Intestinal DMT1 is critical for iron absorption in the mouse but is not required for the absorption of copper or manganese.
Divalent metal-ion transporter-1 (DMT1) is a widely expressed iron-preferring membrane-transport protein that serves a critical role in erythroid iron utilization. We have investigated its role in intestinal metal absorption by studying a mouse model lacking intestinal DMT1 (i.e., DMT1(int/int)). DMT1(int/int) mice exhibited a profound hypochromic-microcytic anemia, splenomegaly, and cardiomegaly. That the anemia was due to iron deficiency was demonstrated by the following observations in DMT1(int/int) mice: 1) blood iron and tissue nonheme-iron stores were depleted; 2) mRNA expression of liver hepcidin (Hamp1) was depressed; and 3) intraperitoneal iron injection corrected the anemia, and reversed the changes in blood iron, nonheme-iron stores, and hepcidin expression levels. We observed decreased total iron content in multiple tissues from DMT1(int/int) mice compared with DMT1(+/+) mice but no meaningful change in copper, manganese, or zinc. DMT1(int/int) mice absorbed (64)Cu and (54)Mn from an intragastric dose to the same extent as did DMT1(+/+) mice but the absorption of (59)Fe was virtually abolished in DMT1(int/int) mice. This study reveals a critical function for DMT1 in intestinal nonheme-iron absorption for normal growth and development. Further, this work demonstrates that intestinal DMT1 is not required for the intestinal transport of copper, manganese, or zinc.


Averting inflammation by targeting the cytokine environment.
Cytokines are key instigators and regulators of immune responses and therefore hold great potential as targets for new therapeutic strategies. However, the selection of which cytokines to target, and in particular the identification of which cytokines regulate the rate-limiting steps of disease pathways, is crucial to the success of such strategies. Moreover, balancing the need for ablating pathological inflammatory responses and simultaneously maintaining the ability to control infectious agents is a key consideration. Recent advances in our understanding of cytokine networks, as well as technical progress in blocking cytokines in vivo, are likely to be a source for new drugs that can control chronic inflammatory diseases.


A clinically integrated curriculum in evidence-based medicine for just-in-time learning through on-the-job training: the EU-EBM project.
Over the last years key stake holders in the healthcare sector have increasingly recognised evidence based medicine (EBM) as a means to improving the quality of healthcare. However, there is considerable uncertainty about the best way to disseminate basic knowledge of EBM. As a result, huge variation in EBM educational provision, setting, duration, intensity, content, and teaching methodology exists across Europe and worldwide. Most courses for health care professionals are delivered outside the work context ('stand alone') and lack adaptation to the specific needs for EBM at the learners' workplace. Courses with modern 'adaptive' EBM teaching that employ principles of effective continuing education might fill that gap. We aimed to develop a course for post-graduate education which is clinically integrated and allows maximum flexibility for teachers and learners. A group of experienced EBM teachers, clinical epidemiologists, clinicians and educationalists from institutions from eight European countries participated. We used an established methodology of curriculum development to design a clinically integrated EBM course with substantial components of e-learning. An independent European steering committee provided input into the process. We defined explicit learning objectives about knowledge, skills, attitudes and behaviour for the five steps of EBM. A handbook guides facilitator and learner through five modules with clinical and e-learning components. Focussed activities and targeted assignments round off the learning process, after which each module is formally assessed. The course is learner-centred, problem-based, integrated with activities in the workplace and flexible. When successfully implemented, the course is designed to provide just-in-time learning through on-the-job-training, with the potential for teaching and learning to directly impact on practice.


In vitro growth and differentiation of rabbit bulbar, fornix, and palpebral conjunctival epithelia. Implications on conjunctival epithelial transdifferentiation and stem cells.
The anterior surface of the eye is covered by several physically contiguous but histologically distinguishable epithelial overlying the cornea, limbus, bulbar conjunctiva, fornix conjunctiva, and palpebral conjunctiva. It is important to determine whether the different phenotypes of these epithelia are the result of intrinsic divergence, extrinsic modulation, or a combination of both. Based on keratin expression and cell kinetic criteria, the authors previously suggested that corneal epithelial stem cells may actually reside in the limbal basal layer. In this article, the relationship between the corneal-limbal epithelial cells and conjunctival epithelial cells was analyzed by comparing their growth and differentiation properties in an identical cell culture environment. Using Dispase instead of trypsin to dissociate the cells, the authors were able to grow all five rabbit ocular surface epithelia in the presence of 3T3 feeder cells. They found that corneal and limbal cells synthesize identical keratins, including large amounts of the K3 and K12 markers of corneal-type differentiation. By contrast, all three conjunctival epithelia shared another keratin pattern, with large amounts of simple epithelial keratins but only minute amounts of K3/K12 keratins. This observation, coupled with previous findings that the "transdifferentiation" of conjunctival epithelial cells to corneal epithelium appears to be both incomplete and reversible, provides strong evidence that (1) the limbal-corneal epithelial cells form a lineage distinct from the conjunctival lineage and (2) conjunctival transdifferentiation actually represents a process of environmental modulation. In addition, of the three types of conjunctival epithelial cells, fornix cells were found to have a much greater proliferative potential than bulbar and palpebral cells. This observation, coupled with recent finding that fornix is enriched in slow-cycling (label-retaining) cells, raises the possibility that conjunctival epithelial stem cells may preferentially reside in the fornix.


Fast food for family meals: relationships with parent and adolescent food intake, home food availability and weight status.
The purpose of the present study was to examine the prevalence of fast-food purchases for family meals and the associations with sociodemographic variables, dietary intake, home food environment, and weight status in adolescents and their parents. This study is a cross-sectional evaluation of parent interviews and adolescent surveys from Project EAT (Eating Among Teens). Subjects included 902 middle-school and high-school adolescents (53% female, 47% male) and their parents (89% female, 11% male). The adolescent population was ethnically diverse: 29% white, 24% black, 21% Asian American, 14% Hispanic and 12% other. Results showed that parents who reported purchasing fast food for family meals at least 3 times per week were significantly more likely than parents who reported purchasing fewer fast-food family meals to report the availability of soda pop and chips in the home. Adolescents in homes with fewer than 3 fast-food family meals per week were significantly more likely than adolescents in homes with more fast-food family meals to report having vegetables and milk served with meals at home. Fast-food purchases for family meals were positively associated with the intake of fast foods and salty snack foods for both parents and adolescents; and weight status among parents. Fast-food purchases for family meals were negatively associated with parental vegetable intake. Fast-food purchases may be helpful for busy families, but families need to be educated on the effects of fast food for family meals and how to choose healthier, convenient family meals.


Quantum-instanton evaluation of the kinetic isotope effects.
A general quantum-mechanical method for computing kinetic isotope effects is presented. The method is based on the quantum-instanton approximation for the rate constant and on the path-integral Metropolis-Monte Carlo evaluation of the Boltzmann operator matrix elements. It computes the kinetic isotope effect directly, using a thermodynamic integration with respect to the mass of the isotope, thus avoiding the more computationally expensive process of computing the individual rate constants. The method should be more accurate than variational transition-state theories or the semiclassical instanton method since it does not assume a single tunneling path and does not use a semiclassical approximation of the Boltzmann operator. While the general Monte Carlo implementation makes the method accessible to systems with a large number of atoms, we present numerical results for the Eckart barrier and for the collinear and full three-dimensional isotope variants of the hydrogen exchange reaction H + H2 --> H2 + H. In all seven test cases, for temperatures between 250 and 600 K, the error of the quantum instanton approximation for the kinetic isotope effects is less than approximately 10%.


The politics of partnerships: a study of police and housing collaboration to tackle anti-social behaviour on Australian public housing estates.
This paper draws on the findings from a research project on partnership arrangements between the police and housing departments on three Australian public housing estates to tackle problems associated with illicit drug activity and anti-social behaviour (ASB). The analysis focused on the setting up of the partnerships and the interactions that followed from these institutional arrangements. The assumption that informs the paper is that when studying partnerships there is a need for a more critically framed analysis. The temptation to posit "a successful model" of what partnership entails and then to judge practices in relation to this model is considerable, but it inevitably falls into the trap of constructing a narrative of partnership success or failure in terms of individual agency (that is, the degree of commitment from individuals). The analysis undertaken in this paper has therefore sought to fathom a more complex set of organizational processes. Rather than confine the discussion to issues of success and failure, the study foregrounds the subjective accounts of individuals who work within partnership and the constraints they encounter. The paper therefore makes explicit the cultural tensions within and across agencies, contestation as to the extent of the policy "problem," and the divergent perspectives on the appropriate modes of intervention.


The amino acid permeases AAP3 and AAP6 are involved in root-knot nematode parasitism of Arabidopsis.
The root-knot nematode, Meloidogyne incognita, is an obligate parasite which depends entirely on the host plant for its nutrition. Root-knot nematodes induce the formation of a highly specialized feeding site consisting of several giant cells surrounded by a network of vascular tissues. Nutrients, including amino acids and sugars, are transferred apoplastically from the vascular tissues to the feeding site. Using Arabidopsis thaliana lacking the vascular-expressed amino acid permeases (AAP) AAP3 or AAP6, we demonstrate that disruption of amino acid transport can affect nematode parasitism. Nematode infestation levels are significantly reduced on the aap3 and aap6 mutants. AAP3 and AAP6 act distinctly in the transport of amino acids to the feeding site, as demonstrated by differences in their carrying capacity profiles. Furthermore, analyses of promoter: β-glucuronidase lines show different expression patterns for AAP3 and AAP6 in infected roots. In the aap3-3 mutant, part of the decrease in infestation is connected to a defect in early infection, where juveniles enter but then leave the root. Both aap3-3 and aap6-1 produce fewer females and produce more adult male nematodes. Additionally, detrimental effects are observed in the nematodes harvested from aap3-3 and aap6-1 mutants, including decreased egg hatching and infectivity and lower levels of lipid reserves. The transport of amino acids by AAP3 and AAP6 is important for nematode infection and success of the progeny.


