. The total effective rate of anti-spasm was 90.9% (40/44) in the group A, which was superior to 73.8% (32/42) in the group B (P < 0.05). After 2 sessions of treatment, Fugl-Meyer score and FIM score were all improved in both groups (all P < 0.05) and the results in the group A were better than those in the group B (all P < 0.05). Additionally, the improvement of FIM score after the 1st session of treatment in the group A was better as compared with the group B, indicating the significant difference (P < 0.05). Acupuncture at key acupoints combined with rehabilitation therapy effectively relieves the spasmodic condition of the patients with post-stroke spasmodic hemiplegia, improves the limb function and the life activity of the patients.


Diabetes risk and provision of diabetes prevention activities in 44 low-income and middle-income countries: a cross-sectional analysis of nationally representative, individual-level survey data.
The global burden of diabetes is rising rapidly, yet there is little evidence on individual-level diabetes prevention activities undertaken by health systems in low-income and middle-income countries (LMICs). Here we describe the population at high risk of developing diabetes, estimate diabetes prevention activities, and explore sociodemographic variation in these activities across LMICs. We performed a pooled, cross-sectional analysis of individual-level data from nationally representative, population-based surveys conducted in 44 LMICs between October, 2009, and May, 2019. Our sample included all participants older than 25 years who did not have diabetes and were not pregnant. We defined the population at high risk of diabetes on the basis of either the presence of impaired fasting glucose (or prediabetes in countries with a haemoglobin A1c available) or overweight or obesity, consistent with the WHO Package of Essential Noncommunicable Disease Guidelines for type 2 diabetes management. We estimated the proportion of survey participants that were at high risk of developing diabetes based on this definition. We also estimated the proportion of the population at high risk that reported each of four fundamental diabetes prevention activities: physical activity counselling, weight loss counselling, dietary counselling, and blood glucose screening, overall and stratified by World Bank income group. Finally, we used multivariable Poisson regression models to evaluate associations between sociodemographic characteristics and these activities. The final pooled sample included 145 739 adults (86 269 [59·2%] of whom were female and 59 468 [40·4%] of whom were male) across 44 LMICs, of whom 59 308 (40·6% [95% CI 38·5-42·8]) were considered at high risk of diabetes (20·6% [19·8-21·5] in low-income countries, 38·0% [37·2-38·9] in lower-middle-income countries, and 57·5% [54·3-60·6] in upper-middle-income countries). Overall, the reach of diabetes prevention activities was low at 40·0% (38·6-41·4) for physical activity counselling, 37·1% (35·9-38·4) for weight loss counselling, 42·7% (41·6-43·7) for dietary counselling, and 37·1% (34·7-39·6) for blood glucose screening. Diabetes prevention varied widely by national-level wealth: 68·1% (64·6-71·4) of people at high risk of diabetes in low-income countries reported none of these activities, whereas 49·0% (47·4-50·7) at high risk in upper-middle-income countries reported at least three activities. Educational attainment was associated with diabetes prevention, with estimated increases in the predicted probability of receipt ranging between 6·5 (3·6-9·4) percentage points for dietary fruit and vegetable counselling and 21·3 (19·5-23·2) percentage points for blood glucose screening, among people with some secondary schooling compared with people with no formal education. A large proportion of individuals across LMICs are at high risk of diabetes but less than half reported receiving fundamental prevention activities overall, with the lowest receipt of these activities among people in low-income countries and with no formal education. These findings offer foundational evidence to inform future global targets for diabetes prevention and to strengthen policies and programmes to prevent continued increases in diabetes worldwide. Harvard T H Chan School of Public Health McLennan Fund: Dean's Challenge Grant Program and the EU's Research and Innovation programme Horizon 2020.


A common export pathway for proteins binding complex redox cofactors?
The precursor polypeptides of periplasmic proteins binding seven types of redox cofactor have unusually long signal sequences bearing a consensus (S/T)-R-R-x-F-L-K motif immediately before the hydrophobic region. Such "double-arginine' signal sequences are not, in general, found on the precursors of other periplasmic proteins. It is suggested that precursor proteins with double-arginine signal sequences share a common specialization in their export pathway. The nature of this specialization, the structure of the double-arginine signal sequences, and the possible relationship with the double-arginine signal peptide-dependent thylakoid import pathway are discussed.


Spillover modes in multiplex games: double-edged effects on cooperation and their coevolution.
In recent years, there has been growing interest in studying games on multiplex networks that account for interactions across linked social contexts. However, little is known about how potential cross-context interference, or spillover, of individual behavioural strategy impact overall cooperation. We consider three plausible spillover modes, quantifying and comparing their effects on the evolution of cooperation. In our model, social interactions take place on two network layers: repeated interactions with close neighbours in a lattice, and one-shot interactions with random individuals. Spillover can occur during the learning process with accidental cross-layer strategy transfer, or during social interactions with errors in implementation. Our analytical results, using extended pair approximation, are in good agreement with extensive simulations. We find double-edged effects of spillover: increasing the intensity of spillover can promote cooperation provided cooperation is favoured in one layer, but too much spillover is detrimental. We also discover a bistability phenomenon: spillover hinders or promotes cooperation depending on initial frequencies of cooperation in each layer. Furthermore, comparing strategy combinations emerging in each spillover mode provides good indication of their co-evolutionary dynamics with cooperation. Our results make testable predictions that inspire future research, and sheds light on human cooperation across social domains.


