Prolonged-release fampridine in multiple sclerosis: Improved ambulation effected by changes in walking pattern.
Prolonged-release fampridine (PR-fampridine, 4-aminopyridine) increases walking speed in the timed 25-foot walk test (T25FW) in some patients (timed-walk responders) with multiple sclerosis (MS). To explore the effects of PR-fampridine on different aspects of walking function and to identify associated gait modifications in subjects with MS. In this prospective, randomized, placebo-controlled, double-blind, phase II study (FAMPKIN; clinicaltrials.gov, NCT01576354), subjects received a 6-week course of oral placebo or PR-fampridine treatment (10 mg, twice daily) before crossing over. Using 3D-motion-analysis, kinematic and kinetic parameters were assessed during treadmill walking (primary endpoint). Clinical outcome measures included T25FW, 6-minute walk test (6MWT), and balance scales. Physical activity in everyday life was measured with an accelerometer device. Data from 55 patients were suitable for analysis. Seventeen subjects were timed-walk responders under PR-fampridine. For the total study population and for responders, a significant increase in walking speed (T25FW) and distance (6MWT) was observed. Gait pattern changes were found at the single-subject level and correlated with improvements in the T25FW and 6MWT. Physical activity was increased in responders. PR-fampridine improves walking speed, endurance, and everyday physical activity in a subset of subjects with MS and leads to individual modifications of the gait pattern.


Who is caring for the underserved? A comparison of primary care physicians and nonphysician clinicians in California and Washington.
Little is known about whether different types of physician and nonphysician primary care clinicians vary in their propensity to care for underserved populations. The objective of this study was to compare the geographic distribution and patient populations of physician and nonphysician primary care clinicians. This study was a cross-sectional analysis of 1998 administrative and survey data on primary care clinicians (family physicians, general internists, general pediatricians, nurse practitioners, physician assistants, and certified nurse-midwives) in California and Washington. For geographic analysis, main outcome measures were practice in a rural area, a vulnerable population area (communities with high proportions of minorities or low-income residents), or a health professions shortage area (HPSA). For patient population analysis, outcomes were the proportions of Medicaid, uninsured, and minority patients in the practice. Physician assistants ranked first or second in each state in the proportion of their members practicing in rural areas and HPSAs, and in California physician assistants also had the greatest proportion of their members working in vulnerable populations areas (P < .001). Compared with primary care physicians overall, nurse practitioners and certified nurse-midwives also tended to have a greater proportion of their members in rural areas and HPSAs (P < .001). Family physicians were much more likely than other primary care physicians to work in rural areas and HPSAs (P < .001). Compared with physicians, nonphysician clinicians in California had a substantially greater proportion of Medicaid, uninsured, and minority patients (P < .001). Nonphysician primary care clinicians and family physicians have a greater propensity to care for underserved populations than do primary care physicians in other specialties. Achieving a more equitable pattern of service to needy populations will require ongoing, active commitment by policy makers, educational institutions, and the professions to a mission of public service and to incentives that support and promote care to the underserved.


Non-transferrin-bound iron (NTBI) uptake by T lymphocytes: evidence for the selective acquisition of oligomeric ferric citrate species.
Iron is an essential nutrient in several biological processes such as oxygen transport, DNA replication and erythropoiesis. Plasma iron normally circulates bound to transferrin. In iron overload disorders, however, iron concentrations exceed transferrin binding capacity and iron appears complexed with low molecular weight molecules, known as non-transferrin-bound iron (NTBI). NTBI is responsible for the toxicity associated with iron-overload pathologies but the mechanisms leading to NTBI uptake are not fully understood. Here we show for the first time that T lymphocytes are able to take up and accumulate NTBI in a manner that resembles that of hepatocytes. Moreover, we show that both hepatocytes and T lymphocytes take up the oligomeric Fe3Cit3 preferentially to other iron-citrate species, suggesting the existence of a selective NTBI carrier. These results provide a tool for the identification of the still elusive ferric-citrate cellular carrier and may also open a new pathway towards the design of more efficient iron chelators for the treatment of iron overload disorders.


Orientation gradient detection exhibits variable coupling between first- and second-stage filtering mechanisms.
We investigated sensitivity to orientation modulation using visual stimuli with bandpass filtered noise carriers. We characterized the relationship between the spatial parameters of the modulator and the carrier using a 2-AFC detection task. The relationship between these two parameters is potentially informative of the underlying coupling between first- and second-stage filtering mechanisms, which, in turn, may bear on the interrelationship between striate and extrastriate cortical processing. Our previous experiments on analogous motion stimuli found an optimum sensitivity when the ratio of the carrier and modulator spatial frequency parameters (r) was approximately ten. The current results do not exhibit an optimum sensitivity at a given value of the ratio r. Previous experiments involving second-order modulation sensitivity show an inconsistent range of estimates of optimum sensitivity at values of r between 5 and 50. Our results, using a complementary approach, confirm these discrepancies, demonstrating that the coupling between carrier and modulator frequency parameters depends on a number of stimulus-specific factors, such as contrast sensitivity, stimulus eccentricity, and absolute values of the carrier and modulator spatial frequency parameters. We show that these observations are true for a stimulus limited in eccentricity and that this orientation-modulated stimulus does not exhibit scale invariance. Such processing can not be modeled by a generic filter-rectify-filter model.


With the future behind them: convergent evidence from aymara language and gesture in the crosslinguistic comparison of spatial construals of time.
Cognitive research on metaphoric concepts of time has focused on differences between moving Ego and moving time models, but even more basic is the contrast between Ego- and temporal-reference-point models. Dynamic models appear to be quasi-universal cross-culturally, as does the generalization that in Ego-reference-point models, FUTURE IS IN FRONT OF EGO and PAST IS IN BACK OF EGO. The Aymara language instead has a major static model of time wherein FUTURE IS BEHIND EGO and PAST IS IN FRONT OF EGO; linguistic and gestural data give strong confirmation of this unusual culture-specific cognitive pattern. Gestural data provide crucial information unavailable to purely linguistic analysis, suggesting that when investigating conceptual systems both forms of expression should be analyzed complementarily. Important issues in embodied cognition are raised: how fully shared are bodily grounded motivations for universal cognitive patterns, what makes a rare pattern emerge, and what are the cultural entailments of such patterns?


Reactions of persulfides with the heme cofactor of oxidized myoglobin and microperoxidase 11: reduction or coordination.
Persulfides of cysteine (CysSSH), glutathione (GSSH) or N-methoxycarbonyl-penicillamine (NAcPenSSH) react with the ferric form of myoglobin (metMb(iii)) to yield the oxy-ferrous (oxyMb(ii)) or deoxy-ferrous (deoxyMb(ii)) forms of myoglobin under aerobic or anaerobic conditions, respectively. Under aerobic conditions, CysSSH and NAcPenSSH react with the hypervalent form of myoglobin (ferrylMb(iv)) to yield oxyMb(ii) as the final product with the formation of metMb(iii) as an intermediate. CysSSH and NAcPenSSH coordinate the ferric form of N-acetylated microperoxidase (NAcMP11(iii)) to yield the disulfanido complex NAcMP11(iii)(NAcPenSS), as shown by UV-vis and EPR spectroscopy. Experiments carried out with various NAcMP11 derivatives demonstrate a redox equilibrium between the ferric/ferrous forms of the heme and the polysulfides/persulfides couple. Our results suggest that persulfides possess uncommon redox properties, analogous to that of dihydrolipoic acid.


Analytical determination and pharmacokinetics of robenacoxib in the dog.
An analytical method was developed and validated for the measurement of the novel analgesic and anti-inflammatory drug robenacoxib in blood and plasma of dogs and cats. To prevent nonreproducible carry-over effects, an initial solid phase extraction procedure was followed by high pressure liquid chromatography analysis for samples with concentrations in the range 500 to 20,000 ng/mL. To improve accuracy, samples of concentration 3 to 100 ng/mL were analyzed by liquid chromatography-mass spectrometry. Applying these methods, blood concentration-time profiles and pharmacokinetic variables of robenacoxib in dogs were determined in a four-phase cross-over study, which compared different routes of administration of the drug, including intravenous (i.v.) injection, oral application with and without feed, and subcutaneous (s.c.) application. After i.v. administration the mean clearance from blood was 0.81 L/kg/h, the volume of distribution was 0.77 L/kg for the elimination phase and 0.24 L/kg for steady-state, and the terminal half-life in blood was 0.63 h. Maximum blood concentrations were obtained in less than 1 h following oral or s.c. application. Absolute bioavailability was 88% after s.c. injection, 84% after oral administration to fasted dogs, but was reduced to 62% when applied orally to fed dogs. In canine and feline plasma the degree of binding of robenacoxib to plasma protein in vitro was greater than 98%. The blood:plasma concentration ratio was 0.44:1 in the dog and 0.65:1 in the cat. In conclusion analytical methods for the quantification of robenacoxib in blood and plasma in the dog and cat were developed and validated. In dogs, robenacoxib has good bioavailability after oral (84%) and subcutaneous (88%) administration.


Kuwanon G Preserves LPS-Induced Disruption of Gut Epithelial Barrier In Vitro.
Defects in the gut epithelial barrier have now been recognized to be responsible for diabetic endotoxemia. In everyday life, Mulberry leaf tea is widely used in Asian nations due to its proposed benefits to health and control of diabetes. Evidence indicates the potential role of Kuwanon G (KWG), a component from Morus alba L., on blocking the gut epithelial barrier. In lipopolysaccharides (LPS)-damaged Caco-2 cells, it was found that KWG increased the viability of cells in a concentration-dependent manner. KWG administration significantly elevated the anti-oxidant abilities via increasing ratio of superoxidase dismutase (SOD)/malondialdehyde (MDA) and decreasing reactive oxygen species (ROS) within the cells. During KWG incubation, pro-inflammatory cytokines including interleukin (IL)-1β and tumor necrosis factor (TNF)-α were significantly reduced, tight junction proteins including zonula occludens (ZO)-1, intercellular adhesion molecule (ICAM)-1 and Occludin were dramatically increased as detected by immunofluorescence assay, trans-epithelial electrical resistance was significantly increased and the transmission of albumin-fluorescein isothiocyanate (FITC) across the barrier was decreased. In conclusion, the present study demonstrated that KWG could ameliorate LPS-induced disruption of the gut epithelial barrier by increasing cell viability and tight junction between cells, and decreasing pro-inflammatory cytokines and oxidative damage.