Influence of Human Leukocyte Antigen (HLA) Alleles and Killer Cell Immunoglobulin-Like Receptors (KIR) Types on Heparin-Induced Thrombocytopenia (HIT).
Heparin-induced thrombocytopenia (HIT) is an unpredictable, life-threatening, immune-mediated reaction to heparin. Variation in human leukocyte antigen (HLA) genes is now used to prevent immune-mediated adverse drug reactions. Combinations of HLA alleles and killer cell immunoglobulin-like receptors (KIR) are associated with multiple autoimmune diseases and infections. The objective of this study is to evaluate the association of HLA alleles and KIR types, alone or in the presence of different HLA ligands, with HIT. HIT cases and heparin-exposed controls were identified in BioVU, an electronic health record coupled to a DNA biobank. HLA sequencing and KIR type imputation using Illumina OMNI-Quad data were performed. Odds ratios for HLA alleles and KIR types and HLA*KIR interactions using conditional logistic regressions were determined in the overall population and by race/ethnicity. Analysis was restricted to KIR types and HLA alleles with a frequency greater than 0.01. The p values for HLA and KIR association were corrected by using a false discovery rate q<0.05 and HLA*KIR interactions were considered significant at p<0.05. Sixty-five HIT cases and 350 matched controls were identified. No statistical differences in baseline characteristics were observed between cases and controls. The HLA-DRB3*01:01 allele was significantly associated with HIT in the overall population (odds ratio 2.81 [1.57-5.02], p=2.1×10-4 , q=0.02) and in individuals with European ancestry, independent of other alleles. No KIR types were associated with HIT, although a significant interaction was observed between KIR2DS5 and the HLA-C1 KIR binding group (p=0.03). The HLA-DRB3*01:01 allele was identified as a potential risk factor for HIT. This class II HLA gene and allele represent biologically plausible candidates for influencing HIT pathogenesis. We found limited evidence of the role of KIR types in HIT pathogenesis. Replication and further study of the HLA-DRB3*01:01 association is necessary.


Alcohol and breast cancer: a cohort study.
The relation between alcohol consumption and several causes of death, including breast cancer, was examined in a population of 581, 321 women enrolled in a prospective study in 1959 and followed for 12 years. Women who drank occasionally had about the same breast cancer mortality rate as nondrinkers; those who drank one to four drinks per day had SMRs 7-26% higher; five drinks per day, 1.89; and six or more drinks per day, 1.65. The two highest-consumption groups' risks were significantly higher than those of nondrinkers after multivariate adjustment for several breast cancer risk factors. Distinctive dose-response relationships were observed for two known alcohol-related conditions: cirrhosis of the liver and cancer of the aero-digestive tract, suggesting that results for other causes are not seriously biased by misclassification of drinking habits. Death rates from all causes combined were elevated for drinkers of three or more drinks per day. Whether or not the association of elevated breast cancer death rates ultimately turns out to be causal, there is ample reason to continue to warn the public against excessive drinking.


Pigments and plastic in limnetic ecosystems: A qualitative and quantitative study on microparticles of different size classes.
Recently, macroplastic (>5 mm) and especially microplastic (<5 mm) particles have been reported as emerging contaminants in marine and limnetic ecosystems. Their coloration is gained by the addition of pigments to the polymer blend which is the major component of the respective product. However, color is also a feature of paint and coatings whereby the pigment is the major component. Once abraded from a surface, paint particles may enter the environment via similar pathways as microplastic particles. So far no detailed studies of microplastic particles (pigmented and non-pigmented) as well as paint particles have been performed focusing on very small microparticles (1-50 μm), in either marine or limnetic ecosystems. Using Raman microspectroscopy with a spatial resolution down to 1 μm, we report a remarkable increase in the occurrence of (pigmented) microplastic particles below 500 μm. Among those, most particles were found at a size of ∼130 μm in a freshwater ecosystem (subalpine Lake Garda, Italy). Moreover, our qualitative and quantitative analyses revealed that the number of paint microparticles significantly increased below the size range of 50 μm due to their brittleness (the smallest detected paint particle had a size of 4 μm). Inductively coupled plasma mass spectrometry measurements showed that both colored particles found in nature as well as virgin particles contain a high variety of metals such as cadmium, lead and copper. These additives may elicit adverse effects in biota ingesting these microparticles, thus paints and associated compounds may act as formerly overlooked contaminants in freshwater ecosystems.


Phase I study of Navitoclax (ABT-263), a novel Bcl-2 family inhibitor, in patients with small-cell lung cancer and other solid tumors.
Resistance to chemotherapy-induced apoptosis represents a major obstacle to cancer control. Overexpression of Bcl-2 is seen in multiple tumor types and targeting Bcl-2 may provide therapeutic benefit. A phase I study of navitoclax, a novel inhibitor of Bcl-2 family proteins, was conducted to evaluate safety, pharmacokinetics, and preliminary efficacy in patients with solid tumors. Patients enrolled to intermittent dosing cohorts received navitoclax on day -3, followed by dosing on days 1 to 14 of a 21-day cycle. Patients on continuous dosing received a 1-week lead-in dose of 150 mg followed by continuous daily administration. Blood samples were collected for pharmacokinetic analyses, biomarker analyses, and platelet monitoring. Forty-seven patients, including 29 with small-cell lung cancer (SCLC) or pulmonary carcinoid, were enrolled between 2007 and 2008, 35 on intermittent and 12 on continuous dosing cohorts. Primary toxicities included diarrhea (40%), nausea (34%), vomiting (36%), and fatigue (34%); most were grade 1 or 2. Dose- and schedule-dependent thrombocytopenia was seen in all patients. One patient with SCLC had a confirmed partial response lasting longer than 2 years, and eight patients with SCLC or carcinoid had stable disease (one remained on study for 13 months). Pro-gastrin releasing peptide (pro-GRP) was identified as a surrogate marker of Bcl-2 amplification and changes correlated with changes in tumor volume. Navitoclax is safe and well tolerated, with dose-dependent thrombocytopenia as the major adverse effect. Preliminary efficacy data are encouraging in SCLC. Efficacy in SCLC and the utility of pro-GRP as a marker of treatment response will be further evaluated in phase II studies.


Pulmonary outcomes following specialized respiratory management for acute cervical spinal cord injury: a retrospective analysis.
Retrospective analysis. To identify multivariate interactions of respiratory function that are sensitive to spinal cord injury level and pharmacological treatment to promote strategies that increase successful liberation from mechanical ventilation. United States regional spinal cord injury (SCI) treatment center. Retrospective chart review of patients consecutively admitted to Santa Clara Valley Medical Center between May 2013 and December 2014 for ventilator weaning with C1-C5 American Spinal Injury Association Impairment Scale (AIS) A or B SCI, <3 months from injury and who had a tracheostomy in place. A nonlinear, categorical principal component analysis (NL-PCA) was performed to test the multivariate interaction of respiratory outcomes from patients (N=36) being weaned off ventilator support after acute SCI with (N=15) or without (N=21) theophylline treatment. In total, 36 patients met inclusion criteria (2 C1, 5 C2, 11 C3, 14 C4 and 4 C5). The NL-PCA returned three independent components that accounted for 95% of the variance in the data set. Multivariate general linear models hypothesis tests revealed a significant syndromic interaction between theophylline treatment and SCI level (Wilks' Lambda, P=0.028, F (12,64)=2.116, η2=0.256, 1-β=0.838), with post hoc testing demonstrating a significant interaction on PC1, explained by a positive correlation between improved forced vital capacity and time it took to reach 16 h of ventilator-free breathing. Thirty-three patients (92%) achieved 16 h of ventilator-free breathing (VFB) and 30 patients (83%) achieved 24 h of VFB. We suspect that some portion of the high success rate of ventilator weaning may be attributable to theophylline use in higher cervical SCI, in addition to our aggressive regimen of high volume ventilation, medication optimization and pulmonary toilet (positive pressure treatments and mechanical insufflation-exsufflation).


Treatment of depression: time to consider folic acid and vitamin B12.
We review the findings in major depression of a low plasma and particularly red cell folate, but also of low vitamin B12 status. Both low folate and low vitamin B12 status have been found in studies of depressive patients, and an association between depression and low levels of the two vitamins is found in studies of the general population. Low plasma or serum folate has also been found in patients with recurrent mood disorders treated by lithium. A link between depression and low folate has similarly been found in patients with alcoholism. It is interesting to note that Hong Kong and Taiwan populations with traditional Chinese diets (rich in folate), including patients with major depression, have high serum folate concentrations. However, these countries have very low life time rates of major depression. Low folate levels are furthermore linked to a poor response to antidepressants, and treatment with folic acid is shown to improve response to antidepressants. A recent study also suggests that high vitamin B12 status may be associated with better treatment outcome. Folate and vitamin B12 are major determinants of one-carbon metabolism, in which S-adenosylmethionine (SAM) is formed. SAM donates methyl groups that are crucial for neurological function. Increased plasma homocysteine is a functional marker of both folate and vitamin B12 deficiency. Increased homocysteine levels are found in depressive patients. In a large population study from Norway increased plasma homocysteine was associated with increased risk of depression but not anxiety. There is now substantial evidence of a common decrease in serum/red blood cell folate, serum vitamin B12 and an increase in plasma homocysteine in depression. Furthermore, the MTHFR C677T polymorphism that impairs the homocysteine metabolism is shown to be overrepresented among depressive patients, which strengthens the association. On the basis of current data, we suggest that oral doses of both folic acid (800 microg daily) and vitamin B12 (1 mg daily) should be tried to improve treatment outcome in depression.


Aneurysm retreatment after Guglielmi detachable coil and nondetachable coil embolization: report of nine cases and review of the literature.
Guglielmi detachable coil embolization of cerebral aneurysms is becoming increasingly used to manage certain intracranial lesions based on aneurysm geometry, patient condition, and patient and surgeon preferences. Aneurysm recurrences or incomplete initial treatments are not uncommon, making repeat treatment necessary using either surgical or endovascular techniques. Between January 1993 and June 1998, 1025 cerebral aneurysms were managed by the authors at a single hospital. One hundred twenty-four of these lesions were treated using Guglielmi detachable coils, and one was managed with nondetachable coils. During the follow-up period, eight patients who underwent embolization at our institution and one who underwent embolization elsewhere received repeat treatment. Five were approached surgically, and four underwent re-embolization. All charts and films were reviewed retrospectively to determine patient outcome and clinical success. No patient in the subgroup of this clinical study suffered a permanent complication from initial aneurysm coiling, no episodes of subsequent bleeding occurred, and no complications resulted from any subsequent therapies. The anatomic results were excellent, and all aneurysms were totally or near totally obliterated. Subtotal initial coil embolization of aneurysms can be managed safely using a variety of surgical and endovascular techniques. Our approach to this predicament, lessons we have learned, and a review of the literature are herein discussed.