Quercus pyrenaica Honeydew Honey With High Phenolic Contents Cause DNA Damage, Apoptosis, and Cell Death Through Generation of Reactive Oxygen Species in Gastric Adenocarcinoma Cells.
Many studies have shown that honey with high phenolic contents prevents cancer formation. Furthermore, recent studies have demonstrated that honey can be used for the treatment of cancer as well as cancer prevention. Antineoplastic effects of honey are often associated with their antioxidant phenolic contents. However, very few studies have dealt with the association of phenolic contents of honeys in terms of antiproliferative effects. The aim of this study was, therefore, to elucidate the cytotoxic, genotoxic, apoptotic, and reactive oxygen species (ROS) generating effects of honey samples on the basis of their phenolic and flavonoid contents. Fourteen different honey varieties were collected from various parts of Turkey, and their characteristics regarding total phenols, flavonoids, and antioxidant contents were determined to test their effects on gastric cancer cells (AGS). For convenience, 2 honey varieties were selected, namely, Ida Mountains Quercus pyrenaica honeydew honey (QPHH-IM) having the highest phenolic and antioxidant content and Canakkale multifloral honey (MFH-C) with the lowest phenolic and antioxidant content. Levels of 11 different phenolic compounds in QPHH-IM and MFH-C samples were determined by LC-MS/MS. AGS cells were incubated with different concentrations of QPHH-IM and MFH-C for 24 hours, then the cell viability, DNA damage, apoptosis, and generation of ROS were determined. We found that QPHH-IM had more cytotoxic, genotoxic, and apoptotic effects than that of MFH-C. We think that these effects are probably related to pro-oxidant activities due to the high phenolic contents present. Therefore, further research on high-phenolic honey may contribute to the future development of cancer therapeutics.


Blockade of immunosuppressive cytokines restores NK cell antiviral function in chronic hepatitis B virus infection.
NK cells are enriched in the liver, constituting around a third of intrahepatic lymphocytes. We have previously demonstrated that they upregulate the death ligand TRAIL in patients with chronic hepatitis B virus infection (CHB), allowing them to kill hepatocytes bearing TRAIL receptors. In this study we investigated whether, in addition to their pathogenic role, NK cells have antiviral potential in CHB. We characterised NK cell subsets and effector function in 64 patients with CHB compared to 31 healthy controls. We found that, in contrast to their upregulated TRAIL expression and maintenance of cytolytic function, NK cells had a markedly impaired capacity to produce IFN-γ in CHB. This functional dichotomy of NK cells could be recapitulated in vitro by exposure to the immunosuppressive cytokine IL-10, which was induced in patients with active CHB. IL-10 selectively suppressed NK cell IFN-γ production without altering cytotoxicity or death ligand expression. Potent antiviral therapy reduced TRAIL-expressing CD56(bright) NK cells, consistent with the reduction in liver inflammation it induced; however, it was not able to normalise IL-10 levels or the capacity of NK cells to produce the antiviral cytokine IFN-γ. Blockade of IL-10 +/- TGF-β restored the capacity of NK cells from both the periphery and liver of patients with CHB to produce IFN-γ, thereby enhancing their non-cytolytic antiviral capacity. In conclusion, NK cells may be driven to a state of partial functional tolerance by the immunosuppressive cytokine environment in CHB. Their defective capacity to produce the antiviral cytokine IFN-γ persists in patients on antiviral therapy but can be corrected in vitro by IL-10+/- TGF-β blockade.


Targeting cancer stem cells with oncolytic virus.
Cancer stem cells (CSCs) represent a distinct subpopulation of cancer cells which are shown to be relatively resistant to conventional anticancer therapies and have been correlated to disease recurrence. Oncolytic viruses utilize methods of cell killing that differ from traditional therapies and thus are able to elude the typical mechanisms that CSCs use to resist current chemotherapies and radiotherapies. Moreover, genetically engineered oncolytic viruses may further augment the oncolytic effects. Here we review the recent data regarding the ability of several oncolytic viruses to eradicate CSCs.


On the brink of isolation: Population estimates of the Araguaian river dolphin in a human-impacted region in Brazil.
Populations of freshwater dolphins are declining in response to increased human pressure, including habitat degradation, overfishing, bycatch, poaching and obstruction of free-flowing river corridors by dams. At least three river dolphin species occur in South America: the Amazonian river dolphin, or boto (Inia geoffrensis), the Bolivian river dolphin (Inia boliviensis) and the tucuxi (Sotalia fluviatilis). A fourth species, the Araguaian boto (Inia araguaiaensis), been proposed for the Tocantins-Araguaia, a large river basin in northern Brazil. Here we show that the Araguaian boto population in the Tocantins River is relatively small (N = 1083, CV = 0.52). During a survey to estimate density and abundance, 138 groups (198 individuals) of botos were observed along a ~600 km stretch of the Tocantins River in five different habitats (river margin, river channel, channel, island margin, and a dam reservoir). Overall, lower densities of the Araguaian boto were registered downstream of the Tucuruí dam, the world's fifth largest hydropower dam. Density was 68% lower in the river margin habitat downstream (0.23 ind./km2, CV = 0.92) than upstream (0.72 ind./km2, CV = 0.53). In addition, density within the Tucuruí reservoir decreases from upstream areas towards the dam. Geographic post-stratification of data into sub-regions (downstream, reservoir, upstream) in relation to the Tucuruí dam helped to reduce CV by ~70%, which illustrates the high variability in the encounter rate in these areas. Our findings suggest that the Araguaian boto population has been impacted by the construction of the Tucuruí dam. The construction of other dams proposed for the Tocantins-Araguaia basin should be planned strategically to minimize overlapping with the Araguaian boto distribution. Coordinated conservation actions are imperative to prevent the Araguaian boto from reaching extinction or near-extinction as some of their Asian counterparts such as the Yangtze, Ganges, and Indus river dolphins.