Quantifying the impacts of defaunation on natural forest regeneration in a global meta-analysis.
Intact forests provide diverse and irreplaceable ecosystem services that are critical to human well-being, such as carbon storage to mitigate climate change. However, the ecosystem functions that underpin these services are highly dependent on the woody vegetation-animal interactions occurring within forests. While vertebrate defaunation is of growing policy concern, the effects of vertebrate loss on natural forest regeneration have yet to be quantified globally. Here we conduct a meta-analysis to assess the direction and magnitude of defaunation impacts on forests. We demonstrate that real-world defaunation caused by hunting and habitat fragmentation leads to reduced forest regeneration, although manipulation experiments provide contrasting findings. The extirpation of primates and birds cause the greatest declines in forest regeneration, emphasising their key role in maintaining carbon stores, and the need for national and international climate change and conservation strategies to protect forests from defaunation fronts as well as deforestation fronts.


Perspectives of Rehabilitation Professionals on Implementing a Validated Home Telerehabilitation Intervention for Older Adults in Geriatric Rehabilitation: Multisite Focus Group Study.
Owing to demographic trends and increasing health care costs, quick discharge with geriatric rehabilitation at home is advised and recommended for older adults. Telerehabilitation has been identified as a promising tool to support rehabilitation at home. However, there is insufficient knowledge about how to implement a validated home telerehabilitation system in other contexts. One of the major challenges for rehabilitation professionals is transitioning to a blended work process in which human coaching is supplemented via digital care. The study aimed to gain an in-depth understanding of the factors that influence the implementation of an evidence-based sensor monitoring intervention (SMI) for older adults by analyzing the perspectives of rehabilitation professionals working in 2 different health ecosystems and mapping SMI barriers and facilitators. We adopted a qualitative study design to conduct 2 focus groups, 1 in person in the Netherlands during winter of 2017 and 1 on the web via Zoom (Zoom Video Communications; owing to the COVID-19 pandemic) in Canada during winter of 2022, to explore rehabilitation providers' perspectives about implementing SMI. Qualitative data obtained were analyzed using thematic analysis. Participants were a group of rehabilitation professionals in the Netherlands who have previously worked with the SMI and a group of rehabilitation professionals in the province of Manitoba (Canada) who have not previously worked with the SMI but who were introduced to the intervention through a 30-minute web-based presentation before the focus group. The participants expressed different characteristics of the telerehabilitation intervention that contributed to making the intervention successful for at-home rehabilitation: focus on future participation goals, technology support provides the rehabilitation professionals with objective and additional insight into the daily functioning of the older adults at home, SMI can be used as a goal-setting tool, and SMI deepens their contact with older adults. The analysis showed facilitators of and barriers to the implementation of the telerehabilitation intervention. These included personal or client-related, therapist-related, and technology-related aspects. Rehabilitation professionals believed that telerehabilitation could be suitable for monitoring and supporting older adults' rehabilitation at home. To better guide the implementation of telerehabilitation in the daily practice of rehabilitation professionals, the following steps are needed: ensuring that technology is feasible for communities with limited digital health literacy and cognitive impairments, developing instruction tools and guidelines, and training and coaching of rehabilitation professionals.


Development of directed and random exploration in children.
Are young children just random explorers who learn serendipitously? Or are even young children guided by uncertainty-directed sampling, seeking to explore in a systematic fashion? We study how children between the ages of 4 and 9 search in an explore-exploit task with spatially correlated rewards, where exhaustive exploration is infeasible and not all options can be experienced. By combining behavioral data with a computational model that decomposes search into similarity-based generalization, uncertainty-directed exploration, and random exploration, we map out developmental trajectories of generalization and exploration. The behavioral data show strong developmental differences in children's capability to exploit environmental structure, with performance and adaptiveness of sampling decisions increasing with age. Through model-based analyses, we disentangle different forms of exploration, finding signature of both uncertainty-directed and random exploration. The amount of random exploration strongly decreases as children get older, supporting the notion of a developmental "cooling off" process that modulates the randomness in sampling. However, even at the youngest age range, children do not solely rely on random exploration. Even as random exploration begins to taper off, children are actively seeking out options with high uncertainty in a goal-directed fashion, and using inductive inferences to generalize their experience to novel options. Our findings provide critical insights into the behavioral and computational principles underlying the developmental trajectory of learning and exploration.


Comparison of the immunogenicity and safety of a split-virion, inactivated, trivalent influenza vaccine (Fluzone®) administered by intradermal and intramuscular route in healthy adults.
The aim of the study was to determine whether reduced doses of trivalent inactivated influenza vaccine (TIV) administered by the intradermal (ID) route generated similar immune responses to standard TIV given intramuscularly (IM) with comparable safety profiles. Recent changes in immunization recommendations have increased the number of people for whom influenza vaccination is recommended. Thus, given this increased need and intermittent vaccine shortages, means to rapidly expand the vaccine supply are needed. Previously healthy subjects 18-64 years of age were randomly assigned to one of four TIV vaccine groups: standard 15 μg HA/strain TIV IM, either 9 μg or 6 μg HA/strain of TIV ID given using a new microinjection system (BD Soluvia™ Microinjection System), or 3 μg HA/strain of TIV ID given by Mantoux technique. All vaccines contained A/New Caledonia (H1N1), A/Wyoming (H3N2) and B/Jiangsu strains of influenza. Sera were obtained 21 days after vaccination and hemagglutination inhibition (HAI) assays were performed and geometric mean titers (GMT) were compared among the groups. Participants were queried immediately following vaccination regarding injection pain and quality of the experience. Local and systemic reactions were collected for 7 days following vaccination and compared. Ten study sites enrolled 1592 subjects stratified by age; 18-49 years [N=814] and 50-64 years [N=778]. Among all subjects, for each of the three vaccine strains, the GMTs at 21 days post-vaccination for both the 9 μg and the 6 μg doses of each strain given ID were non inferior to GMTs generated after standard 15 μg doses/strain IM. However, for the 3 μg ID dose, only the A/Wyoming antigen produced a GMT that was non-inferior to the standard IM dose. Additionally, in the subgroup of subjects 50-64 years of age, the 6μg dose given ID induced GMTs that were inferior to the standard IM TIV for the A/H1N1 and B strains. No ID dose produced a GMT superior to that seen after standard IM TIV. Local erythema and swelling were significantly more common in the ID groups but the reactions were mild to moderate and short-lived. No significant safety issues related to intradermal administration were identified. Participants given TIV ID provided favorable responses to questions about their experiences with ID administration. In conclusion, for the aggregated cohorts of adults 18-64 years of age, reduced doses (6 μg and 9 μg) of TIV delivered ID using a novel microinjection system stimulated comparable HAI antibody responses to standard TIV given IM. The reduced 3 μg dose administered ID by needle and syringe, as well as the 6 μg ID for subjects aged 50-64 years of age generated poorer immune responses as compared to the 15 μg IM dose.


Self-assembled air-stable magnesium hydride embedded in 3-D activated carbon for reversible hydrogen storage.
The rational design of stable, inexpensive catalysts with excellent hydrogen dynamics and sorption characteristics under realistic environments for reversible hydrogen storage remains a great challenge. Here, we present a simple and scalable strategy to fabricate a monodispersed, air-stable, magnesium hydride embedded in three-dimensional activated carbon with periodic synchronization of transition metals (MHCH). The high surface area, homogeneous distribution of MgH2 nanoparticles, excellent thermal stability, high energy density, steric confinement by carbon, and robust architecture of the catalyst resulted in a noticeable enhancement of the hydrogen storage performance. The resulting MHCH-5 exhibited outstanding hydrogen storage performance, better than that of most reported Mg-based hydrides, with a high storage density of 6.63 wt% H2, a rapid kinetics loading in <5 min at 180 °C, superior reversibility, and excellent long-term cycling stability over ∼435 h. The significant reduction of the enthalpy and activation energy observed in the MHCH-5 demonstrated enhancement of the kinetics of de-/hydrogenation compared to that of commercial MgH2. The origin of the intrinsic hydrogen thermodynamics was elucidated via solid state 1H NMR. This work presents a readily scaled-up strategy towards the design of realistic catalysts with superior functionality and stability for applications in reversible hydrogen storage, lithium ion batteries, and fuel cells.


Local induction of patterning and programmed cell death in the developing Drosophila retina.
Local cell signaling can pattern the nervous system by directing cell fates, including programmed cell death. In the developing Drosophila retina, programmed cell death is used to remove excess cells between ommatidia. Cell ablation revealed the source and position of signals required for regulating the pattern of programmed cell death among these interommatidial cells. Two types of signals regulate this patterning event. Notch-mediated signals between interommatidial precursors result in removal of unneeded cells. Cone cells and primary pigment cells oppose this signal by supplying a 'life'-promoting activity; evidence is provided that this signal occurs through localized activation of the EGF Receptor/Ras pathway. Together, these signals refine the highly regular pattern observed in the adult retina.