Structural flexibility of the macrophage dengue virus receptor CLEC5A: implications for ligand binding and signaling.
The human C-type lectin-like molecule CLEC5A is a critical macrophage receptor for dengue virus. The binding of dengue virus to CLEC5A triggers signaling through the associated adapter molecule DAP12, stimulating proinflammatory cytokine release. We have crystallized an informative ensemble of CLEC5A structural conformers at 1.9-Å resolution and demonstrate how an on-off extension to a β-sheet acts as a binary switch regulating the flexibility of the molecule. This structural information together with molecular dynamics simulations suggests a mechanism whereby extracellular events may be transmitted through the membrane and influence DAP12 signaling. We demonstrate that CLEC5A is homodimeric at the cell surface and binds to dengue virus serotypes 1-4. We used blotting experiments, surface analyses, glycan microarray, and docking studies to investigate the ligand binding potential of CLEC5A with particular respect to dengue virus. This study provides a rational foundation for understanding the dengue virus-macrophage interaction and the role of CLEC5A in dengue virus-induced lethal disease.


Heterosexual HIV-1 infectiousness and antiretroviral use: systematic review of prospective studies of discordant couples.
Recent studies have estimated the reduction in HIV-1 infectiousness with antiretroviral therapy (ART), but high-quality studies such as randomized controlled trials, accompanied by rigorous adherence counseling, are likely to overestimate the effectiveness of treatment-as-prevention in real-life settings. We attempted to summarize the effect of ART on HIV transmission by undertaking a systematic review and meta-analysis of HIV-1 infectiousness per heterosexual partnership (incidence rate and cumulative incidence over study follow-up) estimated from prospective studies of discordant couples. We used random-effects Poisson regression models to obtain summary estimates. When possible, the analyses were further stratified by direction of transmission (man-to-woman or woman-to-man) and economic setting (high- or low-income countries). Potential causes of heterogeneity of estimates were explored through subgroup analyses. Fifty publications were included. Nine allowed comparison between ART and non-ART users within studies (ART-stratified studies), in which summary incidence rates were 3.6/100 person-years (95% confidence interval = 2.0-6.5) and 0.2/100 person-years (0.07-0.7) for non-ART- and ART-using couples, respectively (P < 0.001), constituting a 91% (79-96%) reduction in per-partner HIV-1 incidence rate with ART use. The 41 studies that did not stratify by ART use provided estimates with high levels of heterogeneity (I statistic) and few reported levels of ART use, making interpretation difficult. Nevertheless, estimates tended to be lower with ART use. Infectiousness tended to be higher for low-income than high-income settings, but there was no clear pattern by direction of transmission (man-to-woman and woman-to-man). ART substantially reduces HIV-1 infectiousness within discordant couples, based on observational studies, and could play a major part in HIV-1 prevention efforts. However, the non-zero risk from partners receiving ART demonstrates that appropriate counseling and other risk-reduction strategies for discordant couples are still required. Additional estimates of ART effectiveness by adherence level from real-life settings will be important, especially for persons starting treatment early without symptoms.


Diabetic peripheral neuropathy compromises balance during daily activities.
Patients with diabetes with peripheral neuropathy have a well-recognized increased risk of falls that may result in hospitalization. Therefore this study aimed to assess balance during the dynamic daily activities of walking on level ground and stair negotiation, where falls are most likely to occur. Gait analysis during level walking and stair negotiation was performed in 22 patients with diabetic neuropathy (DPN), 39 patients with diabetes without neuropathy (D), and 28 nondiabetic control subjects (C) using a motion analysis system and embedded force plates in a staircase and level walkway. Balance was assessed by measuring the separation between the body center of mass and center of pressure during level walking, stair ascent, and stair descent. DPN patients demonstrated greater (P < 0.05) maximum and range of separations of their center of mass from their center of pressure in the medial-lateral plane during stair descent, stair ascent, and level walking compared with the C group, as well as increased (P < 0.05) mean separation during level walking and stair ascent. The same group also demonstrated greater (P < 0.05) maximum anterior separations (toward the staircase) during stair ascent. No differences were observed in D patients. Greater separations of the center of mass from the center of pressure present a greater challenge to balance. Therefore, the higher medial-lateral separations found in patients with DPN will require greater muscular demands to control upright posture. This may contribute to explaining why patients with DPN are more likely to fall, with the higher separations placing them at a higher risk of experiencing a sideways fall than nondiabetic control subjects.


Epidemiology of Prehospital Care at the San Diego (USA) - Tijuana (Mexico) International Border Crossing.
Introduction: San Diego has one of the busiest international land border crossings in the world. The epidemiology of prehospital care at the San Diego (California, USA)-Tijuana (Baja California, Mexico) border crossings are previously unreported. Investigators sought to describe prehospital care provided at the San Diego border crossings. Methods: This was a cross-sectional, retrospective data collection from April 2014 to March 2017 evaluating prehospital provider (PHP) contacts at 2 international border crossing addresses in San Diego. The 9-1-1 dispatch center and first response were provided by a single municipal fire agency with ambulance transportation provided by a contracting private agency. Patient dispatch data and electronic patient care records were queried for patient demographics, PHP arrival time, incident complaint, assessment narrative, and treatments provided. Natural language processing techniques were applied to map the narrative to the National Library of Medicine's Unified Medical Language System. Descriptive analysis was performed in the R software program. Results: A total of 6,261 PHP patient contacts were made at the 2 border crossings during the study period. 87% of the calls were at the San Ysidro border crossing compared to 13% at Otay Mesa. The population, composed of 50.8% males, had ages ranging from 0 days-103 years old, with a median age of 45 years old. There were 606 (9.7%) pediatric patients (<18 years) and 1,416 (22.6%) geriatric patients (>65 years). The top 3 incident complaints were respiratory distress (830, 10.8%), blunt trauma (827, 10.7%), and abdominal pain (814, 10.6%); and, the top 3 medications provided were oxygen (481, 7.7%), normal saline (393, 6.3%), and ondansetron (352, 5.6%). Conclusion: This was the first study of PHP assessment and care at the San Diego-Tijuana border crossings. We found a large diversity in patient ages. Respiratory distress and blunt trauma were the most frequent complaints and oxygen was the most frequent medication. The busiest day of the week was Sunday, and the busiest month of the year was July. Our newly described findings may assist EMS agencies with optimizing staff, equipment, and training at international border crossings.


Effects of AZD3480 on cognition in patients with mild-to-moderate Alzheimer's disease: a phase IIb dose-finding study.
AZD3480 is a selective agonist of the central α4β2 and α2β2 neuronal nicotinic cholinergic receptors (NNRs). Its effects on cognition were investigated in 567 patients with mild-to-moderate Alzheimer's disease (AD) (Mini Mental State Examination [MMSE] 12-26). Mean baseline MMSE was 21 (SD ± 3.7), with 61% of patients having mild disease (MMSE 21-26). Mean age was 74 (range 58-85) years. Patients were randomized to one of 5 treatment groups: AZD3480 5 mg, 20 mg or 35/100 mg, donepezil 10 mg (active comparator) or placebo, and treated once daily for 12 weeks. The primary outcome measure was change from baseline at Week 12 on the AD Assessment Scale-Cognitive Subscale (ADAS-Cog). Neither AZD3480 nor donepezil showed a statistically significant improvement versus placebo on ADAS-Cog. Improvements in a number of secondary outcome measures (MMSE, AD Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC) and Disability Assessment for Dementia [DAD]) were observed for AZD3480 and for donepezil. A post-hoc analysis on ADAS-Cog, excluding patients with very mild AD (MMSE 25-26) indicated improvement versus placebo for AZD3480 20 mg (-1.4, 95% CI: -3.0; 0.2) and donepezil (-1.0, 95% CI: -2.3; 0.3). AZD3480 was well tolerated. The study did not meet proof of concept criteria: since neither AZD3480 nor donepezil were statistically significantly superior to placebo on ADAS-Cog and was considered to be inconclusive. Further studies are required to determine the therapeutic potential of stimulating α4β2 receptors with NNRs in AD patients.


Motion analyses of adolescent rugby union players: a comparison of training and game demands.
This research described the physiological demands of participation in adolescent rugby union including positional differences and the degree to which training practices replicate game demands. Between 2003 and 2008, 118 male adolescent rugby players aged 14 to 18 years were recruited from 10 teams representing 3 levels of adolescent rugby. Time-motion analyses using global positioning satellite tracking devices (SPI10; GPSports Systems Pty Ltd 2003) and computer-based tracking software (Trak Performance; Sports Tec Pty Ltd) applied to video footage determined player movement patterns 161 times during rugby training sessions and 53 times during rugby games. Compared with rugby training, rugby games were consistently characterized by more time spent jogging (14 vs. 8%), striding (3.2 vs. 1.3%), and sprinting (1.3 vs. 0.1%) (p < 0.001). Players also covered greater distances (4000 ± 500 vs. 2710 ± 770 m) and performed more sprints (21.8 vs. 1) during games compared with training (p < 0.001). The average sprint duration of 2 seconds was similar in games and training; however, the frequency of sprint efforts in training sessions was low (1 per hour). A major finding of this study is the disparity between physical game demands and on-field rugby training practices in adolescent players determined using time-motion analyses. Sprint pattern differences between games and training in particular could have important implications for player performance during competition. Results of this study should assist in the development of game-specific training sessions and drills that provide the kinds of physically demanding experiences observed in games. Additionally, coaches could assist in the management of adolescent players' participation loads by increasing the intensity and specificity and decreasing the volume of training.


Small angle X-ray scattering coupled with in situ electromechanical probing of nanoparticle-based resistive strain gauges.
A comprehensive study on the electromechanical behavior of nanoparticle-based resistive strain gauges in action through normal and grazing incidence small angle X-ray scattering (SAXS/GISAXS) investigations is presented. The strain gauges were fabricated from arrays of colloidal gold nanoparticle (NP) wires assembled on flexible polyethylene terephthalate and polyimide substrates by convective self-assembly. Microstructural changes (mean interparticle distance variations) within these NP wires under uniaxial stretching estimated by SAXS/GISAXS are correlated to their macroscopic electrical resistance variations. SAXS measurements suggest a linear longitudinal extension and transversal contraction of the NP wires with applied strain (0 to ∼ 13%). The slope of this longitudinal variation is less than unity, implying a partial strain transfer from the substrate to the NP wires. The simultaneously measured electrical resistance of the strain gauges shows an exponential variation within the elastic domain of the substrate deformation, consistent with electron tunnelling through the interparticle gaps. A slower variation observed within the plastic domain suggests the formation of new electronic conduction pathways. Implications of transversal contraction of the NP wires on the directional sensitivities of strain gauges are evaluated by simulating electronic conduction in models mimicking a realistic NP arrangement. A loss of directionality of the NP-based strain gauges due to transversal current flow within the NP wires is deduced.