Primary neuron culture for nerve growth and axon guidance studies in zebrafish (Danio rerio).
Zebrafish (Danio rerio) is a widely used model organism in genetics and developmental biology research. Genetic screens have proven useful for studying embryonic development of the nervous system in vivo, but in vitro studies utilizing zebrafish have been limited. Here, we introduce a robust zebrafish primary neuron culture system for functional nerve growth and guidance assays. Distinct classes of central nervous system neurons from the spinal cord, hindbrain, forebrain, and retina from wild type zebrafish, and fluorescent motor neurons from transgenic reporter zebrafish lines, were dissociated and plated onto various biological and synthetic substrates to optimize conditions for axon outgrowth. Time-lapse microscopy revealed dynamically moving growth cones at the tips of extending axons. The mean rate of axon extension in vitro was 21.4±1.2 µm hr(-1) s.e.m. for spinal cord neurons, which corresponds to the typical ∼0.5 mm day(-1) growth rate of nerves in vivo. Fluorescence labeling and confocal microscopy demonstrated that bundled microtubules project along axons to the growth cone central domain, with filamentous actin enriched in the growth cone peripheral domain. Importantly, the growth cone surface membrane expresses receptors for chemotropic factors, as detected by immunofluorescence microscopy. Live-cell functional assays of axon extension and directional guidance demonstrated mammalian brain-derived neurotrophic factor (BDNF)-dependent stimulation of outgrowth and growth cone chemoattraction, whereas mammalian myelin-associated glycoprotein inhibited outgrowth. High-resolution live-cell Ca(2+)-imaging revealed local elevation of cytoplasmic Ca(2+) concentration in the growth cone induced by BDNF application. Moreover, BDNF-induced axon outgrowth, but not basal outgrowth, was blocked by treatments to suppress cytoplasmic Ca(2+) signals. Thus, this primary neuron culture model system may be useful for studies of neuronal development, chemotropic axon guidance, and mechanisms underlying inhibition of neural regeneration in vitro, and complement observations made in vivo.


Informal caregivers in stroke: Life impact, support, and psychological well-being-A Swedish Stroke Register (Riksstroke) study.
Informal caregivers provide a large amount of day-to-day assistance and are crucial for the ability of survivors to recover and adapt to life after stroke. The development of caregiver support programs is limited by lack of large long-term follow-up studies. We present a comprehensive study of Swedish stroke caregivers' life situation in relation to degree of functional dependency of the survivor. In 2016, the Swedish Stroke Register, Riksstroke, conducted a long-term follow-up survey on caregivers to patients with stroke three and five years earlier. Items on psychological well-being were adapted from the 36-item short-form health survey and poor outcome was defined using the 36-item short-form health survey reference material. Survivor degree of dependency was indicated by the caregiver as independent, partially dependent, or completely dependent. A total of 5063 community dwelling dyads were included: 56.5% of survivors were independent, 33.4% partially dependent, and 10.1% completely dependent. Caregiver life impact, need of support, and proportion of poor psychological well-being increased incrementally with survivor degree of dependency. In the completely dependent group where 41.1% of survivors could not be left unattended for more than 1 h, 23.7% of caregivers expressed unmet need of caregiver support; 51.4% reported poor psychological well-being compared to 19.3% in the independent group. The caregiver situation varies greatly with degree of survivor dependency which makes generalizations of caregiver needs difficult. Our results emphasize the need for integrating support aimed specifically at caregivers to survivors of stroke with a large degree of dependency.


Cervical cancer screening rates in the United States and the potential impact of implementation of screening guidelines.
The remarkable success achieved in cervical cancer prevention is largely attributable to cervical cytology screening, also known as the Papanicolaou (Pap) test. The American Cancer Society (ACS) revised screening guidelines for Pap testing in 2002. The impact of these changes on future numbers of Pap tests has not been assessed. Using National Health Interview Survey (NHIS) data to determine historical screening patterns, we extrapolate the numbers of Pap tests that would be performed through 2010, under 5 different scenarios of implementation of screening guidelines. From 1993 to 2003, there was a steady increase in the number of Pap tests, with an estimated 65.6 million Pap tests performed in 2003. Approximately two thirds of women born after 1930 reported having been screened within the previous year, and 85% within the previous 3 years. Fifteen percent of Pap tests were performed in hysterectomized women, most of whom, according to current guidelines, should not be screened. Based on population projections, if screening behavior remains unchanged, 75 million Pap tests will be performed in 2010. Full compliance with ACS guidelines would approximately halve the total number of tests to 34 million. Potentially, with more appropriate allocation of resources according to guidelines, all women could be screened and the total number of Pap tests reduced, despite projected increases in the population.


Stability, Reactivity, Selectivity, Catalysis, and Predictions of 1,3,2,5-Diazadiborinine: Computational Insight into a Boron-Boron Frustrated Lewis Pair.
Recent progress in frustrated Lewis pairs (FLPs) has attracted increasing attention. However, most of the FLPs are composed of Lewis basic phosphines and Lewis acidic boranes. In 2015, Kinjo and co-workers reported the first intramolecular boron-boron FLP, namely, 1,3,2,5-diazadiborinine (1), which showed high regioselectivity in the reactions with methyl trifluoromethansulfonate, phenylacetylene, and CO2. More interestingly, the activation of CO2 was found to be reversible when the temperature was elevated to 90 °C. Here, we performed thorough density functional theory (DFT) calculations to understand the stability, reactivity, and selectivity of 1. The electronic effects and favorable orbital overlap of 1 with substrates play a crucial role in the reactivity and regioselectivity. On the basis of the results, several organic substrates activated by 1, including aldehyde, ketone, imine, hydrogen, ammonia, phosphine, and silane, were explored computationally. The results show that the combination of aldehyde, ketone, imine, and silane are reversible, especially at elevated temperatures. Finally, we modeled the hydrophosphination of Me2PH and phenylacetylene catalyzed by 1. The rate-determining step is the hydrophosphination with an activation barrier of 31.7 kcal/mol, indicating that the process might be possible. Our results may open an avenue to design new organocatalysts.