Improving communication when seeking informed consent: a randomised controlled study of a computer-based method for providing information to prospective clinical trial participants.
To assess the efficacy, with respect to participant understanding of information, of a computer-based approach to communication about complex, technical issues that commonly arise when seeking informed consent for clinical research trials. An open, randomised controlled study of 60 patients with diabetes mellitus, aged 27-70 years, recruited between August 2006 and October 2007 from the Department of Diabetes and Endocrinology at the Alfred Hospital and Baker IDI Heart and Diabetes Institute, Melbourne. Participants were asked to read information about a mock study via a computer-based presentation (n = 30) or a conventional paper-based information statement (n = 30). The computer-based presentation contained visual aids, including diagrams, video, hyperlinks and quiz pages. Understanding of information as assessed by quantitative and qualitative means. Assessment scores used to measure level of understanding were significantly higher in the group that completed the computer-based task than the group that completed the paper-based task (82% v 73%; P = 0.005). More participants in the group that completed the computer-based task expressed interest in taking part in the mock study (23 v 17 participants; P = 0.01). Most participants from both groups preferred the idea of a computer-based presentation to the paper-based statement (21 in the computer-based task group, 18 in the paper-based task group). A computer-based method of providing information may help overcome existing deficiencies in communication about clinical research, and may reduce costs and improve efficiency in recruiting participants for clinical trials.


[Cost-effectiveness of diagnostic strategies of severe bacterial infection in infants with fever without a source].
Serious bacterial infections in infants under 2-years-of-age with fever without a source remains, despite advances in vaccination, a matter of concern for both physicians and parents. Having cost-effectiveness information is relevant to guide decision making in clinical practice in this scenario. Objective: To determine the cost-effectiveness of four different strategies of screening in Argentina for serious bacterial infection in children presenting with fever without a source. Materials and methods: We designed a decision tree to model a hypothetical cohort of 10,000 children with fever without a source. We compared the incremental cost-effectiveness of four strategies to detect serious bacterial infection: Rochester criteria + C reactive protein test, Rochester criteria + procalcitonin test, Rochester criteria, and expectant observation. Results: Rochester criteria + C reactive protein test was the most cost-effective strategy with USD$ 784 for each correctly diagnosed case versus USD$ 839 of Rochester criteria + procalcitonin test, USD$ 1,116 of expectant observation or USD$ 1,193 of Rochester criteria. When the probability of serious bacterial infections was equal or less than 14%, the strategy of choice was expectant observation. Conclusions: The Rochester criteria + C reactive protein test was the most cost-effective strategy to detect serious bacterial infection in one to three months old children with fever without a source. However, in low risk settings for such infection, the strategy of choice is expectant observation.


A comparative evaluation of nebulized dexmedetomidine, nebulized ketamine, and their combination as premedication for outpatient pediatric dental surgery.
This study was designed to evaluate and compare the efficacy and safety of nebulized dexmedetomidine, nebulized ketamine, and the combination of these drugs for premedication in pediatric outpatient dental surgeries. A prospective, randomized, double-blinded study was conducted in the Alexandria Main University Hospital after approval of the Medical Ethics Committee. Informed written consent was obtained from the patient's parents. Sixty children classified as ASA physical status I and II and aged 3 to 6 years were enrolled in the study. Patients were randomly categorized into 3 equal groups. Group K was premedicated with nebulized ketamine (2 mg/kg), group D was premedicated with nebulized dexmedetomidine (2 μg/kg), and group DK was premedicated with combined nebulized dexmedetomidine and ketamine (1 μg/kg + 1 mg/kg). The primary end point was the level of sedation when the child was first seen in the operating room 30 minutes after sedation. The secondary end points were tolerance of mask induction, hemodynamic changes, analgesia, sedation at emergence, and wake-up behavior. Studied groups were comparable as regards demographic data (age, weight, sex, and ASA physical status, all P > 0.226) and patients' acceptance of drugs (P = 0.968). Level of sedation at 30 minutes was significantly greater in group DK than in either group K (P = 0.003) or group D (P = 0.009). Group DK had the briefest recovery times, followed in order by group K and group D with progressively longer recovery times. Recovery times were significantly briefer in group DK than in either group K (P = 0.039) or group D (P < 0.001). Group DK had significantly briefer discharge times than group D (P < 0.001). Postoperative analgesia was significantly better in group DK compared with group K (P = 0.008). A nebulized combination of low-dose ketamine and dexmedetomidine produced more satisfactory sedation and provided a smoother induction of general anesthesia than nebulized ketamine or dexmedetomidine alone, with more rapid recovery and no significant side effects.


Effects of carving plane, level of harvest, and oppositional suturing techniques on costal cartilage warping.
Cartilage warping has plagued reconstructive and cosmetic rhinoplasty since the introduction of extra-anatomical cartilage use. With the present level of knowledge, there is no evidence of the warping properties with respect to cartilage harvest and suture techniques and level of rib harvest. This report aims to improve understanding of costal cartilage warping. The sixth through tenth costal cartilages were harvested from six fresh cadavers aged 54 to 90 years. Warping characteristics were followed with respect to level of harvest (i.e., sixth versus seventh), carving orientation, and oppositional suturing. Digital photography of the specimens was performed at various time points (immediately, 1 hour, and 1 month postoperatively). All specimens showed signs of warping beyond 1 hour of carving that continued in a linear fashion to 1 month. There was no statistical difference in the amount of warping specific to the level of harvest, orientation, or with or without oppositional suturing (p<0.05). Cartilage warping remains a problematic obstacle in nasal reconstruction and revision rhinoplasty, but costal cartilage remains the workhorse graft and is an excellent autologous option. Our findings are the first to be described in the literature regarding warping characteristics of costal cartilage with regard to the level of harvest, orientation of carving, and oppositional suturing techniques in a cadaveric model.


Functional diversity positively affects prey suppression by invertebrate predators: a meta-analysis.
The use of pesticides within agricultural ecosystems has led to wide concern regarding negative effects on the environment. One possible alternative is the use of predators of pest species that naturally occur within agricultural ecosystems. However, the mechanistic basis for how species can be manipulated in order to maximize pest control remains unclear. We carried out a meta-analysis of 51 studies that manipulated predator species richness in reference to suppression of herbivore prey to determine which components of predator diversity affect pest control. Overall, functional diversity (FD) based on predator's habitat domain, diet breadth and hunting strategy was ranked as the most important variable. Our analysis showed that increases in FD in polycultures led to greater prey suppression compared to both the mean of the component predator species, and the most effective predator species, in monocultures. Further analysis of individual traits indicated these effects are likely to be driven by broad niche differentiation and greater resource exploitation in functionally diverse predator communities. A decoupled measure of phylogenetic diversity, whereby the overlap in variation with FD was removed, was not found to be an important driver of prey suppression. Our results suggest that increasing FD in predatory invertebrates will help maximize pest control ecosystem services in agricultural ecosystems, with the potential to increase suppression above that of the most effective predator species.


Diagnosis of Peritoneal Metastases with Abdominal Malignancies: Role of ADC Measurement on Diffusion Weighted MRI.
The purpose of our study was to evaluate the utility of the apparent diffusion coefficient (ADC) value measurement in the diagnosis of peritoneal metastases on diffusion weighted magnetic resonance imaging. Diffusion weighted imaging with conventional magnetic resonance imaging sequences was performed on twenty consecutive oncology patients (group I) with peritoneal metastases. The ADC values of the metastases, the peritoneal fat around the metastases (group I) and the peritoneal fat in patients with no malignancy (group II) at b(0-100), b(0-600), and (b 0-1000) s/mm(2) gradients were measured and compared. The apparent diffusion coefficient values of three gradients in peritoneal metastases (2.27±0.4; 1.67±0.7 and 1.09±0.4×10(-3) mm(2)/s at b 100, 600 and 1000 gradients, respectively) were significantly lower than the ADC values of the peritoneal fat around metastases (3.07±0.4; 2.07±0.4; 1.33±0.3×10(-3) mm(2)/s at b 100, 600 and 1000 gradients, respectively) (p<0.05). There was no significant difference between the ADC values of peritoneal fat in the patients of group I and group II at the 3 diffusion gradients (p>0.05). The measurement of ADC values may be used as a complementary diagnostic method in differentiating peritoneal metastases from peritoneal fat on Diffusion Weighted MRI (DWMRI DWMRI).


The Role of Imaging in COVID-19 Pneumonia Diagnosis and Management: Main Positions of the Experts, Key Imaging Features and Open Answers.
Lung imaging is widely involved in facing the coronavirus disease (COVID-19) pandemic. In fact, the COVID-19 infection may lead to a rapidly evolving and potentially fatal pneumonia. Moreover, computed tomography (CT) can be more sensitive than the COVID-19 reverse transcriptase-polymerase chain reaction test, especially at the beginning of the disease. Only patients with mild features consistent with COVID-19 infection, negative COVID-19 test, or positive COVID-19 test but at low risk for disease progression should avoid imaging. However, imaging becomes mandatory if respiratory symptoms worsen. A CT pattern classification has been designed to help both radiologists and clinicians. The typical pattern of COVID-19 is depicted by multifocal, bilateral, and peripheral ground-glass opacities (with or without consolidations or crazy paving) or findings of organizing pneumonia. Moreover, CT has demonstrated a prognostic role in patients with a diagnosis of COVID-19 pneumonia. Lung ultrasounds (LUS) are an emergent tool in the diagnosis of the disease. The adoption of LUS combined to chest X-rays in COVID-19 in pneumonia diagnosis is an interesting prospect that needs to be confirmed.


Intra-catheter leukocyte culture to monitor hemodialysis catheter colonization. A prospective study to prevent catheter-related bloodstream infections.
The most serious problem related to the use of tunneled catheters in hemodialysis is bacteremia. The aim of this study was to detect hemodialysis catheter colonization and, establish a preemptive therapy based on a catheter antibiotic lock in order to prevent development of catheter-related bloodstream infections. During a 24-month period, all patients with tunneled catheters in our hemodialysis unit were evaluated by extracting a through-catheter leukocyte culture every 15 days.There were 28 episodes of catheter colonization occurring in 13 patients (2.2 colonization episodes per 1000 catheter patient-days). At the time of colonization, catheters had been in place for a mean of 562 days (range: 16 to 1475 days). Coagulase negative staphylococci (CNS) were the most common microorganisms to be isolated. A preemptive therapy consisting in teicoplanin locks (10 mg/mL) for 21 days was able to eradicate catheter colonization in 89% of the cases when CNS were isolated. However, relapse of colonization occurred in 61.2% of these cases. The mean duration of catheter use was 239 days (range: 9 to 483 days) after treatment of a colonization episode. The incidence of catheter-related bloodstream infection in our population was 0.78 episodes per 1000 catheter patient-days (IC 95%: 0.374-1.434). This study shows the utility of intra-catheter leukocyte culture for early detection of hemodialysis catheter colonization. Moreover, it establishes that the eradication of biofilm-related CNS is possible without the removal of the catheter, thus enabling a longer catheter lifespan.


