A genetically encoded toolkit of functionalized nanobodies against fluorescent proteins for visualizing and manipulating intracellular signalling.
Intrabodies enable targeting of proteins in live cells, but generating specific intrabodies against the thousands of proteins in a proteome poses a challenge. We leverage the widespread availability of fluorescently labelled proteins to visualize and manipulate intracellular signalling pathways in live cells by using nanobodies targeting fluorescent protein tags. We generated a toolkit of plasmids encoding nanobodies against red and green fluorescent proteins (RFP and GFP variants), fused to functional modules. These include fluorescent sensors for visualization of Ca2+, H+ and ATP/ADP dynamics; oligomerising or heterodimerising modules that allow recruitment or sequestration of proteins and identification of membrane contact sites between organelles; SNAP tags that allow labelling with fluorescent dyes and targeted chromophore-assisted light inactivation; and nanobodies targeted to lumenal sub-compartments of the secretory pathway. We also developed two methods for crosslinking tagged proteins: a dimeric nanobody, and RFP-targeting and GFP-targeting nanobodies fused to complementary hetero-dimerizing domains. We show various applications of the toolkit and demonstrate, for example, that IP3 receptors deliver Ca2+ to the outer membrane of only a subset of mitochondria and that only one or two sites on a mitochondrion form membrane contacts with the plasma membrane. This toolkit greatly expands the utility of intrabodies and will enable a range of approaches for studying and manipulating cell signalling in live cells.


Experience-dependent neuroplasticity in trained musicians modulates the effects of chronic pain on insula-based networks - A resting-state fMRI study.
Recent resting-state fMRI studies associated extensive musical training with increased insula-based connectivity in large-scale networks involved in salience, emotion, and higher-order cognitive processes. Similar changes have also been found in chronic pain patients, suggesting that both types of experiences can have comparable effects on insula circuitries. Based on these observations, the current study asked the question whether, and if so in what way, different forms of experience-dependent neuroplasticity may interact. Here we assessed insula-based connectivity during fMRI resting-state between musicians and non-musicians both with and without chronic pain, and correlated the results with clinical pain duration and intensity. As expected, insula connectivity was increased in chronic pain non-musicians relative to healthy non-musicians (with cingulate cortex and supplementary motor area), yet no differences were found between chronic pain non-musicians and healthy musicians. In contrast, musicians with chronic pain showed decreased insula connectivity relative to both healthy musicians (with sensorimotor and memory regions) and chronic pain non-musicians (with the hippocampus, inferior temporal gyrus, and orbitofrontal cortex), as well as lower pain-related inferences with daily activities. Pain duration correlated positively with insula connectivity only in non-musicians, whereas pain intensity exhibited distinct relationships across groups. We conclude that although music-related sensorimotor training and chronic pain, taken in isolation, can lead to increased insula-based connectivity, their combination may lead to higher-order plasticity (metaplasticity) in chronic pain musicians, engaging brain mechanisms that can modulate the consequences of maladaptive experience-dependent neural reorganization (i.e., pain chronification).


Triggering comprehensive enhancement in oxygen evolution reaction by using newly created solvent.
Theoretical calculations indicate that the properties of confined liquid water, or liquid water at surfaces, are dramatically different from those of liquid bulk water. Here we present an experimentally innovative strategy on comprehensively efficient oxygen evolution reaction (OER) utilizing plasmon-induced activated water, creating from hot electron decay at resonantly illuminated Au nanoparticles (NPs). Compared to conventional deionized (DI) water, the created water owns intrinsically reduced hydrogen-bonded structure and a higher chemical potential. The created water takes an advantage in OER because the corresponding activation energy can be effectively reduced by itself. Compared to DI water-based solutions, the OER efficiencies at Pt electrodes increased by 69.3%, 21.1% and 14.5% in created water-based acidic, neutral and alkaline electrolyte solutions, respectively. The created water was also effective for OERs in photoelectrochemically catalytic and in inert systems. In addition, the efficiency of OER increased by 47.5% in created water-based alkaline electrolyte solution prepared in situ on a roughened Au electrode. These results suggest that the created water has emerged as an innovative activator in comprehensively effective OERs.


Association between androgenetic alopecia and coronary artery disease in young male patients.
Several studies have demonstrated an association between androgenetic alopecia (AGA) and cardiovascular disease. Still controversies exist regarding the association. Are they truly associated? The purpose of the present study was to assess the prevalence of AGA and establish its association in young (<45 years) Asian Indian Gujarati male patients having coronary artery disease (CAD). Case-control prospective multicentric study was carried out on 424 men. Case group consisted of 212 male subjects having CAD (Group 1) and another 212, either sibling or first degree male relative of the case subjects (having no evidence of CAD) were considered as the control group (Group 2). Age, total cholesterol, incidence of diabetes mellitus, and hypertension were similar in both groups. The degree of alopecia was assessed using the Norwood-Hamilton scale for men. Statistical analysis was performed using Chi-square test. AGA was found in 80 (37.73%) young CAD patients (Group 1), whereas 44 (20.7%) of patients had alopecia in the control group (Group 2). There was statistically significant association between male AGA and CAD (P = 0.001). Odds ratio was 2.70 (95% confidence interval [CI], 1.72 ± 4.26). Statistically significant association was found between high grade baldness (Grades IV-VII) and CAD in young men (P < 0.05). Odds ratio = 2.36 (95% CI, 1.108 ± 5.033). There is statistically significant association of AGA in young Asian Gujarati male with CAD and the prevalence of AGA in young CAD patient is 37.73%. This study implies early onset AGA in male is independently associated with CAD, though mechanisms need to be investigated.


Developmental monitoring using caregiver reports in a resource-limited setting: the case of Kilifi, Kenya.
The main aim of the current study was to evaluate the reliability, validity and acceptability of developmental monitoring using caregiver reports among mothers in a rural African setting. A structured interview for parents of children aged 24 months and less was developed through both participant consultation and a review of literature. The reliability and validity of the schedule was evaluated through a 10-month monitoring programme of 95 children, aged 2-10 months. The acceptability of the process was evaluated by studying retention rates and by organizing focus group discussions with participating mothers. The structured interview 'Developmental Milestones Checklist' consisted of 66 items covering three broad domains of child functioning: motor, language and personal-social development. The interview yielded scores of developmental achievements that showed high internal consistency and excellent test-retest reliability. The results were sensitive to maturational changes and nutritional deficiencies. In addition, acceptable retention rates of approximately 80% were found. Participating mothers reported that they found the procedures both acceptable and beneficial. Developmental monitoring using caregiver report is a viable method to identify and monitor at-risk children in Sub-Saharan Africa.


Pooling of contrast material on computed tomography mandates aggressive management of blunt hepatic injury.
Nonoperative management of blunt hepatic injury is currently a widely accepted treatment modality. Computed tomography (CT) is an important imaging study both for diagnosis and follow-up of these patients. There is, however, no reliable predictor of failure of nonoperative treatment other than the ultimate development of hemodynamic instability. Previous reports mostly were based on the data obtained from low-speed dynamic incremental scanners. The purpose of this study is to evaluate the value of a high-speed helical scanner in predicting the outcome of patients managed nonoperatively. During a 30-month period, 194 patients with blunt hepatic injury were treated, 150 of them were hemodynamically stable after initial resuscitation and underwent abdominal CT examination. All CT scans were performed with the High Speed Advantage Scanner. The CT scans and medical records were reviewed. Nonoperative management was successfully applied to all patients with grade I and II, 93% of grade III, 87% of grade IV, and 67% of grade V liver injuries. Twelve patients required liver-related celiotomy. Pooling of contrast material was detected on the CT scans of 8 patients. Six (75%) of these patients developed hemodynamic instability and required liver-related celiotomy later. Pooling of contrast material can be detected in 50% of the patients receiving liver-related celiotomy. The presence of pooling of contrast material within the hepatic parenchyma indicates free extravasation of blood as a result of active bleeding. In patients with blunt hepatic injury, if this sign is detected, nonoperative treatment should be terminated and angiography or celiotomy undertaken promptly. With the increasing use of high-speed spiral CT scanner and improvement in scanning technique, pooling of contrast material may become a sensitive sign for active bleeding and may be used as a guide for the selection of treatment modality.


Cognitive and behavioural strategies for self-directed weight loss: systematic review of qualitative studies.
We conducted a systematic review of qualitative studies to examine the strategies people employ as part of self-directed weight loss attempts, map these to an existing behaviour change taxonomy and explore attitudes and beliefs surrounding these strategies. Seven electronic databases were searched in December 2015 for qualitative studies in overweight and obese adults attempting to lose weight through behaviour change. We were interested in strategies used by participants in self-directed efforts to lose weight. Two reviewers extracted data from included studies. Thematic and narrative synthesis techniques were used. Thirty one studies, representing over 1,000 participants, were included. Quality of the included studies was mixed. The most commonly covered types of strategies were restrictions, self-monitoring, scheduling, professional support and weight management aids. With the exception of scheduling, for which participant experiences were predominantly positive, participants' attitudes and beliefs surrounding implementation of these groups of strategies were mixed. Two new groups of strategies were added to the existing taxonomy: reframing and self-experimentation. This review demonstrates that at present, interventions targeting individuals engaged in self-management of weight do not necessarily reflect lived experiences of self-directed weight loss.


Are safety data sheets for cleaning products used in Norway a factor contributing to the risk of workers exposure to chemicals?
Cleaning products are considered less hazardous than those used in other sectors. Suppliers and distributors are less conscientious when it comes to informing users on health risks. The aim of the study was to elaborate on the usefulness and clarity of information in the safety data sheets (SDS) for cleaning products, and considering if the use of these SDSs can be seen as a risk factor towards occupational exposure to hazardous chemicals in the sector. Safety data sheets were selected based on the risk level of the product assigned in an industrial sector scheme. 320 SDSs for cleaning products were reviewed. Constituent components found in the products over a given threshold were listed and available information thereof used to assess the perceived non-hazard consideration of the chemicals. The contents of the SDSs was generic and mostly incomplete. Safety measures and health information lacked sufficient specificity despite varying compositions and concentrations of components. There is generally incompatibility between mentioned sections on the suggested non-hazardous nature of the products and health effects. Not all substances used in these products have harmonized classifications, which makes them open to various classification of the products and the suggested safety measures. This results in different companies classifying similar products differently. Risk management measures and suggested personal protective equipment (PPEs) are given haphazardly. Physical properties relevant to risk assessment are not included. The safety data sheets are ambiguous, and they lack relevant and important information. Inadequate information and risk assessment concerning the products can lead to workers being exposed to hazardous chemicals. Underestimation of the hazard contribution of the components of the products and the insufficient, non-objective mention of appropriate control and protective measures are the major contributing elements. There is a need to test the products in order to establish health effects and product specific safety measures.