The AMPAR subunit GluR2: still front and center-stage.
Abnormal influx of Ca(2+) through AMPA-type glutamate receptors (AMPARs) is thought to contribute to the neuronal death associated with a number of brain disorders. AMPARs exist as both Ca(2+)-impermeable and Ca(2+)-permeable channels. AMPARs are encoded by four genes designated GluR1 (GluR-A) through GluR4 (GluR-D). The presence of the GluR2 subunit renders heteromeric AMPA receptor assemblies Ca(2+)-impermeable. Molecular diversity of AMPARs under physiological and pathological conditions is generated by differential spatio-temporal patterns of GluR expression, by alternative RNA splicing and editing and by targeting and trafficking of receptor subunits at dendritic spines. The GluR2 gene is under transcriptional control by the RE1 element specific transcription factor, a gene silencing factor which renders it neuron-specific. GluR2 transcripts are edited by ADAR2 (double-stranded RNA-specific editase 1). AMPAR targeting and trafficking to spines are regulated by synaptic activity and are critical to synaptic plasticity. Recent studies involving animal models of transient forebrain ischemia and epilepsy show that GluR2 mRNA and GluR2 subunit expression are downregulated in vulnerable neurons prior to cell death. Ca(2+) imaging and electrical recording from individual pyramidal neurons in hippocampal slices reveal changes in AMPAR functional properties after ischemia. In slices from post-ischemia animals, CA1 neurons with robust action potentials exhibit greatly enhanced AMPA-elicited rises in intracellular Ca(2+). Excitatory postsynaptic currents in post-ischemic CA1 exhibit an enhanced Ca(2+)-dependent component that appears to be mediated by Ca(2+)-permeable AMPARs. These studies provide evidence for Ca(2+) influx through AMPARs in neurons destined to die. To examine whether acute GluR2 downregulation, even in the absence of a neurological insult, can induce neuronal death, we performed knockdown experiments in rats and gerbils with antisense oligonucleotides targeted to GluR2 mRNA. GluR2 antisense oligonucleotide induced neuronal cell death of pyramidal neurons and enhanced pathogenicity of brief ischemic episodes. These observations provide evidence for Ca(2+) influx through AMPARs in neurons destined to die and implicate Ca(2+)-permeable AMPARs in the pathogenesis of ischemia-induced neuronal death.


A monocyte-TNF-endothelial activation axis in sickle transgenic mice: Therapeutic benefit from TNF blockade.
Elaboration of tumor necrosis factor (TNF) is a very early event in development of ischemia/reperfusion injury pathophysiology. Therefore, TNF may be a prominent mediator of endothelial cell and vascular wall dysfunction in sickle cell anemia, a hypothesis we addressed using NY1DD, S+SAntilles , and SS-BERK sickle transgenic mice. Transfusion experiments revealed participation of abnormally activated blood monocytes exerting an endothelial activating effect, dependent upon Egr-1 in both vessel wall and blood cells, and upon NFκB(p50) in a blood cell only. Involvement of TNF was identified by beneficial impact from TNF blockers, etanercept and infliximab, with less benefit from an IL-1 blocker, anakinra. In therapeutic studies, etanercept ameliorated multiple disturbances of the murine sickle condition: monocyte activation, blood biomarkers of inflammation, low platelet count and Hb, vascular stasis triggered by hypoxia/reoxygenation (but not if triggered by hemin infusion), tissue production of neuro-inflammatory mediators, endothelial activation (monitored by tissue factor and VCAM-1 expression), histopathologic liver injury, and three surrogate markers of pulmonary hypertension (perivascular inflammatory aggregates, arteriolar muscularization, and right ventricular mean systolic pressure). In aggregate, these studies identify a prominent-and possibly dominant-role for an abnormal monocyte-TNF-endothelial activation axis in the sickle context. Its presence, plus the many benefits of etanercept observed here, argue that pilot testing of TNF blockade should be considered for human sickle cell anemia, a challenging but achievable translational research goal.


IC-2 Suppresses Proliferation and Induces Apoptosis of Bladder Cancer Cells via the Wnt/β-Catenin Pathway.
BACKGROUND The Wnt/β-catenin signaling pathway participates in many important tumorigeneses processes, including bladder cancer. The inhibition of abnormal activation of Wnt pathways might provide a new approach to tumor treatment. In the present study, we investigated the role of IC-2, a novel Wnt pathways small molecular inhibitor, in bladder cancer tumorigenesis. MATERIAL AND METHODS Bladder cancer cells were treated with various concentrations of IC-2 (0-5 μM) in vitro. The proliferation ability was measured using colony formation assay and apoptosis was measured using flow cytometry analysis. The protein expression was detected using Western blot analysis. Xenograft in vivo assay was performed to assess tumor growth. RESULTS IC-2 suppressed the proliferation and aggravated the apoptosis of bladder cancer cells in dose-dependent and time-dependent manners in vitro. Moreover, high concentrations of IC-2 inhibited the Wnt pathway-related protein expression levels, including β-catenin, Cyclin D1, and TCF4. In vivo, administration of IC-2 in xenograft mice decreased the β-catenin expression and reduced the tumor volume. CONCLUSIONS Our results validate the tumor-inhibition effect of IC-2 on bladder cancer in vivo and in vitro, providing a novel therapeutic strategy for bladder cancer.


Craniocervical fixation with occipital condyle screws: biomechanical analysis of a novel technique.
A human cadaveric biomechanical study comparing craniocervical fixation techniques. To quantitatively compare the biomechanical stability of a new technique for occipitocervical fixation using the occipital condyles with an established method for craniocervical spine fusion. Stabilization of the occipitocervical junction remains a challenge. The occiput does not easily accommodate instrumentation because of access and spatial constraints. In fact, the area available for the implant fixation is limited and can be restricted further when a suboccipital craniectomy has been performed, posing a challenge to current fixation techniques. Occipital screws are also associated with the potential for intracranial complications. Six fresh frozen cadaveric specimens occiput-C4 were tested intact, after destabilization and after fixation as follows: (1) occipital plate with C1 lateral mass screws and C2 pars screws and (2) occipital condyle screws with C1 lateral mass screws and C2 pars screws. Specimens were loaded in a custom spine testing apparatus and subjected to the following tests, all performed under 50-N unconstrained axial preload: flexion, extension, lateral bending, and axial rotation at 1.5 Nm. The constructs were statistically compared with a one-way analysis of variance and compared with the intact condition. Motions were reduced by approximately 80% compared with the intact condition for both configurations under all motions. There were no statistically significant differences in the range of motion (ROM) between the 2 instrumentation conditions. The mean values indicated decreased ROM with the novel occipital condyle screw construct in comparison with the standard occipital plate and rod system. Craniocervical stabilization using occipital condyle screws as the sole cephalad fixation point is biomechanically equivalent with regard to the modes tested (ROM and stiffness) to the standard occipital plate construct.