Ex vivo expanded mesenchymal stromal cell minimal quality requirements for clinical application.
Mesenchymal stromal cells (MSCs), as advanced therapy products, must satisfy all the requirements for human use of medicinal products, aiming to maintain the quality and safety of the cells. The MSC manufacturing process for clinical use should comply with the principles of Good Manufacturing Practice (GMP). This ensures that cell preparations are produced and controlled, from the collection and manipulation of raw materials, through the processing of intermediate products, to the quality controls, storage, labeling and packaging, and release. The objective of this document is to provide the minimal quality requirements for the MSC production and its delivery for clinical use, so that the safety of the final cell therapy product will not be compromised. For this purpose, the document evaluates the most important steps of GMP-compliant MSC production: the isolation and expansion process; the validation phase of the process, including all quality controls for the characterization, functionality, potency, and safety of MSCs; and the quality control at the batch release to guarantee the safety of patient infusion.


A yeast tRNA mutant that causes pseudohyphal growth exhibits reduced rates of CAG codon translation.
In Saccharomyces cerevisiae, the SUP70 gene encodes the CAG-decoding tRNA(Gln)(CUG). A mutant allele, sup70-65, induces pseudohyphal growth on rich medium, an inappropriate nitrogen starvation response. This mutant tRNA is also a UAG nonsense suppressor via first base wobble. To investigate the basis of the pseudohyphal phenotype, 10 novel sup70 UAG suppressor alleles were identified, defining positions in the tRNA(Gln)(CUG) anticodon stem that restrict first base wobble. However, none conferred pseudohyphal growth, showing altered CUG anticodon presentation cannot itself induce pseudohyphal growth. Northern blot analysis revealed the sup70-65 tRNA(Gln)(CUG) is unstable, inefficiently charged, and 80% reduced in its effective concentration. A stochastic model simulation of translation predicted compromised expression of CAG-rich ORFs in the tRNA(Gln)(CUG)-depleted sup70-65 mutant. This prediction was validated by demonstrating that luciferase expression in the mutant was 60% reduced by introducing multiple tandem CAG (but not CAA) codons into this ORF. In addition, the sup70-65 pseudohyphal phenotype was partly complemented by overexpressing CAA-decoding tRNA(Gln)(UUG), an inefficient wobble-decoder of CAG. We thus show that introducing codons decoded by a rare tRNA near the 5' end of an ORF can reduce eukaryote translational expression, and that the mutant tRNA(CUG)(Gln) constitutive pseudohyphal differentiation phenotype correlates strongly with reduced CAG decoding efficiency.


Effects of intravenous administration of pimobendan on hemodynamic indices and indices of left atrial longitudinal strain derived from speckle-tracking echocardiography in healthy dogs.
To determine the effects of IV administration of pimobendan on hemodynamic indices and indices of left atrial (LA) longitudinal strain by speckle-tracking echocardiography in healthy dogs. 6 healthy Beagles. After the dogs were anesthetized, the right heart was catheterized and 2-D conventional and speckle-tracking echocardiography were performed before and after IV administration of 0.15 mg of pimobendan/kg. Speckle-tracking echocardiography was performed to assess the 3 LA phasic functions through LA deformation (longitudinal strain and strain rate) and volumetric analyses. Pimobendan significantly increased stroke volume and cardiac output and decreased systemic vascular resistance. Pimobendan significantly improved left ventricular function assessed by the Tei index and LA booster pump function assessed by LA longitudinal strain and change in fractional volume during atrial systole. Indices of LA reservoir and conduit function were unchanged. In healthy dogs, IV administration of pimobendan improved the indices of LA booster pump function but had no effect on the indices of LA reservoir and conduit functions. Further studies are needed to validate whether these results can be extrapolated to dogs with myxomatous mitral valve disease.


Resolution limits of extrinsic Fabry-Perot interferometric displacement sensors utilizing wavelength scanning interrogation.
The factors limiting the resolution of displacement sensors based on the extrinsic Fabry-Perot interferometer were studied. An analytical model giving the dependency of extrinsic Fabry-Perot interferometric (EFPI) resolution on the parameters of an optical setup and a sensor interrogator was developed. The proposed model enables one to either estimate the limit of possible resolution achievable with a given setup, or derive the requirements for optical elements and/or a sensor interrogator necessary for attaining the desired sensor resolution. An experiment supporting the analytical derivations was performed, demonstrating a large dynamic measurement range (with cavity length from tens of microns to 5 mm), a high baseline resolution (from 14 pm), and good agreement with the model.


Structure of the periplasmic component of a bacterial drug efflux pump.
Multidrug resistance among Gram-negative bacteria is conferred by three-component membrane pumps that expel diverse antibiotics from the cell. These efflux pumps consist of an inner membrane transporter such as the AcrB proton antiporter, an outer membrane exit duct of the TolC family, and a periplasmic protein known as the adaptor. We present the x-ray structure of the MexA adaptor from the human pathogen Pseudomonas aeruginosa. The elongated molecule contains three linearly arranged subdomains; a 47-A-long alpha-helical hairpin, a lipoyl domain, and a six-stranded beta-barrel. In the crystal, hairpins of neighboring MexA monomers pack side-by-side to form twisted arcs. We discuss the implications of the packing of molecules within the crystal. On the basis of the structure and packing, we suggest a model for the key periplasmic interaction between the outer membrane channel and the adaptor protein in the assembled drug efflux pump.


Interleukin-6 receptor-mediated activation of signal transducer and activator of transcription-3 (STAT3) promotes choroidal neovascularization.
Interleukin (IL)-6, a potent proinflammatory cytokine, is suggested to be a risk factor for choroidal neovascularization (CNV) because of its increased levels in the serum of patients with age-related macular degeneration; however, the role of IL-6 in CNV has not been defined. The present study reveals the critical contribution of IL-6 signaling and its downstream STAT3 pathway to the murine model of laser-induced CNV. CNV induction by laser treatment stimulated IL-6 expression in the retinal pigment epithelium-choroid complex, and antibody-based blockade of IL-6 receptor or genetic ablation of IL-6 led to significant suppression of CNV. CNV generation was accompanied by STAT3 activation in choroidal endothelial cells and macrophages, and IL-6 receptor blockade resulted in selectively inhibited phosphorylation of STAT3 but not extracellular signal-regulated kinase 1/2. Consistently, pharmacological blockade of STAT3 pathway also suppressed CNV. In addition, IL-6 receptor neutralization led to significant inhibition of the in vivo and in vitro expression of inflammation-related molecules including monocyte chemotactic protein, intercellular adhesion molecule-1, and vascular endothelial growth factor, and of macrophage infiltration into CNV. These results indicate the significant involvement of IL-6 receptor-mediated activation of STAT3 inflammatory pathway in CNV generation, suggesting the possibility of IL-6 receptor blockade as a therapeutic strategy to suppress CNV associated with age-related macular degeneration.


Capicua DNA-binding sites are general response elements for RTK signaling in Drosophila.
RTK/Ras/MAPK signaling pathways play key functions in metazoan development, but how they control expression of downstream genes is not well understood. In Drosophila, it is generally assumed that most transcriptional responses to RTK signal activation depend on binding of Ets-family proteins to specific cis-acting sites in target enhancers. Here, we show that several Drosophila RTK pathways control expression of downstream genes through common octameric elements that are binding sites for the HMG-box factor Capicua, a transcriptional repressor that is downregulated by RTK signaling in different contexts. We show that Torso RTK-dependent regulation of terminal gap gene expression in the early embryo critically depends on Capicua octameric sites, and that binding of Capicua to these sites is essential for recruitment of the Groucho co-repressor to the huckebein enhancer in vivo. We then show that subsequent activation of the EGFR RTK pathway in the neuroectodermal region of the embryo controls dorsal-ventral gene expression by downregulating the Capicua protein, and that this control also depends on Capicua octameric motifs. Thus, a similar mechanism of RTK regulation operates during subdivision of the anterior-posterior and dorsal-ventral embryonic axes. We also find that identical DNA octamers mediate Capicua-dependent regulation of another EGFR target in the developing wing. Remarkably, a simple combination of activator-binding sites and Capicua motifs is sufficient to establish complex patterns of gene expression in response to both Torso and EGFR activation in different tissues. We conclude that Capicua octamers are general response elements for RTK signaling in Drosophila.


Methyl-deficient diet promotes colitis and SIRT1-mediated endoplasmic reticulum stress.
Methyl donor deficiency (MDD) aggravates experimental colitis in rats and increases endoplasmic reticulum (ER) stress through decreased sirtuin 1 (SIRT1) in neuronal cells and myocardium. ER stress plays a key role in IBD pathogenesis. We investigated whether the influence of MDD on colitis resulted from an ER stress response triggered by decreased SIRT1 expression. The unfolded protein response (UPR), chaperones proteins, heat shock factor protein 1 (HSF1) and SIRT1 were examined in rats with MDD and dextran sulfate sodium (DSS)-induced colitis in a Caco-2 cell model with stable expression of transcobalamin-oleosin (TO) chimera, which impairs cellular availability of vitamin B12, and in IBD. The effects of SIRT1 activation were studied both in vitro and in vivo. MDD aggravated DSS-induced colitis clinically, endoscopically and histologically. MDD activated ER stress pathways, with increased phosphorylate-PKR-like ER kinase, P-eiF-2α, P-IRE-1α, activating transcription factor (ATF)6, XBP1-S protein and ATF4 mRNA expression levels in rats. This was accompanied by reduced SIRT1 expression level and greater acetylation of HSF1, in relation with a dramatic decrease of chaperones (binding immunoglobulin protein (BIP), heat shock protein (HSP)27 and HSP90). Adding either vitamin B12, S-adenosylmethionine or an SIRT1 activator (SRT1720) reduced the UPR in vitro. In rats, SIRT1 activation by SRT1720 prevented colitis by reducing HSF1 acetylation and increasing expression of BIP, HSP27 and HSP90. Immunohistochemistry showed impaired expression of SIRT1 in the colonic epithelium of patients with IBD. SIRT1 is a master regulator of ER stress and severity of experimental colitis in case of MDD. It could deserve further interest as a therapeutic target of IBD.