Math anxiety: Brain cortical network changes in anticipation of doing mathematics.
Following our previous work regarding the involvement of math anxiety (MA) in math-oriented tasks, this study tries to explore the differences in the cerebral networks' topology between self-reported low math-anxious (LMA) and high math-anxious (HMA) individuals, during the anticipation phase prior to a mathematical related experiment. For this reason, multichannel EEG recordings were adopted, while the solution of the inverse problem was applied in a generic head model, in order to obtain the cortical signals. The cortical networks have been computed for each band separately, using the magnitude square coherence metric. The main graph theoretical parameters, showed differences in segregation and integration in almost all EEG bands of the HMAs in comparison to LMAs, indicative of a great influence of the anticipatory anxiety prior to mathematical performance.


Biology and Sources of Inoculum of Geotrichum candidum Causing Sour Rot of Peach and Nectarine Fruit in California.
Geotrichum candidum causes sour rot of fresh-market stone fruit such as peach and nectarine. Since 2001, the incidence of sour rot has increased in California, a semi-arid production area, which is considered atypical for the occurrence of the disease. In this study, sour rot developed at significantly higher incidence on wounded fruit compared with unwounded fruit, and disease severity increased as fruit matured. In packinghouse surveys, sour rot was found on up to 4% of non-fungicide- treated peach and nectarine fruit. In laboratory assays, sour rot developed when fruit were inoculated with a minimum of 20 conidia per inoculation site. Inoculum of G. candidum could be detected in California orchard soils at depths of up to 10 cm. The amount of inoculum in the soil was positively correlated with that on leaf and fruit surfaces of trees growing at a specific site. Moreover, inoculum of G. candidum was detected at different areas of packing lines in seven packinghouses. There were significant differences among the packinghouses evaluated, and these differences could be attributed, in part, to different sanitation practices used. Nitidulid beetles and fruit flies were found to play a role in disease transmission. These insects acquired sour rot inoculum in the orchard, and 25% of nitidulid beetles and 26% of fruit flies collected were positive for the pathogen. Spore survival in the soil over a 12-month period decreased significantly when soil depth increased from 10 to 20 cm. This study identified sources of inoculum of G. candidum in orchards and packinghouses, and provides information to guide development of disease management programs under the semi-arid conditions of California.


Triboelectric nanogenerators as new energy technology and self-powered sensors - principles, problems and perspectives.
Triboelectrification is one of the most common effects in our daily life, but it is usually taken as a negative effect with very limited positive applications. Here, we invented a triboelectric nanogenerator (TENG) based on organic materials that is used to convert mechanical energy into electricity. The TENG is based on the conjunction of triboelectrification and electrostatic induction, and it utilizes the most common materials available in our daily life, such as papers, fabrics, PTFE, PDMS, Al, PVC etc. In this short review, we first introduce the four most fundamental modes of TENG, based on which a range of applications have been demonstrated. The area power density reaches 1200 W m(-2), volume density reaches 490 kW m(-3), and an energy conversion efficiency of ∼50-85% has been demonstrated. The TENG can be applied to harvest all kinds of mechanical energy that is available in our daily life, such as human motion, walking, vibration, mechanical triggering, rotation energy, wind, a moving automobile, flowing water, rain drops, tide and ocean waves. Therefore, it is a new paradigm for energy harvesting. Furthermore, TENG can be a sensor that directly converts a mechanical triggering into a self-generated electric signal for detection of motion, vibration, mechanical stimuli, physical touching, and biological movement. After a summary of TENG for micro-scale energy harvesting, mega-scale energy harvesting, and self-powered systems, we will present a set of questions that need to be discussed and explored for applications of the TENG. Lastly, since the energy conversion efficiencies for each mode can be different although the materials are the same, depending on the triggering conditions and design geometry. But one common factor that determines the performance of all the TENGs is the charge density on the two surfaces, the saturation value of which may independent of the triggering configurations of the TENG. Therefore, the triboelectric charge density or the relative charge density in reference to a standard material (such as polytetrafluoroethylene (PTFE)) can be taken as a measuring matrix for characterizing the performance of the material for the TENG.


Screening for autism in Mexico.
In order to conduct the screening phase of the first epidemiological survey of autism spectrum disorders (ASDs) in Mexico, we needed a screening tool to detect autistic symptomatology in a large sample of school-age children. We used the Spanish version of the Social Responsiveness Scale (SRS). We recruited a clinical sample of 200 children (81% males; mean age: 7.4 years) with a confirmed diagnosis of ASDs and a sample of 363 control children (59.5% males; mean age: 8.5 years) without ASDs. Three-way analyses of variance (ANOVAs) identified a main effect of clinical status (ASDs vs. controls) for both parent and teacher scales, but no gender or age effect. The mean total and subscale raw scores were significantly different between the clinical and control groups for the parent and for the teacher SRS (P < 0.001). The internal consistency of the SRS was excellent. Receiver operating characteristic (ROC) analyses showed excellent discriminant validity of the SRS in the Mexican sample (area under the curve: 0.962 for the parent, 0.960 for the teacher). ROC curves were also used to determine which cutoff would provide the best trade-off between sensitivity and specificity. Mexican SRS scores were significantly higher than in the U.S. and German population for typically developing children but comparable for clinically referred subjects. The SRS is an acceptable screening instrument for epidemiological studies of ASDs in Mexico. Its psychometric properties are excellent and comparable to those derived from North American and other samples.


Broviac catheterization in low birth weight infants: incidence and treatment of associated complications.
Fifty-two Broviac catheters were inserted in 40 preterm and eight term infants for 1733 days of catheter use. Thirty-six (69%) catheters were associated with complications of infection and/or thrombosis, a complication rate of 1/48 catheter days. The patients who developed complications were of a significantly lower gestational age and had a lower mean birth weight when compared with those who developed no complications. The incidence of catheter-related sepsis was 69% in the very low birth weight infants and only 20% in the infants with birth weights over 1500 g. Eighteen of the 26 catheter-associated infections were treated with antibiotics without catheter removal. Successful resolution of the infections with retention of the catheter occurred in 14 of the 18 episodes. Infections with Staphylococcus aureus constituted three of four treatment failures. Urokinase infusion was successful in causing thrombolysis in eight of the nine cases. Broviac catheters in neonates, and especially in preterm infants under 1500 g, are associated with a high incidence of complications. Our experience indicates that some complications can be selectively managed without sacrificing the venous access.


High-sensitivity sequencing reveals multi-organ somatic mosaicism causing DICER1 syndrome.
Somatic mosaicism is being increasingly recognised as an important cause of non-Mendelian presentations of hereditary syndromes. A previous whole-exome sequencing study using DNA derived from peripheral blood identified mosaic mutations in DICER1 in two children with overgrowth and developmental delay as well as more typical phenotypes of germline DICER1 mutation. However, very-low-frequency mosaicism is difficult to detect, and thus, causal mutations can go unnoticed. Highly sensitive, cost-effective approaches are needed to molecularly diagnose these persons. We studied four children with multiple primary tumours known to be associated with the DICER1 syndrome, but in whom germline DICER1 mutations were not detected by conventional mutation detection techniques. We observed the same missense mutation within the DICER1 RNase IIIb domain in multiple tumours from different sites in each patient, raising suspicion of somatic mosaicism. We implemented three different targeted-capture technologies, including the novel HaloPlex(HS) (Agilent Technologies), followed by deep sequencing, and confirmed that the identified mutations are mosaic in origin in three patients, detectable in 0.24-31% of sequencing reads in constitutional DNA. The mosaic origin of patient 4's mutation remains to be unequivocally established. We also discovered likely pathogenic second somatic mutations or loss of heterozygosity (LOH) in tumours from all four patients. Mosaic DICER1 mutations are an important cause of the DICER1 syndrome in patients with severe phenotypes and often appear to be accompanied by second somatic truncating mutations or LOH in the associated tumours. Furthermore, the molecular barcode-containing HaloPlex(HS) provides the sensitivity required for detection of such low-level mosaic mutations and could have general applicability.


Fluorescent imaging of nitric oxide production in neuronal varicosities associated with intraparenchymal arterioles in rat hippocampal slices.
The fluorescent indicator 4,5-diaminofluorescein (DAF-2) has been used to investigate the production of nitric oxide in the vicinity of intraparenchymal cerebral blood vessels. Slices of rat hippocampus 300-350 microm thick, were loaded with 5 microM DAF-2 diacetate. On exposure to light of 450-490 nm wavelength, point sources of fluorescence, 1.8+/-0.2 microm in diameter (mean+/-SEM), were observed in close apposition to the outer surface of the vascular smooth muscle wall of 10/15 arterioles. In fixed slices, resectioned and processed for nicotinamide adenine dinucleotide phosphate-dependent diaphorase, stained varicose fibres were also seen in close association with the smooth muscle wall of small arterioles. These findings suggest that tonic activity in perivascular nitrergic nerve fibres lying in close proximity to intraparenchymal microvessels may be a source of dilator tone within the parenchyma.


Ultrahigh capacity 2 × 2 MIMO RoF system at 60 GHz employing single-sideband single-carrier modulation.
This article proposes and experimentally demonstrates a radio-over-fiber system employing single-sideband single-carrier (SSB-SC) modulation at 60 GHz. SSB-SC modulation has a lower peak-to-average-power ratio than orthogonal frequency division multiplex (OFDM) modulation; therefore, the SSB-SC signals provide superior nonlinear tolerance, compared to OFDM signals. Moreover, multiple-input multiple-output (MIMO) technology was used extensively to enhance spectral efficiency. A least-mean-square-based equalizer was implemented, including MIMO channel estimation, frequency response equalization, and I/Q imbalance compensation to recover the MIMO signals. Thus, using 2×2 MIMO technology and 64-QAM SSB-SC signals, we achieved the highest data rate of 84 Gbps with 12 bit/s/Hz spectral efficiency using the 7-GHz license-free band at 60 GHz.