Ince-Gaussian beams in a quadratic-index medium.
The propagation of Ince-Gaussian beams in media where the refractive index varies quadratically with the distance from the optical axis is studied. Explicit expressions for the complex beam parameter and the longitudinal phase shift are derived and discussed. Ince-Gaussian eigenmodes with constant width can be obtained by satisfying a relation between the beam width and the quadratic-medium coefficient. The derivation has included the possibility of propagation of Ince-Gaussian beams in complex lenslike media having quadratic transverse variations of the index of refraction and the gain or loss.


The identification of combat survivability tasks associated with naval vessel damage in maritime environments.
Effective Navy personnel have the physical ability to perform combat survivability tasks commensurate with their unique physical requirements due to the distinctive characteristics of naval platforms. The aim of this investigation was to identify the physically demanding whole-of-ship tasks that are performed by Navy personnel while at sea. A mixed method design was used to identify tasks, inclusive of focus groups and field observations. From a series of ten focus groups, nine tasks were deemed to be physically demanding whole-of-ship tasks. A subsequent field observation of a combat survivability training course resulted in a refined and expanded 33-item list of physically demanding whole-of-ship tasks across six categories, including; replenishment at sea, emergency response, firefighting, leak stop and repair, toxic hazard and casualty evacuation. The findings from this study provide the basis for the development of physical employment standards for whole-of-ship tasks within the Royal Australian Navy.


Control in chronic condition self-care management: how it occurs in the health worker-client relationship and implications for client empowerment.
To examine health worker-client interactions during care planning to understand processes that foster client empowerment and disempowerment. It is unclear how health worker-client exchanges and information sharing through chronic condition care planning currently operate in primary health care. Moreover, it is unclear how control in these exchanges either enhances collaborative decision-making, partnership and client empowerment, or works to create client disempowerment and dependency on workers and health services. Critical discourse analysis of qualitative data from ethnographic observations and audio-taped worker-client consultations. Multidisciplinary teams in two Australian community-based primary healthcare sites participated. This included nurses, general practitioners and allied health workers and their clients who had a chronic condition care plan. Nineteen worker-client consultations were observed/recorded in 2011. Control was expressed through multiple processes inherent in the worker role and in their interactions with clients. When workers exercised disproportionate control and clients relinquished their own control, client disempowerment and dependency were evident. Clients' attempts to gain control and workers' attempts to relinquish control alleviated clients' disempowerment and dependency. However, structural features of information sharing systems and workers' care planning behaviours diminished such efforts. Worker awareness of their communication style and the power of their role must improve for client chronic condition self-care management to be achieved. Training on the impacts of control in worker communication and systems where they work must be provided if unbeneficial forms of client dependency are to be overcome and true self-care management is to be realized.


suddengains: An R package to identify sudden gains in longitudinal data.
Sudden gains are large and stable improvements in an outcome variable between consecutive measurements, for example during a psychological intervention with multiple assessments. Researching these occurrences could help understand individual change processes in longitudinal data. Three criteria are generally used to identify sudden gains in psychological interventions. However, applying these criteria can be time consuming and prone to errors if not fully automated. Adaptations to these criteria and methodological decisions such as how multiple gains are handled vary across studies and are reported with different levels of detail. These problems limit the comparability of individual studies and make it hard to understand or replicate the exact methods used. The R package suddengains provides a set of tools to facilitate sudden gains research. This article illustrates how to use the package to identify sudden gains or sudden losses and how to extract descriptive statistics as well as exportable data files for further analysis. It also outlines how these analyses can be customised to apply adaptations of the standard criteria. The suddengains package therefore offers significant scope to improve the efficiency, reporting, and reproducibility of sudden gains research.


Distribution of volumes of individual glomeruli in kidneys at autopsy: association with age, nephron number, birth weight and body mass index.
Glomerular hypertrophy occurs in a number of normal and pathological states. Glomerular volume in kidneys at autopsy is usually indirectly derived from estimates of total glomerular mass and nephron number, and provides only a single value per kidney, with no indication of the range of volumes of glomeruli within the kidney of any given subject. We review findings of the distribution of volumes of different glomeruli within subjects without kidney disease, and their correlations with age, nephron number, birth weight and body mass index (BMI). The study describes findings from autopsy kidneys of selected adult white males from the Southeast USA who had unexpected deaths, and who did not have renal scarring or renal disease. Total glomerular (nephron) number and total glomerular volume were estimated using the disector/fractionator combination, and mean glomerular volume (Vglom) was derived. The volumes of 30 individual glomeruli (IGV) in each subject were determined using the disector/Cavalieri method. IGV values were compared by categories of age, nephron number, birth weight and BMI. There was substantial variation in IGV within subjects. Older age, lower nephron number, lower birth weight and gross obesity were associated with higher mean IGV and with greater IGV heterogeneity. High Vglom and high IGVs were associated with more glomerulosclerosis. However, amongst the generally modest numbers of sclerosed glomeruli, the pattern was uniformly of ischemic collapse of the glomerular tuft. There was no detectable focal segmental glomerular tuft injury. In this series of people without overt renal disease, greater age, nephron deficit, lower birth weight and obesity were marked by glomerular enlargement and greater glomerular volume heterogeneity within individuals.