Unesterified cholesterol accumulation in late endosomes/lysosomes causes neurodegeneration and is prevented by driving cholesterol export from this compartment.
While unesterified cholesterol (C) is essential for remodeling neuronal plasma membranes, its role in certain neurodegenerative disorders remains poorly defined. Uptake of sterol from pericellular fluid requires processing that involves two lysosomal proteins, lysosomal acid lipase, which hydrolyzes C esters, and NPC1 (Niemann-Pick type C1). In systemic tissues, inactivation of either protein led to sterol accumulation and cell death, but in the brain, inactivation of only NPC1 caused C sequestration and neurodegeneration. When injected into the CNS of the npc1(-/-) mouse, 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), a compound known to prevent this C accumulation, diffused throughout the brain and was excreted with a t(½) of 6.5 h. This agent caused suppression of C synthesis, elevation of C esters, suppression of sterol regulatory-binding protein 2 (SREBP2) target genes, and activation of liver X receptor-controlled genes. These findings indicated that HP-β-CD promoted movement of the sequestered C from lysosomes to the metabolically active pool of C in the cytosolic compartment of cells in the CNS. The ED(50) for this agent in the brain was ∼0.5 mg/kg, and the therapeutic effect lasted >7 d. Continuous infusion of HP-β-CD into the ventricular system of npc1(-/-) animals between 3 and 7 weeks of age normalized the biochemical abnormalities and completely prevented the expected neurodegeneration. These studies support the concept that neurons continuously acquire C from interstitial fluid to permit plasma membrane turnover and remodeling. Inactivation of NPC1 leads to lysosomal C sequestration and neurodegeneration, but this is prevented by the continuous, direct administration of HP-β-CD into the CNS.


Incorporating Uncertainty Into Parallel Analysis for Choosing the Number of Factors via Bayesian Methods.
A number of psychometricians have suggested that parallel analysis (PA) tends to yield more accurate results in determining the number of factors in comparison with other statistical methods. Nevertheless, all too often PA can suggest an incorrect number of factors, particularly in statistically unfavorable conditions (e.g., small sample sizes and low factor loadings). Because of this, researchers have recommended using multiple methods to make judgments about the number of factors to extract. Implicit in this recommendation is that, when the number of factors is chosen based on PA, uncertainty nevertheless exists. We propose a Bayesian parallel analysis (B-PA) method to incorporate the uncertainty with decisions about the number of factors. B-PA yields a probability distribution for the various possible numbers of factors. We implement and compare B-PA with a frequentist approach, revised parallel analysis (R-PA), in the contexts of real and simulated data. Results show that B-PA provides relevant information regarding the uncertainty in determining the number of factors, particularly under conditions with small sample sizes, low factor loadings, and less distinguishable factors. Even if the indicated number of factors with the highest probability is incorrect, B-PA can show a sizable probability of retaining the correct number of factors. Interestingly, when the mode of the distribution of the probabilities associated with different numbers of factors was treated as the number of factors to retain, B-PA was somewhat more accurate than R-PA in a majority of the conditions.


Have CT--will travel: to boldly go where no scan has gone before.
In 1962, a new English rock-and-roll band named The Beatles signed a recording contract with Electric & Music Industries (EMI). The Beatles were so financially successful that EMI was able to fund research and development in other divisions of the company; in particular, the work of an enterprising young engineer named Godfrey Hounsfield. His groundbreaking work in x-ray imaging produced the first commercially available head-only CT scanner, and a Nobel Prize.


Prioritizing IVF patients according to the number of existing children--a proposed refinement to the current guideline.
The National Health Service is poised to offer at least one cycle of IVF treatment to couples free of charge in the UK, provided that certain clinical criteria are met. At a local level, therefore, funders are faced with the problem of prioritizing patients and establishing waiting lists. It is generally accepted that preference should be given to those with the 'greatest need', but it is uncertain what the criterion is for greatest need. We propose two recommendations as a modification to the current guideline that stresses the need to favour 'couples who do not have any children living with them'. First, we suggest that parental responsibility rather than shared residence be used as a criterion for eligibility. Second, we present a need-based approach that can be used coherently to distribute publicly funded treatment. In an attempt to achieve a fair way of prioritizing patients, we make distinctions between different family arrangements, based on the number of existing children. Our approach treats like cases alike, and ranks different cases relative to each other in a manner that is equitable and charitable.


Rapid accumulation of inositol trisphosphate reveals that agonists hydrolyse polyphosphoinositides instead of phosphatidylinositol.
The agonist-dependent hydrolysis of inositol phospholipids was investigated by studying the breakdown of prelabelled lipid or by measuring the accumulation of inositol phosphates. Stimulation of insect salivary glands with 5-hydroxytryptamine for 6 min provoked a rapid disappearance of [3H]phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] and [3H]phosphatidylinositol 4-phosphate (PtdIns4P) but had no effect on the level of [3H]phosphatidylinositol (PtdIns). The breakdown of PtdIns(4,5)P2 was associated with a very rapid release of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], which reached a peak 5 1/2 times that of the resting level after 5 s of stimulation. This high level was not maintained but declined to a lower level, perhaps reflecting the disappearance of PtdIns(4,5)P2. 5-Hydroxytryptamine also induced a rapid and massive accumulation of inositol 1,4-bisphosphate [Ins(1,4)P2]. The fact that these increases in Ins(1,4,5)P3 and Ins(1,4)P2 precede in time any increase in the level of inositol 1-phosphate or inositol provides a clear indication that the primary action of 5-hydroxytryptamine is to stimulate the hydrolysis of PtdIns(4,5)P2 to yield diacylglycerol and Ins(1,4,5)P3. The latter is then hydrolysed by a series of phosphomonoesterases to produce Ins(1,4)P2, Ins1P and finally inositol. The very rapid agonist-dependent increases in Ins(1,4,5)P3 and Ins(1,4)P2 suggests that they could function as second messengers, perhaps to control the release of calcium from internal pools. The PtdIns(4,5)P2 that is used by the receptor mechanism represents a small hormone-sensitive pool that must be constantly replenished by phosphorylation of PtdIns. Small changes in the size of this small energy-dependent pool of polyphosphoinositide will alter the effectiveness of the receptor mechanism and could account for phenomena such as desensitization and super-sensitivity.


Paired Associative Stimulation as a Tool to Assess Plasticity Enhancers in Chronic Stroke.
The potential for adaptive plasticity in the post-stroke brain is difficult to estimate, as is the demonstration of central nervous system (CNS) target engagement of drugs that show promise in facilitating stroke recovery. We set out to determine if paired associative stimulation (PAS) can be used (a) as an assay of CNS plasticity in patients with chronic stroke, and (b) to demonstrate CNS engagement by memantine, a drug which has potential plasticity-modulating effects for use in motor recovery following stroke. We examined the effect of PAS in fourteen participants with chronic hemiparetic stroke at five time-points in a within-subjects repeated measures design study: baseline off-drug, and following a week of orally administered memantine at doses of 5, 10, 15, and 20 mg, comprising a total of seventy sessions. Each week, MEP amplitude pre and post-PAS was assessed in the contralesional hemisphere as a marker of enhanced or diminished plasticity. Strength and dexterity were recorded each week to monitor motor-specific clinical status across the study period. We found that MEP amplitude was significantly larger after PAS in baseline sessions off-drug, and responsiveness to PAS in these sessions was associated with increased clinical severity. There was no observed increase in MEP amplitude after PAS with memantine at any dose. Motor threshold (MT), strength, and dexterity remained unchanged during the study. Paired associative stimulation successfully induced corticospinal excitability enhancement in chronic stroke subjects at the group level. However, this response did not occur in all participants, and was associated with increased clinical severity. This could be an important way to stratify patients for future PAS-drug studies. PAS was suppressed by memantine at all doses, regardless of responsiveness to PAS off-drug, indicating CNS engagement.


Corneal epithelial and aqueous humor acidification during in vivo contact lens wear in rabbits.
Based on contact lens-induced stromal acidification of the cornea, it has been suggested that the corneal epithelial and endothelial cells also become acidotic during contact lens wear. This alleged acidification may have a role in altered cell appearance and metabolism during contact lens wear. This study investigated the effects of anoxia, carbon dioxide retention, and contact lens gas transmissibility on the epithelial and aqueous humor pH in living rabbits. Epithelial intracellular pH (pHi) and aqueous humor pH were fluorophotometrically measured with a pH sensitive-dye (BCECF) during contact lens wear or exposure to various gas mixtures. Polymethylmethacrylate (PMMA) lens wear acidified epithelial cells by preventing CO2 efflux and by inducing hypoxia. Increasing lens oxygen transmissibility decreased epithelial acidification. After initiation of rigid, gas-permeable (RGP) lens wear or CO2-air exposure, pHi dropped transiently and then recovered partially. This recovery of pHi was not observed during anoxia, whether induced by PMMA lens wear or exposure to 100% N2. The aqueous humor also acidified during PMMA lens wear, a phenomenon not observed during RGP lens wear. Changes in aqueous pH were smaller, slower, and delayed when compared to their epithelial counterparts. Hypoxic contact lens wear acidifies the corneal epithelium and aqueous humor. The aqueous humor pH change indicates a probable endothelial acidification during hypoxic contact lens wear; the pH changes are caused by two separate and additive effects, CO2 retention and hypoxic acidosis. Increases in the oxygen transmissibility of the lens decrease the cellular acidosis, which might minimize cellular complications arising from contact lens wear. We estimate that a lens with an oxygen transmissibility (Dk/L) of 300 x 10(-11) (cm/sec)(ml O2/ml x mm Hg) is needed to prevent epithelial pHi changes in the open eye. In contrast, lenses with Dk/L as low as 18 x 10(-9) (cm/sec)(ml O2/ml x mm Hg) can prevent aqueous humor pH changes.


[Structural modifications of the esophagus linked to osmoregulation in eels].
In comparison with the fresh-water Eel (Anguilla anguilla L.), the oesophagus of the sea-water Eel displays an almost complete regression of the multilayered epithelium and consequently the disappearance of mucous cells. In contrast, a large development of a columnaŕ epithelium is associated with a tremendous proliferation of the subepithelial vascularization. These modifications enable one to interpret the changes in ionic permeability of the oesophagus during transfer from freshwater to seawater.