Critical limb ischemia.
Critical limb ischemia (CLI) continues to be a significantly morbid disease process for the aging population. Rigid guidelines for the management of patients with CLI are inappropriate due to the complexities that are involved in optimally treating these patients. A thin line exists in the decision process between medical management vs surgical management by revascularization or amputation, and the perception of "success" in this patient population is evolving. This review explores these issues and examines the challenges the treating physician will face when managing the care of patients with CLI. The epidemiology and natural history of CLI is discussed, along with the pathophysiology of the disease process. A review of the literature in regards to the different treatment modalities is presented to help the physician optimize therapy for patients with CLI. New scoring systems to help predict outcomes in patients with CLI undergoing revascularization or amputation are discussed, and an overview of the current status of patient-oriented outcomes is provided. Finally, we briefly examine emerging therapies for the treatment of CLI and provide an algorithm to help guide the practicing physician on how to approach the critically ischemic limb with regard to the complicated issues surrounding these patients.


AARC Clinical Practice Guideline. Surfactant replacement therapy: 2013.
We searched the MEDLINE, CINAHL, and Cochrane Library databases for English-language randomized controlled trials, systematic reviews, and articles investigating surfactant replacement therapy published between January 1990 and July 2012. By inspection of titles, references having no relevance to the clinical practice guideline were eliminated. The update of this clinical practice guideline is based on 253 clinical trials and systematic reviews, and 12 articles investigating surfactant replacement therapy. The following recommendations are made following the Grading of Recommendations Assessment, Development, and Evaluation scoring system: 1: Administration of surfactant replacement therapy is strongly recommended in a clinical setting where properly trained personnel and equipment for intubation and resuscitation are readily available. 2: Prophylactic surfactant administration is recommended for neonatal respiratory distress syndrome (RDS) in which surfactant deficiency is suspected. 3: Rescue or therapeutic administration of surfactant after the initiation of mechanical ventilation in infants with clinically confirmed RDS is strongly recommended. 4: A multiple surfactant dose strategy is recommended over a single dose strategy. 5: Natural exogenous surfactant preparations are recommended over laboratory derived synthetic suspensions at this time. 6: We suggest that aerosolized delivery of surfactant not be utilized at this time.


Search for microbial signatures within human and microbial calcifications using soft x-ray spectromicroscopy.
The origin of advanced arterial and renal calcification remains poorly understood. Self-replicating, calcifying entities have been detected and isolated from calcified human tissues, including blood vessels and kidney stones, and are referred to as nanobacteria. However, the microbiologic nature of putative nanobacteria continues to be debated, in part because of the difficulty in discriminating biomineralized microbes from minerals nucleated on anything else (eg, macromolecules, cell membranes). To address this controversy, the use of techniques capable of characterizing the organic and mineral content of these self-replicated structures at the submicrometer scale would be beneficial. Calcifying gram-negative bacteria (Caulobacter crescentus, Ramlibacter tataouinensis) used as references and self-replicating calcified nanoparticles cultured from human samples of calcified aneurysms were examined using a scanning transmission x-ray microscope (STXM) at the Advanced Light Source at Lawrence Berkeley National Laboratory. This microscope uses a monochromated and focused synchrotron x-ray beam (80-2,200 eV) to yield microscopic and spectroscopic information on both organic compounds and minerals at the 25 nm scale. High-spatial and energy resolution near-edge x-ray absorption fine structure (NEXAFS) spectra indicative of elemental speciation acquired at the C K-edge, N K-edge, and Ca L(2,3)-edge on a single-cell scale from calcified C. crescentus and R. tataouinensis displayed unique spectral signatures different from that of nonbiologic hydroxyapatite (Ca(10)(PO(4))(6)(OH)(2)). Further, preliminary NEXAFS measurements of calcium, carbon, and nitrogen functional groups of cultured calcified nanoparticles from humans revealed evidence of organics, likely peptides or proteins, specifically associated with hydroxyapatite minerals. Using NEXAFS at the 25 nm spatial scale, it is possible to define a biochemical signature for cultured calcified bacteria, including proteins, polysaccharides, nucleic acids, and hydroxyapatite. These preliminary studies suggest that nanoparticles isolated from human samples share spectroscopic characteristics with calcified proteins.


Artemisinins target the SERCA of Plasmodium falciparum.
Artemisinins are extracted from sweet wormwood (Artemisia annua) and are the most potent antimalarials available, rapidly killing all asexual stages of Plasmodium falciparum. Artemisinins are sesquiterpene lactones widely used to treat multidrug-resistant malaria, a disease that annually claims 1 million lives. Despite extensive clinical and laboratory experience their molecular target is not yet identified. Activated artemisinins form adducts with a variety of biological macromolecules, including haem, translationally controlled tumour protein (TCTP) and other higher-molecular-weight proteins. Here we show that artemisinins, but not quinine or chloroquine, inhibit the SERCA orthologue (PfATP6) of Plasmodium falciparum in Xenopus oocytes with similar potency to thapsigargin (another sesquiterpene lactone and highly specific SERCA inhibitor). As predicted, thapsigargin also antagonizes the parasiticidal activity of artemisinin. Desoxyartemisinin lacks an endoperoxide bridge and is ineffective both as an inhibitor of PfATP6 and as an antimalarial. Chelation of iron by desferrioxamine abrogates the antiparasitic activity of artemisinins and correspondingly attenuates inhibition of PfATP6. Imaging of parasites with BODIPY-thapsigargin labels the cytosolic compartment and is competed by artemisinin. Fluorescent artemisinin labels parasites similarly and irreversibly in an Fe2+-dependent manner. These data provide compelling evidence that artemisinins act by inhibiting PfATP6 outside the food vacuole after activation by iron.


If at first you don't succeed: Assessing influences associated with mothers' reoffering of vegetables to preschool age children.
Repeatedly offering vegetables has been shown to be one of the most effective methods for increasing acceptance and subsequent intake in young children. In order to increase successful offerings of vegetables and resultant consumption amongst young children, it is necessary to consider the influences on maternal reoffering of vegetables. This study aimed to investigate the relationships between mothers' tendency to reoffer vegetables and a range of demographic factors and psychological variables. A cross-sectional design was used, where mothers completed questionnaires assessing how often they reoffer rejected vegetables, concerns for economic factors, and a range of possible child and maternal influences. Mothers of preschool children were recruited from toddler groups across Leicestershire, UK, as well as online. Spearman's correlations were run to look for associations between demographic and psychological factors with maternal reoffering of vegetables. Significantly associated factors were then entered into a stepwise regression to predict maternal reoffering of vegetables. Mothers were significantly less likely to reoffer rejected vegetables if they were concerned about time, money, and waste, were influenced by their child's mood, or were concerned about their child having tantrums. Moreover, mothers who consumed more vegetables themselves reoffered vegetables more frequently. Regression analyses revealed that mothers' concern about food waste and tantrums, as well as maternal vegetable consumption, all significantly predicted mothers' reoffering of vegetables. With these findings in mind, mothers should be educated and supported with how to tackle and minimise children's tantrums during feeding, as well as being made aware of effective methods for avoiding food waste. Moreover, given that mothers' own vegetable consumption is associated with lower reoffering of vegetables to their child, interventions which seek to increase familial vegetable consumption should be pursued.


In vitro cytotoxicity of different desensitizers under simulated pulpal flow conditions.
To evaluate the cytotoxicity of three desensitizers, one nonrinse, and one etch-and-rinse adhesive system applied on dentin specimens of different thickness. The test materials (A: Admira Protect, B: Gluma Desensitizer, C: Seal&Protect, D: Clearfil Protect Bond, E: Optibond FL) and a positive control (35% H2O2) were applied on 1.0-, 1.5-, and 2.5-mm-thick bovine dentin specimens (each subgroup n = 5) in a dentin barrier test device. The experiments were performed with perfusion (2 ml/h) of the pulpal part of the chamber. The eluates were obtained before (baseline) and 15, 30, 45, 60, and 120 min after application of the adhesives and pipetted onto L-929 fibroblasts. Cytotoxicity of the materials was determined in relation to the baseline value using the MTT assay (succinic dehydrogenase activity). Statistical analysis was performed using ANOVA and Student's t-test. With regard to 1.0-mm dentin specimens, application of Clearfil Protect Bond (D) decreased enzyme activity significantly, while test materials A to C and E were not cytotoxic. However, cytotoxicity of D was limited to up to 15 min and decreased thereafter. Application of the test materials A to E on 1.5- and 2.5-mm dentin samples exhibited no significant cytotoxic effects within 120 min. Generally, ANOVA found significant interactions between the test materials and dentin thickness. However, only for Admira Protect was a significant increase of enzyme activity with increasing dentin thickness observed. Desensitizing agents might exhibit cytotoxic potential when applied on dentin less than 1.0 mm thick. The tested desensitizers and the adhesive systems caused similar effects, in which cytotoxicty might be influenced by the duration of perfusion and dentin thickness.


Inflammation and Nutritional Science for Programs/Policies and Interpretation of Research Evidence (INSPIRE).
An increasing recognition has emerged of the complexities of the global health agenda—specifically, the collision of infections and noncommunicable diseases and the dual burden of over- and undernutrition. Of particular practical concern are both 1) the need for a better understanding of the bidirectional relations between nutritional status and the development and function of the immune and inflammatory response and 2) the specific impact of the inflammatory response on the selection, use, and interpretation of nutrient biomarkers. The goal of the Inflammation and Nutritional Science for Programs/Policies and Interpretation of Research Evidence (INSPIRE) is to provide guidance for those users represented by the global food and nutrition enterprise. These include researchers (bench and clinical), clinicians providing care/treatment, those developing and evaluating programs/interventions at scale, and those responsible for generating evidence-based policy. The INSPIRE process included convening 5 thematic working groups (WGs) charged with developing summary reports around the following issues: 1) basic overview of the interactions between nutrition, immune function, and the inflammatory response; 2) examination of the evidence regarding the impact of nutrition on immune function and inflammation; 3) evaluation of the impact of inflammation and clinical conditions (acute and chronic) on nutrition; 4) examination of existing and potential new approaches to account for the impact of inflammation on biomarker interpretation and use; and 5) the presentation of new approaches to the study of these relations. Each WG was tasked with synthesizing a summary of the evidence for each of these topics and delineating the remaining gaps in our knowledge. This review consists of a summary of the INSPIRE workshop and the WG deliberations.