Regulation and Levels of the Thylakoid K+/H+ Antiporter KEA3 Shape the Dynamic Response of Photosynthesis in Fluctuating Light.
Crop canopies create environments of highly fluctuating light intensities. In such environments, photoprotective mechanisms and their relaxation kinetics have been hypothesized to limit photosynthetic efficiency and therefore crop yield potential. Here, we show that overexpression of the Arabidopsis thylakoid K+/H+ antiporter KEA3 accelerates the relaxation of photoprotective energy-dependent quenching after transitions from high to low light in Arabidopsis and tobacco. This, in turn, enhances PSII quantum efficiency in both organisms, supporting that in wild-type plants, residual light energy quenching following a high to low light transition represents a limitation to photosynthetic efficiency in fluctuating light. This finding underscores the potential of accelerating quenching relaxation as a building block for improving photosynthetic efficiency in the field. Additionally, by overexpressing natural KEA3 variants with modification to the C-terminus, we show that KEA3 activity is regulated by a mechanism involving its lumen-localized C-terminus, which lowers KEA3 activity in high light. This regulatory mechanism fine-tunes the balance between photoprotective energy dissipation in high light and maximum quantum yield in low light, likely to be critical for efficient photosynthesis in fluctuating light conditions.


The relation between major histocompatibility complex (MHC) restriction and the capacity of Ia to bind immunogenic peptides.
The capacity of purified I-Ad, I-Ed, I-Ak, and I-Ek to bind to protein derived peptides that have been previously reported to be T cell immunogens has been examined. For each of the 12 peptides studied strong binding to the relevant Ia restriction element was observed. All the peptides bound more than one Ia molecule; however, for 11 of 12 peptides, the dominant binding was to the restriction element, whereas in one instance the dominant binding was to a nonrestriction element. When the peptides were used to inhibit the presentation of antigen by prefixed accessory cells to T cells, an excellent correlation was found between the capacity of a peptide to inhibit the binding of an antigen to purified Ia and the capacity of the peptide to inhibit accessory cell presentation of the antigen. Thus, the binding of peptide to purified Ia is immunologically relevant, and Ia seems to be the only saturable molecule on the surface of the accessory cell involved in antigen presentation. Inhibition analysis also indicated that all peptides restricted to a particular Ia molecule competitively inhibited one another, suggesting that each Ia restriction element has a single binding site for antigen. Cross-linking of labeled peptides to Ia followed by electrophoretic analysis and autoradiography suggested that this single binding site is made up of portions of both alpha and beta chains of Ia.


Simultaneous Micropatterning of Fibrous Meshes and Bioinks for the Fabrication of Living Tissue Constructs.
Fabrication of biomimetic tissues holds much promise for the regeneration of cells or organs that are lost or damaged due to injury or disease. To enable the generation of complex, multicellular tissues on demand, the ability to design and incorporate different materials and cell types needs to be improved. Two techniques are combined: extrusion-based bioprinting, which enables printing of cell-encapsulated hydrogels; and melt electrowriting (MEW), which enables fabrication of aligned (sub)-micrometer fibers into a single-step biofabrication process. Composite structures generated by infusion of MEW fiber structures with hydrogels have resulted in mechanically and biologically competent constructs; however, their preparation involves a two-step fabrication procedure that limits freedom of design of microfiber architectures and the use of multiple materials and cell types. How convergence of MEW and extrusion-based bioprinting allows fabrication of mechanically stable constructs with the spatial distributions of different cell types without compromising cell viability and chondrogenic differentiation of mesenchymal stromal cells is demonstrated for the first time. Moreover, this converged printing approach improves freedom of design of the MEW fibers, enabling 3D fiber deposition. This is an important step toward biofabrication of voluminous and complex hierarchical structures that can better resemble the characteristics of functional biological tissues.


Tumor Mutation Burden and Prognosis in Patients with Colorectal Cancer Treated with Adjuvant Fluoropyrimidine and Oxaliplatin.
Recent sequencing studies revealed that a subset of colorectal cancer harbors a significantly higher number of somatic mutations. These hypermutated tumors show distinct clinicopathologic features. However, the prognostic impact of the hypermutated tumors is not clearly established. We analyzed tumor mutation burden (TMB) from targeted next-generation sequencing data of 40 major genes in 516 patients with colorectal cancer. TMB was defined as total number of nonsynonymous mutations per tumor. Cutoff value for TMB-high was chosen by which best discriminated relapse-free survival (RFS) using the Contal and O'Quigley method. In the TCGA data, mutation count of the selected 40 genes reflected the whole exome mutation burden (Pearson correlation = 0.873, P < 0.001). In our patient cohort, 8 or more mutations in the 40 genes was defined as TMB-high, which best discriminated RFS. A total of 55 patients (10.7%) had TMB-high. TMB-high tumors were more frequently found in a proximal location (63.6%) and had a higher proportion of N0 disease (30.9%) and MSI-H (49.1%) compared with TMB-low. Most importantly, TMB-high was associated with better 5-year RFS compared with TMB-low (96.3% vs. 79.8%, P = 0.005). Although there was significant overlap between TMB-high and MSI-H, MSI-H status was not significantly associated with RFS. Multivariate analysis revealed TMB-high as an independent positive prognostic factor for RFS [adjusted HR, 0.16 (95% confidence interval, 0.04-0.66), P = 0.011]. TMB-high is associated with better prognosis in patients with colorectal cancer treated with curative surgery followed by adjuvant fluoropyrimidine and oxaliplatin chemotherapy.