Can a Corn-Derived Biosurfactant Improve Colour Traits of Wine? First Insight on Its Application during Winegrape Skin Maceration versus Oenological Tannins.
In winemaking, oenological tannins are used to preserve wine colour by enhancing the antioxidant activity, taking part in copigmentation, and forming polymeric pigments with anthocyanins. As a novel processing aid, in this study, a biosurfactant extract was evaluated as a solubilizing and stabilizing agent of anthocyanins in red wine. The biosurfactant extract under evaluation was obtained from a fermented residual stream of the corn milling industry named corn steep liquor (CSL). Two red winegrape varieties (Vitis vinifera L. cv. Aglianico and Cabernet sauvignon) were studied for anthocyanin content and profile, and colour traits, during simulated skin maceration for 7 days at 25 °C, as well as polymerization and copigmentation at the end of maceration. A model wine solution was used as a control, which was added either with the CSL biosurfactant or with four different oenological tannins (from grape skin, grape seed, quebracho, and acacia). The results showed that CSL biosurfactant addition improved the colour properties of skin extracts by the formation of more stable compounds mainly through copigmentation interactions. These preliminary results highlighted that the effectiveness of CSL biosurfactant is variety-dependent; however, there is no significant protection of individual anthocyanin compounds as observed for delphinidin and petunidin forms using quebracho tannin.


Neurotoxic mechanisms caused by the Alzheimer's disease-linked Swedish amyloid precursor protein mutation: oxidative stress, caspases, and the JNK pathway.
Autosomal dominant forms of familial Alzheimer's disease (FAD) are caused by mutations of the amyloid precursor protein (APP) gene and by mutations of the genes encoding for presenilin 1 or presenilin 2. Simultaneously, evidence is provided that increased oxidative stress might play a crucial role in the rapid progression of the Swedish FAD. Here we investigated the effect of the Swedish double mutation (K670M/N671L) in the beta-amyloid precursor protein on oxidative stress-induced cell death mechanisms in PC12 cells. Western blot analysis and cleavage studies of caspase substrates revealed an elevated activity of the executor caspase 3 after treatment with hydrogen peroxide in cells containing the Swedish APP mutation. This elevated activity is the result of the enhanced activation of both intrinsic and extrinsic apoptosis pathways, including activation of caspase 2 and caspase 8. Furthermore, we observed an enhanced activation of JNK pathway and an attenuation of apoptosis by SP600125, a JNK inhibitor, through protection of mitochondrial dysfunction and reduction of caspase 9 activity. Our findings provide evidence that the massive neurodegeneration in early age of FAD patients could be a result of an increased vulnerability of neurons through activation of different apoptotic pathways as a consequence of elevated levels of oxidative stress.


Sympathetic cooling of a membrane oscillator in a hybrid mechanical-atomic system.
Sympathetic cooling with ultracold atoms and atomic ions enables ultralow temperatures in systems where direct laser or evaporative cooling is not possible. It has so far been limited to the cooling of other microscopic particles, with masses up to 90 times larger than that of the coolant atom. Here, we use ultracold atoms to sympathetically cool the vibrations of a Si3N4 nanomembrane, the mass of which exceeds that of the atomic ensemble by a factor of 10(10). The coupling of atomic and membrane vibrations is mediated by laser light over a macroscopic distance and is enhanced by placing the membrane in an optical cavity. We observe cooling of the membrane vibrations from room temperature to 650 ± 230 mK, exploiting the large atom-membrane cooperativity of our hybrid optomechanical system. With technical improvements, our scheme could provide ground-state cooling and quantum control of low-frequency oscillators such as nanomembranes or levitated nanoparticles, in a regime where purely optomechanical techniques cannot reach the ground state.


Geo-spatial aspects of acceptance of illegal hunting of large carnivores in Scandinavia.
Human-carnivore conflicts are complex and are influenced by: the spatial distribution of the conflict species; the organisation and intensity of management measures such as zoning; historical experience with wildlife; land use patterns; and local cultural traditions. We have used a geographically stratified sampling of social values and attitudes to provide a novel perspective to the human - wildlife conflict. We have focused on acceptance by and disagreements between residents (measured as Potential Conflict Index; PCI) towards illegal hunting of four species of large carnivores (bear, lynx, wolf, wolverine). The study is based on surveys of residents in every municipality in Sweden and Norway who were asked their opinion on illegal hunting. Our results show how certain social values are associated with acceptance of poaching, and how these values differ geographically independent of carnivore abundance. Our approach differs from traditional survey designs, which are often biased towards urban areas. Although these traditional designs intend to be representative of a region (i.e. a random sample from a country), they tend to receive relatively few respondents from rural areas that experience the majority of conflict with carnivores. Acceptance of poaching differed significantly between Norway (12.7-15.7% of respondents) and Sweden (3.3-4.1% of respondents). We found the highest acceptance of illegal hunting in rural areas with free-ranging sheep and strong hunting traditions. Disagreements between residents (as measured by PCI) were highest in areas with intermediate population density. There was no correlation between carnivore density and either acceptance of illegal hunting or PCI. A strong positive correlation between acceptance of illegal hunting and PCI showed that areas with high acceptance of illegal hunting are areas with high potential conflict between people. Our results show that spatially-stratified surveys are required to reveal the large scale patterns in social dynamics of human-wildlife conflicts.


Dopamine D2 Receptors Dimers: How can we Pharmacologically Target Them?
Dopamine D2 and D3 receptors can form homo- and heterodimers and are important targets in Schizophrenia and Parkinson's. Recently, many efforts have been made to pharmacologically target these receptor complexes. This review focuses on various strategies to act specifically on dopamine receptor dimers, that are transiently formed. Various binding and functional assays were reviewed to study the properties of bivalent ligands, particularly for the dualsteric compound SB269,652. The dimerization of D2 and D3 receptors were analyzed by using single particle tracking microscopy. The specific targeting of dopamine D2 and D3 dimers can be achieved with bifunctional ligands, composed of two pharmacophores binding the two orthosteric sites of the dimeric complex. If the target is a homodimer, then the ligand is homobivalent. Instead, if the target is a heterodimer, then the ligand is heterobivalent. However, there is some concern regarding pharmacokinetics and binding properties of such drugs. Recently, a new generation of bitopic compounds with dualsteric properties have been discovered that bind to the orthosteric and the allosteric sites in one monomeric receptor. Regarding dopamine D2 and D3 receptors, a new dualsteric molecule SB269,652 was shown to have selective negative allosteric properties across D2 and D3 homodimers, but it behaves as an orthosteric antagonist on receptor monomer. Targeting dimers is also complicated as they are transiently formed with varying monomer/dimer ratio. Furthermore, this ratio can be altered by administering an agonist or a bifunctional antagonist. Last 15 years have witnessed an explosive amount of work aimed at generating bifunctional compounds as a novel strategy to target GPCR homo- and heterodimers, including dopamine receptors. Their clinical use is far from trivial, but, at least, they have been used to validate the existence of receptor dimers in-vitro and in-vivo. The dualsteric compound SB269, 652, with its peculiar pharmacological profile, may offer therapeutic advantages and a better tolerability in comparison with pure antagonists at D2 and D3 receptors and pave the way for a new generation of antipsychotic drugs.


A global survey of changing patterns of food allergy burden in children.
While food allergies and eczema are among the most common chronic non-communicable diseases in children in many countries worldwide, quality data on the burden of these diseases is lacking, particularly in developing countries. This 2012 survey was performed to collect information on existing data on the global patterns and prevalence of food allergy by surveying all the national member societies of the World Allergy Organization, and some of their neighbouring countries. Data were collected from 89 countries, including published data, and changes in the health care burden of food allergy. More than half of the countries surveyed (52/89) did not have any data on food allergy prevalence. Only 10% (9/89) of countries had accurate food allergy prevalence data, based on oral food challenges (OFC). The remaining countries (23/89) had data largely based on parent-reporting of a food allergy diagnosis or symptoms, which is recognised to overestimate the prevalence of food allergy. Based on more accurate measures, the prevalence of clinical (OFC proven) food allergy in preschool children in developed countries is now as high as 10%. In large and rapidly emerging societies of Asia, such as China, where there are documented increases in food allergy, the prevalence of OFC-proven food allergy is now around 7% in pre-schoolers, comparable to the reported prevalence in European regions. While food allergy appears to be increasing in both developed and developing countries in the last 10-15 years, there is a lack of quality comparative data. This survey also highlights inequities in paediatric allergy services, availability of adrenaline auto-injectors and standardised National Anaphylaxis Action plans. In conclusion, there remains a need to gather more accurate data on the prevalence of food allergy in many developed and developing countries to better anticipate and address the rising community and health service burden of food allergy.


Coffee and the risk of hepatocellular carcinoma and chronic liver disease: a systematic review and meta-analysis of prospective studies.
An inverse association has been reported between coffee drinking and the risk of hepatocellular carcinoma (HCC) and chronic liver disease (CLD), but its magnitude is still unclear. Thus, we carried out a systematic review and meta-analysis of prospective cohort studies that investigated the association between coffee consumption and the risk of HCC or CLD. We separately estimated the relative risk (RR) of the two conditions, for regular, low, and high consumption compared with no or occasional coffee consumption; we also calculated the summary RR for an increment of one cup of coffee per day. Twelve studies on HCC (3414 cases) and six studies on CLD (1463 cases) were identified. The summary RRs for HCC were 0.66 [95% confidence interval (CI): 0.55-0.78] for regular, 0.78 (95% CI: 0.66-0.91) for low, and 0.50 (95% CI: 0.43-0.58) for high coffee consumption, respectively. The summary RR for an increment of one cup per day was 0.85 (95% CI: 0.81-0.90). The summary RRs for CLD were 0.62 (95% CI: 0.47-0.82) for regular, 0.72 (95% CI: 0.59-0.88) for low, 0.35 (95% CI: 0.22-0.56) for high, and 0.74 (95% CI: 0.65-0.83) for an increment of one cup per day. The present meta-analysis provides a precise quantification of the inverse relation between coffee consumption and the risk of HCC, and adds evidence to the presence of an even stronger negative association with CLD.