Ants on Clerodendrum infortunatum: Disentangling Effects of Larceny and Herbivory.
Nectar larcenists extract nectar from flowers without pollinating them. A reasonable expectation is that any form of nectar larceny should have a detrimental effect on the plant's reproductive success. However, studies reveal an entire range of effects, from highly negative to highly positive. This variation in effect may be partly explained by additional, unmeasured, effects of nectar larcenists on plants. In a study system where two ant species Tapinoma melanocephalum (Fabr.) (Hymenoptera: Formicidae) and Trichomyrmex destructor (Jerd.) (Hymenoptera: Formicidae) act as nectar larcenists, we examined the effect of larceny on the female reproductive success of Clerodendrum infortunatum Gaertn. (Lamiales: Lamiaceae) in rain forest fragments of the Western Ghats, India. This was done through a combination of field observations and a series of field experiments looking at the effects of excluding ants from inflorescences. We found that T. destructor reduces fruit set considerably. Rather than this being a consequence of nectar larceny, however, our experiments show that the negative effect arises instead from the herbivorous behavior of the ant. At a population level, both ant species prefer edges over interiors of forest patches, spatially concentrating the interaction zone to forest edges. Simultaneously considering multiple ecological interactions and disentangling their relative contributions might explain the large variation across species in the observed effect of larceny. The overall population effect of nectar larceny and herbivory is likely to depend on the spatial structuring of plants and ants.


The learning curve for colorectal stent insertion for the treatment of malignant colorectal obstruction.
We aimed to assess the effectiveness of self-expanding metal stent (SEMS) insertion by evaluating the learning curve in relation to the experience of an endoscopist. We retrospectively analyzed the outcomes of 120 SEMS insertion procedures performed by one endoscopist in patients with malignant colorectal obstruction. We compared the technical and clinical success rates, complication rates, and duration of the procedures by quartiles. The mean age of the patients (76 men and 44 women) was 64.6 years. The overall technical success rate was 95.0% (114/120), and the clinical success rate was 90.0% (108/120). The median procedure duration was 16.2 minutes (range, 3.4 to 96.5 minutes). From the first to the last quartile, the technical success rates were 90.0%, 96.7%, 96.7%, and 96.7% (p=0.263), and the clinical success rates were 90.0%, 90.0%, 96.7%, and 83.3% (p=0.588), respectively. Procedure-related complications were observed in 28 patients (23.3%). The complication rates for SEMS insertion when patients were divided by quartiles were 26.7%, 23.3%, 10.0%, and 33.3% (p=0.184), respectively. Moreover, the number of stents per procedure was 1.13, 1.03, 1.00, and 1.00 (p=0.029), respectively. The median duration of SEMS insertion decreased significantly, 20.9 to 14.8 minutes after the first 30 procedures (p=0.005). An experienced endoscopist was able to perform the SEMS insertion procedure easily and effectively after performing 30 SEMS insertions.


Regulatory B10 cell development and function.
B cells are known to instigate and promulgate immune responses by producing antibodies and presenting antigens to T cells. However, a rare but potent B-cell subset in both humans and mice is capable of inhibiting immune responses through the production of the anti-inflammatory cytokine IL-10. Regulatory B cells do not express any unique combination of surface markers but instead represent a small population of B cells that have acquired the unique ability to produce IL-10. This numerically rare B-cell subset is therefore functionally referred to as 'B10 cells' to reflect both their molecular program and the fact that their anti-inflammatory effects in models of autoimmunity, infection and cancer are solely attributable to IL-10 production. As with most B cells, B10 cell development and function appear to be predominantly, if not exclusively, driven by antigen-receptor signals. Once generated, B10 cells respond to both innate and adaptive immune signals, with a requirement for antigen-specific local interactions with T cells to induce IL-10 production and to provide optimal immune suppression in mouse models of autoimmune disease. B10 cells therefore provide an antigen-specific mechanism for delivering IL-10 locally to sites of immune activation and inflammation. The ability of B10 cells to regulate innate and adaptive immune responses makes them an ideal therapeutic target for the treatment of many immune-related disorders.


Alternative Splicing of the Amelogenin Gene in a Caudate Amphibian, Plethodon cinereus.
As the major enamel matrix protein contributing to tooth development, amelogenin has been demonstrated to play a crucial role in tooth enamel formation. Previous studies have revealed amelogenin alternative splicing as a mechanism for amelogenin heterogeneous expression in mammals. While amelogenin and its splicing forms in mammalian vertebrates have been characterized, splicing variants of amelogenin gene still remains largely unknown in non-mammalian species. Here, using PCR and sequence analysis we discovered two novel amelogenin transcript variants in tooth organ extracts from a caudate amphibian, the salamander Plethodoncinereus. The one was shorter -S- (416 nucleotides including untranslated regions, 5 exons) and the other larger -L- (851 nt, 7 exons) than the previously published "normal" gene in this species -M- (812 nucleotides, 6 exons). This is the first report demonstrating the amelogenin alternative splicing in amphibian, revealing a unique exon 2b and two novel amelogenin gene transcripts in Plethodoncinereus.


Sex differences in sleep and influence of the menstrual cycle on women's sleep in junior endurance athletes.
Previous research shows that female athletes sleep better according to objective parameters but report worse subjective sleep quality than male athletes. However, existing sleep studies did not investigate variations in sleep and sleep stages over longer periods and have, so far, not elucidated the role of the menstrual cycle in female athletes' sleep. To address these methodological shortcomings, we investigated sex differences in sleep and sleep stages over 61 continuous days in 37 men and 19 women and examined the role of the menstrual cycle and its phases in 15 women. Sleep was measured by a non-contact radar, and menstrual bleeding was self-reported. Associations were investigated with multilevel modeling. Overall, women tended to report poorer subjective sleep quality (p = .057), but objective measurements showed that women obtained longer sleep duration (p < .001), more light (p = .013) and rapid eye movement sleep (REM; hours (h): p < .001, %: p = .007), shorter REM latency (p < .001), and higher sleep efficiency (p = .003) than men. R2 values showed that sleep duration, REM and REM latency were especially affected by sex. Among women, we found longer time in bed (p = .027) and deep sleep (h: p = .036), and shorter light sleep (%: p = .021) during menstrual bleeding vs. non-bleeding days; less light sleep (h: p = .040), deep sleep (%: p = .013) and shorter REM latency (p = .011) during the menstrual than pre-menstrual phase; and lower sleep efficiency (p = .042) and more deep sleep (%: p = .026) during the follicular than luteal phase. These findings indicate that the menstrual cycle may impact the need for physiological recovery, as evidenced by the sleep stage variations. Altogether, the observed sex differences in subjective and objective sleep parameters may be related to the female athletes' menstrual cycle. The paper provides unique data of sex differences in sleep stages and novel insights into the role of the menstrual cycle in sleep among female athletes.


A prospective study of the impact of musculoskeletal pain and radiographic osteoarthritis on health related quality of life in community dwelling older people.
Pain and radiographic changes are common in persons with osteoarthritis, but their relative contributions to quality of life are unknown. Prospective cohort study of 1098 men and women aged 50-80 years, randomly selected from the electoral roll. Participants were interviewed at baseline and approximately 2.6 and five years later. Participants self-reported prior diagnosis of arthritis and presence of joint pain. Joint space narrowing (JSN) and osteophytes at the hip and knee were assessed by X-ray. Quality of life (QoL) was assessed using the Assessment of QoL (AQoL) instrument. Data was analysed using linear regression and mixed modelling. The median AQoL score at baseline was 7.0, indicating very good QoL. Prevalence of pain ranged from 38-62%. Over five years of observation, pain in the neck, shoulders, back, hips, hands, knees and feet were all independently and negatively associated with QoL, in a dose-response relationship. Diagnosed osteoarthritis at all sites was associated with poorer QoL but after adjustment for pain, this only remained significant at the back. Radiographic OA was not associated with QoL. While AQoL scores declined over five years, there was no evidence of an interaction between pain and time. Pain is common in older adults, is stable over time, and the strongest musculoskeletal correlate of QoL. It also mediates the association between diagnosed OA and QoL. Since the same factors were associated with quality of life over time as at baseline, this suggests that quality of life tracks over a five year period.


Development of Three Different NK Cell Subpopulations during Immune Reconstitution after Pediatric Allogeneic Hematopoietic Stem Cell Transplantation: Prognostic Markers in GvHD and Viral Infections.
Natural killer (NK) cells play an important role following allogeneic hematopoietic stem cell transplantation (HSCT) exerting graft-versus-leukemia/tumor effect and mediating pathogen-specific immunity. Although NK cells are the first donor-derived lymphocytes reconstituting post-HSCT, their distribution of CD56++CD16- (CD56bright), CD56++CD16+ (CD56intermediate=int), and CD56+CD16++ (CD56dim) NK cells is explicitly divergent from healthy adults, but to some extent comparable to the NK cell development in early childhood. The proportion of CD56bright/CD56int/CD56dim changed from 15/8/78% in early childhood to 6/4/90% in adults, respectively. Within this study, we first compared the NK cell reconstitution post-HSCT to reference values of NK cell subpopulations of healthy children. Afterward, we investigated the reconstitution of NK cell subpopulations post-HSCT in correlation to acute graft versus host disease (aGvHD) and chronic graft versus host disease (cGvHD) as well as to viral infections. Interestingly, after a HSCT follow-up phase of 12 months, the distribution of NK cell subpopulations largely matched the 50th percentile of the reference range for healthy individuals. Patients suffering from aGvHD and cGvHD showed a delayed reconstitution of NK cells. Remarkably, within the first 2 months post-HSCT, patients suffering from aGvHD had significantly lower levels of CD56bright NK cells compared to patients without viral infection or without graft versus host disease (GvHD). Therefore, the amount of CD56bright NK cells might serve as an early prognostic factor for GvHD development. Furthermore, a prolonged and elevated peak in CD56int NK cells seemed to be characteristic for the chronification of GvHD. In context of viral infection, a slightly lower CD56 and CD16 receptor expression followed by a considerable reduction in the absolute CD56dim NK cell numbers combined with reoccurrence of CD56int NK cells was observed. Our results suggest that a precise analysis of the reconstitution of NK cell subpopulations post-HSCT might indicate the occurrence of undesired events post-HSCT such as severe aGvHD.