"Oh, I'm Not Alone": Experiences of HIV-Positive Men Who Have Sex With Men in a Health Navigation Program to Promote Timely Linkage to Care in Guatemala City.
Men who have sex with men (MSM) are disproportionately affected by HIV in Guatemala and may benefit from health navigation programs designed to support timely linkage to HIV care. We conducted qualitative in-depth interviews with MSM (n = 19) linked to care within days of their diagnosis with the support of a health navigator. We used narrative analysis and systematic coding to identify themes related to HIV diagnosis and experiences with navigators. Participants experienced strong feelings of fear upon receiving a positive HIV test result. This fear led to social isolation and limited reliance on family and friend networks for support. Health navigators developed strong relationships with participants by providing the emotional support they were lacking and supporting them to overcome their fear and other structural barriers to HIV care, including stigma. Findings support the important role of navigation for MSM in Guatemala and could be transferable to other settings in Latin America.


Serotonin, Amygdala and Fear: Assembling the Puzzle.
The fear circuitry orchestrates defense mechanisms in response to environmental threats. This circuitry is evolutionarily crucial for survival, but its dysregulation is thought to play a major role in the pathophysiology of psychiatric conditions in humans. The amygdala is a key player in the processing of fear. This brain area is prominently modulated by the neurotransmitter serotonin (5-hydroxytryptamine, 5-HT). The 5-HT input to the amygdala has drawn particular interest because genetic and pharmacological alterations of the 5-HT transporter (5-HTT) affect amygdala activation in response to emotional stimuli. Nonetheless, the impact of 5-HT on fear processing remains poorly understood.The aim of this review is to elucidate the physiological role of 5-HT in fear learning via its action on the neuronal circuits of the amygdala. Since 5-HT release increases in the basolateral amygdala (BLA) during both fear memory acquisition and expression, we examine whether and how 5-HT neurons encode aversive stimuli and aversive cues. Next, we describe pharmacological and genetic alterations of 5-HT neurotransmission that, in both rodents and humans, lead to altered fear learning. To explore the mechanisms through which 5-HT could modulate conditioned fear, we focus on the rodent BLA. We propose that a circuit-based approach taking into account the localization of specific 5-HT receptors on neurochemically-defined neurons in the BLA may be essential to decipher the role of 5-HT in emotional behavior. In keeping with a 5-HT control of fear learning, we review electrophysiological data suggesting that 5-HT regulates synaptic plasticity, spike synchrony and theta oscillations in the BLA via actions on different subcellular compartments of principal neurons and distinct GABAergic interneuron populations. Finally, we discuss how recently developed optogenetic tools combined with electrophysiological recordings and behavior could progress the knowledge of the mechanisms underlying 5-HT modulation of fear learning via action on amygdala circuits. Such advancement could pave the way for a deeper understanding of 5-HT in emotional behavior in both health and disease.


Adult obesity, disease and longevity in Mexico.
To obtain estimates of the effects of overweight and obesity on the incidence of type 2 diabetes (T2D) and adult mortality. We use three waves (2000, 2002, 2012) of the Mexican Health and Aging Survey (MHAS).We employ parametric hazard models to estimate mortality and conventional logistic models to estimate incidence of T2D. Obesity and overweight have a strong effect on the incidence of T2D;this, combined with the large impact of diabetes on adult mortality, generates increases in mortality that translate into losses of 2 to 3 years of life expectancy at age 50. If increasing trends in obesity in Mexico continue as in the past, progress in adult survival may be slowed down considerably and the incidence of T2D will continue to increase.


Nucleophosmin suppresses oncogene-induced apoptosis and senescence and enhances oncogenic cooperation in cells with genomic instability.
Cells from patients with genomic instability syndromes have high predisposition to cancer. However, little is known about whether these mutant cells have high susceptibility to oncogenic transformation. We have tested the hypothesis that a defect in maintaining genome integrity is necessary but not sufficient alone for oncogenic transformation and needs to collaborate with other signals in order to produce full oncogenic transformation. Using genetically matched primary cells deficient for the Fanconi complementation group C gene (Fancc) and the ataxia telangiectasia mutated gene (Atm), we found that certain forms of oncogenic activation and cooperation require a combination of genomic instability with increased expression of nucleophosmin (NPM) to prevent oncogenic stress-induced apoptosis or senescence. Intriguingly, co-expression of c-Myc and NPM leads to a synergistic increase in the proliferation rate in Fancc-/- or Atm-/- cells. Analysis of p53 stabilization and activation by c-Myc demonstrates that over-expression of NPM significantly reduces the accumulation of the activated p53 but not the stability of p53. Moreover, NPM is shown to enhance transforming activity of co-expressed Myc and Ras in wild-type and, to a greater degree, in Fancc-/- or Atm-/- cells, suggesting a role in oncogenic cooperation. Finally, a partial knockdown of NPM is sufficient to cause massive apoptosis in Fancc-/- or Atm-/- cells co-expressing c-Myc and Ras while sparing untransformed cells. Our study demonstrates a novel mechanism of NPM tumorigenesis by establishing NPM as a crucial inhibitor of oncogene-induced apoptosis and senescence.


Removal of Silicone Oil From Intraocular Lens Using Novel Surgical Materials.
To design, fabricate, and evaluate novel materials to remove silicone oil (SiO) droplets from intraocular lenses (IOL) during vitreoretinal surgery. Three different designs were fabricated using soft lithography of polydimethylsiloxane (PDMS), three-dimensional (3D) inverse PDMS fabrication using water dissolvable particles, and atomic layer deposition (ALD) of alumina (Al2O3) on surgical cellulose fibers. Laboratory tests included static and dynamic contact angle (CA) measurements with water and SiO, nondestructive x-ray microcomputer tomography (micro-CT), and microscopy. SiO removal was performed in vitro and ex vivo using implantable IOLs and explanted porcine eyes. All designs exhibited enhanced hydrophobicity and oleophilicity. Static CA measurements with water ranged from 131° to 160° and with SiO CA approximately 0° in 120 seconds following exposure. Nondestructive x-ray analysis of the 3D PDMS showed presence of interconnected polydispersed porosity of 100 to 300 μm in diameter. SiO removal from IOLs was achieved in vitro and ex vivo using standard 20-G vitrectomy instrumentation. Removal of SiO from IOLs can be achieved using materials with lower surface energy than that of the IOLs. This can be achieved using appropriate surface chemistry and surface topography. Three designs, with enhanced hydrophobic properties, were fabricated and tested in vitro and ex vivo. All materials remove SiO within an aqueous environment. Preliminary ex vivo results were very promising, opening new possibilities for SiO removal in vitreoretinal surgeries. This is the first report of an instrument that can lead to successful removal of SiO from the surface of IOL. In addition to the use of this instrument/material in medicine it can also be used in the industry, for example, retrieval of oil spills from bodies of water.


Erythema nodosum induced by kerion celsi of the scalp in a child: a case report and mini-review of literature.
A 9-year-old girl, presented with a 4-week history of an inflammatory suppurative plaque, 8 cm in diameter, localised in the occipital area of the scalp. Mycological direct examination showed ectoendothrix invasion of the hair and Trichophyton mentagrophytes was isolated. Oral therapy with griseofulvin 25 mg kg(-1) day(-1) was prescribed, but after 2 weeks of treatment appeared multiple erythematous subcutaneous nodules localised in the legs. Erythema nodosum (EN) was confirmed by histological examination of a nodule and then we combined therapy with 1 mg Kg(-1) day(-1) of prednisone. At the remission of the panniculitis, which occurred in about 10 days, the steroid therapy was suspended, while the orally administered griseofulvin continued for 6 weeks until full recovery. EN is the most frequent clinical form of acute nodular panniculitis and it is considered an epiphenomenon relative to various infectious and non-infectious stimuli. The association of EN with dermatophytosis of the scalp is infrequent, with only 15 cases reported in the Literature.


Secretagogin is a Ca2+-binding protein specifying subpopulations of telencephalic neurons.
The Ca(2+)-binding proteins (CBPs) parvalbumin, calbindin, and calretinin are phenotypic markers of terminally differentiated neurons in the adult brain. Although subtle phylogenetic variations in the neuronal distribution of these CBPs may occur, morphologically and functionally diverse subclasses of interneurons harbor these proteins in olfactory and corticolimbic areas. Secretagogin (scgn) is a recently cloned CBP from pancreatic beta and neuroendocrine cells. We hypothesized that scgn is expressed in the mammalian brain. We find that scgn is a marker of neuroblasts commuting in the rostral migratory stream. Terminally differentiated neurons in the olfactory bulb retain scgn expression, with scgn being present in periglomerular cells and granular layer interneurons. In the corticolimbic system, scgn identifies granule cells distributed along the dentate gyrus, indusium griseum, and anterior hippocampal continuation emphasizing the shared developmental origins, and cytoarchitectural and functional similarities of these neurons. We also uncover unexpected phylogenetic differences in scgn expression, since this CBP is restricted to primate cholinergic basal forebrain neurons. Overall, we characterize scgn as a neuron-specific CBP whose distribution identifies neuronal subtypes and hierarchical organizing principles in the mammalian brain.