Attachment sites of four tick species (Acari: Ixodidae) parasitizing humans in Georgia and South Carolina.
From June 1995 through January 1998, 677 tick specimens were submitted by 521 humans from 14 states. Analysis was limited to specimens originating in Georgia and South Carolina, representing 87.3% of total submissions. Attachment sites were specified in 367 specimens (62.3%). The American dog tick, Dermacentor variabilis (Say), a vector of the agent of Rocky Mountain spotted fever, favored the head and neck in 59% of attached specimens. The lone star tick, Amblyomma americanum (L.), a strongly implicated vector of the agent of human monocytic ehrlichiosis, favored the lower extremities, buttocks, and groin in 54% of specimens. The blacklegged tick, Ixodes scapularis Say, the main eastern vector of the Lyme disease spirochete, had widely distributed attachment sites with no apparent site preference. The Gulf Coast tick, A. maculatum Koch, parasitized humans in too few instances for analysis. In the southeastern United States, prevention of tick bites and tickborne illnesses such as Rocky Mountain spotted fever, ehrlichiosis, and Lyme disease may be enhanced by personal practices and public health measures based on knowledge of preferred attachment sites of potentially infectious tick species.


Sociodemographic correlates of exclusive and concurrent use of smokeless and smoked tobacco products among Nigerian men.
This study sought to determine the sociodemographic correlates of exclusive and concurrent use of smokeless tobacco (SLT) and smoking (i.e., dual use) and to explore the association between SLT use and the amount of cigarettes smoked per day. Data were obtained from a nationally representative sample of Nigerian men aged 15-59 years (N = 15,453) who participated in the 2008 Nigerian Demographic and Health Survey (NDHS). The NDHS used an interviewer-administered questionnaire to collect data on participants' sociodemographic characteristics and tobacco use status. Taking account of the multistage sampling used in the NDHS, data analyses included descriptive statistics, chi-square analysis, and multivariable adjusted multinomial logistic regression analysis. Of the respondents, 12.2% (n = 1,842) were tobacco users, out of which 24.5% (n = 477) were exclusive SLT users, 69% (n = 1,236) were exclusive smokers, and 6.5% (n = 129) were dual users. Both SLT use and smoking were most prevalent among the Igbo ethnic group and among the least educated men. SLT use was most prevalent among those in the southeast (9.75%) and north-central (7.71%) regions, where smoking was also common. The number of cigarettes smoked per day was not significantly different among dual users when compared with exclusive smokers (7.3 vs. 5.6; p = .088). Dual users were also more likely to self-identify as traditionalists (RRR = 6.03; 95% CI = 2.96-12.28) compared with self-identifying as practicing Islam or Christianity. There are distinctive ethnic and regional differences in tobacco use patterns among Nigerian men, and SLT use was not associated with reduced smoking intensity among dual users.


A mouse model of osteochondromagenesis from clonal inactivation of Ext1 in chondrocytes.
We report a mouse model of multiple osteochondromas (MO), an autosomal dominant disease in humans, also known as multiple hereditary exostoses (MHE or HME) and characterized by the formation of cartilage-capped osseous growths projecting from the metaphyses of endochondral bones. The pathogenesis of these osteochondromas has remained unclear. Mice heterozygous for Ext1 or Ext2, modeling the human genotypes that cause MO, occasionally develop solitary osteochondroma-like structures on ribs [Lin et al. (2000) Dev Biol 224(2):299-311; Stickens et al. (2005) Development 132(22):5055-5068]. Rather than model the germ-line genotype, we modeled the chimeric tissue genotype of somatic loss of heterozygosity (LOH), by conditionally inactivating Ext1 via head-to-head loxP sites and temporally controlled Cre-recombinase in chondrocytes. These mice faithfully recapitulate the human phenotype of multiple metaphyseal osteochondromas. We also confirm homozygous disruption of Ext1 in osteochondroma chondrocytes and their origin in proliferating physeal chondrocytes. These results explain prior modeling failures with the necessity for somatic LOH in a developmentally regulated cell type.


Mini dental implants retaining mandibular overdentures: A dental practice-based retrospective analysis.
The purpose of this study was to assess the survival of mini dental implants (MDI) and to measure prosthetic maintenance needs in a dental practice-based setting. Patients with mandibular removable dentures were provided with MDI to improve denture retention. Complications and maintenance were analyzed by use of patient records and evaluated with Kaplan-Meier curves and the log rank test at a significance level of 0.05. Ninety-nine MDI were placed in 25 patients (mean age: 72 years). Two MDI fractured during placement and eight implants failed during the first weeks. No more implants were lost for up to seven years, resulting in 92% survival. Implant survival differed significantly depending on whether the maxilla was provided with complete dentures (94.9%) or with partial dentures (81%). All prostheses were in use at the time of data extraction. Denture base fractures were observed in six cases, an incidence of fractures of 24%. Some minor intervention was necessary: one resin tooth fractured, retention rings were changed in five cases, and repeated relining was required for 16% of the dentures. After mid-term observation, survival of MDI was good. However, the incidence of denture base fractures and of minor prosthetic complications should not be under-estimated.


Clinical Characteristics of COVID-19 from the Otorhinolaryngologist's Perspective: First Report from Oman, Middle East.
To study the otorhinolaryngological clinical characteristics of COVID-19 positive patients. A prospective cross sectional study on sixty five patients who were SARS-CoV-2 PCR positive, and completed 14 days of isolation period were surveyed with a questionnaire. The responses were evaluated and assessed. Sixty five SARS-Cov-2 PCR positive cases were included in the study. There were 57 (87.6%) males and 8 (12.3%) females. Thirty five (53.8%) were in home isolation, whereas, 30 (46.2%) were under institutional care. Forty five patients (72.6%) presented with mild symptoms, and 4 (6.4%) developed moderate symptoms. Thirteen (21%) were asymptomatic. Overall, 46 patients (70.7%) presented with upper airway symptoms with or without general symptoms. More than half of the patients experienced pharyngodynia or sorethroat, smell and taste dysfunction as common symptoms (66.7%, 61.4% and 50.7% respectively). Severe headache was noticed by eighteen (27.7%) patients. Other respiratory symptoms such as nasal congestion, rhinorrhoea, sneezing, facial pain, etc. were present with less frequency. In more than half of the patients (61.5%), all the symptoms recovered within 5 days, in 12 (18.5%) between 5 and 8 days, and in 9 (13.8%), between 9 and14 days. However, in four patients, symptoms lasted for 28-30 days. In seven patients (10.7%), symptoms recurred after the period of isolation, however, the retest was negative. Fever, cough and or shortness of breath are the commonly reported prominent symptoms of COVID-19, however, there is a changing trend of clinical presentation towards variable otorhinolaryngologic manifestations. Pharyngodynia, taste and smell dysfunctions are common in patients with COVID-19, and could represent potential characters.


The effect of poultry manure application rate and AlCl(3) treatment on bacterial fecal indicators in runoff.
Increasing costs associated with inorganic fertilizer have led to widespread use of broiler litter. Proper land application, typically limiting nutrient loss, is essential to protect surface water. This study was designed to evaluate litter-borne microbial runoff (heterotrophic plate count bacteria, staphylococci, Escherichia coli, enterococci, and Clostridium perfringens) while applying typical nutrient-control methods. Field studies were conducted in which plots with high and low litter rates, inorganic fertilizer, AlCl(3)-treated litter, and controls were rained on five times using a rain generator. Overall, microbial runoff from poultry litter applied plots was consistently greater (2-5 log(10) plot(-1)) than controls. No appreciable effect on microbial runoff was noted from variable litter application rate or AlCl(3) treatments, though rain event, not time, significantly affected runoff load. C. perfringens and staphylococci runoff were consistently associated with poultry litter application, during early rain events, while other indicators were unreliable. Large microbial runoff pulses were observed, ranging from 10(2) to 10(10) CFU plot(-1); however, only a small fraction of litter-borne microbes were recoverable in runoff. This study indicated that microbial runoff from litter-applied plots can be substantial, and that methods intended to reduce nutrient losses do not necessarily reduce microbial runoff.


Applicability of the CALUX bioassay for screening of dioxin levels in human milk samples.
The CALUX (chemically activated luciferase expression) bioassay based on rat hepatoma (H4IIE) cells is a sensitive assay for the detection of Ah receptor agonists like 2,3,7,8-substituted chlorinated dibenzo-p-dioxins and dibenzofurans and related PCBs. In this paper, the assay was optimized and applied for monitoring levels of dioxins in human milk samples. Combination effects of dioxin-like compounds were evaluated by testing potential mechanisms of interaction between seven of the major dioxin-like compounds in human milk using the isobole method. Results showed that the compounds acted additively, indicating that the usual assumption of additivity in the risk assessment process is valid. In general the relative potencies (REPs) of the single agents were in accordance with their TEFs assigned by the World Health Organisation, except for the mono-ortho-substituted PCB118 that had a 40-fold lower REP in CALUX. The total dioxin-like activity was determined in 16 Danish human milk samples and was in the range 20.5-55.8 pg TEQ g(-1) fat. These values were compared with TEQs obtained from GC/MS analysis (range 14.8-43.6 pg TEQ-g(-1) fat) that overall were a little lower than CALUX TEQs. The results obtained with the bioassay when testing milk extracts fractionated into dioxins/furans, non-ortho PCB and mono/di-ortho PCB fractions indicated that the correlation between the bioassay and the chemical analyses depends primarily on the Ah receptor activity observed in the mono/di-ortho PCB fraction.


TLR signaling is required for Salmonella typhimurium virulence.
Toll-like receptors (TLRs) contribute to host resistance to microbial pathogens and can drive the evolution of virulence mechanisms. We have examined the relationship between host resistance and pathogen virulence using mice with a functional allele of the nramp-1 gene and lacking combinations of TLRs. Mice deficient in both TLR2 and TLR4 were highly susceptible to the intracellular bacterial pathogen Salmonella typhimurium, consistent with reduced innate immune function. However, mice lacking additional TLRs involved in S. typhimurium recognition were less susceptible to infection. In these TLR-deficient cells, bacteria failed to upregulate Salmonella pathogenicity island 2 (SPI-2) genes and did not form a replicative compartment. We demonstrate that TLR signaling enhances the rate of acidification of the Salmonella-containing phagosome, and inhibition of this acidification prevents SPI-2 induction. Our results indicate that S. typhimurium requires cues from the innate immune system to regulate virulence genes necessary for intracellular survival, growth, and systemic infection.