Functional outcomes of acetabular fractures.
The Musculoskeletal Function Assessment is a validated, well-designed, self-administered questionnaire that is useful for determining health status. The modified Merle d'Aubigné clinical hip score is the most generally accepted clinical grading system for evaluating the results of acetabular fracture treatment. The purpose of the present study was to evaluate the relationship between the Musculoskeletal Function Assessment and modified Merle d'Aubigné scores in evaluating the results of acetabular fracture treatment. One hundred and fifty patients with an acetabular fracture met the criteria for inclusion in the present study, which included a minimum of two years of postoperative follow-up, a complete physical examination with use of the Merle d'Aubigné score, and successful completion of the Musculoskeletal Function Assessment questionnaire. Patients were stratified according to a number of factors, including the type of fracture and whether the fracture was an isolated injury or was part of a multiple-injury complex. The mean duration of follow-up was five years (range, two to seventeen years). The mean modified Merle d'Aubigné score was 16.8 (range, 9 to 18), and the mean Musculoskeletal Function Assessment score was 24.9 (range, 0 to 79). The Spearman correlation coefficient between the Merle d'Aubigné score and the Musculoskeletal Function Assessment score was -0.61 (p < 0.0001). Stratification of the patients did not alter these overall results. However, the presence or absence of associated injuries was a significant factor for the Merle d'Aubigné score (p = 0.03). In addition, the Merle d'Aubigné score data were asymmetric, demonstrating a ceiling effect. The Musculoskeletal Function Assessment scores for these patients were relatively high compared with those for the normal population, indicating that complete return to a preinjury functional level is uncommon despite a good-to-excellent Merle d'Aubigné clinical score. Furthermore, the ceiling effect demonstrated by the Merle d'Aubigné score (despite its high correlation with the Musculoskeletal Function Assessment score) limits its usefulness as a method for evaluating the outcome of treatment of acetabular fractures. There have been few published clinical studies in which the Musculoskeletal Function Assessment score has been used as an outcome measure, and reference values are lacking. The present study provides initial guideline reference values for use in the evaluation of patients following an acetabular fracture.


Interventions based on early intensive applied behaviour analysis for autistic children: a systematic review and cost-effectiveness analysis.
Early intensive applied behaviour analysis-based interventions are intensive interventions for autistic children that are often delivered on a one-to-one basis for 20-50 hours per week. To evaluate the clinical effectiveness and cost-effectiveness of early intensive applied behaviour analysis-based interventions for autistic children, based on current evidence. A systematic review and individual participant data meta-analysis were conducted to evaluate the clinical effectiveness of an early intensive applied behaviour analysis-based intervention for autistic children. An economic analysis included a review of existing analyses and the development of a new model. Twenty studies were included in the clinical review. Individual participant data were retrieved from 15 of these studies. Results favoured the interventions when assessing adaptive behaviour after 2 years compared with treatment as usual/eclectic interventions (mean difference 7.00, 95% confidence interval 1.95 to 12.06). In analyses of cognitive ability (intelligence quotient), results favoured the interventions by approximately 10 points after 1 year (mean difference 9.16, 95% confidence interval 4.38 to 13.93) and 2 years (mean difference 14.13, 95% confidence interval 9.16 to 19.10). Evidence for other outcomes was limited and meta-analyses were generally inconclusive. There was no evidence that the effect of the interventions varied with characteristics of the children, but data were limited. Adopting a £30,000 per quality-adjusted life-year threshold, the results of the cost-effectiveness analysis indicate that early intensive applied behaviour analysis-based interventions would need to generate larger benefits or cost savings to be cost-effective. Adopting a public sector perspective and making pessimistic assumptions about long-term effects, the incremental cost-effectiveness ratio for early intensive applied behaviour analysis-based therapy compared with treatment as usual is £189,122 per quality-adjusted life-year. When optimistic assumptions are made, the incremental cost-effectiveness ratio is £46,768 per quality-adjusted life-year. Scenario analyses indicated that these interventions can potentially be cost-effective if long-term improvements persist into adulthood, or if they have significant impact on educational placement. Care should be taken when interpreting these scenarios owing to the limited data. All included studies were at risk of bias, there was substantial heterogeneity and effects varied considerably across studies. The effect of intervention on autism symptom severity, language development and school placement remains uncertain because of the limited data. The long-term effects are unclear owing to a lack of follow-up data. This review found limited evidence that early intensive applied behaviour analysis-based interventions may improve cognitive ability and adaptive behaviour, but the long-term impact of the interventions remains unknown. The economic analysis is constrained by the limited effectiveness evidence, but suggests that these interventions are unlikely to be cost-effective unless clear long-term benefits, or a substantial change in which schools children attend, can be identified. Further studies into the effectiveness of early intensive applied behaviour analysis-based interventions may be warranted if they include well-defined, alternative interventions as comparators and collect relevant outcomes. Consideration should be given to future studies that not only address whether or not early intensive applied behaviour analysis-based interventions are clinically effective, but also aim to identify which components of early intensive applied behaviour analysis-based interventions might drive effectiveness. This study is registered as PROSPERO CRD42017068303. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 24, No. 35. See the NIHR Journals Library website for further project information.


Catechol O-methyltransferase Val158Met polymorphism is associated with cognitive performance in nondemented adults.
The catechol O-methyltransferase (COMT) gene is essential in the metabolic degradation of dopamine in the prefrontal cortex. In the present study, we examined the effect of a Val158Met polymorphism in the COMT gene on individual differences and changes in cognition (executive functions and visuospatial ability) in adulthood and old age. The participants were 292 nondemented men (initially aged 35-85 years) from a random sample of the population (i.e., the Betula study) tested at two occasions with a 5-year interval. Confirmatory factor analyses were used to test the underlying structure of three indicators of executive functions (verbal fluency, working memory, and Tower of Hanoi). Associations between COMT, age, executive functioning, and visuospatial (block design) tasks were examined using repeated-measures analyses of variance. Carriers of the Val allele (with higher enzyme activity) compared with carriers of the Met/Met genotype (with low enzyme activity) performed worse on executive functioning and visuospatial tasks. Individuals with the Val/Val genotype declined in executive functioning over the 5-year period, whereas carriers of the Met allele remained stable in performance. An Age x COMT interaction for visuospatial ability located the effect for middle-aged men only. This COMT polymorphism is a plausible candidate gene for executive functioning and fluid intelligence in nondemented middle-aged and older adults.


Child-care effect sizes for the NICHD Study of Early Child Care and Youth Development.
This report summarizes findings from the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development as effect sizes for exclusive maternal care and--for children in child care--type, quality, and quantity of care. Children (n = 1,261) were recruited at birth and assessed at 15, 24, 36, and 54 months. Exclusive maternal care did not predict child outcomes, but multiple features of child-care experience were modestly to moderately predictive. Higher quality child care was related to advanced cognitive, language, and preacademic outcomes at every age and better socioemotional and peer outcomes at some ages. More child-care hours predicted more behavior problems and conflict, according to care providers. More center-care time was related to higher cognitive and language scores and more problem and fewer prosocial behaviors, according to care providers. Child-care effect sizes are discussed from 3 perspectives: (a) absolute effect sizes, reflecting established guidelines; (b) relative effect sizes, comparing child-care and parenting effects; and (c) possible individual and collective implications for the large numbers of children experiencing child care.


Digital nanoliter to milliliter flow rate sensor with in vivo demonstration for continuous sweat rate measurement.
Microfluidic flow rate sensors have constraints in both detection limits and dynamic range, and are not often easily integrated into lab-on-chip or wearable sensing systems. We constructed a flow rate sensor that easily couples to the outlet of a microfluidic channel, and measures the flow rate by temporarily shorting periodic droplets generated between two electrodes. The device was tested in a dynamic range as low as 25 nL min-1 and as high as 900 000 nL min-1 (36 000× range). It was tested to continuously operate up to ∼200 hours. The device is also simple to fabricate, requiring inexpensive parts, and is small enough to be integrated into wearable devices. The required input pressure is as low as 370 Pascals. An ultra-low flow rate application was demonstrated for wearable sweat biosensing where sweat generation rates (nL min-1 per gland) were accurately measured in human subjects. The digital nanoliter device provides real-time flow rates for sweat rates and may have other applications for low flow rates in microfluidic devices.


A phosphatidylinositol 3-kinase-Pax3 axis regulates Brn-2 expression in melanoma.
Deregulation of transcription arising from mutations in key signaling pathways is a hallmark of cancer. In melanoma, the most aggressive and lethal form of skin cancer, the Brn-2 transcription factor (POU3F2) regulates proliferation and invasiveness and lies downstream from mitogen-activated protein kinase (MAPK) and Wnt/β-catenin, two melanoma-associated signaling pathways. In vivo Brn-2 represses expression of the microphthalmia-associated transcription factor, MITF, to drive cells to a more stem cell-like and invasive phenotype. Given the key role of Brn-2 in regulating melanoma biology, understanding the signaling pathways that drive Brn-2 expression is an important issue. Here, we show that inhibition of phosphatidylinositol 3-kinase (PI3K) signaling reduces invasiveness of melanoma cells in culture and strongly inhibits Brn-2 expression. Pax3, a transcription factor regulating melanocyte lineage-specific genes, directly binds and regulates the Brn-2 promoter, and Pax3 expression is also decreased upon PI3K inhibition. Collectively, our results highlight a crucial role for PI3K in regulating Brn-2 and Pax3 expression, reveal a mechanism by which PI3K can regulate invasiveness, and imply that PI3K signaling is a key determinant of melanoma subpopulation diversity. Together with our previous work, the results presented here now place Brn-2 downstream of three melanoma-associated signaling pathways.