The Effects of Piper Guineense versus Sesamum Indicum Aqueous Extracts on Lipid Metabolism and Antioxidants in Hypercholesterolemic Rats.
Piper guineense (PG) and Sesamum indicum (SI) have been shown to be rich sources of antioxidants and other health benefits; hence, we evaluated the impact of its consumption in hypercholesterolemic model on lipid metabolism. Forty-eight animals were divided into eight groups of six rats each. Rats were given cholesterol (40 mg/0.3ml), PG and SI extract (100 and 200 mg/kg), and Questran (0.26 g/kg) orally, five times a week for 28 days. Lipid profile, hepatic antioxidant status, biomarkers of liver toxicity, and tissue histopathology were examined. Data were analyzed using one-way ANOVA and P<0.05 were considered statistically significant. Cholesterol feeding caused 100% gain in weight, significantly increased AST, LPO (P=0.41 and 0.002) but significantly decreased SOD (P=0.003) compared to control. CHPG(1)/(2) and CHSI(1)/(2) caused a significant decrease (P=0.01, 0.005, 0.003, and 0.023) in cholesterol-induced body-weight gain and decreased serum total cholesterol by 20-30% compared to untreated-hypercholesterolemic rats. Triglyceride and LDL-c decreased with extract administration and specifically HDL-c increased significantly (P<0.001) by CHSI(1) compared to untreated-hypercholesterol rats. Furthermore, an increase in HDL-c was higher (P=0.04 and 0.002) by SI compared to PG at both doses. These results indicate that PG and SI exerts a hypolipidemic effect, reduces cholesterol intake induced body weight gain, and increases the body's antioxidant defense system in experimental hypercholesterolemia.


Melamine contamination in nutritional supplements--Is it an alarm bell for the general consumer, athletes, and 'Weekend Warriors'?
Nutritional supplements are used or experimented with by consumers, notably these are; competitive and recreational athletes of all ages, and 'weekend warriors'. As a consequence the supplement industry has grown to meet the increasing demand. A Global Industry Analysts Inc. report indicates that the herbal supplement market has not declined during the worldwide recession, but in fact exhibited steady growth over the period 2008 to 2009. It is anticipated that the market will reach US$93.15 billion by the year 2015. These supplements may contain adulterated substances that may potentially have harmful short - and long-term health consequences to the consumer. "Scrap Melamine" is such an example, which has been implicated in the kidney failure and death of several cats, dogs and pigs. In China in 2008, reports described very severe health effects in infants and young children. At the time over 294,000 infants were screened and diagnosed with urinary tract stones and sand-like calculi associated with melamine in milk products, of which 50,000 infants were hospitalised, and at least six associated deaths, recorded. The extent that melamine contamination occurs in nutritional supplements is not known. Therefore, the aim of this study was to determine whether commercially available nutritional and traditional supplement products contain melamine, even though they are not declared by the manufacturer on the product label. A total of 138 nutritional supplements products were obtained from (i) direct purchases from shops, pharmacies and outlets, (ii) directly from consumers, and (iii) from suppliers, manufacturers and distributors. The products were laboratory analysed for melamine, using Tandem Liquid Chromatography Mass Spectrometry. Forty-seven % of all the products (n = 138) tested positive for melamine. Eight-two % of the South African produced products (n = 27) tested positive and 58 % of the products imported into South Africa (n = 50) tested positive. The median concentration estimate for melamine in the products tested were, 6.0 μg/g for the 138 supplements tested, 8.9 μg/g for South African produced products, and 6.9 μg/g for products imported into South Africa. The melamine (undeclared on product label) levels detected in the nutritional supplements products investigated were within the Tolerable Daily intake (TDI) limit guidelines of 200 μg/g as set by WHO and others. Melamine over exposure within the context of the nutritional supplements consumption in the products investigated should not be of concern to the consumer provided the recommended guidelines of daily product use are adhered to. Further investigation is warranted to determine, (i) the link of melamine as (part) substitute for the perceived total declared protein content on the product label, (ii) cyanuric and uric acid presence in the supplement products that could form chemical-complex formation with melamine and/or analogues that could cause adverse health effects.


The ADDITION-Cambridge trial protocol: a cluster -- randomised controlled trial of screening for type 2 diabetes and intensive treatment for screen-detected patients.
The increasing prevalence of type 2 diabetes poses a major public health challenge. Population-based screening and early treatment for type 2 diabetes could reduce this growing burden. However, the benefits of such a strategy remain uncertain. The ADDITION-Cambridge study aims to evaluate the effectiveness and cost-effectiveness of (i) a stepwise screening strategy for type 2 diabetes; and (ii) intensive multifactorial treatment for people with screen-detected diabetes in primary care. 63 practices in the East Anglia region participated. Three undertook the pilot study, 33 were allocated to three groups: no screening (control), screening followed by intensive treatment (IT) and screening plus routine care (RC) in an unbalanced (1:3:3) randomisation. The remaining 27 practices were randomly allocated to IT and RC. A risk score incorporating routine practice data was used to identify people aged 40-69 years at high-risk of undiagnosed diabetes. In the screening practices, high-risk individuals were invited to take part in a stepwise screening programme. In the IT group, diabetes treatment is optimised through guidelines, target-led multifactorial treatment, audit, feedback, and academic detailing for practice teams, alongside provision of educational materials for newly diagnosed participants. Primary endpoints are modelled cardiovascular risk at one year, and cardiovascular mortality and morbidity at five years after diagnosis of diabetes. Secondary endpoints include all-cause mortality, development of renal and visual impairment, peripheral neuropathy, health service costs, self-reported quality of life, functional status and health utility. Impact of the screening programme at the population level is also assessed through measures of mortality, cardiovascular morbidity, health status and health service use among high-risk individuals. ADDITION-Cambridge is conducted in a defined high-risk group accessible through primary care. It addresses the feasibility of population-based screening for diabetes, as well as the benefits and costs of screening and intensive multifactorial treatment early in the disease trajectory. The intensive treatment algorithm is based on evidence from studies including individuals with clinically diagnosed diabetes and the education materials are informed by psychological theory. ADDITION-Cambridge will provide timely evidence concerning the benefits of early intensive treatment and will inform policy decisions concerning screening for type 2 diabetes. Current Controlled trials ISRCTN86769081.


Developing a Process for Returning Medically Actionable Genomic Variants to Latino Patients in a Federally Qualified Health Center.
To develop a process for returning medically actionable genomic variants to Latino patients receiving care in a Federally Qualified Health Center. Prior to recruitment, researchers met with primary care providers to (1) orient clinicians to the project, (2) establish a process for returning actionable and nonactionable results to participants and providers through the electronic health record, and (3) develop a process for offering clinical decision support for follow-up education and care. A Community Advisory Board was engaged to provide input on recruitment strategies and materials for conveying results to participants. Participants in the Sangre Por Salud (Blood for Health) Biobank with hyperlipidemia or colon polyps represented the pool of potentially eligible participants. A total of 1,621 individuals were invited to participate and 710 agreed to an in- person consenting visit (194 no-showed and 16 declined). Over 12-months, 500 participants were enrolled. Participants were primarily Spanish-speaking (81.6%), female (74.2%), and enrolled because of hyperlipidemia (95.4%). All but 2 participants opted to receive primary (i.e., related to enrollment phenotypes) as well as secondary actionable results. Efforts to bring precision medicine to community-based health centers serving minority patients may require multilevel engagement activities to include individuals, providers, health systems, and the community.


Regulation of fluid intake in dehydrated humans: role of oropharyngeal stimulation.
We examined the effect of oropharyngeal stimulation on thirst, secretion of arginine vasopressin ([AVP]p), and fluid intake in six healthy adults after dehydration (28.6 +/- 1.4 ml/kg water loss) induced by mild exercise in the heat (2 h, 38 degrees C, relative humidity < 30%). Subjects performed three identical dehydration protocols followed by 75 min of rehydration at 27 degrees C consisting of 1) ad libitum drinking (Con), 2) infusion of a similar volume of water directly into the stomach via a nasogastric tube (Inf) during the first 25 min followed by combined Inf and ad libitum drinking during the remaining 50 min of rehydration; or 3) ad libitum drinking with simultaneous extraction of ingested fluid via a nasogastric tube (Ext). Plasma osmolality (Posm), [AVP]p, fluid intake, and thirst perceptions were measured throughout. On average, for all three protocols, Posm increased 7.8 +/- 0.6 mosmol/kgH2O and plasma volume decreased 4.7 +/- 1.3%, whereas thirst ratings and [AVP]p increased 7.6 +/- 1.3 cm and 3.1 +/- 0.4 pg/ml, respectively. Reflex inhibition of [AVP]p and thirst occurred within 5 min of rehydration in Con and Ext (P < 0.05) but not during Inf, supporting the hypothesis that oropharyngeal reflexes modulate osmotically stimulated thirst and [AVP]p. However, the reduction in [AVP]p during the first 5 min of Ext (-1.1 +/- 0.3 pg/ml) was less than that seen during Con (-2.1 +/- 0.4 pg/ml), suggesting that oropharyngeal stimulation is not the only factor contributing to the rapid reduction in [AVP]p during the first 5 min of drinking. During Con, subjects ingested 20.0 +/- 2.0 ml/kg of water but only drank 15% more (31.3 +/- 7.1 ml/kg) during Ext, demonstrating a clear role of oropharyngeal metering in limiting total fluid intake in humans in the presence of a persistently high dipsogenic drive.


Xanthomonas citri: breaking the surface.
SUMMARY Taxonomy: Bacteria; Proteobacteria, Gammaproteobacteria; Xanthomonadales; Xanthomonadaceae, Xanthomonas. Microbiological properties: Gram-negative, obligately aerobic, straight rods, motile by a single polar flagellum, yellow pigment. Related species: X. campestris, X. axonopodis, X. oryzae, X. albilineans. Affects Rutaceous plants, primarily Citrus spp., Fortunella spp., and Poncirus spp., world-wide. Quarantined pathogen in many countries. Economically important hosts are cultivated orange, grapefruit, lime, lemon, pomelo and citrus rootstock. Disease symptoms: On leaves, first appearance is as oily looking, 2-10 mm, similarly sized, circular spots, usually on the abaxial surface. On leaves, stems, thorns and fruit, circular lesions become raised and blister-like, growing into white or yellow spongy pustules. These pustules then darken and thicken into a light tan to brown corky canker, which is rough to the touch. On stems, pustules may coalesce to split the epidermis along the stem length, and occasionally girdling of young stems may occur. Older lesions on leaves and fruit tend to have more elevated margins and are at times surrounded by a yellow chlorotic halo (that may disappear) and a sunken centre. Sunken craters are especially noticeable on fruit, but the lesions do not penetrate far into the rind. Defoliation and premature abscission of affected fruit occurs on heavily infected trees. <http://www.biotech.ufl.edu/PlantContainment/canker.htm>; <http://cancer.lbi.ic.unicamp.br/xanthomonas/>


Solitarylike Raman states in massive wavelength-division-multiplexed systems.
We show that solitarylike stimulated Raman excitations are possible in massive wavelength-division-multiplexed fiber-optic systems. These self-similar states can propagate across channels at a constant rate that depends on the total power launched in the system and the wave's spectral width. The evolution of these excitations under the action of noise and during collisions is investigated in detail.