Physiological and molecular responses to drought in Petunia: the importance of stress severity.
Plant responses to drought stress vary depending on the severity of stress and the stage of drought progression. To improve the understanding of such responses, the leaf physiology, abscisic acid (ABA) concentration, and expression of genes associated with ABA metabolism and signalling were investigated in Petunia × hybrida. Plants were exposed to different specific substrate water contents (θ = 0.10, 0.20, 0.30, or 0.40 m(3)·m(-3)) to induce varying levels of drought stress. Plant responses were investigated both during the drying period (θ decreased to the θ thresholds) and while those threshold θ were maintained. Stomatal conductance (g(s)) and net photosynthesis (A) decreased with decreasing midday leaf water potential (Ψ(leaf)). Leaf ABA concentration increased with decreasing midday Ψ(leaf) and was negatively correlated with g(s) (r = -0.92). Despite the increase in leaf ABA concentration under drought, no significant effects on the expression of ABA biosynthesis genes were observed. However, the ABA catabolism-related gene CYP707A2 was downregulated, primarily in plants under severe drought (θ = 0.10 m(3)•m(-3)), suggesting a decrease in ABA catabolism under severe drought. Expression of phospholipase Dα (PLDα), involved in regulating stomatal responses to ABA, was enhanced under drought during the drying phase, but there was no relationship between PLDα expression and midday Ψ(leaf) after the θ thresholds had been reached. The results show that drought response of plants depends on the severity of drought stress and the phase of drought progression.


Mononuclear and Polynuclear Chemistry of Rhenium (III): Its Pronounced Homophilicity.
Extensive chemical, spectrophotometric, and x-ray structural studies have shown that trivalent rhenium is strongly homophilic-that is, it tends to form bonds to other Re(111) atoms-and it forms at least three different series of [ReX(4)](n)(n-) complexes. The mononuclear, square complex, [ReBr(4)](-), adds two water molecules to give trans-[ReBr(4)(H(2)O)(2)](-). The binuclear complexes [Re(2)Cl(8)](2-) and [Re(2)Br(8)](2-) have strong Re-Re bonds, unsupported by halide bridges. The trinuclear species, [Re(3)X(12)](3-) or Re(3)X(9)L(3), contain the triangular Re(3)X(9) clusters. Use of ReCI(3) appears always to lead directly to products containing Re(3)Cl(9); this unit exists in ReCl(3) itself and does not appear to be kinetically labile. The [Re(2)X(8)](2-) ions are obtained by reduction of ReO(4)(-) in aqueous HCl or HBr. Salts of [ReBr(4)(H(2)O)(2)](-) can be obtained directly from solutions of ReBr(3) in HBr along with numerous other compounds, some containing trinuclear clusters.


Global expression studies in baker's yeast reveal target genes for the improvement of industrially-relevant traits: the cases of CAF16 and ORC2.
Recent years have seen a huge growth in the market of industrial yeasts with the need for strains affording better performance or to be used in new applications. Stress tolerance of commercial Saccharomyces cerevisiae yeasts is, without doubt, a trait that needs improving. Such trait is, however, complex, and therefore only in-depth knowledge of their biochemical, physiological and genetic principles can help us to define improvement strategies and to identify the key factors for strain selection. We have determined the transcriptional response of commercial baker's yeast cells to both high-sucrose and lean dough by using DNA macroarrays and liquid dough (LD) model system. Cells from compressed yeast blocks display a reciprocal transcription program to that commonly reported for laboratory strains exposed to osmotic stress. This discrepancy likely reflects differences in strain background and/or experimental design. Quite remarkably, we also found that the transcriptional response of starved baker's yeast cells was qualitatively similar in the presence or absence of sucrose in the LD. Nevertheless, there was a set of differentially regulated genes, which might be relevant for cells to adapt to high osmolarity. Consistent with this, overexpression of CAF16 or ORC2, two transcriptional factor-encoding genes included in this group, had positive effects on leavening activity of baker's yeast. Moreover, these effects were more pronounced during freezing and frozen storage of high-sucrose LD. Engineering of differentially regulated genes opens the possibility to improve the physiological behavior of baker's yeast cells under stress conditions like those encountered in downstream applications.


Patterns of Care and Clinical Outcomes of HER2-positive Metastatic Breast Cancer Patients With Newly Diagnosed Stage IV or Recurrent Disease Undergoing First-line Trastuzumab-based Therapy: A Multicenter Retrospective Cohort Study.
The aim of the study was to compare the patterns of care and clinical outcomes of HER2-positive metastatic breast cancer (MBC) patients with de novo or recurrent disease who underwent first-line trastuzumab-based therapy. This was a multicenter retrospective cohort study including consecutive patients with HER2-positive MBC who received first-line trastuzumab-based therapy. Analyses on treatment response and effectiveness were conducted according to type of metastatic presentation (ie, de novo vs. recurrent disease). Exploratory analyses were used to evaluate whether the use of surgery of the primary tumor in the de novo cohort influenced patients' survival. From January 2000 to December 2013, 416 patients were included in the study, 113 (27.2%) presented with de novo MBC and 303 (72.8%) with recurrent disease. Compared with patients in the recurrence cohort, those in the de novo cohort had worse baseline characteristics, received more aggressive first-line treatments, and showed better survival, with an adjusted hazard ratio (HR) for progression-free survival (PFS) of 0.65 (95% confidence interval [CI], 0.43-0.97; P = .035) and for overall survival (OS) of 0.53 (95% CI, 0.30-0.95; P = .034). In the de novo cohort, the 54 patients (47.8%) who underwent surgery of the primary tumor had significantly better PFS (adjusted HR, 0.44; 95% CI, 0.26-0.72; P = .001) and OS (adjusted HR, 0.49; 95% CI, 0.26-0.93; P = .029) than those who did not undergo surgery. Patients with de novo HER2-positive MBC showed significantly better survival outcomes than those with recurrent disease. In this population, surgery of the primary breast tumor was associated with better outcomes.