Effects of Charge State on the Structures of Serum Albumin Ions in the Gas Phase: Insights from Cation-to-Anion Proton-Transfer Reactions, Ion Mobility, and Mass Spectrometry.
Understanding the structures of proteins in the gas phase is essential for using gas-phase measurements to infer the properties of proteins in solution. Using serum albumin as a model, this study aims to expand our understanding of this relationship for a larger (66 kDa), multidomain protein that contains 17 internal disulfide bonds. Gas-phase ions were generated from five solutions that preserve varying extents of the native structure. Ion mobility (IM) mass spectrometry, cation-to-anion proton-transfer-reactions (CAPTR), and energy-dependent IM were used to probe the relationship between structure, charge, and solution. Ions generated from increasingly disruptive conditions exhibited higher charge states and larger collision cross-section values. The collision cross-sections of all CAPTR products depend on the original solution and to varying extents the charge state of the product and the precursor. For example, the collision cross-sections of CAPTR products from denaturing conditions are all significantly larger than those of the original native-like ions. Results from energy-dependent experiments show that the structures of the original ions from electrospray ionization and their CAPTR products are a consequence of kinetic trapping and depend on higher-order structure and disulfide bonding in solution. This study builds on our understanding of the relationship between solution condition, disulfide bonding, collision cross-section, and charge for a larger, multidomain protein, which may be applicable for future characterization of biotherapeutics that share these structural features.


A Novel Method for Inducing Amastigote-To-Trypomastigote Transformation In Vitro in Trypanosoma cruzi Reveals the Importance of Inositol 1,4,5-Trisphosphate Receptor.
Trypanosoma cruzi is a parasitic protist that causes Chagas disease, which is prevalent in Latin America. Because of the unavailability of an effective drug or vaccine, and because about 8 million people are infected with the parasite worldwide, the development of novel drugs demands urgent attention. T. cruzi infects a wide variety of mammalian nucleated cells, with a preference for myocardial cells. Non-dividing trypomastigotes in the bloodstream infect host cells where they are transformed into replication-capable amastigotes. The amastigotes revert to trypomastigotes (trypomastigogenesis) before being shed out of the host cells. Although trypomastigote transformation is an essential process for the parasite, the molecular mechanisms underlying this process have not yet been clarified, mainly because of the lack of an assay system to induce trypomastigogenesis in vitro. Cultivation of amastigotes in a transformation medium composed of 80% RPMI-1640 and 20% Grace's Insect Medium mediated their transformation into trypomastigotes. Grace's Insect Medium alone also induced trypomastigogenesis. Furthermore, trypomastigogenesis was induced more efficiently in the presence of fetal bovine serum. Trypomastigotes derived from in vitro trypomastigogenesis were able to infect mammalian host cells as efficiently as tissue-culture-derived trypomastigotes (TCT) and expressed a marker protein for TCT. Using this assay system, we demonstrated that T. cruzi inositol 1,4,5-trisphosphate receptor (TcIP3R)-an intracellular Ca2+ channel and a key molecule involved in Ca2+ signaling in the parasite-is important for the transformation process. Our findings provide a new tool to identify the molecular mechanisms of the amastigote-to-trypomastigote transformation, leading to a new strategy for drug development against Chagas disease.


Children's looking preference for biological motion may be related to an affinity for mathematical chaos.
Recognition of biological motion is pervasive in early child development. Further, viewing the movement behavior of others is a primary component of a child's acquisition of complex, robust movement repertoires, through imitation and real-time coordinated action. We theorize that inherent to biological movements are particular qualities of mathematical chaos and complexity. We further posit that this character affords the rich and complex inter-dynamics throughout early motor development. Specifically, we explored whether children's preference for biological motion may be related to an affinity for mathematical chaos. Cross recurrence quantification analysis (cRQA) was used to investigate the coordination of gaze and posture with various temporal structures (periodic, chaotic, and aperiodic) of the motion of an oscillating visual stimulus. Children appear to competently perceive and respond to chaotic motion, both in rate (cRQA-percent determinism) and duration (cRQA-maxline) of coordination. We interpret this to indicate that children not only recognize chaotic motion structures, but also have a preference for coordination with them. Further, stratification of our sample (by age) uncovers the suggestion that this preference may become refined with age.


Differences in epitope restriction of autoantibodies to native human insulin (IAA) and antibodies to heterologous insulin (IA).
The binding profiles of insulin autoantibodies (IAA) and insulin antibodies (IA) to highly purified species variants and fragments of insulin were studied in direct immunospecific enzyme-linked immunosorbent assay (ELISA) and indirect absorption experiments with insulins covalently coupled to Sepharose beads. Five of 10 IAA-containing sera from insulin-naive nondiabetics that bound whole human (H) insulin did not bind whole porcine (P) or whole bovine (B) insulins. These sera bound H insulin B-chain but not P B-chain or desalanated P insulin, suggesting they were dependent on the presence of threonine B30. The other 5 IAA-containing sera bound H, P, B, and desalanated porcine insulins, but only 1 bound isolated B-chains. All 10 IA-containing sera from insulin-treated diabetics bound H, P, B, and desalanated P insulins, but only 1 bound to human (and porcine) B-chain. Further binding studies with ovine, rabbit, rat, and guinea pig insulins confirmed the H (threonine B30) specificity of the 5 IAA-containing sera. B30 residues do not appear to be dominant, however, when insulin is administered exogenously. Instead, IA bind predominantly to A-chain or conformational determinants involving both chains. Scatchard analysis of a representative H insulin-specific IAA serum suggested that it contained a single binding affinity, whereas analysis of a representative IA-diabetic serum suggested it contained several different affinities.


Preoperative chemoradiation and pancreaticoduodenectomy for adenocarcinoma of the pancreas.
Chemoradiation prior to pancreaticoduodenectomy ensures that all patients who undergo resection complete multimodality therapy, avoids resection in patients with rapidly progressive disease, and allows radiation therapy to be delivered to well-oxygenated cells before surgical devascularization. Twenty-eight patients with cytologic or histologic proof of localized adenocarcinoma of the pancreatic head received preoperative chemoradiation (fluorouracil, 300 mg/m2 per day, and 50.4 Gy) with the intent of proceeding to resection; all 28 completed this preoperative therapy. Hospital admission because of gastrointestinal toxic effects was required in nine patients, yet no patient experienced a delay in operation. Restaging was performed 4 to 5 weeks after completion of chemoradiation, and five patients were found to have metastatic disease; the 23 patients without evidence of progressive disease underwent laparotomy. At laparotomy, three patients were found to have unsuspected metastatic disease, three patients had unresectable locally advanced disease, and 17 patients were able to undergo pancreaticoduodenectomy. One perioperative death resulted from myocardial infarction, and perioperative complications occurred in three patients. Histologic evidence of tumor cell injury was present in all resected specimens. Our results suggest that pancreaticoduodenectomy can be performed with a low incidence of complications after chemoradiation for localized adenocarcinoma of the pancreas.


An interactive videogame for arm and hand exercise in people with Parkinson's disease: A randomized controlled trial.
People with Parkinson's disease (PD) have difficulty performing upper extremity (UE) activities. The aim of this study was to investigate if exergames targeting the UE improve arm and hand activities and impairments and to establish the acceptability and feasibility of these games in people with PD. Two tablet-based exergames were developed which were controlled with finger movements or unimanual whole arm movements. Participants with PD were randomized to an exergame (n = 19) or control (n = 19) group. The exergame group performed UE exergames at home, 3 times per week for 12 weeks. The primary outcome measure was the nine hole peg test. Secondary outcomes included measures of UE activities and impairments, including the tapping test [speed (taps/60s), and error (weighted error score/speed)]. There were no between group differences in the nine hole peg test, or in any secondary outcome measures except for the tapping test. Horizontal tapping test results showed that exergame participants improved their speed (mean difference = 10.9 taps/60s, p < 0.001) but increased error (mean difference = 0.03, p = 0.03) compared to the control group. Participants enjoyed the games and improved in their ability to play the games. There were no adverse events. The UE exergames were acceptable and safe, but did not translate to improvement in functional activities. It is likely that the requirement of the games resulted in increased movement speed at the detriment of accuracy. The design of exergames should consider task specificity.


Thyroid hormone may regulate mRNA abundance in liver by acting on microRNAs.
MicroRNAs (miRNAs) are extensively involved in diverse biological processes. However, very little is known about the role of miRNAs in mediating the action of thyroid hormones (TH). Appropriate TH levels are known to be critically important for development, differentiation and maintenance of metabolic balance in mammals. We induced transient hypothyroidism in juvenile mice by short-term exposure to methimazole and perchlorate from post natal day (PND) 12 to 15. The expression of miRNAs in the liver was analyzed using Taqman Low Density Arrays (containing up to 600 rodent miRNAs). We found the expression of 40 miRNAs was significantly altered in the livers of hypothyroid mice compared to euthyroid controls. Among the miRNAs, miRs-1, 206, 133a and 133b exhibited a massive increase in expression (50- to 500-fold). The regulation of TH on the expression of miRs-1, 206, 133a and 133b was confirmed in various mouse models including: chronic hypothyroid, short-term hyperthyroid and short-term hypothyroid followed by TH supplementation. TH regulation of these miRNAs was also confirmed in mouse hepatocyte AML 12 cells. The expression of precursors of miRs-1, 206, 133a and 133b were examined in AML 12 cells and shown to decrease after TH treatment, only pre-mir-206 and pre-mir-133b reached statistical significance. To identify the targets of these miRNAs, DNA microarrays were used to examine hepatic mRNA levels in the short-term hypothyroid mouse model relative to controls. We found transcripts from 92 known genes were significantly altered in these hypothyroid mice. Web-based target predication software (TargetScan and Microcosm) identified 14 of these transcripts as targets of miRs-1, 206, 133a and 133b. The vast majority of these mRNA targets were significantly down-regulated in hypothyroid mice, corresponding with the up-regulation of miRs-1, 206, 133a and 133b in hypothyroid mouse liver. To further investigate target genes, miR-206 was over-expressed in AML 12 cells. TH treatment of cells over-expressing miR-206 resulted in decreased miR-206 expression, and a significant increase in two predicted target genes, Mup1 and Gpd2. The results suggest that TH regulation of these genes may occur secondarily via miR-206. These studies provide new insight into the role of miRNAs in mediating TH regulation of gene expression.