The phylogenetic utility and functional constraint of microRNA flanking sequences.
MicroRNAs (miRNAs) have recently risen to prominence as novel factors responsible for post-transcriptional regulation of gene expression. miRNA genes have been posited as highly conserved in the clades in which they exist. Consequently, miRNAs have been used as rare genome change characters to estimate phylogeny by tracking their gain and loss. However, their short length (21-23 bp) has limited their perceived utility in sequenced-based phylogenetic inference. Here, using reference taxa with established phylogenetic relationships, we demonstrate that miRNA sequences are of high utility in quantitative, rather than in qualitative, phylogenetic analysis. The clear orthology among miRNA genes from different species makes it straightforward to identify and align these sequences from even fragmentary datasets. We also identify significant sequence conservation in the regions directly flanking miRNA genes, and show that this too is of utility in phylogenetic analysis, as well as highlighting conserved regions that will be of interest to other fields. Employing miRNA sequences from 12 sequenced drosophilid genomes, together with a Tribolium castaneum outgroup, we demonstrate that this approach is robust using Bayesian and maximum-likelihood methods. The utility of these characters is further demonstrated in the rhabditid nematodes and primates. As next-generation sequencing makes it more cost-effective to sequence genomes and small RNA libraries, this methodology provides an alternative data source for phylogenetic analysis. The approach allows rapid resolution of relationships between both closely related and rapidly evolving species, and provides an additional tool for investigation of relationships within the tree of life.


A novel achiral seco-amino-cyclopropylindoline (CI) analog of CC-1065 and the duocarmycins: design, synthesis and biological studies.
The design, synthesis and DNA binding properties of a novel achiral and amino-containing seco-cyclopropylindoline analog (seco-amino-CI-TMI, 1) of the duocarmycins are described. Thermal induced DNA cleavage studies on pUC18 DNA revealed compound 1 to preferentially bind in the minor groove and to covalently react with AT-rich sequences, particularly at the underlined adenine-N3 group of 5'-AAAAA(865)-3'. This sequence specificity is similar to adozelesin and CC-1065. Using a 4-day continuous exposure, compound 1 inhibited the growth of K562 human chronic myeloid leukemia cells in culture. Compound 1 has appreciable cytotoxicity (IC50 value of 1.30 microM) relative to compound 2 (0.15 microM), the corresponding racemic and hydroxy-seco-CI-TMI analog. These results indicate that the aminophenethyl chloride group present in compound 1 has similar sequence specific and cytotoxic properties to the hydroxy-containing seco-precursors of CC-1065 and the duocarmycins. Moreover, the results suggest that the chiral center present in the natural products is not absolutely necessary for biological activity. The novel aminophenethyl halide moiety is, therefore, a useful template from which to develop future achiral analogs of CC-1065 and the duocarmycins.


Childhood Illness Prevalence and Health Seeking Behavior Patterns in Rural Tanzania.
This paper identifies factors influencing differences in the prevalence of diarrhea, fever and acute respiratory infection (ARI), and health seeking behavior among caregivers of children under age five in rural Tanzania. Using cross-sectional survey data collected in Kilombero, Ulanga, and Rufiji districts, the analysis included 1,643 caregivers who lived with 2,077 children under five years old. Logistic multivariate and multinomial regressions were used to analyze factors related to disease prevalence and to health seeking behavior. One quarter of the children had experienced fever in the past two weeks, 12.0 % had diarrhea and 6.7 % experienced ARI. Children two years of age and older were less likely to experience morbidity than children under one year [ORfever = 0.77, 95 % CI 0.61-0.96; ORdiarrhea = 0.26, 95 % CI 0.18-0.37; ORARI = 0.60 95 % CI 0.41-0.89]. Children aged two and older were more likely than children under one to receive no care or to receive care at home, rather than to receive care at a facility [RRRdiarrhea = 3.47, 95 % CI 1.19-10.17 for "No care"]. Children living with an educated caregiver were less likely to receive no care or home care rather than care at a facility as compared to those who lived with an uneducated caregiver [RRRdiarrhea = 0.28, 95 % CI 1.10-0.79 for "No care"]. Children living in the wealthiest households were less likely to receive no care or home care for fever as compared to those who lived poorest households. Children living more than 1 km from health facility were more likely to receive no care or to receive home care for diarrhea rather than care at a facility as compared to those living less than 1 km from a facility [RRRdiarrhea = 3.50, 95 % CI 1.13-10.82 for "No care"]. Finally, caregivers who lived with more than one child under age five were more likely to provide no care or home care rather than to seek treatment at a facility as compared to those living with only one child under five. Our results suggest that child age, caregiver education attainment, and household wealth and location may be associated with childhood illness and care seeking behavior patterns. Interventions should be explored that target children and caregivers according to these factors, thereby better addressing barriers and optimizing health outcomes especially for children at risk of dying before the age of five.


Amelioration of Coagulation Disorders and Inflammation by Hydrogen-Rich Solution Reduces Intestinal Ischemia/Reperfusion Injury in Rats through NF-κB/NLRP3 Pathway.
Intestinal ischemia/reperfusion (I/R) injury often causes inflammatory responses and coagulation disorders, which is further promoting the deterioration of the disease. Hydrogen has anti-inflammatory, antioxidative, and antiapoptotic properties against various diseases. However, the effect of hydrogen on coagulation dysfunction after intestinal I/R and the underlying mechanism remains unclear. The purpose of this study was to explore whether hydrogen-rich solution (HRS) could attenuate coagulation disorders and inflammation to improve intestinal injury and poor survival following intestinal I/R. The rat model of intestinal I/R injury was established by clamping the superior mesenteric artery for 90 min and reperfusion for 2 h. HRS (10 or 20 mL/kg) or 20 mL/kg 0.9% normal saline was intravenously injected at 10 min before reperfusion, respectively. The samples were harvested at 2 h after reperfusion for further analyses. Moreover, the survival rate was observed for 24 h. The results showed that HRS improved the survival rate and alleviated serum diamine oxidase activities, intestinal injury, edema, and apoptosis. Interestingly, HRS markedly improved intestinal I/R-mediated coagulation disorders as evidenced by abnormal conventional indicators of coagulation and thromboelastography. Additionally, HRS attenuated inflammatory responses and the elevated tissue factor (TF) and inhibited nuclear factor kappa beta (NF-κB) and nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation in peripheral blood mononuclear cells. Moreover, inflammatory factors and myeloperoxidase were closely associated with TF level. This study thus emphasized upon the amelioration of coagulation disorders and inflammation by HRS as a mechanism to improve intestinal I/R-induced intestinal injury and poor survival, which might be partially related to inhibition of NF-κB/NLRP3 pathway.


Improving the management of post-operative acute pain: priorities for change.
Poor management of post-operative acute pain can contribute to medical complications including pneumonia, deep vein thrombosis, infection and delayed healing, as well as the development of chronic pain. It is therefore important that all patients undergoing surgery should receive adequate pain management. However, evidence suggests this is not currently the case; between 10% and 50% of patients develop chronic pain after various common operations, and one recent US study recorded >80% of patients experiencing post-operative pain. At the first meeting of the acute chapter of the Change Pain Advisory Board, key priorities for improving post-operative pain management were identified in four different areas. Firstly, patients should be more involved in decisions regarding their own treatment, particularly when fateful alternatives are being considered. For this to be meaningful, relevant information should be provided so they are well informed about the various options available. Good physician/patient communication is also essential. Secondly, better professional education and training of the various members of the multidisciplinary pain management team would enhance their skills and knowledge, and thereby improve patient care. Thirdly, there is scope for optimizing treatment. Examples include the use of synergistic analgesia to target pain at different points along pain pathways, more widespread adoption of patient-controlled analgesia, and the use of minimally invasive rather than open surgery. Fourthly, organizational change could provide similar benefits; introducing acute pain services and increasing their availability towards the 24 hours/day ideal, greater adherence to protocols, increased use of patient-reported outcomes, and greater receptivity to technological advances would all help to enhance performance and increase patient satisfaction. It must be acknowledged that implementing these recommendations would incur a considerable cost that purchasers of healthcare may be unwilling or unable to finance. Nevertheless, change is under way and the political will exists for it to continue.


Phenyl Ether- and Aniline-Containing 2-Aminoquinolines as Potent and Selective Inhibitors of Neuronal Nitric Oxide Synthase.
Excess nitric oxide (NO) produced by neuronal nitric oxide synthase (nNOS) is implicated in neurodegenerative disorders. As a result, inhibition of nNOS and reduction of NO levels is desirable therapeutically, but many nNOS inhibitors are poorly bioavailable. Promising members of our previously reported 2-aminoquinoline class of nNOS inhibitors, although orally bioavailable and brain-penetrant, suffer from unfavorable off-target binding to other CNS receptors, and they resemble known promiscuous binders. Rearranged phenyl ether- and aniline-linked 2-aminoquinoline derivatives were therefore designed to (a) disrupt the promiscuous binding pharmacophore and diminish off-target interactions and (b) preserve potency, isoform selectivity, and cell permeability. A series of these compounds was synthesized and tested against purified nNOS, endothelial NOS (eNOS), and inducible NOS (iNOS) enzymes. One compound, 20, displayed high potency, selectivity, and good human nNOS inhibition, and retained some permeability in a Caco-2 assay. Most promisingly, CNS receptor counterscreening revealed that this rearranged scaffold significantly reduces off-target binding.