Large and Small Dendritic Spines Serve Different Interacting Functions in Hippocampal Synaptic Plasticity and Homeostasis.
The laying down of memory requires strong stimulation resulting in specific changes in synaptic strength and corresponding changes in size of dendritic spines. Strong stimuli can also be pathological, causing a homeostatic response, depressing and shrinking the synapse to prevent damage from too much Ca(2+) influx. But do all types of dendritic spines serve both of these apparently opposite functions? Using confocal microscopy in organotypic slices from mice expressing green fluorescent protein in hippocampal neurones, the size of individual spines along sections of dendrite has been tracked in response to application of tetraethylammonium. This strong stimulus would be expected to cause both a protective homeostatic response and long-term potentiation. We report separation of these functions, with spines of different sizes reacting differently to the same strong stimulus. The immediate shrinkage of large spines suggests a homeostatic protective response during the period of potential danger. In CA1, long-lasting growth of small spines subsequently occurs consolidating long-term potentiation but only after the large spines return to their original size. In contrast, small spines do not change in dentate gyrus where potentiation does not occur. The separation in time of these changes allows clear functional differentiation of spines of different sizes.


Forcing the reversibility of a mechanochemical reaction.
Mechanical force modifies the free-energy surface of chemical reactions, often enabling thermodynamically unfavoured reaction pathways. Most of our molecular understanding of force-induced reactivity is restricted to the irreversible homolytic scission of covalent bonds and ring-opening in polymer mechanophores. Whether mechanical force can by-pass thermodynamically locked reactivity in heterolytic bimolecular reactions and how this impacts the reaction reversibility remains poorly understood. Using single-molecule force-clamp spectroscopy, here we show that mechanical force promotes the thermodynamically disfavored SN2 cleavage of an individual protein disulfide bond by poor nucleophilic organic thiols. Upon force removal, the transition from the resulting high-energy unstable mixed disulfide product back to the initial, low-energy disulfide bond reactant becomes suddenly spontaneous, rendering the reaction fully reversible. By rationally varying the nucleophilicity of a series of small thiols, we demonstrate how force-regulated chemical kinetics can be finely coupled with thermodynamics to predict and modulate the reversibility of bimolecular mechanochemical reactions.


Voronoi dipole moments for the simulation of bulk phase vibrational spectra.
We present the Voronoi tessellation of electron density data to obtain molecular dipole moments in bulk phase ab initio molecular dynamics simulations for the calculation of vibrational spectra. Opposed to the established scheme of maximally localized Wannier functions, this approach does not rely on computationally demanding localization procedures. Nevertheless, we show at the examples of methanol, benzene, and phenol that it provides infrared and Raman spectra of similar quality and is even superior in specific cases like the Raman spectra of benzene and phenol. We have also applied the Voronoi method to a mixture of the ionic liquid 1-ethyl-3-methylimidazolium acetate with water, and show that it is advantageous in systems with significant charge transfer.


Compensation of magnetic disturbances improves inertial and magnetic sensing of human body segment orientation.
This paper describes a complementary Kalman filter design to estimate orientation of human body segments by fusing gyroscope, accelerometer, and magnetometer signals from miniature sensors. Ferromagnetic materials or other magnetic fields near the sensor module disturb the local earth magnetic field and, therefore, the orientation estimation, which impedes many (ambulatory) applications. In the filter, the gyroscope bias error, orientation error, and magnetic disturbance error are estimated. The filter was tested under quasi-static and dynamic conditions with ferromagnetic materials close to the sensor module. The quasi-static experiments implied static positions and rotations around the three axes. In the dynamic experiments, three-dimensional rotations were performed near a metal tool case. The orientation estimated by the filter was compared with the orientation obtained with an optical reference system Vicon. Results show accurate and drift-free orientation estimates. The compensation results in a significant difference (p < 0.01) between the orientation estimates with compensation of magnetic disturbances in comparison to no compensation or only gyroscopes. The average static error was 1.4 degrees (standard deviation 0.4) in the magnetically disturbed experiments. The dynamic error was 2.6 degrees root means square.


Enhancement of two photon processes in quantum dots embedded in subwavelength metallic gratings.
We show a large enhancement of two-photon absorption processes in nanocrystal quantum dots and of light upconversion efficiency from the IR to the near-IR spectral regime, using a hybrid optical device in which near-IR emitting InAs quantum dots were embedded on top a metallic nanoslit array. The resonant enhancement of these nonlinear optical processes is due to the strong local electromagnetic field enhancements inside the nanoslit array structure at the extraordinary transmission resonances. A maximal two-photon absorption enhancement of more than 20 was inferred. Different high field regions were identified for different polarizations, which can be used for designing and optimizing efficient nonlinear processes in such hybrid structures. Combining nanocrystal quantum dots with subwavelength metallic nanostructures is therefore a promising way for a range of possible nonlinear optical devices.


Zopiclone as positive control in studies examining the residual effects of hypnotic drugs on driving ability.
Zopiclone (7.5 mg) is frequently used as a positive control in studies that examine the residual effects of hypnotic drugs on driving ability and related skills. This review summarizes studies examining the effects of zopiclone, and discusses its usefulness as a comparator drug for investigations of residual effects of novel sleep medication. A literature review (Pubmed and Embase) was conducted searching for studies that tested zopiclone on driving. Cross references were checked for additional papers. Eight studies utilizing the standardized on-the-road driving test consistently showed that in the morning following bedtime administration zopiclone (7.5 mg) significantly impaired driving performance. A total of 191 healthy volunteers were tested after placebo and zopiclone (7.5 mg). Meta analyses showed no significant differences in driving performance after zopiclone (7.5 mg) between adult and elderly healthy volunteers. The combined effect size (ES) and 95% confidence interval (95%CI) for healthy volunteers was 0.782 (0.620, 0.944). Relative to placebo, an average increment of 3.0 cm in Standard Deviation of Lateral Position (SDLP) was observed when treated with zopiclone (7.5 mg). This deviation was higher than the increment in SDLP reported for drivers with a blood alcohol concentration of 0.05% (+2.4 cm). Results from driving simulators and psychometric tests are consistent with the on-road driving test results. In conclusion, zopiclone (7.5 mg) is a reliable positive control, that consistently shows significant and meaningful impairment on the on-the-road driving test.