Intracellular distribution of amyloid beta peptide and its relationship to the lysosomal system.
Amyloid beta peptide (Aβ) is the main component of extraneuronal senile plaques typical of Alzheimer's disease (AD) brains. Although Aβ is produced by normal neurons, it is shown to accumulate in large amounts within neuronal lysosomes in AD. We have recently shown that under normal conditions the majority of Aβ is localized extralysosomally, while oxidative stress significantly increases intralysosomal Aβ content through activation of macroautophagy. It is also suggested that impaired Aβ secretion and resulting intraneuronal increase of Aβ can contribute to AD pathology. However, it is not clear how Aβ is distributed inside normal neurons, and how this distribution is effected when Aβ secretion is inhibited. Using retinoic acid differentiated neuroblastoma cells and neonatal rat cortical neurons, we studied intracellular distribution of Aβ by double immunofluorescence microscopy for Aβ40 or Aβ42 and different organelle markers. In addition, we analysed the effect of tetanus toxin-induced exocytosis inhibition on the intracellular distribution of Aβ. Under normal conditions, Aβ was found in the small cytoplasmic granules in both neurites and perikarya. Only minor portion of Aβ was colocalized with trans-Golgi network, Golgi-derived vesicles, early and late endosomes, lysosomes, and synaptic vesicles, while the majority of Aβ granules were not colocalized with any of these structures. Furthermore, treatment of cells with tetanus toxin significantly increased the amount of intracellular Aβ in both perikarya and neurites. Finally, we found that tetanus toxin increased the levels of intralysosomal Aβ although the majority of Aβ still remained extralysosomally. Our results indicate that most Aβ is not localized to Golgi-related structures, endosomes, lysosomes secretory vesicles or other organelles, while the suppression of Aβ secretion increases intracellular intra- and extralysosomal Aβ.


Protective effect of carbon monoxide inhalation on lung injury after hemorrhagic shock/resuscitation in rats.
Hemorrhagic shock and resuscitation (HSR) induces pulmonary inflammation that leads to acute lung injury. Carbon monoxide (CO), a by-product of heme catalysis, was shown to have potent cytoprotective and anti-inflammatory effects. The aim of this study was to examine the effects of CO inhalation at low concentration on lung injury induced by HSR in rats. Rats were subjected to HSR by bleeding to achieve mean arterial pressure of 30 mm Hg for 60 minutes followed by resuscitation with shed blood and saline as needed to restore blood pressure. HSR animals were either maintained in room air or were exposed to CO at 250 ppm for 1 hour before and 3 hours after HSR. HSR caused an increase in the DNA binding activity of nuclear factor-kappaB and activator protein-1 in the lung followed by the up-regulation of pulmonary gene expression of tumor necrosis factor-alpha, inducible nitric oxide synthase, and interleukin (IL)-10. HSR also resulted in an increase in myeloperoxidase activity and wet weight to dry weight ratio in the lung, and more prominent histopathologic changes including congestion, edema, cellular infiltration, and hemorrhage. In contrast, CO inhalation significantly ameliorated these inflammatory events as judged by fewer histologic changes, less up-regulation of inflammatory mediators, and less activation of nuclear factor-kappaB and activator protein-1. Interestingly, the protective effects against lung injury afforded by CO were associated with further increases in mRNA expression of IL-10 in the lung. These findings suggest that inhaled CO at a low concentration ameliorated HSR-induced lung injury and attenuated inflammatory cascades by up-regulation of anti-inflammatory IL-10.


Behavioral and socioeconomic correlates of dental problem experience and patterns of health care-seeking.
To recognize and better understand barriers to dental care, the authors studied behavioral and socioeconomic factors specific to low-income and minority populations that contribute to patterns of professional health care-seeking. A stratified random sample of 27,002 Maryland households participated in a cross-sectional telephone survey. The authors targeted respondents who were low-income non-Hispanic white, non-Hispanic black or Hispanic who had experienced a dental problem or injury during the previous 12 months, as well as similar people with higher incomes for comparison. A total of 94.8 percent (401 of 423) of eligible people completed the survey. Logistic regression modeling (95 percent confidence interval) revealed that the respondents' race or ethnicity and sex were associated with the number of dental problems experienced during the previous 10 years, and that having a regular source of medical care and the type of dental problem experienced were associated with earlier health care-seeking. The first choice of treatment site (physician's office, emergency department or dentist's office) was associated with the respondents' income, pain level, number of problems experienced in the past 10 years and the degree to which their problem was a burden. The authors found that predisposing, enabling and need-based factors were associated with dental problem experience and patterns of professional health care service use.


Occupational inequalities in self-rated health and non-communicable diseases in different regions of Europe: findings from the European Social Survey (2014) special module on the social determinants of health.
Socioeconomic inequalities in the prevalence of non-communicable diseases (NCDs) are evident across European populations. Several previous studies have addressed the question of whether occupational inequalities in health differ across European regions. It is uncertain however, the degree to which occupational inequalities in NCDs are similar or dissimilar across different European regions. Using 2014 European Social Survey data from 20 countries, this article examines occupational inequalities in poor self-rated health (SRH) and 14 self-reported NCDs separately for women and men, by European region: heart/circulatory problems, high blood pressure, back pain, arm/hand pain, foot/leg pain, allergies, breathing problems, stomach/digestion problems, skin conditions, diabetes, severe headaches, cancer, obesity and depression. Age-controlled adjusted risk ratios were calculated and separately compared a working class and intermediate occupational group with a salariat group. Working class Europeans appear to have the highest risk of reporting poor SRH and a number of NCDs. We find inequalities in some NCDS to be the largest in the Northern region, suggesting further evidence of a Nordic paradox. Like some previous work, we did not find larger inequalities in poor SRH in the Central/East region. However, we did find the largest inequalities in this region for some NCDs. Our results do not align completely with previous work which finds smaller health inequalities in Southern Europe. This work provides a first look at occupational inequalities across a range of NCDs for European men and women by region. Future work is needed to identify the underlying determinants behind regional differences.


Transition of mesothelial cell to fibroblast in peritoneal dialysis: EMT, stem cell or bystander?
Long-term peritoneal dialysis (PD) can lead to fibrotic changes in the peritoneum, characterized by loss of mesothelial cells (MCs) and thickening of the submesothelial area with an accumulation of collagen and myofibroblasts. The origin of myofibroblasts is a central question in peritoneal fibrosis that remains unanswered at present. Numerous clinical and experimental studies have suggested that MCs, through epithelial-mesenchymal transition (EMT), contribute to the pool of peritoneal myofibroblasts. However, recent work has placed significant doubts on the paradigm of EMT in organ fibrogenesis (in the kidney particularly), highlighting the need to reconsider the role of EMT in the generation of myofibroblasts in peritoneal fibrosis. In particular, selective cell isolation and lineage-tracing experiments have suggested the existence of progenitor cells in the peritoneum, which are able to switch to fibroblast-like cells when stimulated by the local environment. These findings highlight the plastic nature of MCs and its contribution to peritoneal fibrogenesis. In this review, we summarize the key findings and caveats of EMT in organ fibrogenesis, with a focus on PD-related peritoneal fibrosis, and discuss the potential of peritoneal MCs as a source of myofibroblasts.


OptiTope--a web server for the selection of an optimal set of peptides for epitope-based vaccines.
Epitope-based vaccines (EVs) have recently been attracting significant interest. They trigger an immune response by confronting the immune system with immunogenic peptides derived from, e.g. viral- or cancer-related proteins. Binding of these peptides to proteins from the major histocompatibility complex (MHC) is crucial for immune system activation. However, since the MHC is highly polymorphic, different patients typically bind different repertoires of peptides. Furthermore, economical and regulatory issues impose strong limitations on the number of peptides that can be included in an EV. Hence, it is crucial to identify the optimal set of peptides for a vaccine, given constraints such as MHC allele probabilities in the target population, peptide mutation rates and maximum number of selected peptides. OptiTope aims at assisting immunologists in this critical task. With OptiTope, we provide an easy-to-use tool to determine a provably optimal set of epitopes with respect to overall immunogenicity in a specific individual (personalized medicine) or a target population (e.g. a certain ethnic group). OptiTope is available at http://www.epitoolkit.org/optitope.


Engineering biofunctional in vitro vessel models using a multilayer bioprinting technique.
Recent advances in the field of bioprinting have led to the development of perfusable complex structures. However, most of the existing printed vascular channels lack the composition or key structural and physiological features of natural blood vessels or they make use of more easily printable but less biocompatible hydrogels. Here, we use a drop-on-demand bioprinting technique to generate in vitro blood vessel models, consisting of a continuous endothelium imitating the tunica intima, an elastic smooth muscle cell layer mimicking the tunica media, and a surrounding fibrous and collagenous matrix of fibroblasts mimicking the tunica adventitia. These vessel models with a wall thickness of up to 425 µm and a diameter of about 1 mm were dynamically cultivated in fluidic bioreactors for up to three weeks under physiological flow conditions. High cell viability (>83%) after printing and the expression of VE-Cadherin, smooth muscle actin, and collagen IV were observed throughout the cultivation period. It can be concluded that the proposed novel technique is suitable to achieve perfusable vessel models with a biofunctional multilayer wall composition. Such structures hold potential for the creation of more physiologically relevant in vitro disease models suitable especially as platforms for the pre-screening of drugs.