Association Between Usage of an App to Redeem Prescribed Food Benefits and Redemption Behaviors Among the Special Supplemental Nutrition Program for Women, Infants, and Children Participants: Cross-Sectional Study.
The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) is one of the most important food assistance programs in the United States, serving 6.4 million low-income, eligible women, infants, and children under 5 years of age in 2019. In the program, participants are prescribed a list of food benefits, which can be redeemed in WIC-authorized stores. However, there are multiple behavioral barriers in the program and the stores that prevent participants from redeeming the benefits fully. This study aims to examine the relationship between the use of a widely used mobile phone app, WICShopper, and the redemption of the prescribed food packages. WIC administrative data were obtained from West Virginia for the period January 2019 to January 2020 and included 30,440 WIC households that had received food benefits in that period. The redemption rates of 18 WIC food benefits were compared between app users and nonapp users, that is, those who never used the app in the study period. The use behaviors were defined for the app users, including the number of active use benefit cycles, active benefit cycle rates, number of active use days in the cycle, and proportion rates of daytime use. Panel linear regressions were applied to examine how the redemption rates were related to these behaviors over time. App users consistently had higher average redemption rates than nonapp users; the difference ranged from 3.6% (4.8% relative) for infant formula to 14.3% (40.7% relative) for fish. After controlling for sociodemographics, the coefficients of app use were significantly positive for all benefit categories except for WIC-eligible nutritionals. More active cycles and active days in the cycle were significantly related to redemption rates for all categories, except for frozen juice (coefficient=-0.002, P=.09). Daytime app access was positively associated with redemption rates for most food benefits except only a few, such as infant formula (coefficient=-0.03, P<.001). Use of the WIC app was significantly related to higher redemption rates across food benefits, although the association varied across benefit categories. More active days were positively related to benefit redemptions across food categories, and the app's daytime use was positively associated with the redemption of most benefit categories. These findings suggest that the WIC app can be an important tool for the promotion of benefit redemption among WIC participants.


Photocatalytic oxidation of methane over SrCO3 decorated SrTiO3 nanocatalysts via a synergistic effect.
Because of the high C-H bond energy as well as the non-polar feature of CH4 molecules, oxidation of methane under mild conditions remains a challenging task for both C1 utilization and atmospheric environmental cleansing. Here we report that by using a sol-gel method SrCO3 decorated SrTiO3 nanocatalysts (SrTiO3-S) with an average particle size of ∼25 nm can be readily prepared, which surprisingly show efficient performance for photocatalytic oxidation of methane with the activity close to fourfold of P25, a benchmark photocatalyst. Further investigation revealed a synergistic effect between SrCO3 and SrTiO3 when combined together into a composite material as both of which are totally inactive for methane oxidation if used alone. Gas adsorption characterization disclosed that the SrCO3 can adsorb methane and cannot adsorb carbon dioxide, whereas the SrTiO3 will preferentially adsorb CO2 instead of CH4. Photocurrent and photoluminescence measurements indicate that SrCO3 exhibits a negligible photocurrent response relative to the SrTiO3 semiconductor under simulated solar light illumination but the formation of the SrCO3/SrTiO3 junction structure (SrTiO3-S) helps reduce surface recombination of the photogenerated electrons and holes. All these results refer to the synergistic mechanism in which the SrCO3 acts as a trapping agent to adsorb methane and weaken its C-H bond while the SrTiO3 acts as a photocatalyst to activate and oxidize methane under light illumination. The underlying photooxidation mechanism is further investigated with the aid of in situ electron paramagnetic resonance and infrared spectroscopy.


Isolation and characterization of the plant immune-priming compounds Imprimatin B3 and -B4, potentiators of disease resistance in Arabidopsis thaliana.
Plant activators are chemical crop protectants that fortify the immune system in plants. Unlike pesticides that target pathogens, plant activators provide durable effects against a broad spectrum of diseases, which have not been overcome by pathogenic microbes. Plant activators are not only useful agrochemicals, but can also help to elucidate the details of the plant immune system. Using an established high-throughput screening procedure, we previously identified 5 compounds, designated as Imprimatins, which prime plant immune response. These compounds increased disease resistance against pathogenic Pseudomonas bacteria in Arabidopsis plants by inhibiting 2 salicylic acid (SA) glucosyltransferases (SAGTs), resulting in accumulation of the phytohormone SA. Here, we report the isolation of 2 additional Imprimatins, B3 and B4, which are structurally similar to Imprimatin B1 and B2. Because these compounds did not have strong inhibitory effects on SAGTs in vitro, they may exert their function after metabolic conversion in vivo.


Neuroprotective Effects of Etidronate and 2,3,3-Trisphosphonate Against Glutamate-Induced Toxicity in PC12 Cells.
Etidronate is one of the best known bisphosphonates (BP) derivatives. It is often used as a reference drug in research related to hypercalcaemia and other common bone diseases. 2,3,3-trisphosphonate (TrisPP) is brand new analogue of BP, that also contains a 'germinal bisphosphonate' unit with an additional phosphoryl group attached in proximity to the BP unit. It is known that BPs bind to calcium by chemisorptions to form Ca-BP complexes through (O)P-C-P(O) moiety and hydrogen coordinations, and so they suppress calcium flow by interfering with Ca(2+) channel operations. The mechanistic actions of BP, involving interactions and regulations of Ca(2+), are somewhat similar to the pathogenesis of well-known neurodegenerative disorders, such as Alzheimer's disease, Parkinson's disease and Huntington's disease. To investigate if neuroprotective effects are exhibited by the compounds of interests, we used a rat adrenal pheochromocytoma cell line (PC12) as our in vitro model to observe any occurrence of neuron inter-reflection. We pre-treated these PC12 cells with etidronate and TrisPP before challenging the cells with a high concentration of the neurotoxin, glutamate. Our data showed that pre-treatment with 100 μM etidronate partially ameliorated the glutamate-induced decrease in cell viability (47 %), whereas pre-treating cells with 10-100 μM TrisPP showed remarkable cell protection (78-86 %). Moreover, pre-treatments of the cells with etidronate or TrisPP attenuated cell apoptosis, reactive oxygen species generation, Ca(2+) overloading and caspase-3 protein expression, which were associated with a remarkable increase in superoxide dismutase activity in our glutamate-injured PC12 cells. Therefore, this study supports the notion that etidronate and TrisPP may be promising neuroprotective agents.


Criteria for the systematic review of health promotion and public health interventions.
Systematic reviews of public health interventions are fraught with challenges. Complexity is inherent; this may be due to multi-component interventions, diverse study populations, multiple outcomes measured, mixed study designs utilized and the effect of context on intervention design, implementation and effectiveness. For policy makers and practitioners to use systematic reviews to implement effective public health programmes, systematic reviews must include this information, which seeks to answer the questions posed by decision makers, including recipients of programmes. This necessitates expanding the traditional evaluation of evidence to incorporate the assessment of theory, integrity of interventions, context and sustainability of the interventions and outcomes. Unfortunately however, the critical information required for judging both the quality of a public health intervention and whether or not an intervention is worthwhile or replicable is missing from most public health intervention studies. When the raw material is not available in primary studies the systematic review process becomes even more challenging. Systematic reviews, which highlight these critical gaps, may act to encourage better reporting in primary studies. This paper provides recommendations to reviewers on the issues to address within a public health systematic review and, indirectly, provides advice to researchers on the reporting requirements of primary studies for the production of high quality systematic reviews.


Does obesity affect outcomes in hip arthroscopy? A matched-pair controlled study with minimum 2-year follow-up.
Hip arthroscopy has gained popularity over the past decade, and its indications have broadened as newer techniques have been developed. However, there has been a paucity of literature evaluating the outcomes of hip arthroscopy in obese patients. To compare 2-year clinical outcomes of obese patients undergoing primary hip arthroscopy with matched nonobese controls. Cohort study; Level of evidence, 3. From February 2008 to February 2012, data were collected prospectively on all obese patients undergoing primary hip arthroscopy. A matched-pair nonobese control group was selected at a 1:2 ratio. All patients were assessed pre- and postoperatively with 4 patient-reported outcome (PRO) measures: the modified Harris Hip Score, Non-Arthritic Hip Score, Hip Outcome Score-Activities of Daily Living, and Hip Outcome Score-Sport Specific Subscale. Pain was estimated on the visual analog scale, and satisfaction was measured on a scale from 0 to 10. Sixty-two hips (62 patients) were included in the obese group and 124 hips (124 patients) in the control group. At preoperative baseline, the obese group had significantly lower PRO scores when compared with the control group. Both groups demonstrated statistically significant postoperative improvement in all scores (P < .05). Absolute scores were significantly lower in the obese group for all PRO measures, pre- and postoperatively. However, the improvement (delta) in PRO scores from pre- to postoperative time was not significantly different between groups. The rate of conversion to total hip arthroplasty, the rate of revision, and the complication rate were not significantly different between the 2 groups; however, rates of conversion to total hip arthroplasty and revision tended to be twice as high in the obese patients, but the study was not powered for these 2 outcomes. Overall, obese patients had lower absolute PRO scores preoperatively and at 2-year follow-up. Both obese and nonobese patients demonstrated significant improvement in all PRO scores, and the change in scores were similar between groups. These results indicate that while obese patients may not have similar absolute scores after hip arthroscopy, they may show similar gains in improvement when compared with baseline. Hip arthroscopy appears to be a viable treatment option in the obese patient as long as expectations are adjusted accordingly.


Use of recombinant human bone morphogenetic protein for revision cervical spine fusion in children with Down syndrome: a case series.
Patients with trisomy 21 (Down syndrome; DS) often have atlantoaxial instability (AAI), which, if severe, causes myelopathy and neurological deterioration. Children with DS and AAI who undergo cervical spine fusion have a high rate of nonunion requiring revision surgery. Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a TGF-β growth factor that is used to induce bone formation in spine fusion. Although previous studies in the adult population have reported no reduction in pseudarthrosis rates with the use of rhBMP-2, there is a lack of literature in the pediatric DS population. This study describes the use of rhBMP-2 in children with DS and AAI during revision to treat nonunion. A retrospective review of a cervical spine fusion database (n = 175) was conducted. This database included all cervical spine fusions using modern instrumentation at the authors' institution from 2002 to 2019. Patients with DS who underwent a revision utilizing rhBMP-2 were included in the study. The number of prior fusions, use of rhBMP-2 in fusions, length of stay, halo use, and surgical data were collected. Postoperative complications and length of follow-up were also recorded. Eight patients (75% female) met the inclusion criteria. The average age at revision with rhBMP-2 was 11 years (range 3-19 years). All patients were diagnosed with nonunion after an initial cervical fusion. All revisions were posterior fusions of C1-2 (n = 2) or occiput to cervical (n = 6). All revisions included implant revisions, iliac crest bone grafting, and rhBMP-2 use. One patient required irrigation and debridement of an rhBMP-induced seroma. Another patient required return to the operating room to repair a dural tear. There were no neurological, infectious, airway, or implant-related complications. Revision utilizing rhBMP-2 achieved fusion in 100% (n = 8) of patients. The average length of follow-up was 42.6 months. All patients demonstrated solid fusion mass on the last radiograph. This is the first case series reporting the successful use of rhBMP-2 to facilitate cervical spine fusion in patients with DS after previous nonunion. In addition, few rhBMP-2-related postoperative complications occurred.