Occurrence and Ecotoxicological Effects of Free, Conjugated, and Halogenated Steroids Including 17α-Hydroxypregnanolone and Pregnanediol in Swiss Wastewater and Surface Water.
Apart from estrogens, the occurrence and ecotoxicity of steroids in aquatic environments is poorly known. Here, we analyzed 33 steroids, including estrogens, androgens, progestins, and glucocorticoids, in hospital wastewaters, river water, and municipal wastewater treatment plant (WTP) influents and effluents at different sites in Switzerland. In addition, wastewater from different treatment steps of two WTPs with advanced treatment, such as ozonation or pulverized activated carbon, were analyzed to study the steroid's behavior during treatment. Considerable levels of different steroids occurred in hospital and raw municipal wastewater, but they were low (lower than 1 ng/L) or below the detection level in effluents of WTPs and river water. In WTP influents, estrogens (estrone, 17β-estradiol, and estriol), androgens (androstenedione, androsterone, trans-androsterone, and testosterone), progestins and metabolites (progesterone, medroxyprogesterone acetate, megestrol acetate, mifepristone, pregnanediol, 17α-hydroxypregnanolone, 17α-hydroxyprogesterone, and 21α-hydroxyprogesterone) were detected and removed effectively during biological treatment. Ozonation further removed the steroids. Exposure of zebrafish embryos demonstrated negligible effects of pregnanediol and 17α-hydroxypregnanolone, while mixtures that mimic wastewater and river water composition affected embryo development and led to the alteration of steroidogenesis gene transcripts at nanogram per liter concentrations. Although steroid concentrations are low in Swiss rivers, the possibility of additive effects may be of concern.


The experiences of ICU patients during follow-up sessions--a qualitative study.
Evidence supports the recommendation for follow-up session(s) for patients after discharge from an intensive care unit (ICU). The aim of these follow-up sessions is to allow patients to express and discuss their experiences and problems following their time in an ICU. To optimise the knowledge gained from the follow-up session experience, it is necessary to describe how patients experience these sessions. The aim of this study was to describe how ICU-patients, experience a follow-up session. This study adopted a qualitative design utilising semi-structured interviews, and which examined the experiences of seven men and five women. Qualitative content analysis was utilised. The participants stated that the information gained from these sessions, which had previously seemed unclear to some of them, was, on the whole, now clarified and confirmed. A discernible difference was found between participants who were cared for on a general ward and those who were cared for on a rehabilitation ward and also were offered a meeting with a counsellor, following discharge their from the ICU. The findings also indicated that participants who were not offered psychosocial support showed a greater need for a follow-up session. This study has highlighted the need for increasing collaboration between intensive care staff and staff in other units to provide support to this patient group in order to reduce their suffering post intensive care experience.


Qualitative process evaluation from a complex systems perspective: A systematic review and framework for public health evaluators.
Public health evaluation methods have been criticized for being overly reductionist and failing to generate suitable evidence for public health decision-making. A "complex systems approach" has been advocated to account for real world complexity. Qualitative methods may be well suited to understanding change in complex social environments, but guidance on applying a complex systems approach to inform qualitative research remains limited and underdeveloped. This systematic review aims to analyze published examples of process evaluations that utilize qualitative methods that involve a complex systems perspective and proposes a framework for qualitative complex system process evaluations. We conducted a systematic search to identify complex system process evaluations that involve qualitative methods by searching electronic databases from January 1, 2014-September 30, 2019 (Scopus, MEDLINE, Web of Science), citation searching, and expert consultations. Process evaluations were included if they self-identified as taking a systems- or complexity-oriented approach, integrated qualitative methods, reported empirical findings, and evaluated public health interventions. Two reviewers independently assessed each study to identify concepts associated with the systems thinking and complexity science traditions. Twenty-one unique studies were identified evaluating a wide range of public health interventions in, for example, urban planning, sexual health, violence prevention, substance use, and community transformation. Evaluations were conducted in settings such as schools, workplaces, and neighborhoods in 13 different countries (9 high-income and 4 middle-income). All reported some utilization of complex systems concepts in the analysis of qualitative data. In 14 evaluations, the consideration of complex systems influenced intervention design, evaluation planning, or fieldwork. The identified studies used systems concepts to depict and describe a system at one point in time. Only 4 evaluations explicitly utilized a range of complexity concepts to assess changes within the system resulting from, or co-occurring with, intervention implementation over time. Limitations to our approach are including only English-language papers, reliance on study authors reporting their utilization of complex systems concepts, and subjective judgment from the reviewers relating to which concepts featured in each study. This study found no consensus on what bringing a complex systems perspective to public health process evaluations with qualitative methods looks like in practice and that many studies of this nature describe static systems at a single time point. We suggest future studies use a 2-phase framework for qualitative process evaluations that seek to assess changes over time from a complex systems perspective. The first phase involves producing a description of the system and identifying hypotheses about how the system may change in response to the intervention. The second phase involves following the pathway of emergent findings in an adaptive evaluation approach.


High frequency (900 MHz) low amplitude (5 V m-1) electromagnetic field: a genuine environmental stimulus that affects transcription, translation, calcium and energy charge in tomato.
Using an especially-designed facility, the Mode Stirred Reverberation Chamber, we exposed tomato plants (Lycopersicon esculentum Mill. VFN8) to low level (900 MHz, 5 V m(-1)) electromagnetic fields for a short period (10 min) and measured changes in abundance of three specific mRNA soon after exposure. Within minutes of electromagnetic stimulation, stress-related mRNA (calmodulin, calcium-dependent protein kinase and proteinase inhibitor) accumulated in a rapid, large and 3-phase manner typical of an environmental stress response. Accumulation of these transcripts into the polysomal RNA also took place (indicating that the encoded proteins were translated) but was delayed (indicating that newly-synthesized mRNA was not immediately recruited into polysomes). Transcript accumulation was maximal at normal Ca(2+) levels and was depressed at higher Ca(2+), especially for those encoding calcium-binding proteins. Removal of Ca(2+) (by addition of chelating agents or Ca(2+) channel blocker) led to total suppression of mRNA accumulation. Finally, 30 min after the electromagnetic treatment, ATP concentration and adenylate energy charge were transiently decreased, while transcript accumulation was totally prevented by application of the uncoupling reagent, CCCP. These responses occur very soon after exposure, strongly suggesting that they are the direct consequence of application of radio-frequency fields and their similarities to wound responses strongly suggests that this radiation is perceived by plants as an injurious stimulus.


Long-term homeostasis and wound healing in an in vitro epithelial stem cell niche model.
Cultures of epithelial cells are limited by the proliferative capacity of primary cells and cell senescence. Herein we show that primary human epithelial cell sheets cultured without dermal equivalents maintained homeostasis in vitro for at least 1 year. Transparency of these sheets enabled live observation of pigmented melanocytes and Fluorescent Ubiquitination-based Cell Cycle Indicator (FUCCI) labeled epithelial cells during wound healing. Cell turn over and KRT15 expression pattern stabilized within 3 months, when KRT15 bright clusters often associated with niche-like melanocytes became apparent. EdU labels were retained in a subset of epithelial cells and melanocytes after 6 months chasing, suggesting their slow cell cycling property. FUCCI-labeling demonstrated robust cell migration and proliferation following wounding. Transparency and long-term (1 year) homeostasis of this model will be a powerful tool for the study of wound healing and cell linage tracing.


Comparative genome analysis provides insights into the evolution and adaptation of Pseudomonas syringae pv. aesculi on Aesculus hippocastanum.
A recently emerging bleeding canker disease, caused by Pseudomonas syringae pathovar aesculi (Pae), is threatening European horse chestnut in northwest Europe. Very little is known about the origin and biology of this new disease. We used the nucleotide sequences of seven commonly used marker genes to investigate the phylogeny of three strains isolated recently from bleeding stem cankers on European horse chestnut in Britain (E-Pae). On the basis of these sequences alone, the E-Pae strains were identical to the Pae type-strain (I-Pae), isolated from leaf spots on Indian horse chestnut in India in 1969. The phylogenetic analyses also showed that Pae belongs to a distinct clade of P. syringae pathovars adapted to woody hosts. We generated genome-wide Illumina sequence data from the three E-Pae strains and one strain of I-Pae. Comparative genomic analyses revealed pathovar-specific genomic regions in Pae potentially implicated in virulence on a tree host, including genes for the catabolism of plant-derived aromatic compounds and enterobactin synthesis. Several gene clusters displayed intra-pathovar variation, including those encoding type IV secretion, a novel fatty acid biosynthesis pathway and a sucrose uptake pathway. Rates of single nucleotide polymorphisms in the four Pae genomes indicate that the three E-Pae strains diverged from each other much more recently than they diverged from I-Pae. The very low genetic diversity among the three geographically distinct E-Pae strains suggests that they originate from a single, recent introduction into Britain, thus highlighting the serious environmental risks posed by the spread of an exotic plant pathogenic bacterium to a new geographic location. The genomic regions in Pae that are absent from other P. syringae pathovars that infect herbaceous hosts may represent candidate genetic adaptations to infection of the woody parts of the tree.


Quantifying the unmet needs of caregivers of people with dementia: a critical review of the quality of measures.
The array of demanding tasks carried out by caregivers of people with dementia have significant negative impacts on their physical, mental and social well-being. Needs assessment allows individuals to indicate the extent to which their needs across different areas have or have not been met, allowing for estimations of the prevalence of needs and the extent to which help is required. This approach is extremely valuable in a clinical context, as it enables identification of the areas with which caregivers report a particular desire for help and allows targeting of support and resources to those who identify high levels of unmet needs. This systematic review aimed to critically examine the psychometric properties of measures that assess unmet needs of caregivers of people with dementia. Medline, Embase, PsycINFO and Cochrane electronic databases were searched between January 1990 and August 2015 for English-language publications describing the development or validation of measures assessing the unmet needs of adult caregivers of people with dementia. The psychometric properties of included measures were assessed against standard criteria for psychometric quality. Four measures met the inclusion criteria. Only half of the indices of psychometric quality were tested across measures. Three measures had adequate internal consistency reliability, of which one also showed adequate test-retest reliability. Two measures reported adequate construct validity, while criterion validity was not assessed for any measure. There is a clear need to develop a psychometrically rigorous instrument to identify the unmet needs of caregivers of people with dementia. Copyright © 2016 John Wiley & Sons, Ltd.