Speech sound errors of Mandarin-speaking preschool children with co-occurring speech sound disorder and developmental language disorder.
This study examined the occurrence and types of speech sound errors in Mandarin-speaking children and compared those with co-occurring developmental language disorder (DLD) and speech sound disorders (SSDs) to those with SSD only and those with no DLD or SSD. The participants were 64 four-year-old Mandarin-speaking children, including 20 age-matched children with co-occurring SSD and LI (SSD + DLD), 20 with SSD only, and 24 with no DLD or SSD (typical development [TD]). Speech samples from a list of 20 pictures of common objects and animals were elicited in a picture-naming task. One speech-language therapist transcribed and analysed consonant production accuracy and coded speech errors into the types of substitution, omission, distortion, and addition and phonological processes into typical and atypical sound changes. The interrater reliability of consonant production analysis was checked by another experienced speech-language pathologist. Children with co-occurring SSD and DLD demonstrated the least accuracy of speech production among the three groups of children. Children with co-occurring SSD and DLD showed more frequent use of substitutions, omissions, typical sound changes (i.e. stopping of affricates, stopping of fricatives, deaspiration), and atypical sound changes (i.e. initial consonant deletion, fronting, affrication of stops) than children with SSD only, while no significant difference between the SSD and TD groups was observed in omission error types and atypical sound changes. These results suggest that the disorders in speech sound production in children with SSD and DLD may be associated with underlying phonological-linguistic processing.


Body representation alterations in personal but not in extrapersonal neglect patients.
There is still a matter of debate around the nature of personal neglect. Is it an attention disorder or a body representation disorder? Here we investigate the presence of body representation deficits (i.e., the visuo-spatial body map) in right and left brain-damaged patients and in particular in those affected by personal neglect. 23 unilateral brain-damaged patients (5 left-brain-damaged and 18 right-brain-damaged patients) and 15 healthy controls took part in the study. The visuo-spatial body map was assessed by means of the "Frontal body-evocation subtest (FBE)," in which participants have to put tiles representing body parts on a small wooden board where only the head is depicted as a reference point. In order to compare performance on the FBE with performance on an inanimate object that had well-defined right and left sides, participants also performed the "Car test." Group statistical analysis shows that the performance of patients with personal neglect is significantly worse than that of the controls and patients without personal neglect in the FBE but not in the Car test. Single case analyses of the five patients with pure personal neglect confirm the results of group analysis. Our data supports the hypothesis that personal neglect is a pervasive body representation disorder.


Visualizing medium and biodistribution in complex cell culture bioreactors using in vivo imaging.
There is a dearth of technology and methods to aid process characterization, control and scale-up of complex culture platforms that provide niche micro-environments for some stem cell-based products. We have demonstrated a novel use of 3d in vivo imaging systems to visualize medium flow and cell distribution within a complex culture platform (hollow fiber bioreactor) to aid characterization of potential spatial heterogeneity and identify potential routes of bioreactor failure or sources of variability. This can then aid process characterization and control of such systems with a view to scale-up. Two potential sources of variation were observed with multiple bioreactors repeatedly imaged using two different imaging systems: shortcutting of medium between adjacent inlet and outlet ports with the potential to create medium gradients within the bioreactor, and localization of bioluminescent murine 4T1-luc2 cells upon inoculation with the potential to create variable seeding densities at different points within the cell growth chamber. The ability of the imaging technique to identify these key operational bioreactor characteristics demonstrates an emerging technique in troubleshooting and engineering optimization of bioreactor performance.


Nanotemplating for two-dimensional molecular imprinting.
A new 2D molecular imprinting technique based on nanotemplating and soft-lithography techniques is reported. This technique allows the creation of target-specific synthetic recognition sites on different substrates using a uniquely oriented and immobilized template and the attachment of a molecularly imprinted polymer on a substrate. The molecularly imprinted polymer was characterized by AFM, fluorescence microscopy, and ATR-FTIR. We evaluated the rebinding ability of the sites with theophylline (the target molecule). The selectivity of the molecularly imprinted polymer was determined for the theophylline-caffeine couple. The molecularly imprinted polymer exhibited selectivity for theophylline, as revealed by competitive rebinding experiments. Fluorescence microscopy experiments provided complementary proof of the selectivity of the molecularly imprinted polymer surfaces toward theophylline. These selective molecularly imprinted polymers have the potential for chemical sensor applications. Because of its 2D nature, this novel chemical sensor technology can be integrated with many existing high-sensitivity multichannel detection technologies.


Comparative molecular analysis of ovine and bovine Streptococcus uberis isolates.
Streptococcus uberis causes clinical and subclinical mastitis in cattle and sheep, but it is unknown whether the composition of Strep. uberis populations differs between host species. To address this, we characterized a collection of bovine and ovine Strep. uberis isolates with shared geographical and temporal origins by means of an expanded multilocus sequence typing scheme. Among 14 ovine and 35 bovine isolates, 35 allelic profiles were detected. Each allelic profile was associated with a single host species and all but one were new to the multilocus sequence typing database. The median number of new alleles per isolate was higher for ovine isolates than for bovine isolates. None of the ovine isolates belonged to the global clonal complexes 5 or 143, which are commonly associated with bovine mastitis and which have a wide geographical distribution. Ovine isolates also differed from bovine isolates in carriage of plasminogen activator genes, with significantly higher prevalence of pauB in ovine isolates. Isolates that were negative for yqiL, one of the targets of multilocus sequence typing, were found among ovine and bovine isolates and were not associated with a specific sequence type or global clonal complex. One bovine isolate carried a gapC allele that was probably acquired through lateral gene transfer, most likely from Streptococcus salivarius. We conclude that ovine isolates are distinct from bovine isolates of Strep. uberis, and that recombination between isolates from different host species or bacterial species could contribute to changes in virulence gene profiles with relevance for vaccine development.


Strength and Pain Threshold Handheld Dynamometry Test Reliability in Patellofemoral Pain.
Patellofemoral pain syndrome (PFPS), characterized by peri- and retropatellar pain, is a common disorder in young, active people. The etiology is unclear; however, quadriceps strength seems to be a contributing factor, and sensitization might play a role. The study purpose is determining the inter-rater reliability of handheld dynamometry to test both quadriceps strength and pressure pain threshold (PPT), a measure for sensitization, in patients with PFPS. This cross-sectional case-control study comprises 3 quadriceps strength and one PPT measurements performed by 2 independent investigators in 22 PFPS patients and 16 matched controls. Inter-rater reliability was analyzed using intraclass correlation coefficients (ICC) and Bland-Altman plots. Inter-rater reliability of quadriceps strength testing was fair to good in PFPS patients (ICC=0.72) and controls (ICC=0.63). Bland-Altman plots showed an increased difference between assessors when average quadriceps strength values exceeded 250 N. Inter-rater reliability of PPT was excellent in patients (ICC=0.79) and fair to good in controls (ICC=0.52). Handheld dynamometry seems to be a reliable method to test both quadriceps strength and PPT in PFPS patients. Inter-rater reliability was higher in PFPS patients compared to control subjects. With regard to quadriceps testing, a higher variance between assessors occurs when quadriceps strength increases.


Genetic susceptibility, dietary antioxidants, and long-term incidence of age-related macular degeneration in two populations.
To examine effect modification between genetic susceptibility to age-related macular degeneration (AMD) and dietary antioxidant or fish consumption on AMD risk. Pooled data analysis of population-based cohorts. Participants from the Blue Mountains Eye Study (BMES) and Rotterdam Study (RS). Dietary intakes of antioxidants (lutein/zeaxanthin [LZ], β-carotene, and vitamin C), long-chain omega-3 polyunsaturated fatty acids, and zinc were estimated from food frequency questionnaires. The AMD genetic risk was classified according to the number of risk alleles of CFH (rs1061170) or ARMS2 (rs10490924) as low (no or 1 risk allele) or high (≥ 2 risk alleles). Interactions between dietary intake and genetic risk levels were assessed. Associations between dietary intake and AMD risk were assessed comparing the highest with the 2 lower intake tertiles by genetic risk subgroups using discrete logistic regression, conducted in each study separately and then using pooled data. Participants without AMD lesions at any visit were controls. We adjusted for age and sex in analyses of each cohort sample and for smoking status and study site in pooled-data analyses. All 15-year incident late AMD cases were confirmed by chief investigators of the Beaver Dam Eye Study, BMES, and RS. Intergrader reproducibility was assessed in an early AMD subsample, with 86.4% agreement between BMES and RS graders, allowing for a 1-step difference on a 5-step AMD severity scale. In pooled data analyses, we found significant interaction between AMD genetic risk status and LZ intake (P=0.0009) but nonsignificant interactions between genetic risk status and weekly fish consumption (P=0.05) for risk of any AMD. Among participants with high genetic risk, the highest intake tertile of LZ was associated with a >20% reduced risk of early AMD, and weekly consumption of fish was associated with a 40% reduced risk of late AMD. No similar association was evident among participants with low genetic risk. No interaction was detected between β-carotene or vitamin C and genetic risk status. Protection against AMD from greater LZ and fish consumption in persons with high genetic risk based on 2 major AMD genes raises the possibility of personalized preventive interventions.


Does price reveal poor-quality drugs? Evidence from 17 countries.
Focusing on 8 drug types on the WHO-approved medicine list, we constructed an original dataset of 899 drug samples from 17 low- and median-income countries and tested them for visual appearance, disintegration, and analyzed their ingredients by chromatography and spectrometry. Fifteen percent of the samples fail at least one test and can be considered substandard. After controlling for local factors, we find that failing drugs are priced 13.6-18.7% lower than non-failing drugs but the signaling effect of price is far from complete, especially for non-innovator brands. The look of the pharmacy, as assessed by our covert shoppers, is weakly correlated with the results of quality tests. These findings suggest that consumers are likely to suspect low quality from market price, non-innovator brand and the look of the pharmacy, but none of these signals can perfectly identify substandard and counterfeit drugs.