PopART-IBM, a highly efficient stochastic individual-based simulation model of generalised HIV epidemics developed in the context of the HPTN 071 (PopART) trial.
Mathematical models are powerful tools in HIV epidemiology, producing quantitative projections of key indicators such as HIV incidence and prevalence. In order to improve the accuracy of predictions, such models need to incorporate a number of behavioural and biological heterogeneities, especially those related to the sexual network within which HIV transmission occurs. An individual-based model, which explicitly models sexual partnerships, is thus often the most natural type of model to choose. In this paper we present PopART-IBM, a computationally efficient individual-based model capable of simulating 50 years of an HIV epidemic in a large, high-prevalence community in under a minute. We show how the model calibrates within a Bayesian inference framework to detailed age- and sex-stratified data from multiple sources on HIV prevalence, awareness of HIV status, ART status, and viral suppression for an HPTN 071 (PopART) study community in Zambia, and present future projections of HIV prevalence and incidence for this community in the absence of trial intervention.


A 12-year retrospective survey of management of patients with malignant neoplasms in the orbital cavity in a brazilian cancer hospital.
The aim of this study was to investigate the distribution of different types of malignant tumors in the orbital cavity, surgical treatments, and subsequent prosthetic rehabilitation. A retrospective sample of 269 charts of patients attending a public reference cancer hospital in Goiania, Goias, Brazil, between 1998 and 2009 was selected. Study variables included demographic data, clinical and histopathological features, and patient rehabilitation. Frequency analysis, chi-square test and multiple logistic regression were used for data analysis. Patients' age ranged from 1 to 92 years and 50.2% were female. The most prevalent tumors were basal cell carcinoma (58%), retinoblastoma (17.1%), squamous cell carcinoma of the conjunctiva (7.1%), malignant melanoma (6.7%), and rhabdomyosarcoma (4.5%). Surgical treatment was performed in 250 patients (92.3%), but only 29 patients (10.8%) received ocular prosthetic rehabilitation. Lower age (p<0.001) and enucleation surgery (p<0.001) were significantly associated with prosthetic rehabilitation. Results revealed high occurrence of mutilating interventions and low provision of prosthetic care, limited to younger patients who had undergone enucleation surgery. The high unmet demand for rehabilitation emphasizes the need to incorporate appropriate care for ocular and orbital deformities into the Brazilian public health system.


A Patient-Matched Entire First Metacarpal Prosthesis in Treatment of Giant Cell Tumor of Bone.
Giant cell tumor of the bones occurring in the first metacarpals frequently requires entire metacarpal resection due to the aggressive nature and high rate of recurrence. Bone reconstruction can be performed with autogenous bone grafts. Here we describe a new technique of reconstruction using a patient-matched three-dimensional printed titanium first metacarpal prosthesis. This prosthesis has a special design for ligament reconstruction in the proximal and distal portions. Good hand function and aesthetic appearance were maintained at a 24-month follow-up visit. This reconstructive technique can avoid donor-site complications and spare the autogenous bone grafts for revision options.


Vasculoprotective effect of U50,488H in rats exposed to chronic hypoxia: role of Akt-stimulated NO production.
Impairment of pulmonary endothelium function in the pulmonary artery is a direct result of chronic hypoxia. This study is to investigate the vasculoprotective effects of U50,488H (a selective κ-opioid receptor agonist) and its underlying mechanism in hypoxia-induced pulmonary artery endothelial functional injury. Chronic hypoxia was simulated by exposing the rats to 10% oxygen for 2 wk. After hypoxia, right ventricular pressure (RVP) and right ventricular hypertrophy index (RVHI) were measured. The pulmonary vascular dysfunction, effect of nitric oxide synthase inhibitor (l-NAME) on the relaxation of U50,488H, and level of nitric oxide (NO) were determined. In vitro, the signaling pathway involved in the anti-apoptotic effect of U50,488H was investigated. Cultured endothelial cells were subjected to simulated hypoxia, and cell apoptosis was determined by TUNEL staining. U50,488H (1.25 mg/kg) significantly reduced RVP and RVHI in hypoxia. U50,488H markedly improved both pulmonary endothelial function (maximal vasorelaxation in response to ACh: 74.9 ± 1.8%, n = 6, P <0.01 vs. hypoxia for 2 wk group) and increased total NO production (1.65 fold). U50,488H relaxed the pulmonary artery rings of the hypoxic rats. This effect was partly abolished by l-NAME. In cells, U50,488H both increased NO production and reduced hypoxia-induced apoptosis. Moreover, pretreatment with nor-binaltorphimine (nor-BNI, a selective κ-opioid receptor antagonist), PI3K inhibitor, Akt inhibitor or l-NAME almost abolished anti-apoptotic effect exerted by U50,488H. U50,488H resulted in increases in Akt and eNOS phosphorylation. These results demonstrate that pretreatment with U50,488H attenuates hypoxia-induced pulmonary vascular endothelial dysfunction in an Akt-dependent and NO-mediated fashion.


Differences in growth and survival between cod Gadus morhua and herring Clupea harengus early stages co-reared at variable prey concentrations.
It was hypothesized that the survival and growth strategies of herring Clupea harengus, displaying a flexible reproductive activity, are adapted to coping with longer periods of prey deprivation (i.e. more variable prey availability), in contrast to cod Gadus morhua, which are adapted to match growth and survival at high prey concentrations. Experimental larval growth and survival data for the two naturally co-occurring species reared either in separate tanks or in combination are presented to test this hypothesis. Natural zooplankton was supplied either ad libitum or in a periodically restricted manner to mimic natural suboptimal conditions. Periodically restricted feeding significantly reduced initial growth of G. morhua larvae co-reared with C. harengus, while no such initial effect was seen for co-reared C. harengus. Overall survival of G. morhua was higher when reared together with C. harengus (32 v. 24%), while C. harengus had higher survival without the presence of G. morhua (59 v. 44%), indicating that both species were affected by higher densities of G. morhua larvae. Furthermore, the final survival in G. morhua was inversely related to average final size, while in C. harengus an opposite trend was observed. How potential behavioural interactions may drive the present results are discussed and contended that a better insight into field vital rates may be gained from further exploration of co-rearing experiments.


Quality use of medicines within universal health coverage: challenges and opportunities.
Medicines are a major driver of quality, safety, equity, and cost of care in low and middle-income country health systems. Universal health coverage implementers must explicitly address appropriate use of medicines to realize the health benefits of medicines, avoid wasting scarce resources, and sustain the financial viability of universal health coverage schemes. Medicines are major contributors to the health and well-being of individuals and populations when used appropriately, and they waste resources and endanger health when used unnecessarily or incorrectly. Stakeholders need to balance inherently competing objectives in the pharmaceutical sector. Emerging and expanding UHC schemes provide potential levers to balance competing system objectives.To use these levers, sustainable universal coverage programs will require a) information systems that can track medicines utilization, expenditures, and quality of medicines use; b) routine monitoring of indicators of medicines availability, access, affordability, and use; c) policies and programs that facilitate appropriate medicines use by prescribers, dispensers, and patients; d) transparency in setting priorities for medicines coverage under resource constraints; and e) a system perspective to engage diverse actors.As they operationalize paths toward universal health coverage and include targeted medicines coverage policies and programs, systems can build on, and innovate, pharmaceutical policy frameworks and management tools from different countries' settings. Ensuring that medicines which achieve important health outcomes are available, accessible to all, used appropriately, and sustainably affordable is essential for realizing universal health coverage. Stakeholder cooperation and use of information and financing system levers provide opportunities to work toward this goal.


1α,25 dihydroxyvitamin D3 enhances cellular defences against UV-induced oxidative and other forms of DNA damage in skin.
DNA damage induced by ultraviolet radiation is the key initiator for skin carcinogenesis since mutations may arise from the photoproducts and it also contributes to photoimmune suppression. The active vitamin D hormone, 1α,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) reduces thymine dimers, the major photoproduct found in human skin after UV exposure, and suppresses the accumulation of nitric oxide derivatives that lead to more toxic reactive nitrogen species (RNS). We examined whether other forms of DNA damage are reduced by 1,25(OH)(2)D(3), and hypothesized that photoprotection by 1,25(OH)(2)D(3) is, in part, due to the suppression of various forms of promutagenic DNA damage, including thymine dimers, through a reduction of genotoxic RNS. Different forms of UV-induced DNA damage were investigated in irradiated skin cells treated with or without 1,25(OH)(2)D(3), or inhibitors of metabolism and inducible nitric oxide synthase. Keratinocytes were also treated with nitric oxide donors in the absence of UV light. DNA damage was assessed by comet assay incorporating site specific DNA repair endonucleases, and by immunohistochemistry using antibodies to thymine dimers or 8-oxo-7,8-dihydro-2'-deoxyguanosine, and quantified by image analysis. Strand breaks in T4 endonuclease V, endonuclease IV and human 8-oxoguanine DNA glycosylase digests increased more than 2-fold in UV irradiated human keratinocytes, and were reduced by 1,25(OH)(2)D(3) treatment after UV exposure, and also by low temperature, sodium azide and an inhibitor of inducible nitric oxide synthase. Conversely, nitric oxide donors induced all three types of DNA damage in the absence of UV. We present data to show that 1,25(OH)(2)D(3) protects skin cells from at least three forms of UV-induced DNA damage, and provide further evidence to support the proposal that a reduction in RNS by 1,25(OH)(2)D(3) is a likely mechanism for its photoprotective effect against oxidative and nitrative DNA damage, as well as cyclobutane pyrimidine dimers.